RU2774881C2 - System for determining interaction between test atmosphere and respiratory tract model (options) and its use (options), method for modeling such an interaction (options), pump (options), connecting structure, aerosol generating device and method for determining test atmosphere impact (options) - Google Patents

Abstract

FIELD: tobacco industry.

SUBSTANCE: group of inventions relates to a system for determining the interaction between a test atmosphere and a respiratory tract model, a pump for mixing a gas volume, a piston pump for mixing a gas volume, a connecting structure, a method for modeling the interaction between a test atmosphere and a respiratory tract model, the use of the specified systems, a method for determining a test atmosphere impact on a respiratory tract model, and an aerosol generating device. The system contains: (a) the first pump containing: (i) a chamber made with the possibility of containing the first gas volume containing the test atmosphere; (ii) the first channel made with the possibility of receiving and releasing gas and containing a valve for regulating a gas flow passing through the first channel, wherein the specified valve is made with the possibility of moving between open and closed positions, wherein, in the open position, the specified valve is capable of opening in the direction of the test atmosphere or ambient air; (iii) the second channel made with the possibility of receiving and releasing gas and containing a valve for regulating a gas flow passing through the second channel, wherein the specified valve is made with the possibility of moving between open and closed positions; (iv) a piston plate in the chamber, wherein the specified piston plate contains one or more openings for gas entry or inflow into the chamber, while one or more or each of openings includes a valve made with the possibility of moving between open and closed positions and capable of regulating the gas entry or inflow; (b) the second pump containing: (i) a chamber made with the possibility of containing the second gas volume, while the first and the second gas volumes differ from each other; (ii) a channel made with the possibility of receiving and releasing gas; and (iii) an engine to control the operation of the second pump; (c) a connecting structure used to transfer gas from the first pump to the second pump; and (d) one or more holes in the first pump, or the second pump, or walls of the connecting structure, or in combination of two or more of the specified options, wherein the specified holes are capable of accommodating a module for containing a cell cultivation medium, or for monitoring conditions in the chamber, or for sampling gas, or for determining gas characteristics.

EFFECT: obtaining a system for determining the interaction between a test atmosphere and a respiratory tract model.

16 cl, 3 dwg

Description

FIELD OF TECHNOLOGY

The present invention relates, inter alia, to devices, systems, methods and applications related to the modeling of the structure and/or functions of the airways and their respiratory characteristics. The present invention is applicable to the study of interactions between test substances (eg inhaled substances in test atmospheres) and the respiratory tract. In particular, the present invention relates to the simulation of the human respiratory tract.

BACKGROUND OF THE INVENTION

The respiratory system runs from the nose and upper respiratory tract to the alveolar surface of the lungs, where gas exchange occurs. Inhaled aerosols move from the mouth through the upper respiratory tract and eventually enter the alveoli. As the aerosol travels deeper into the airways, more soluble gases are adsorbed and the aerosol particles settle deep in the airways and alveoli. In particular, one aerosol of interest to us in the context of the present invention is smoke - such as tobacco smoke - since it is possible to study the effect of smoke on or through a respiratory tract model.

Existing aerosol exposure systems (see, for example, WO 2016118935 A1) most often rely on continuous, unidirectional aerosol flow or passive settling. Continuous aerosol streams are generated by positive or negative pressure and the aerosols are directed perpendicular to the biological testing system or run parallel to the plane of the biological testing system. The efficiency of dose delivery in this mode of exposure mainly depends on the geometric shape of the exposure chamber and the flow rate of the test aerosol. However, the mouth, breath-hold time, and dynamic flow patterns encountered during human breathing (i.e., inhalation and exhalation) cannot be modeled with current aerosol exposure systems. Therefore, there is little chance of achieving dose delivery typical of the in vivo situation, especially the relative delivery of particulate and gas aerosol constituents. In addition, the filtering action of the conducting airways cannot be modeled in a characteristic manner. Thus, when using modern aerosol exposure systems, an aerosol with the same size distribution of aerosol particles is used, for example, with cultures of bronchial cells and nasal cultures. However, in living organisms, larger aerosol particles are mainly interacted with and removed from the aerosol in the upper respiratory tract, while the deeper regions of the respiratory tract are mainly exposed to smaller aerosol particles and gaseous constituents. In addition, in continuous flow treatment systems, dilution of the aerosol is achieved by continuously adding dilution air to the aerosol upstream in the vicinity of the exposure chambers. However, if it is necessary to simulate the characteristics of smoking as in a biological organism, this dilution regimen is not organism-specific because it does not take into account the retention period in the mouth, during which the high-density aerosol is allowed to linger for a few seconds before being drawn into the respiratory tract. together with a large volume of dilution air.

In passive aerosol deposition, the test aerosol is injected into a chamber at the bottom of which the biological test systems are located. After injection of the aerosol, the aerosol is allowed to settle onto the test system, usually by gravity. The electrostatic attraction of aerosol particles to the test system can be used to increase the deposition of aerosol particles. Nanosized aerosol particles, in particular, may require electrostatic attraction due to their low deposition efficiency. The effects of complex aerosols containing aerosol particles with different sizes and densities, as well as gaseous components, cannot be carried out in systems with passive aerosol deposition, since the differential relative delivery of large aerosol particles, small aerosol particles and gaseous aerosol components is not characteristic of processes occurring in living organism.

There is still a need in the art for improved airway simulation systems.

BRIEF DESCRIPTION OF THE INVENTION

In a first aspect, a system is provided for determining the interaction between a test atmosphere and a model airway, said system comprising: (a) a first pump comprising: (i) a chamber configured to contain a first volume of gas containing the test atmosphere; (ii) a first channel capable of receiving and discharging gas and containing a valve for adjusting the flow of gas passing through the first channel, wherein said valve is movable between open and closed positions, while in the open position, said valve is able to open in the direction test atmosphere or ambient air; (iii) a second channel configured to receive and release gas and containing a valve for adjusting the flow of gas passing through the second channel, and the specified valve is movable between open and closed positions; (iv) a piston plate in the chamber, said piston plate having one or more lumens for gas entry or inflow into the chamber, wherein one or more, or each, of the lumen includes a valve movable between open and closed positions and capable of regulating the supply or inflow of gas; and (v) a motor for controlling the operation of the first pump; (b) a second pump comprising: (i) a chamber configured to contain a second volume of gas, wherein the first and second volumes of gas are different from each other; (ii) a channel configured to receive and release gas; and (iii) a motor for controlling the operation of the second pump; (c) a connection structure used to transfer gas from the first pump to the second pump; and (d) one or more openings in the first pump or the second pump or the walls of the connecting structure, or a combination of two or more of these options, and these openings are capable of receiving a module for containing cell culture medium or configured to store cell culture medium , or a module for monitoring conditions in a chamber or configured to monitor conditions in a chamber, or a module for sampling a gas or configured to sample a gas, or a module for determining the characteristics of a gas or configured to characterize a gas.

Suitably, the pumps are piston pumps comprising a piston plate and a base.

Suitably, the connecting structure is hollow.

Suitably, the connecting structure is branched.

Suitably, one end branch of the connecting structure is connected to the second pump and one or more other end branches are connected to a separate pump, each separate pump comprising: (i) a chamber for containing a volume of gas, this volume of gas being identical to the second volume of gas in the second pump ; (ii) a conduit for receiving and discharging gas and for connection to the connecting structure; and (iii) a motor for driving the pump.

Suitably, each individual pump is identical to the second pump.

Suitably, the system is contained in a housing, suitably in a temperature controlled housing.

Suitably, the jacket temperature is controlled by a thermostat.

Suitably, the temperature in the casing is approximately 37°C.

Suitably, the different volumes of the first and second pumps represent the internal volume of different parts of the airways, suitably of the human airways.

Suitably, the volumes of the individual pumps represent the internal volume of the various sections of the airways, suitably the human airways.

Suitably, the working volume of the first and second pumps is at least equal to the maximum achievable inlet volume in the corresponding section of the respiratory tract.

Suitably, the working volume of the individual pumps is at least equal to the maximum achievable inlet volume in the corresponding section of the respiratory tract.

Suitably, the discharge pressure of the motor or pump may be at atmospheric pressure, or above or below atmospheric pressure.

Suitably, the working volume of the first pump is from about 0 to 100 ml, or from about 1 to 100 ml.

Suitably, the working volume of the second pump is from about 0 to 4000 ml, or from about 1 to about 4000 ml.

Suitably, the pumps comprise stainless steel.

Suitably, the chamber is a cylinder.

Suitably, the chamber comprises glass.

Suitably, the volume of the chamber of the first pump is less than that of the chamber of the second pump.

Suitably, the chamber volume of the first pump represents the volume of the oral cavity and oropharynx.

Suitably, the volume of the chamber of the second pump represents the volume of the cavity of the lung or part thereof.

Suitably, the volume of the connecting structure represents the volume of the conducting airways of the lung, suitably the human lung.

Suitably, the system further comprises a computer controller capable of synchronizing the operation of the system.

Suitably, one or more of the first pump or second pump or connection structure comprises one or more modules containing a quartz crystal microbalance.

Suitably, the openings in the first pump or the second pump or the walls of the connecting structure are threaded or non-threaded.

Suitably, one or more holes contain the module.

Suitably, one or more modules are adapted to contain cell culture medium, or are configured to store cell culture medium, or are adapted to monitor chamber conditions, or are adapted to monitor chamber conditions, or are adapted to take gas samples, or adapted to characterize the gas.

Suitably, the modules are located on the basis of the piston plate of the first and/or second pump and/or in the walls of the connecting structure.

Suitably, one or more modules contain a cell culture medium.

Suitably, the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

Suitably, modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected thereto.

Suitably, the modules are located in a horizontal plane in one or more of the following: the first pump or the second pump, or the walls of the connecting structure.

Suitably, the connecting structure comprises stainless steel.

Suitably, the first pump chamber has a volume of approximately 100 ml.

Suitably, the chamber of the second pump has a volume of from about 1 liter to about 4 liters.

In a further aspect, a gas volume displacement pump is provided, comprising: (i) a chamber configured to contain a gas volume and comprising a base and one or more openings capable of receiving one or more modules for containing or storing cell culture medium, or for monitoring the conditions in the chamber, or for sampling the gas, or for characterizing the gas; (ii) a first channel for receiving and discharging gas when it is contained in the chamber, and containing a first valve for adjusting the flow of gas passing through the first channel, wherein said first valve is movable between open and closed positions, and in the open position, said the valve may open towards the test atmosphere or ambient air; and (iii) a second passage for receiving and discharging gas when contained in the chamber and containing a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions; and (iv) a piston plate in the chamber, said piston plate having one or more lumens for gas entry or flow into the chamber, wherein one or more, or each, of the lumen includes a valve movable between open and closed positions. and capable of regulating the flow or inflow of gas.

Suitably, the pump is a piston pump.

Suitably, one or more openings in the chamber are threaded or non-threaded.

Suitably, one or more holes contain the module.

Suitably, the module is threaded or threadless.

Suitably, one or more modules are adapted to contain cell culture medium, or are configured to store cell culture medium, or are adapted to monitor chamber conditions, or are adapted to monitor chamber conditions, or are adapted to take gas samples, or adapted to characterize the gas.

Suitably, the modules are located on the base of the pump.

Suitably, one or more modules contain a cell culture medium.

Suitably, the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

Suitably, modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected thereto.

Suitably, the modules are located in a horizontal plane in one or more of the following: the first pump, or the second pump, or the walls of the connecting structure.

Suitably, the module contains a microbalance on a quartz crystal.

Suitably, the pump further comprises a motor.

Suitably, the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

Suitably, the pump displacement is from about 0 to 100 ml, or from about 1 to about 100 ml.

Suitably, the pump comprises stainless steel.

Suitably, the chamber is a cylinder.

Suitably, the chamber comprises glass.

Suitably, the pump chamber has a volume of approximately 100 ml.

In a further aspect, a piston pump is provided for displacing a volume of gas, comprising: (i) a chamber configured to contain a volume of gas and containing a piston plate containing one or more gaps for gas inflow or inflow into the chamber, with one or more, or each , of the lumens include a valve movable between open and closed positions and capable of controlling the flow or flow of gas; (ii) a first channel for receiving gas, and containing a first valve for adjusting the flow of gas passing through the first channel, and the specified first valve is movable between open and closed positions; and (iii) a second passage for venting gas when contained in the chamber and comprising a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions. Suitably, the chamber includes a base and includes one or more openings.

Suitably, the holes are threaded or non-threaded.

Suitably, the holes contain the module in one or more holes.

Suitably, the module is threaded or threadless.

Suitably, the module is adapted to contain or store the cell culture medium, or to monitor the conditions in the chamber, or to sample the gas, or to characterize the gas.

Suitably, one or more modules contain a cell culture medium.

Suitably, the cell culture medium comprises a cell culture, suitably a 2- or 3-dimensional cell culture.

Suitably, modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected thereto.

Suitably, the module contains a microbalance on a quartz crystal.

Suitably, the connecting structure is connected to the second channel.

Suitably, the connecting structure is hollow.

Suitably, the pump further comprises a motor.

Suitably, the pump discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

Suitably, the pump displacement is from about 0 to 100 ml, or from about 1 to about 100 ml.

Suitably, the pump comprises stainless steel.

Suitably, the chamber is a cylinder.

Suitably, the chamber comprises glass.

Suitably, the pump chamber has a volume of approximately 100 ml.

In a further aspect, a gas volume displacement pump is provided, comprising: (i) a chamber configured to contain a gas volume, said chamber comprising a base and one or more modules for containing or storing cell culture medium, or for monitoring conditions in the chamber , or for gas sampling, or for gas characterization; and (ii) a conduit used to receive and release the gas.

Suitably, the pump is a piston pump.

Suitably, the modules are located at the base of the chamber.

Suitably, the modules are threaded or non-threaded.

Suitably, the modules are adapted to contain cell culture medium, or are configured to store cell culture medium, or are adapted to monitor chamber conditions, or are adapted to monitor chamber conditions, or are adapted to gas sampling, or are adapted to determination of gas characteristics.

Suitably, one or more modules contain a cell culture medium.

Suitably, the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

Suitably, modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected thereto.

Suitably, the module contains a microbalance on a quartz crystal.

Suitably, the connecting structure is connected to the channel.

Suitably, the connecting structure is hollow.

Suitably, the pump further comprises a motor.

Suitably, the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

Suitably, the pump displacement is from about 0 to 1000 ml, or from about 1 to about 100 ml.

Suitably, the pump comprises stainless steel.

Suitably, the chamber is a cylinder.

Suitably, the chamber comprises glass.

Suitably, the volume of the chamber represents the volume of the cavity of the lung or part thereof.

In a further aspect, a connection structure is provided adapted to connect at least two pumps for transferring gas between them, said connection structure comprising a hollow channel and one or more threaded or non-threaded holes in the walls of the connection structure.

Suitably, the threaded holes comprise a threaded module in one or more holes, said module being adapted to contain the cell culture medium, or configured to store the cell culture medium, or adapted to monitor the conditions in the chamber, or adapted to monitor the conditions in the cell. chamber, or adapted for gas sampling, or adapted for gas characterization.

Suitably, one or more modules contain a cell culture medium.

Suitably, the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

Suitably, modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected thereto.

Suitably, the connecting structure is hollow.

Suitably, the connecting structure is branched.

Suitably, each terminal branch of the connection structure may be connected to a separate pump.

Suitably, the connecting structure represents the volume of the conducting airways of the lung, suitably the human lung.

Suitably, the modules are arranged in a horizontal plane in the walls of the connecting structure.

Suitably, the modules are adapted to contain a cell culture, suitably a 2- or 3-dimensional cell culture.

Suitably, the module is a cell culture chamber, said chamber comprising a microfluidic channel and optionally a microfluidic pump connected thereto.

Suitably, the modules are adapted to monitor conditions in the connection structure and/or to take gas samples and/or to characterize the gas.

Suitably, the connecting structure comprises stainless steel.

Also described is a system containing the pumps described in this document.

Suitably, the system further comprises a connecting structure as described herein.

Suitably, the pumps are connected by a connecting structure.

In a further aspect, a method is provided for modeling the interaction between a test atmosphere and a model airway, including using the system described herein.

Also describes the use of the system described in this document to simulate the interaction between the test atmosphere and the model of the respiratory tract.

Also described is a method for determining the effect of the test atmosphere on a cell culture contained in a model respiratory tract, including the use of the system described in this document.

Also described is the use of the system described herein to determine the effect of the test atmosphere on a cell culture contained in a model airway.

In a further aspect, a method for determining the effect of a test atmosphere on a cell culture contained in a model airway is disclosed, comprising the steps of: (a) providing the system described herein, the system comprising the cell culture in one or more modules; and (b) comparing the cell culture before and/or after exposure to the test atmosphere, wherein the difference between the cell culture before and/or after exposure of the cells to the test atmosphere is indicative of whether the test atmosphere is affecting the cell culture.

A further aspect relates to a method for modeling the interaction between a test atmosphere and a model airway in the system described herein, comprising the steps of: (a) with the first valve of the first pump open and the second valve of the first pump closed, providing a gas containing the test atmosphere in the first pump through the first channel; (b) closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; (c) actuating the second pump to draw the test atmosphere into the connection structure and blowing the first pump chamber and connection structure with ambient air; (d) opening the first valve of the first pump towards ambient air and creating a tight connection between the first channel and the second channel of the first pump; and (e) after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump.

Additionally, a method is described for determining the effect of a test atmosphere on a model airway in the system described herein, comprising the following steps: (a) with the first valve of the first pump open and the second valve of the first pump closed, providing a gas containing the test atmosphere in the first pump through the first channel; (b) closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; (c) actuating the second pump to draw the test atmosphere through the connecting structure and blowing the first pump chamber and the connecting structure with ambient air; (d) opening the first valve of the first pump towards the ambient air and creating a tight connection between the first channel and the second channel of the first pump; and (e) after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump; wherein the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options, and this method includes the additional step of determining the effect of the test atmosphere on cell culture, wherein the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture.

Suitably, the modules are adapted to monitor system conditions and/or to take gas samples and/or to characterize the gas, and said method comprises obtaining one or more measurements from the modules.

Also disclosed is a method for simulating the interaction between a test atmosphere and a model airway, comprising the steps of: (a) providing a test atmosphere in a first pump chamber; (b) venting the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; (c) blowing the first pump and at least part of the connection structure with ambient air; (d) holding the test atmosphere in the second pump and connection structure for a specified period of time; (e) displacement of the test atmosphere into the connecting structure and the first pump by means of the second pump; and (f) performing one or more cycles of pumping ambient air into the second pump; while the test atmosphere comes into contact with the cell culture located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options.

Suitably, step (d) includes holding the test atmosphere in the second pump and the part of the connecting structure that still contains the test atmosphere for a certain period of time.

Suitably, the pumps are piston pumps comprising a piston plate and a base.

Suitably, the first pump is as defined in the embodiments of the present invention.

Suitably, the second pump is as defined in the embodiments of the present invention.

Suitably, the connection structure is as defined in the embodiments of the present invention.

Suitably, the process is carried out in a casing, suitably in a temperature controlled casing.

Suitably, the jacket temperature is controlled by a thermostat.

Suitably, the temperature in the casing is approximately 37°C.

Suitably, the different volumes of the first and second pumps represent the internal volume of different parts of the airways, suitably of the human airways.

Suitably, the working volume of the first and second pumps is at least equal to the maximum achievable inlet volume in the corresponding section of the respiratory tract.

Suitably, the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

Suitably, the displacement of the first pump is from about 0 to 100 ml, or from about 1 to about 100 ml.

Suitably, the working volume of the second pump is from about 0 to 4000 ml, or from about 1 to about 4000 ml.

Suitably, the chamber volume of the first pump is smaller than that of the second pump chamber.

Suitably, the chamber volume of the first pump represents the volume of the oral cavity and oropharynx, suitably of the human oral cavity and oropharynx.

Suitably, the volume of the chamber of the second pump represents the volume of the cavity of the lung or part thereof, suitably the cavity of the human lung or part thereof.

Suitably, the connecting structure represents the volume of the conducting airways of the lung, suitably the human lung.

Suitably, the cell culture is located at the base of the piston plate of the first and/or second pump and/or in the walls of the connecting structure.

Suitably, the cell culture is a 2- or 3-dimensional culture.

Suitably, said method further comprises monitoring and/or gas sampling and/or characterizing the gas in one or more of the first pump or second pump or connection structure using one or more of the modules contained therein.

Suitably, the cell culture chamber further comprises a microfluidic channel and optionally a microfluidic pump connected to it.

Suitably, the connecting structure comprises stainless steel.

Suitably, the first pump chamber has a volume of approximately 100 ml.

Suitably, the chamber of the second pump has a volume of from about 1 liter to about 4 liters.

In a further aspect, a method for determining the effect of a test atmosphere on a simulated airway is described, comprising the steps of: (a) providing a test atmosphere in a first pump chamber; (b) withdrawing the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; (c) blowing the first pump and at least part of the connection structure with ambient air; (d) holding the test atmosphere in the second pump and connection structure for a specified period of time; (e) displacement of the test atmosphere through the connecting structure and the first pump with the help of the second pump; and (f) performing one or more cycles of pumping ambient air into the second pump; wherein the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options, and this method includes the additional step of determining the effect of the test atmosphere on cell culture, wherein the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture.

Suitably, the modules are adapted to monitor system conditions and/or to take gas samples and/or to characterize the gas, and said method comprises obtaining one or more measurements from the modules.

Also described is a device capable of or adapted to carry out the method(s) described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

In FIG. 1 shows a system 10 according to one embodiment of the present invention, also showing: a first pump 40 according to one embodiment of the present invention; a second pump 80 according to one embodiment of the present invention; and a connecting structure 50 according to one embodiment of the present invention.

In FIG. 2 and 3 show modules 113, 213 according to embodiments of the present invention.

ADVANTAGES OF THE INVENTION

Because the processes that affect the physicochemical properties of the airway test atmosphere, as well as their mode of interaction (e.g., dose delivery) are modeled using the physical and functional properties of the system, according to embodiments of the present invention, dosage information of clinical relevance can be obtained. .

The effect of one or more test atmospheres on one or more airways can be studied in accordance with embodiments of the present invention.

The effect of one or more test atmospheres on one or more airways can be studied simultaneously or in stages, if necessary, according to embodiments of the present invention.

The modules that can be used in the system offer flexibility in terms of an open system for testing, the expected results of experiments, and the exposure parameters that need to be monitored according to embodiments of the present invention.

Modules can be redesigned or modified according to specific requirements, without the need to change the overall design and function of the system according to embodiments of the present invention.

For many applications, aerosol generation is driven by the system itself, which means that aerosol generators/smoking machines are not required according to embodiments of the present invention. This can simplify the design of the system.

In some embodiments, the system may be modular in nature. This means that various components such as pumps and connection structure, for example, can be easily replaced individually if necessary. This can facilitate partial redesign, improvement or replacement of parts of the system according to specific requirements.

The mouth, breath holding time, and dynamic flow patterns encountered during breathing may be modeled according to embodiments of the present invention.

The present invention, in some embodiments, may allow for a holding period in the mouth during which the high density aerosol is allowed to linger for a few seconds before being drawn into the airways along with a large volume of dilution air.

DETAILED DESCRIPTION OF THE INVENTION

In the practice of the present invention, conventional techniques of engineering, microbiology, cell biology, and biochemistry are used, in certain embodiments. Biological techniques are fully explained in literature such as Molecular Cloning: A Laboratory Manual, Second Edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (MJ. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J. E. CeIMs, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J. P. Mather and P. E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, IB. Griffiths, and D. G. Newell, eds., 1993-8) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994). Procedures that use commercially available kits and reagents will generally be performed according to the protocols specified by the manufacturer, unless otherwise specified.

The technical terms and expressions used in this document should be given the meaning normally applied to them in the art. All definitions of terms used in this text apply to the entire content of this application.

The term "comprising" does not exclude other elements or steps.

The singular form does not exclude the plural.

The term "and/or" means, for example, (a) or (b) or both (a) and (b).

The terms “comprising,” “comprises,” and “comprises” as used herein are synonymous with “comprising,” “comprises,” or “comprising,” “comprises,” and are inclusive or non-limiting and do not exclude additional , not listed representatives, elements or steps of the method. The term "consisting of" means that the additional components are excluded and there are only the mentioned elements and nothing else.

The term "approximately" as used herein in relation to a measurable value, such as a parameter, amount, time period, etc., is intended to cover the variability of the specified value and deviations from it, in particular deviations of ± 10% or less, preferably ±5% or less, more preferably ±1% or less, and even more preferably ±0.1% or less of the specified value, if such changes are acceptable in the practice of the present invention. It should be understood that the meaning to which the modifier "about" refers is also specifically and preferably disclosed on its own.

Before a more detailed discussion of the embodiments, a first overview will be provided. Embodiments provide devices and methods that may have a variety of uses for studying the respiratory tract. For example, embodiments find use in studying the deposition and/or condensation of one or more constituents present in the test atmosphere on the internal surfaces of the device, including their re-evaporation. Embodiments also provide an evaluation of the test atmospheres that can be examined during their passage through the device in order to study changes in aerosol concentration and/or growth of aerosol particles and/or reduction of aerosol particles. The effect of test atmospheres on biological test systems present within the device/system can be studied in embodiments of the present invention.

System

In one aspect, a system 10 is described for determining the interaction between a test atmosphere and a model airway, said system comprising: (a) a first pump comprising: (i) a chamber configured to contain a first volume of gas containing the test atmosphere; (ii) a first passage capable of receiving and discharging gas and comprising a valve for adjusting the flow of gas passing through the first passage, said valve being movable between open and closed positions, said valve being capable of opening towards the atmosphere in the open position test or ambient air; (iii) a second channel configured to receive and release gas and containing a valve for adjusting the flow of gas passing through the second channel, and the specified valve is movable between open and closed positions; and (iv) a motor for controlling the operation of the first pump; a second pump comprising: (i) a chamber configured to contain a second gas volume, wherein the first and second gas volumes are different from each other; (ii) a channel configured to receive and release gas; (iii) a piston plate in the chamber, said piston plate having one or more lumens for gas entry or flow into the chamber, wherein one or more, or each, of the lumens includes a valve movable between open and closed positions, and able to regulate the flow or inflow of gas; and (iv) a motor for controlling the operation of the second pump; (c) a connection structure used to transfer gas from the first pump to the second pump; and (d) one or more openings in the first pump or the second pump or the walls of the interconnecting structure, or a combination of two or more of these options, which openings are capable of receiving a module for containing or storing cell culture medium, or for monitoring conditions in chamber, or for gas sampling, or for gas characterization.

The gas may be the test atmosphere or it may contain the test atmosphere.

In FIG. 1 depicts a system 10 according to one embodiment of the present invention. System 10 includes at least two pumps 40, 80. Two or more pumps 40, 80 are connected to each other. In some embodiments, two or more pumps 40, 80 are connected to each other via a branched hollow structure 50. Each pump 40, 80 may be driven by its own separate motor 41, 81 or two or more pumps may be driven by the same motor. 41, 81, if necessary. The entire system 10 may be housed in a climate enclosure 11 fitted with a thermostat 12 to control the temperature in the enclosure 11. The pump chambers 40, 80 may be configured to represent the internal volumes of different airways, such as those of a human or an animal. These may be configured to provide a working volume that is at least equal to the maximum achievable inlet volume in the relevant section of the respiratory tract. In particular, one (first) pump 40 may represent the volume of the oral cavity and oropharynx, such as the oral cavity and oropharynx of a human or animal. The other second pump 80 may represent the volume of a cavity of a lung or parts thereof, such as the cavity of individual lobes of a lung or smaller elements, in particular the cavity of a human or animal lung or parts thereof. The branched hollow connecting structure 50 may represent the dimensions of the airways, such as one or more of the nasopharyngeal cavity, pharynx, larynx, trachea, bronchi, and bronchiolar structures to respiratory bronchioles, in particular, human or animal airways. The branched hollow connecting structure 50 may represent the dimensions of the airways, including the nasopharynx, pharynx, larynx, trachea, bronchi, and bronchiolar structures to the respiratory bronchioles. Dimensions, such as diameter and length, as well as the branching pattern of various auxiliary parts of the connecting structure 50 may resemble a tree structure of the airways.

As shown in FIG. 1, a branched hollow structure 50 may be connected to a central opening 43, 83 in the base 44, 84 of the chamber 42, 82 of each pump 40, 80. chambers) 42, 82, which can be arranged symmetrically around the central opening 43, 83. Located directly at the junction between the pump 40 and the connecting structure 50, the valve 44a can be used to seal the pump 40 from all other parts of the system.

The pump 40 representing the oral cavity may have one or more openings 43 through which the test atmosphere(s) and dilution air can exit the pump 40 to the branched hollow structure 50. The point of entry and exit of the test atmosphere 90 is typically located on the plate 45 of the pump piston. 40, suitably at its center. It may pass through the hollow shaft 46 of the piston, on top of which may be a valve 44a, such as a three-way valve. In embodiments, valve 44a may be closed or opened towards the source of the test atmosphere or ambient air. A group of one or more (eg, multiple) openings 47 through which ambient air can enter the system are located on the piston plate 45 and are not necessarily radial. One or more valves 48 (e.g., a plurality) at one or more, or all, of the lumens may be used to cause one or more of these lumens 47 to open or close. In some embodiments, each lumen 47 is controlled by a valve 48. In some embodiments, a group one or more (eg, multiple) openings through which ambient air may enter the system may be located on the piston plate 84 of the second pump, optionally in a radial arrangement. One or more valves (eg, a plurality) may be used to allow one or more of these lumens to open or close. In some embodiments, each lumen is controlled by a valve. In some embodiments, a group of one or more (eg, a plurality) of lumen through which ambient air may enter the system may be located on the piston plate of the first and second pump, optionally in a radial arrangement. Advantageously, the branched hollow structure 50 can be detached from the pumps 40, 80. Advantageously, the branched hollow structure 50 can be disassembled into its component parts. This may provide easy access for placing or removing test systems and/or for cleaning.

The bases 44, 84 of the pumps 40, 80 can be removed for placement/removal of test systems and for cleaning.

In the base 44, 84 of each pump 40, 80, as well as in various parts of the connecting structure 50, holes, recesses or sockets 51, such as threaded or threadless holes, threaded or threadless recesses, or threaded or threadless sockets 113, 213, can be located. Holes, recesses or pockets 51 may have different locations, for example, on the bases 44, 84 of one or more pumps 40, 80, or they can be located around the central hole 43, 83 or in a branched hollow structure 50 at various selected locations, as appropriate. on the underside of the branched hollow structure 50, or any combination of the above.

Holes, recesses or sockets 51 may be used to allow mounting of various modules 112, 212 or devices in or on them, which may be used to monitor the operation of the system 10 and/or to conduct experiments and/or to collect samples, etc. Examples of such modules 112, 212 or fixtures are shown in FIG. 2 and FIG. 3 and described in this document.

Advantageously, the pumps 40, 80 used in the system 10 can thus function not only to transport test atmospheres, but they can also function as exposure chambers.

In FIG. 2 and 3 depict an embodiment of the present invention in which holes, recesses or pockets 51 are optionally in the form of threaded holes, recesses or pockets 112, 212. One or more holes, recesses or pockets 51 may include one or more modules 112, 212. Use The threads facilitate easy installation and replacement of the module(s) 112, 212. The use of threads in the holes, recesses or sockets 51 is optional as the modules can be configured to be inserted or pushed into the holes, recesses or sockets 51 to form an airtight interaction between them. Modules may have a tight fit. Sealing can be achieved by using O-rings, etc.

The modules 112, 212 used can be adapted for various purposes depending on the requirements of the configured system. For example, modules 112, 212 may be adapted to contain or store cell culture media, or to monitor conditions in chamber 42, 82, or to sample a gas or liquid, or to characterize a gas, etc. Modules 112, 212 may be located on the base 44, 84 of the first 40 and/or the second 80 pump, and/or in the walls of the connecting structure 50. In one particular embodiment, one or more modules 112, 212 may be configured to contain or store the cell culture medium. According to this embodiment, one or more modules 112, 212 may be containers capable of holding a liquid or solution. The cell culture medium may contain or be contacted with a cell culture, such as a 2- or 3-dimensional cell culture. In some embodiments, the module(s) 113, 213 adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected thereto. Modules 113, 213 will generally be located in a horizontal plane in one or more of the following: the first pump 40, or the second pump 80, or the walls of the connecting structure 50.

As seen in FIG. 2 and 3, holes, recesses or sockets 51 may optionally be configured to contain a threaded recess or socket 112, 212. fixture 116, 216 can be used to monitor the internal conditions of the system or to characterize the test atmosphere or to take samples, etc. The operation of fixture(s) 116, 216 can be controlled by computer 117, 217. Threaded recess or socket 113, 213 and/ or modules 112, 212 can be adapted for use as a culture chamber 115, 215 in which biological testing systems (e.g., organotypic cultures of human airway epithelial cells as described herein) can be placed to be exposed to the test atmosphere. Threaded holes, recesses or sockets 113, 213 may be adapted to contain trapping means in which samples of the test atmosphere may be taken for analysis. Sampling of the cell culture medium or trapping agent during exposure to the test atmosphere can be obtained in a variety of ways, including by using the microfluidic pump system 211. Modules 112, 212 containing a quartz crystal microbalance (QCM, 219) may be used. Modules 112, 212, which can carry sensors 214, chambers 215 or QCM 219, can be inserted into any chamber 42, 82 or the interior of the branched hollow structure 50 of the system. The operation of one or more QCMs 219 may be controlled by computer 220. System 10 may be wholly or partially controlled by computer 13, as desired. System 10 may be partially or fully automated.

System 10 may include one or more (eg, multiple) first pumps. System 10 may include one or more (eg, multiple) second pumps. System 10 may include one or more (eg, multiple) first pumps and one or more second pumps.

First pump

In one aspect, a pump, suitably a piston pump, for displacing a volume of gas is described, comprising: (i) a chamber configured to contain a volume of gas and comprising a base and one or more openings capable of accommodating a module for containing or storing cell culture medium, or to monitor the conditions in the chamber, or to take samples of the gas, or to characterize the gas; (ii) a first channel for receiving and discharging gas when it is contained in the chamber, and containing a first valve for adjusting the flow of gas passing through the first channel, wherein said first valve is movable between open and closed positions, and in the open position, said the valve may open towards the test atmosphere or ambient air; (iii) a second passage for receiving and discharging gas when contained in the chamber and containing a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions; and (iv) a piston plate in the chamber, said piston plate having one or more lumens for gas entry or flow into the chamber, wherein one or more, or each, of the lumen includes a valve movable between open and closed positions. and capable of regulating the supply or inflow of gas.

Also described is a piston pump for displacing a volume of gas, comprising: (i) a chamber configured to contain a volume of gas and containing a piston plate containing one or more gaps for the entry or inflow of gas into the chamber, with one or more, or each, of the lumens include a valve movable between open and closed positions and capable of controlling the flow or flow of gas; (ii) a first gas receiving passage comprising a first valve for adjusting the flow of gas passing through the first passage, said first valve being movable between open and closed positions; and (iii) a second passage for venting gas when contained in the chamber and comprising a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions.

As shown in FIG. 1, system 10 may include a first pump 40 for displacing a volume of gas. The first pump 40 is described herein as a separate aspect of the present invention and its use is not limited to use in the system 10 described herein.

The first pump may be the primary pump, so named because of its location in the system as the entry point for the gas. It comprises a chamber 42 (e.g. a cylinder) configured to contain a volume of gas and includes a base 44 and one or more openings 43 capable of receiving a module, such as a threaded or threadless module 113, 213, in a recess or seat 112, 212, as shown in FIG. 2 and 3 and as described herein. It also includes a first passage 90 for receiving and discharging gas when contained in chamber 42 and includes a first valve 44a, such as a three-way valve for adjusting the flow of gas passing through the first passage 90, said first valve 44a being movable between open and closed positions, wherein in the open position the valve 44a may open towards the test atmosphere or ambient air. It also includes a second channel 43 for receiving and discharging gas when it is contained in the chamber 42. Suitably, the second channel 43 is in the form of an opening. At the junction between the pump 40 and the connecting structure 50, a second valve 49 located at the second channel 43 allows the pump 40 to be sealed from all other parts of the system. The second valve 49 may be used to control the flow of gas through the second passage 43, said second valve 49 being movable between open and closed positions. As seen in FIG. 1, pump 40 may be a piston pump including piston plate 45. One or more of the holes, recesses or sockets 51 in the chamber 42 may be threaded 112, 212 or threadless. One or more of the openings, recesses or sockets 51 in the chamber 42 may contain a module 113, 213, such as a threaded or threadless module 113, 213, as described herein. The pump 40 further comprises a motor 41 for controlling the operation of the pump. The discharge pressure of the engine 41 may be at atmospheric pressure, or above or below atmospheric pressure, as desired. In some embodiments, the delivery pressure of engine 41 may be above or below atmospheric pressure to bias the test atmosphere. The displacement of pump 40 may be from about 0 to 100 ml, or from about 1 to about 100 ml. Chamber 42 of pump 40 may have a volume of up to about 100 ml. Pump 40 may be made from various materials known in the art, such as stainless steel. Suitably, chamber 42 is a cylinder. Suitably, chamber 42 may be made of glass. The plate 45 of the piston of the pump 40 contains one or more gaps 47 for the entry or flow of gas. One or more openings 47 may include a valve 48 movable between open and closed positions and able to control the flow or flow of gas.

As also shown in FIG. 1, pump 40 may be a piston pump for displacing a volume of gas. The piston pump includes a chamber 42 (for example, a cylinder, such as a glass cylinder) configured to contain a volume of gas and containing a piston plate 45 containing one or more lumens 47 for gas to enter or flow into the chamber 42. One or more of the lumens 47 or each of the lumen 47 contains a valve 48 to control the flow or flow of gas through the lumens 47.

It may include a first gas receiving passage 90 and a first valve 44a - such as a three-way valve - for controlling gas flow through the first passage 90. The first valve 44a is movable between open and closed positions. It also includes a second conduit 43 for venting gas when contained in chamber 42, the second conduit 43 optionally including a second valve for adjusting the flow of gas through the second conduit 43, the valve being movable between open and closed positions. The chamber 42 may include a base 44a and one or more openings 43. The base may further include one or more openings, recesses or sockets 51, which may be threaded 119 or threadless and/or they may contain a module 113, 213, as described in this document. The connection structure 50 - such as a hollow connection structure - may be connected to the second channel 43. The pump 40 may further comprise a motor 41 in which the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure. The displacement of pump 40 may be from about 0 to 100 ml, or from about 1 to about 100 ml. Chamber 42 of pump 40 may have a volume of approximately 100 ml.

A system is envisaged comprising two or more first pumps. It is also contemplated to use two or more first pumps in the system to determine the interaction between the test atmosphere and the airway model.

Also provided is a method for culturing the cell in the first pump. The use of the first cell culture pump is also described.

Second pump

In a further aspect, a gas volume displacement pump is described, comprising: (i) a chamber configured to contain a gas volume, said chamber comprising a base and one or more modules for containing or storing cell culture medium, or for monitoring conditions in the chamber. , or for gas sampling, or for gas characterization; and (ii) a conduit used to receive and release the gas. As shown in FIG. 1, the system may comprise a second gas volume displacement pump 80 comprising a chamber 82—such as a cylinder—capable of containing a gas volume, said chamber 82 comprising a base 84 and one or more modules 113, 213 for containing or storing the culture medium. cells, or to monitor conditions in chamber 82, or to sample the gas, or to characterize the gas, etc. The second pump may be a secondary pump.

The second pump 80 is described herein as a separate aspect of the present invention and its use is not limited to use in the system 10 described herein.

The second pump 80 further includes a conduit 83 used to receive and discharge gas. As shown in FIG. 1, the second pump 80 may be a piston pump including a piston plate 83. The piston plate 83 may not contain any gaps or holes. Modules 113, 213, such as threaded or threadless modules 113, 213, as described herein, may be located in the base 84 of the chamber 82. A connection structure 50 - such as a hollow connection structure - can be connected to the channel 83. The second pump 80 further comprises a motor 81. The discharge pressure of the second pump 80 will generally correspond to atmospheric pressure or pressure above or below atmospheric pressure. The displacement of the pump may be from about 0 to about 1000 ml, or from about 0 to about 4000 ml, or from about 1 to about 1000 ml, or from about 1 to about 4000 ml. The volume of the chamber 82 may represent the volume of the cavity of the lung or part of it. In some embodiments, a group of one or more (eg, multiple) openings through which ambient air may enter the system may be located on the piston plate 84, optionally in a radial arrangement. One or more valves (eg, a plurality) may be used to allow one or more of these lumens to open or close. In some embodiments, each lumen is controlled by a valve.

A system comprising two or more second pumps is contemplated. It is also contemplated to use two or more second pumps in the system to determine the interaction between the test atmosphere and the airway model.

A method for culturing a cell in a second pump is also provided. The use of a second cell culture pump is also described.

Connecting structure

Also described is a connection structure used to transfer or transport gas between two or more pumps. The connecting structure may be a pipe, or tube, or conduit, or the like, through which gas can be conducted or transported. The connection structure may be adapted to connect at least two pumps for the purpose of transferring or transporting gas between them. The connection structure may connect the first pump at the second passage of the first pump described herein and the passage of the second pump described herein. The connection structure may include a hollow channel and one or more holes - such as threaded or non-threaded holes - in the walls of the connection structure. The connection structure may be used in the systems and methods described herein. The system may include a connecting structure adapted to connect at least two pumps to transfer gas between them.

One embodiment of the connecting structure 50 is shown in FIG. 1. The connection structure will generally comprise a hollow channel. It will usually be branched. In embodiments, each terminal branch of the connecting structure may be connected to individual pumps 40, 80 when the system includes one. The connection structure 50 may include one or more holes - in particular, threaded or non-threaded holes - in the walls of the connection structure. Threaded or non-threaded ports may comprise a module 112, 212. The module 112, 212 may be adapted to contain the culture medium and/or to monitor system conditions and/or to take gas samples and/or to characterize the gas, such as described in this document. In some embodiments, the implementation of the connecting structure may be branched with two or more branches. Each end branch of the connecting structure can be connected to a separate pump. The connecting structure may represent the volume of the conducting airways of the lung. The connecting structure 50 can be made from various materials. In some embodiments, stainless steel is preferred.

Also provided is a method for culturing a cell, containing a connecting structure. The use of a connecting structure for cell culture is also described.

Operation and function of the system

The system can be fully or partially controlled by a computer, as required. This may allow the operation of some or all of the pumps and valves to be synchronized. This may allow the operation of some or all elements of the system to be synchronized. The computer can be used to set the stroke length of one or more pumps and/or the stroke speed of one or more pumps. The computer can be used to adjust the system temperature.

Next, the operation of the embodiment of the system 10 shown in FIG. 1. At rest, the piston of the first pump 40 is in the down position, while the second pump 80 is in position to hold a certain volume of gas in the chamber 82, and the valves 48, 49 are closed. The test atmosphere is drawn into pump 40, which may represent the oral cavity. This incoming atmosphere may be driven by the pump 40 and may enter the chamber 42 through the hollow shaft 46 of the piston. The hollow shaft 46 of the piston can be directly connected to the source of the test atmosphere. When the pump 40 has completed its upward movement, the valve 44a, which may be a three-way valve, in the central hole on the top of the hollow shaft 46 of the piston closes, and the valves 48 that control the flow of ambient air through the gaps 47 in the plate 45 of the piston and the valve 49 at the inlet connecting structure 50 are opened.

The test atmosphere is pumped out of the chamber 42, which may represent the oral cavity, through the connecting structure 50, which may represent the conductive airways. This process may be driven by an upward movement of the second pump 80, which may represent the cavity of the lung or parts thereof. Since the total displacement of the second pump 80 may be a multiple of the volume of the pump 40, the chambers 42 of the pump 40, as well as at least part of the connecting structure 50, can be purged with ambient air, which may enter the first pump 40 through gaps 47 in the piston plate 45.

In the first pump 40, the valve 44a on the top of the piston shaft 46 may open towards the environment. The valves 48 on the piston plate 45 can close and the pump 40 lowers the piston. In the lower position of the piston, this pump 40 may form a seal between the hollow axis 46 of the piston and the hole 43 in the base 44. The seal may be obtained using a gasket 53. The gasket 53 may be located on the base 44 or on the plate 45 of the piston. After a certain "breath-hold period", the second pump 80 may perform a piston down, thereby displacing the test atmosphere through the connecting structure 50 and through the piston shaft 46 of the first pump 40, directly into the environment. When the piston of pump 40 remains in the down position and valve 44a remains open towards the environment, the second pump(s) 80 may perform one or more (e.g., multiple) cycles of breathing ambient air before valve 44a opens again towards the environment. test atmosphere source and the next test atmosphere inhalation cycle will begin.

In a further aspect, a method is provided comprising the steps of: (a) providing a pump - such as the first pump described herein - comprising a chamber; (b) venting gas - such as the test atmosphere - from the pump to a connection structure connecting the pump to another pump - such as the second pump described herein; (c) purging the pump provided in step (a) and at least a portion of the connection structure with ambient air; (d) holding the gas in another pump and connection structure for a predetermined period of time; (e) displacing the gas into the connection structure and the pump provided in step (a) using a different pump; and (f) performing one or more cycles of pumping ambient air into another pump.

In a further aspect, a method is provided comprising the steps of: (a) providing a gas - such as a test atmosphere - to a pump chamber - such as the first pump described herein; (b) diverting gas from the pump described in step (a) to a connection structure connecting the pump to another pump, such as the second pump described herein; (c) purging the pump provided in step (a) and at least a portion of the connecting structure with ambient air; (d) holding the gas in another pump and connection structure for a predetermined period of time; (e) displacing the gas through the connection structure and the pump provided in step (a) using a different pump; and (f) performing one or more cycles of pumping ambient air into another pump.

Test atmosphere generation

Test atmospheres - such as aerosols - to be studied using the described system 10 and methods, can be generated in various ways. For many applications, such as testing tobacco products or conventional medical inhalers, etc., the generation of the test atmosphere can be driven by the system itself, i.e. the primary or secondary pumps themselves provide the negative pressure needed to generate and extract the test aerosol . which means that the use of aerosol generators/smoking machines is advantageously not required.

The test atmosphere may be a sample of gases or aerosols taken from the environment, for example, to monitor indoor air quality, work-related exposures, or environmental pollution near industrial sites. In this particular case, the test atmosphere is not generated but sampled by the system.

The test atmosphere may be an aerosol such as smoke, or it may be derived from smoke. As used herein, the term "smoke" is used to describe the type of aerosol produced by smoking articles such as cigarettes or by burning an aerosol-forming material. The smoke contains various agents, which, if necessary, can be provided in the form of separate compounds for research. Examples of such agents include nicotine-free dry particulate matter, carbon monoxide, formaldehyde, acetaldehyde, acetone, acrolein, propionaldehyde, crotonaldehyde, methyl ethyl ketone, butyraldehyde, benz[a]pyrene, phenol, m-cresol, o-cresol, p -cresol, catechol, resorcinol, hydroquinone, 1,3-butadiene, isoprene, acrylonitrile, benzene, toluene, pyridine, quinoline, styrene, N'-nitrosonornicotine (NNN), N'-nitrosoanatabine (NAT), N'-nitrosoanabasine ( NAB), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), 1-aminonaphthalene, 2-aminonaphthalene, 3-aminobiphenyl, 4-aminobiphenyl, nitric monoxide (NO), nitric oxide (NOx ), hydrocyanic acid, ammonia, arsenic, cadmium, chromium, lead, nickel, selenium and mercury.

When the aerosol is smoke, the system 10 may optionally be connected to a smoking machine. Suitably, the smoking machine holds and ignites cigarettes, the pumps being provided by the system or being the pumps of the present invention. You can use a certain number of puffs per cigarette and a certain number of puffs per minute of exposure, and vary the number of cigarettes to accommodate exposure time. Reference cigarettes, such as reference 3R4F cigarettes, can be used as a source of smoke and smoked on a smoking machine in accordance with the basic smoking regime established by the International Organization for Standardization (ISO 2000).

It is also envisaged to use a control atmosphere, such as an atmosphere that does not contain a test atmosphere. The use of a control atmosphere can help determine the effect of the test atmosphere versus the control atmosphere.

The system 10 may be connected to the smoking machine via a suitable conduit providing a path for smoke to flow into the system 10. The smoke may be conveyed through the conduit with or without a carrier gas such as air. When using a carrier gas, the conduit preferably includes an inlet for introducing the conductor gas into the conduit to mix it with the smoke stream. The conduit may include at least one inlet for introducing or injecting a standard reference into system 10, such as nicotine, for calibration purposes. The flow of smoke will typically be controlled by the system or pumps of the present invention.

The smoking machine may be a linear or rotary smoking machine. Suitably, the smoking machine is used to smoke a plurality of smoking articles at the same time, so that the total smoke from the plurality of smoking articles can be collected and analyzed. Smoking machines suitable for use in the present invention are well known to those skilled in the art.

The system 10 and method described herein can be used to perform an analysis of the mainstream smoke generated by a smoking article during a smoking test. "Mainstream smoke" refers to the smoke that is drawn through the smoking article and that is inhaled by the consumer during use.

The source of the test atmosphere may be an "aerosol generating device", which is a device that interacts with an aerosol generating substrate to generate an aerosol. An example of an aerosol is smoke. The aerosol generating substrate may be part of the aerosol generating article. The aerosol generating device may comprise one or more components suitable for generating an aerosol from an aerosol generating substrate. The aerosol generating device may be an electrically heated aerosol generating device, which is an aerosol generating device comprising a heater that, by energizing, heats the aerosol generating substrate of the aerosol generating articles to generate the aerosol. The aerosol generating device may be a gas heated aerosol generating device, a device heated by a carbonaceous heat source, another exothermic chemical reaction, or a heat sink device. Other methods suitable for generating an aerosol are well known in the art. The aerosol generating device may be a device that interacts with the aerosol generating substrate of the aerosol generating article to generate an aerosol that can be directly inhaled into the user's lungs through the user's mouth.

Another example of an "aerosol generating device" is an inhalation device (inhaler) which is commonly used to deliver an aerosol containing an active ingredient, such as a compound with an active medical effect. Such an inhalation device is typically used to deliver aerosolized medicaments to the respiratory tract. They can be used to treat respiratory and other ailments. Such inhalers are well known in the art and are generally of the pressurized metered dose type, dry powder type, or nebulizers. Typically, the medicament is in the form of a pressurized formulation containing fine particles of one or more medicinal compounds suspended in a liquefied nebulizer, or in the form of a solution of one or more compounds dissolved in a nebulizer and co-solvent system. Such formulations are well known in the art.

In the context of this document, the term "aerosol-forming substrate" refers to a substrate capable of releasing volatile compounds that can form an aerosol. Such volatile compounds can be released by heating the aerosol-forming substrate. The aerosol-forming substrate may be applied to the substrate or support by adsorption, coating, impregnation, or otherwise. The aerosol generating substrate may conveniently be part of an aerosol generating or smoking article. In some applications, the aerosol generating substrate is contained in an aerosol generating article, for example a rod-shaped aerosol generating article such as a heated aerosol generating article or a heated cigarette. The aerosol generating article is of a suitable size and shape to interact with the aerosol generating device so as to contact the aerosol generating substrate with the heater.

The aerosol-forming substrate may contain compounds with an active medical effect or drugs, such as antibiotics or anti-inflammatory agents, which can be delivered to the patient through the respiratory tract. Numerous inhalation devices (inhalers) are known and prescribed on an ongoing basis for the treatment of various diseases related or not related to the respiratory tract.

The aerosol forming substrate may contain nicotine. The aerosol forming substrate may contain tobacco. The aerosol-forming substrate may comprise, for example, a tobacco-containing material containing volatile tobacco flavor compounds that are released from the aerosol-forming substrate upon heating. In some embodiments, the aerosol forming substrate may comprise a homogenized tobacco material, such as a molded tobacco sheet. In the context of this document, the term "homogenized tobacco material" refers to a material formed by agglomeration of tobacco in the form of particles. The homogenized tobacco may be in the form of a leaf. The aerosolizing agent content of the homogenized tobacco material may be greater than 5% on a dry weight basis. Alternatively, the aerosolizing agent content of the homogenized tobacco material may be from 5% to 30% on a dry weight basis. Sheets of homogenized tobacco material may be formed by agglomerating particulate tobacco obtained by grinding or otherwise grinding one or both of the tobacco leaf layers and tobacco leaf stems. Alternatively or in addition, the sheets of homogenized tobacco material may contain one or more of the following: tobacco dust, tobacco fines, and other particulate tobacco waste generated, for example, during processing, handling and shipping of tobacco. Sheets of homogenized tobacco material may contain one or more intrinsic binders, ie, tobacco endogenous binders, one or more external binders, ie, tobacco exogenous binders, or a combination thereof, which aids in agglomeration of bulk tobacco; alternatively or additionally, the homogenized tobacco material sheets may contain other additives, including, but not limited to, tobacco and non-tobacco fibers, aerosolizers, humectants, plasticizers, flavorants, fillers, aqueous and non-aqueous solvents, and combinations thereof.

Cell cultures

Cell cultures for use in the present invention include 2-dimensional and 3-dimensional cell cultures. As described herein, a cell culture will generally be contained or cultured in one or more modules of one or more pumps and/or a connector structure. The cell culture will be exposed to the test atmosphere so that the effect of the test atmosphere on the cell culture can be determined. Suitably, two or more cell cultures will be located at different locations around one or more pumps and/or connectors and/or system so that the effect of the test atmosphere on the cell cultures at these different locations that mimic the respiratory tract can be determined. 2D cell cultures involve growing cells in flat layers on plastic surfaces, which allows the study of several aspects of cell physiology and responses to stimuli such as test atmosphere(s), but they do not reflect the actual structure and architecture of an organ. In 2-dimensional monolayers, the extracellular matrix, intercellular interactions, and cell-matrix interactions that are essential for differentiation, proliferation, and cellular functions are lost. In 3-dimensional culture systems, functional tissue with characteristics similar to those observed in vivo can be formed. Compared to 2D culture systems, 3D cell culture allows cells to interact with their environment in all three dimensions and is physiologically more significant. Such cells can show improved viability, proliferation, differentiation, morphological characteristics, responses to stimuli, drug metabolism, gene expression and protein synthesis, etc. During 3D cell culture, specific tissue-like structures can form and mimic the functions and responses of real cells. tissues is physiologically more significant than in traditional 2-dimensional cell monolayers. Several 3D tissues that mimic human organs are commercially available. For example, 3-dimensional organotypic tissues of the lung, which are of particular interest in the context of the present invention, can be obtained using primary human cells grown at the fluid-air interface (ALI), where these cells will differentiate and form a functional tissue. These 3-dimensional tissues are morphologically and metabolically similar to human bronchial tissues. They consist of basal, goblet, and ciliated cells arranged in a pseudostratified structure. Like the lung, actively beating cilia are present, allowing their function and activity to be studied. Similar levels of mRNA encoding enzymes that metabolize xenobiotics were found in these 3D ALI cultures compared to human lungs. In addition, these tissues can be maintained in vitro for an extended period of time. This 3D lung tissue model is a suitable model for investigating the effects of test atmospheres and the like according to the present invention.

The term "3-dimensional cell culture" includes any method that provides cell culture in 3 dimensions, with or without the use of a matrix or substrate. A number of different 3D cell culture methods have been developed, including spheroid cultures and organotypic cultures.

spheroids

The term "spheroid" has the meaning commonly understood in the art, which is represented by a single cell dividing to form a 3-dimensional spheroid structure, or a 3-dimensional aggregate of multiple cells with or without the use of a matrix or support to support cell growth in 3 dimensions. in a spheroid. The 3-dimensional spheroid can be an adhesive spheroid or a suspension-growing spheroid. Several different systems are available for culturing spheroids for use in the present invention, including spheroids grown as aggregates, for example, on NanoCulture plates, in suspension culture, on gels, on poly-HEMA-coated plastic material, by cell encapsulation, or in the form of aggregates using the "hanging drop" system. Other methods include the use of shake bottles, rotary systems, concave well plate methods, and liquid culture under non-adhesive conditions. Bioreactors can also be adapted for use in 3D spheroid cell culture. In one embodiment, the method employed is a hanging drop system, such as the GravityPLUS hanging drop system (InSphero). This method involves the use of a GravityTRAP ULA plate, which is a non-adhesive coated microtiter plate designed to produce spheroids. Spheroid maturation usually occurs within 2–5 days after seeding, depending on cell type and culture conditions. The spheroids are preferably cultured in a volume of 100 µl or more, or 200 µl or more, or 300 µl or more. The spheroids are preferably cultured in Corning microplates.^® for spheroids.

3D cell culture arrays or supports can be used to culture spheroids. They are often porous substrates that can support cell growth and differentiation in 3 dimensions. A variety of materials have been developed to obtain 3D substrates, differing in appearance, porosity, permeability, mechanical characteristics, and nanoscale surface morphological characteristics. Examples of such materials include collagen gels, sponge materials or biogels; fibrin; fibronectin; laminin; alginates, hydrogels; cross-linked glycosaminoglycan; polymer substrates, synthetic substrates; peptide supports; and chitosan composite supports.

3D spheroids more closely resemble in vivo tissue in terms of their cellular communication and formation of extracellular matrices. These matrices help cells move within the spheroid, similar to how cells move in living tissue. Thus, spheroids represent much improved models of differentiation, survival, cell migration, cell polarization, gene expression, and growth.

The spheroids can be collected and studied using various methods well known in the art, including colorimetric, fluorescent and luminescent assays with measurements using a plate reader, or they can be easily observed using microscopy. Additional techniques include Western, Northern, or Southern blotting, histological techniques (e.g., immunohistochemistry, in situ hybridization , immunofluorescence), and the like. Optical imaging techniques such as inverted brightfield microscopy, fluorescence, single photon emission computed tomography (SPECT), positron emission tomography (PET), magnetic resonance imaging (MRI), and Cherenkov luminescent imaging (CLI).

Applications for 3D spheroids include the study of cell and tissue proliferation in vitro in an environment that is closest to that found in vivo , screening of compounds and test atmospheres, toxicological assays and clinical trials, etc.

The use of spheroids in 3D cell culture is generally reviewed in Expert Opin. Drug Discov . (2015) 10, 519-540. In vitro , lung spheroid cells can proliferate and can form alveolar-like structures and acquire mature lung epithelial phenotypes.

Cell Sources

Lung cells and cell lines for use in the present invention can be isolated from tissue or fluid using methods well known in the art. They can be obtained by differentiation from stem cells, such as embryonic stem cells or induced pluripotent stem cells, or directly obtained by differentiation from somatic cells. Cells and cell lines can be derived from human or animal subjects, or from human or animal cells, including any of a number of mammalian species, preferably human, but including rat, mouse, pig, rabbit, and primates other than human, etc., or originate from them. Cells and cell lines can be obtained from commercial sources. In some embodiments, it is desirable to use human cells.

One cell type of interest is lung cells, including lung epithelial cells. Bronchial and/or respiratory epithelial cells are particularly useful in the present invention. Human bronchial epithelial cells can be collected by brushing a donor lung biopsy during a bronchoscopy procedure. In one embodiment, the lung cells are normal human bronchial epithelial cells (NHBE). Lung epithelial cells can be cultured as a monolayer of undifferentiated cells, or they can be further formed into an organotypic tissue similar to lung epithelium at the interface between liquid and air. It is possible to obtain established cell lines at the liquid-air interface using the following technique. Briefly, epithelial cells can be cultured in a flask to increase cell numbers. After the incubation period, the cells are separated from the flask, counted, and plated on inserts. On these inserts, cells are incubated with media on the apical and basal sides. This phase ensures that the cells will divide and completely cover the liner to form the epithelium. The apical medium is then removed, the basal medium is retained and replaced with a more complete medium. Cultures are incubated in this manner for an additional period of time. During this time, cells differentiate into 3 cell types: basal, goblet, and ciliated cells. At the end of maturation cultures are ready for use. The use of an air-liquid interface for culturing human nasal epithelial cells is described in J Vis Exp. 2013; (80): 50646.

Lung epithelial cells can be obtained from human or animal subjects with various pathologies, including subjects classified as smokers or non-smokers.

Research

The present invention can be used for various applications to study the effect of the test atmosphere(s) on the airway model. For example, the present invention can be used in an in vitro study of inhalation toxicology, a study of aerosol dynamics in the respiratory tract (e.g., deposition of aerosol particles and absorption of gases in cell cultures), or a study of metabolic activity or transport of atmosphere(s) tests (e.g., aerosol molecules) through the epithelium of the respiratory tract. The present invention can be used to test the effect of aerosol(s), smoke or tobacco products, or the effect of inhalers such as medical inhalers. The present invention can be used to test the effect of aerosol(s), smoke or tobacco products, or the effect of medical inhalers on cells in one or more parts of the respiratory tract.

One aspect relates to a method for determining the exposure of a test atmosphere to a cell culture—such as one or more cell cultures—contained in a model airway, comprising the steps of: (a) providing the system described herein, the system comprising the cell culture in one or more modules; and (b) comparing the cell culture before and/or after exposure to the test atmosphere, wherein the difference between the cell culture before and/or after exposure of the cells to the test atmosphere is indicative of whether the test atmosphere is affecting the cell culture.

In an embodiment where cell culture differences are determined after cells are exposed to the test atmosphere, the cell culture exposed to the test atmosphere can be compared to a cell culture that has not been exposed to the test atmosphere, or to a cell culture that has been exposed to a control atmosphere, such as an atmosphere containing no test atmosphere. According to this embodiment, the difference between a cell culture exposed to a test atmosphere and a cell culture not exposed to a test atmosphere, or a difference between a cell culture exposed to a test atmosphere and a cell culture exposed to a control atmosphere such as an atmosphere, is not containing the test atmosphere - is an indication that the test atmosphere affects the cell culture.

Another aspect relates to a method for modeling the interaction between a test atmosphere and a model airway in the system described herein, comprising the steps of: (a) with the first valve of the first pump open and the second valve of the first pump closed, providing a gas containing the test atmosphere in the first pump through the first channel; (b) closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; (c) actuating the second pump to draw the test atmosphere into the connection structure and blowing the first pump chamber and connection structure with ambient air; (d) opening the first valve of the first pump towards the ambient air and creating a tight connection between the first channel and the second channel of the first pump; and (e) after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump.

Another aspect relates to a method for determining the effect of a test atmosphere on a simulated airway in the system described herein, comprising the steps of: (a) with the first valve of the first pump open and the second valve of the first pump closed, providing a gas containing the test atmosphere to the first pump through the first channel; (b) closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; (c) actuating the second pump to draw the test atmosphere through the connecting structure and blowing the first pump chamber and the connecting structure with ambient air; (d) opening the first valve of the first pump towards the ambient air and creating a tight connection between the first channel and the second channel of the first pump; and (e) after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump; wherein the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options, and this method includes the additional step of determining the effect of the test atmosphere on cell culture, wherein the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture.

In an embodiment where cell culture differences are determined after cells are exposed to the test atmosphere, the cell culture exposed to the test atmosphere can be compared to a cell culture that has not been exposed to the test atmosphere, or to a cell culture that has been exposed to a control atmosphere, such as an atmosphere containing no test atmosphere. According to this embodiment, the difference between a cell culture exposed to a test atmosphere and a cell culture not exposed to a test atmosphere, or a difference between a cell culture exposed to a test atmosphere and a cell culture exposed to a control atmosphere such as an atmosphere, is not containing the test atmosphere - is an indication that the test atmosphere affects the cell culture.

An additional aspect relates to a method for modeling the interaction between the test atmosphere and the airway model, including the following steps: (a) providing a test atmosphere in the first pump chamber; (b) withdrawing the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; (c) blowing the first pump and at least part of the connection structure with ambient air; (d) holding the test atmosphere in the second pump and connection structure for a specified period of time; (e) displacement of the test atmosphere into the connecting structure and the first pump by means of the second pump; and (f) performing one or more cycles of pumping ambient air into the second pump; while the test atmosphere comes into contact with the cell culture located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options.

An additional aspect relates to a method for determining the effect of the test atmosphere on the airway model, including the following steps: (a) providing a test atmosphere in the chamber of the first pump; (b) withdrawing the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; (c) blowing the first pump and at least part of the connection structure with ambient air; (d) holding the test atmosphere in the second pump and connection structure for a specified period of time; (e) displacement of the test atmosphere through the connecting structure and the first pump with the help of the second pump; and (f) performing one or more cycles of pumping ambient air into the second pump; wherein the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options, and this method contains the additional step of determining the effect of the test atmosphere on cell culture, wherein the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture. In an embodiment where cell culture differences are determined after cells are exposed to the test atmosphere, the cell culture exposed to the test atmosphere can be compared to a cell culture that has not been exposed to the test atmosphere, or to a cell culture that has been exposed to a control atmosphere, such as an atmosphere containing no test atmosphere. According to this embodiment, the difference between a cell culture exposed to a test atmosphere and a cell culture not exposed to a test atmosphere, or a difference between a cell culture exposed to a test atmosphere and a cell culture exposed to a control atmosphere such as an atmosphere, is not containing the test atmosphere - is an indication that the test atmosphere affects the cell culture.

The effect of the test atmosphere(s) may be studied in the presence of one or more agents. The agent(s) may include, without limitation, a drug, toxin, pathogen, protein, nucleic acid, antigen, antibody, and chemical compound, etc. Examples of effects that can be measured include oxygen uptake, carbon dioxide production, cell viability, protein expression, enzyme activity, permeation, permeation barrier function, surfactant production, cytokine response, transporter functions, cytochrome P450 expression, albumin secretion, toxicology, and etc.

Several assays can be run in parallel with different concentrations of test atmosphere and/or agent to obtain a differential response to different concentrations.

The agent may be any test compound of interest and includes small organic compounds, polypeptides, peptides, relatively high molecular weight carbohydrates, polynucleotides, fatty acids and lipids, an aerosol or one or more components of an aerosol, etc. Test compounds can be subjected to screening individually or in groups or combinatorial libraries of compounds. Test compounds can be obtained from a wide variety of sources, including libraries of synthetic or natural compounds. You can use libraries of natural compounds in the form of bacterial, fungal, plant and animal extracts. Natural or synthetically derived libraries and compounds modified by conventional chemical, physical, and biochemical methods can be used to generate combinatorial libraries. Known pharmacological agents can be subjected to targeted or random chemical modifications, such as acylation, alkylation, esterification, conversion to an acid form, to obtain structural analogues for screening.

One or more variables that can be measured include cell elements, subcellular material, subcellular components, or cellular products. As an example, the toxicology of the test atmosphere can be measured. Airway aerosol dynamics (eg, deposition of aerosol particles and absorption of gases into cell cultures) can be measured. As a further example, metabolic activity and/or transport of molecules across the airway epithelium can be studied.

A computer

Also disclosed is a computer program that, when executed by a computer/processor, can be used to control a computer to perform one or more of the methods described herein.

One skilled in the art will readily appreciate that the steps of the various methods described above may be performed by programmed computers. In this context, it is contemplated that some embodiments also include a program storage device, such as digital storage media, that can be read by machines or computers and may contain machine-executable or computer-executable program instructions, said instructions executing one or all steps in the above methods. Program storage devices may be, for example, digital storage devices, magnetic storage devices such as magnetic disks and magnetic tapes, hard disks or optical digital data carriers. It is also contemplated that embodiments include computers programmed to perform some or all of the steps of the methods described above.

The functions of various components, including processors or logic circuits, can be provided by using dedicated hardware as well as hardware capable of running software in combination with suitable software. When the functions are provided by a processor, they may be provided by a single dedicated processor, a single shared processor, or multiple separate processors, some of which may be shared. Moreover, explicit use of the term "processor" or "controller" or "logic circuit" should not be construed as referring solely to hardware capable of executing software, and may implicitly include, without limitation, a Digital Signal Processor (DSP). ), a network processor, an application specific integrated circuit (ASIC), a field programmable gate array (FPGA), a read only memory (ROM) for storing software, a random access memory (RAM), and a non-volatile data storage device. Other hardware, conventional and/or custom, may also be included. Likewise, all switches depicted in the figures are only concepts. Their function can be performed through the use of software logic components, dedicated logic components, through the interaction of program control and a dedicated logic component, or even manually, and the particular technique can be chosen by its implementer, as in particular can be understood from the context.

Additional aspects of the present invention are set forth in the following numbered paragraphs:

1. A system for determining the interaction between the test atmosphere and the model of the respiratory tract, and the specified system contains: (a) the first pump containing: (i) a chamber configured to contain the first volume of gas containing the test atmosphere; (ii) a first channel capable of receiving and discharging gas and containing a valve for adjusting the flow of gas passing through the first channel, wherein said valve is movable between open and closed positions, while in the open position, said valve is able to open in the direction test atmosphere or ambient air; (iii) a second channel configured to receive and release gas and containing a valve for adjusting the flow of gas passing through the second channel, and the specified valve is movable between open and closed positions; (iv) a piston plate containing a plurality of holes made in it, and one or more of the holes contain valves adapted to regulate the flow of gas passing through the holes; and (v) a motor for controlling the operation of the first pump; (b) a second pump comprising: (i) a chamber configured to contain a second volume of gas, wherein the first and second volumes of gas are different from each other; (ii) a channel configured to receive and release gas; and (iii) a motor for controlling the operation of the second pump; (c) a connection structure used to transfer gas from the first pump to the second pump; and (d) one or more openings in the first pump or the second pump or the walls of the connecting structure, or a combination of two or more of these options, and these openings are capable of receiving a module for containing cell culture medium or configured to store cell culture medium , or to monitor the conditions in the chamber, or configured to monitor the conditions in the chamber, or for gas sampling or configured to take gas samples, or to characterize the gas, or configured to characterize the gas.

2. The system according to claim. 1, characterized in that the pumps are piston pumps containing a piston plate and a base.

3. The system according to claim 1 or claim 1, characterized in that the connecting structure is hollow.

4. The system according to any one of the preceding claims, characterized in that the connecting structure is branched.

5. The system according to claim 4, characterized in that one end branch of the connecting structure is connected to a second pump and one or more other end branches are connected to a separate pump, each individual pump comprising: (i) a chamber for containing a volume of gas, this the volume of gas is identical to the second volume of gas in the second pump; (ii) a conduit for receiving and discharging gas and for connection to the connecting structure; and (iii) a motor for driving the pump.

6. System according to claim 5, characterized in that each individual pump is identical to the second pump.

7. A system according to any one of the preceding claims, characterized in that the system is contained in a housing, suitably in a temperature controlled housing.

8. The system according to claim. 7, characterized in that the temperature of the casing is regulated by a thermostat.

9. System according to claim 8, characterized in that the temperature in the casing is approximately 37 °C.

10. The system according to any one of the preceding claims, characterized in that the different volumes of the first and second pumps represent the internal volume of different parts of the respiratory tract, suitably the human respiratory tract.

11. The system according to any one of paragraphs. 6-10, characterized in that the volumes of the individual pumps represent the internal volume of different sections of the respiratory tract, suitably the human respiratory tract.

12. The system according to any one of the preceding claims, characterized in that the working volume of the first and second pumps is at least equal to the maximum achievable inlet volume in the corresponding section of the respiratory tract.

13. The system according to any one of paragraphs. 5-12, characterized in that the working volume of the individual pumps is at least equal to the maximum achievable inlet volume in the corresponding section of the respiratory tract.

14. The system according to any one of the preceding claims, characterized in that the discharge pressure of the motor or pump can be at atmospheric pressure or above or below atmospheric pressure.

15. The system according to any one of the preceding claims, characterized in that the working volume of the first pump is from about 0 to 100 ml or from about 1 to 100 ml.

16. A system according to any one of the preceding claims, characterized in that the working volume of the second pump is from about 0 to 4000 ml, or from about 1 to about 4000 ml.

17. System according to any one of the preceding claims, characterized in that the pumps comprise or consist of stainless steel.

18. System according to any one of the preceding claims, characterized in that the chamber is a cylinder.

19. System according to any one of the preceding claims, characterized in that the chamber contains glass.

20. The system according to any one of the preceding claims, characterized in that the volume of the chamber of the first pump is less than that of the chamber of the second pump.

21. The system according to any of the preceding claims, characterized in that the volume of the chamber of the first pump represents the volume of the oral cavity and oropharynx.

22. The system according to any one of the preceding claims, characterized in that the volume of the chamber of the second pump represents the volume of the cavity of the lung or part thereof.

23. The system according to any one of the preceding claims, characterized in that the volume of the connecting structure represents the volume of the conducting airways of the lung, suitably the human lung.

24. The system according to any one of the preceding claims, characterized in that the piston plate of the first pump contains one or more gaps for the entry or flow of gas.

25. The system according to any one of the preceding claims, characterized in that each of the one or more lumens includes a valve movable between open and closed positions and capable of controlling the flow or flow of gas.

26. The system according to any one of the preceding claims, characterized in that the system further comprises a computer controller capable of synchronizing the operation of the system.

27. A system according to any one of the preceding claims, characterized in that one or more of the first pump or the second pump or connection structure comprises one or more modules containing a quartz crystal microbalance.

28. The system according to any one of the preceding claims, characterized in that the openings in the first pump or the second pump or the walls of the connecting structure are threaded or non-threaded.

29. System according to any one of the preceding claims, characterized in that one or more holes contain a module.

30. The system of claim 29, wherein the one or more modules are adapted to contain the cell culture medium, or are configured to store the cell culture medium, or are adapted to monitor conditions in the chamber, or are adapted to monitor conditions in the chamber, or adapted for gas sampling or adapted for gas characterization.

31. The system according to claim 29 or 30, characterized in that the modules are located on the basis of the piston plate of the first and/or second pump and/or in the walls of the connecting structure.

32. The system according to any one of paragraphs. 29-31, characterized in that one or more modules contain a cell culture medium.

33. The system according to any one of the preceding claims, characterized in that the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

34. The system according to any one of paragraphs. 29-32, characterized in that the modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and an optional microfluidic pump connected to it.

35. The system according to any one of paragraphs. 29-34, characterized in that the modules are located in a horizontal plane in one or more of the following: the first pump or the second pump, or the walls of the connecting structure.

36. A system according to any one of the preceding claims, characterized in that the connecting structure comprises stainless steel.

37. A system according to any one of the preceding claims, characterized in that the first pump chamber has a volume of approximately 100 ml.

38. A system according to any one of the preceding claims, characterized in that the second pump chamber has a volume of from about 1 liter to about 4 liters.

39. A pump for displacing a volume of gas, comprising: (i) a chamber configured to contain a volume of gas and containing a base and one or more openings capable of accommodating one or more modules for containing or storing cell culture medium, or for monitoring conditions in a chamber, or for gas sampling, or for gas characterization; (ii) a first channel for receiving and discharging gas when it is contained in the chamber, and containing a first valve for adjusting the flow of gas passing through the first channel, wherein said first valve is movable between open and closed positions, and in the open position, said the valve may open towards the test atmosphere or ambient air; and (iii) a second passage for receiving and discharging gas when contained in the chamber and containing a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions; and (iv) a piston plate containing a plurality of holes made therein, wherein one or more of the holes contain valves adapted to control the flow of gas passing through the holes.

40. The pump according to claim 39, characterized in that the pump is a piston pump containing a piston plate.

41. The pump according to claim 39 or 40, characterized in that one or more holes in the chamber are threaded or threadless.

42. The pump according to any one of paragraphs. 39-41, characterized in that one or more holes contain a module.

43. The pump according to claim 42, characterized in that the module is threaded or threadless.

44. The pump according to claim. 42 or 43, characterized in that one or more modules are adapted to contain the cell culture medium or are configured to store the cell culture medium, or adapted to monitor conditions in the chamber or adapted to monitor conditions in chamber, or adapted for gas sampling or adapted for gas characterization.

45. The pump according to any one of paragraphs. 42-44, characterized in that the modules are located on the basis of the pump.

46. The pump according to any one of paragraphs. 42-45, characterized in that one or more modules contain a medium for culturing cells.

47. The pump according to claim 46, characterized in that the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

48. The pump according to any one of paragraphs. 44-47, characterized in that the modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected to it.

49. The pump according to any one of paragraphs. 42-48, characterized in that the modules are located in a horizontal plane in one or more of the following: the first pump or the second pump, or the walls of the connecting structure.

50. The pump according to any one of paragraphs. 43-49, characterized in that the module contains a microbalance on a quartz crystal.

51. The pump according to any one of paragraphs. 39-50, characterized in that the pump additionally contains a motor.

52. The pump according to any one of paragraphs. 39-51, characterized in that the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

53. The pump according to any one of paragraphs. 39-52, characterized in that the working volume of the pump is from about 0 to 100 ml, or from about 1 to about 100 ml.

54. The pump according to any one of paragraphs. 39-53, characterized in that the pump contains stainless steel.

55. The pump according to any one of paragraphs. 39-54, characterized in that the chamber is a cylinder.

56. The pump according to any one of paragraphs. 39-55, characterized in that the chamber contains glass.

57. The pump according to any one of paragraphs. 39-56, characterized in that the pump chamber has a volume of approximately 100 ml.

58. The pump according to any one of paragraphs. 39-57, characterized in that the pump piston plate contains one or more gaps for gas inflow or inflow.

59. The pump of claim. 58, characterized in that one or more gaps include a valve that is movable between open and closed positions and is able to regulate the flow or flow of gas.

60. A piston pump for displacing a volume of gas, comprising: (i) a chamber configured to contain a volume of gas and containing a piston plate containing one or more gaps for the entry or flow of gas into the chamber, with one or more, or each, of the lumens include a valve movable between open and closed positions and capable of controlling the flow or flow of gas; (ii) a first channel for receiving gas, and containing a first valve for adjusting the flow of gas passing through the first channel, and the specified first valve is movable between open and closed positions; and (iii) a second passage for venting gas when contained in the chamber and comprising a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions.

61. The pump according to claim 60, characterized in that the chamber includes a base and contains one or more holes.

62. The pump according to claim 61, characterized in that the holes are threaded or threadless.

63. The pump according to claim 60 or 61, characterized in that the holes contain a module in one or more holes.

64. The pump according to claim 63, characterized in that the module is threaded or threadless.

65. The pump according to claim. 63 or 64, characterized in that the module is adapted to contain or store the medium for culturing cells, or to monitor conditions in the chamber, or for gas sampling, or for characterizing the gas

66. The pump according to any one of paragraphs. 63-65, characterized in that one or more modules contain a medium for cell cultivation.

67. The pump according to claim 66, wherein the cell culture medium contains a cell culture, suitably a 2- or 3-dimensional cell culture.

68. The pump according to any one of paragraphs. 63-67, characterized in that the modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected to it.

69. The pump according to any one of paragraphs. 63-67, characterized in that the module contains a microbalance on a quartz crystal.

70. The pump according to any one of paragraphs. 60-69, characterized in that the connecting structure is connected to the second channel.

71. The pump according to any one of paragraph 70, characterized in that the connecting structure is hollow.

Suitably, the pump further comprises a motor.

72. The pump according to any one of paragraphs. 60-71, characterized in that the pump discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

73. The pump according to any one of paragraphs. 60-72, characterized in that the working volume of the pump is from about 0 to 100 ml or from about 1 to about 100 ml.

74. The pump according to any one of paragraphs. 60-73, characterized in that the pump contains stainless steel.

75. The pump according to any one of paragraphs. 60-74, characterized in that the chamber is a cylinder.

76. The pump according to any one of paragraphs. 60-75, characterized in that the chamber contains glass.

77. The pump according to any one of paragraphs. 60-76, characterized in that the pump chamber has a volume of approximately 100 ml.

78. A gas volume displacement pump, comprising: (i) a chamber configured to contain a gas volume, said chamber comprising a base and one or more modules for containing or storing cell culture medium, or for monitoring conditions in the chamber, or for gas sampling, or for gas characterization; and (ii) a conduit used to receive and release the gas.

79. The pump according to claim 78, characterized in that the pump is a piston pump containing a piston plate.

80. The pump according to claim 79, characterized in that the piston plate does not contain any gaps or holes.

81. The pump according to any one of paragraphs. 78-80, characterized in that the modules are located at the base of the chamber.

82. The pump according to any one of paragraphs. 78-81, characterized in that the modules are threaded or threadless.

83. The pump according to any one of paragraphs. 78-82, characterized in that the modules are adapted to contain the cell culture medium, or are configured to store the cell culture medium, or are adapted to monitor the conditions in the chamber, or are adapted to monitor the conditions in the chamber, or are adapted to take gas samples, or adapted to characterize the gas.

84. The pump according to any one of paragraphs. 78-83, characterized in that one or more modules contain a medium for culturing cells.

85. The pump according to claim 84, characterized in that the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

86. The pump according to claim 84 or 85, characterized in that the modules adapted to contain or store the cell culture medium further comprise a microfluidic channel and optionally a microfluidic pump connected to it.

87. The pump according to any one of paragraphs. 78-82, characterized in that the module contains a microbalance on a quartz crystal.

88. The pump according to any one of paragraphs. 78-87, characterized in that the connecting structure is connected to the channel.

89. The pump according to any one of paragraphs. 78-88, characterized in that the connecting structure is hollow.

90. The pump according to any one of paragraphs. 78-89, characterized in that the pump additionally contains a motor.

91. The pump according to any one of paragraphs. 78-90, characterized in that the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

92. The pump according to any one of paragraphs. 78-91, characterized in that the displacement of the pump is from about 0 to 1000 ml, or from about 1 to about 100 ml.

93. The pump according to any one of paragraphs. 78-92, characterized in that the pump contains stainless steel.

94. The pump according to any one of paragraphs. 78-93, characterized in that the chamber is a cylinder.

95. The pump according to any one of paragraphs. 78-94, characterized in that the chamber contains glass.

96. The pump according to any one of paragraphs. 78-95, characterized in that the volume of the chamber represents the volume of the cavity of the lung or part thereof.

97. A connection structure adapted to connect at least two pumps for transferring gas between them, said connection structure comprising a hollow channel and one or more threaded or non-threaded holes in the walls of the connection structure.

98. The connecting structure according to claim 97, characterized in that the threaded holes contain a threaded module in one or more holes, and the specified module is adapted to contain the cell culture medium or is configured to store the cell culture medium, or is adapted to monitor conditions in the chamber, or adapted to monitor the conditions in the chamber, or adapted to take gas samples, or adapted to characterize the gas.

99. The connection structure according to claim 98, characterized in that one or more modules contain a cell culture medium.

100. The connector structure according to claim 99, characterized in that the cell culture medium contains or is in contact with the cell culture, suitably a 2- or 3-dimensional cell culture.

101. Connecting structure according to any one of paragraphs. 98-100, characterized in that the modules adapted to contain or store the cell culture medium additionally contain a microfluidic

102. Connecting structure according to any one of paragraphs. 97-102, characterized in that the channel and optionally a microfluidic pump are attached to it.

103. Connecting structure according to any one of paragraphs. 97-102, characterized in that the connecting structure is hollow.

104. Connecting structure according to any one of paragraphs. 97-103, characterized in that the connecting structure is branched.

105. The connecting structure according to claim 104, characterized in that each end branch of the connecting structure can be connected to a separate pump.

106. Connecting structure according to any one of paragraphs. 97-105, characterized in that the connecting structure represents the volume of the conducting airways of the lung, suitably the human lung.

107. Connecting structure according to any one of paragraphs. 98-106, characterized in that the modules are located in a horizontal plane in the walls of the connecting structure.

108. Connecting structure according to any one of paragraphs. 98-107, characterized in that the modules are adapted to contain a cell culture, suitably a 2- or 3-dimensional cell culture.

109. Connecting structure according to any one of paragraphs. 98-108, characterized in that the module is a chamber for containing a cell culture, and said chamber contains a microfluidic channel and optionally a microfluidic pump connected to it.

110. Connecting structure according to any one of paragraphs. 98-103, characterized in that the modules are adapted to monitor conditions in the connecting structure and/or to take gas samples and/or to characterize the gas.

111. Connecting structure according to any one of paragraphs. 97-110, characterized in that the connecting structure contains stainless steel.

112. A system containing one or more pumps, as defined in any of paragraphs. 39-96.

113. The system according to p. 112, characterized in that the system further comprises a connecting structure, defined in any of paragraphs. 97-111.

114. The system according to item 112 or item 112, characterized in that the pumps are connected using a connecting structure.

115. A method for modeling the interaction between the test atmosphere and the model of the respiratory tract, including the use of a system according to any one of paragraphs. 1-38.

116. The use of the system according to any one of paragraphs. 1-38 to simulate the interaction between the test atmosphere and the airway model.

117. A method for determining the impact of the test atmosphere on a cell culture contained in a model of the respiratory tract, including the use of a system according to any one of paragraphs. 1-38.

118. The use of the system according to any one of paragraphs. 1-38 to determine the effect of the test atmosphere on the cell culture contained in the airway model

119. A method for determining the effect of a test atmosphere on a cell culture contained in a model respiratory tract, comprising the following steps: (a) providing the system described herein, the system containing the cell culture in one or more modules; and (b) comparing the cell culture before and/or after exposure to the test atmosphere, wherein the difference between the cell culture before and/or after exposure of the cells to the test atmosphere is indicative of whether the test atmosphere is affecting the cell culture.

120. A method for modeling the interaction between a test atmosphere and a model airway in the system described herein, comprising the following steps: (a) with the first valve of the first pump open and the second valve of the first pump closed, providing gas containing the test atmosphere to the first pump through first channel; (b) closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; (c) actuating the second pump to draw the test atmosphere into the connection structure and blowing the first pump chamber and connection structure with ambient air; (d) opening the first valve of the first pump towards the ambient air and creating a tight connection between the first channel and the second channel of the first pump; and (e) after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump.

121. A method for determining the effect of a test atmosphere on a model airway in the system described herein, comprising the following steps: (a) with the first valve of the first pump open and the second valve of the first pump closed, providing gas containing the test atmosphere in the first pump through the first channel; (b) closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; (c) actuating the second pump to draw the test atmosphere through the connecting structure and blowing the first pump chamber and the connecting structure with ambient air; (d) opening the first valve of the first pump towards ambient air and creating a tight connection between the first channel and the second channel of the first pump; and (e) after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump; wherein the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options, and this method includes the additional step of determining the effect of the test atmosphere on cell culture, wherein the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture.

122. The method of claim 121, wherein the modules are adapted to monitor system conditions and/or to take gas samples and/or to characterize the gas, and said method comprises obtaining one or more measurements from the modules.

123. A method for modeling the interaction between the test atmosphere and the model of the respiratory tract, including the following steps: (a) providing a test atmosphere in the first pump chamber; (b) withdrawing the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; (c) blowing the first pump and at least part of the connection structure with ambient air; (d) holding the test atmosphere in the second pump and connection structure for a specified period of time; (e) displacement of the test atmosphere in the connecting structure and the first pump by means of the second pump; and (f) performing one or more cycles of pumping ambient air into the second pump; while the test atmosphere comes into contact with the cell culture located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options.

124. The method of claim. 123, wherein step (d) includes holding the test atmosphere in the second pump and the part of the connecting structure that still contains the test atmosphere for a certain period of time.

125. The method according to claim 123 or 124, characterized in that the pumps are piston pumps containing a piston plate and a base.

126. The method according to any one of paragraphs. 123-125, characterized in that the first pump is the pump described in any one of paragraphs. 39-77.

127. The method according to any one of paragraphs. 123-126, characterized in that the second pump is the pump described in any one of paragraphs. 78-96.

128. The method according to any one of paragraphs. 123-127, characterized in that the connecting structure is a connecting structure described in any of paragraphs. 97-111.

129. The method according to any one of paragraphs. 123-128, characterized in that the method is carried out in a casing, suitably in a temperature controlled casing.

130. The method according to any one of paragraphs. 123-129, characterized in that the temperature of the casing is controlled by a thermostat.

131. The method according to any one of paragraphs. 123-130, characterized in that the temperature in the casing is approximately 37 °C.

132. The method according to any one of paragraphs. 123-131, characterized in that the different volumes of the first and second pumps represent the internal volume of different sections of the respiratory tract, suitably the human respiratory tract.

133. The method according to any one of paragraphs. 123-132, characterized in that the working volume of the first and second pumps is at least equal to the maximum achievable inlet volume in the corresponding section of the respiratory tract.

134. The method according to any one of paragraphs. 123-133, characterized in that the discharge pressure corresponds to atmospheric pressure or pressure above or below atmospheric pressure.

135. The method according to any one of paragraphs. 123-134, characterized in that the working volume of the first pump is from about 0 to 100 ml, or from about 1 to about 100 ml.

136. The method according to any one of paragraphs. 123-135, characterized in that the working volume of the second pump is from about 0 to 4000 ml or from about 1 to about 4000 ml.

137. The method according to any one of paragraphs. 123-136, characterized in that the volume of the chamber of the first pump is less than the volume of the chamber of the second pump.

138. The method according to any one of paragraphs. 123-137, characterized in that the volume of the chamber of the first pump represents the volume of the oral cavity and oropharynx, suitably of the human oral cavity and oropharynx.

139. The method according to any one of paragraphs. 123-138, characterized in that the volume of the chamber of the second pump represents the volume of the cavity of the lung or part thereof, suitably the cavity of the human lung or part thereof.

140. The method according to any one of paragraphs. 123-139, characterized in that the connecting structure represents the volume of the conducting airways of the lung, suitably the human lung.

141. The method according to any one of paragraphs. 123-140, characterized in that the piston plate contains one or more gaps for the entry or flow of gas into the chamber.

142. The method according to any one of paragraphs. 123-141, characterized in that each of the one or more lumens includes a valve movable between open and closed positions and capable of controlling the flow or flow of gas.

143. The method according to any one of paragraphs. 123-142, characterized in that the cell culture is located on the basis of the piston plate of the first and/or second pump and/or in the walls of the connecting structure.

143. The method according to any one of paragraphs. 122-142, characterized in that the cell culture is a 2- or 3-dimensional culture.

144. The method according to any one of paragraphs. 122-143, characterized in that the method further comprises monitoring conditions and/or gas sampling and/or gas characterization in one or more of the first pump or second pump or connection structure using one or more modules contained therein.

145. The method according to any one of paragraphs. 122-144, characterized in that the chamber containing the cell culture further comprises a microfluidic channel and optionally a microfluidic pump connected to it.

145. The method according to any one of paragraphs. 122-145, characterized in that the connecting structure contains stainless steel.

146. The method according to any one of paragraphs. 122-145, characterized in that the first pump chamber has a volume of approximately 100 ml.

147. The method according to any one of paragraphs. 122-146, characterized in that the chamber of the second pump has a volume of from about 1 liter to about 4 liters.

148. The method of determining the impact of the test atmosphere on the model of the respiratory tract, including the following steps: (a) providing a test atmosphere in the chamber of the first pump; (b) venting the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; (c) blowing the first pump and at least part of the connection structure with ambient air; (d) holding the test atmosphere in the second pump and connection structure for a specified period of time; (e) displacement of the test atmosphere through the connecting structure and the first pump with the help of the second pump; and (f) performing one or more cycles of pumping ambient air into the second pump; wherein the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump or connecting structure, or the second pump, or a combination of two or more of these options, and this method includes the additional step of determining the effect of the test atmosphere on cell culture, wherein the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture.

149. The method according to p. 148, characterized in that the modules are adapted to monitor the conditions of the system and / or to sample the gas, and / or to characterize the gas, and this method includes obtaining one or more measurements from the modules.

Any publication cited or described in this document provides relevant information disclosed prior to the filing date of this application. The statements made herein are not to be construed as an admission that the inventors of the present invention have no basis for setting it apart from such disclosures as before. All publications mentioned in the above description are incorporated into this document by reference. Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been described in connection with specific preferred embodiments, it should be understood that the claimed invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the described embodiments of the present invention, which are obvious to experts in the relevant field of technology, should be included in the scope of the following claims.

Claims (64)

1. A system for determining the interaction between the test atmosphere and the airway model, comprising: first pump containing: a chamber configured to contain a first volume of gas containing the test atmosphere; - the first channel, made with the possibility of receiving and releasing gas and containing a valve for adjusting the flow of gas passing through the first channel, moreover, the specified valve is made movable between open and closed positions, while in the open position, the specified valve is made with the possibility of opening in the direction test atmosphere or ambient air; - the second channel, made with the possibility of receiving and discharging gas and containing a valve for adjusting the flow of gas passing through the second channel, and the specified valve is movable between open and closed positions; - a piston plate in the chamber, containing one or more gaps for the entry or flow of gas into the chamber, while one or more, or each, of the gaps includes a valve configured to move between open and closed positions and regulate the entry or flow of gas ; and - an engine to control the operation of the first pump; second pump containing: a chamber configured to contain a second volume of gas, the first and second volumes of gas being different from each other; - a channel configured to receive and release gas; and - an engine for controlling the operation of the second pump; a connection structure configured to transfer gas from the first pump to the second pump; and one or more openings in the first pump, or the second pump, or the walls of the connecting structure, or a combination of two or more of them, and these openings contain a module for containing cell culture medium, or for monitoring conditions in the chamber, or for sampling gas, or to characterize the gas. 2. A gas displacement pump, comprising: a chamber configured to hold a volume of gas and comprising a base and one or more openings containing one or more modules for holding the cell culture medium, or for monitoring the conditions in the chamber, or for sampling the gas, or for characterizing the gas; the first channel, configured to receive and release gas when it is contained in the chamber, and containing the first valve for adjusting the flow of gas passing through the first channel, and the specified first valve is configured to move between open and closed positions, while in the open position said valve is configured to open towards the test atmosphere or ambient air; a second passage configured to receive and discharge gas when contained in the chamber and comprising a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions; and a piston plate in the chamber containing one or more lumens for gas inflow or inflow into the chamber, wherein one or more, or each, of the lumens includes a valve configured to move between open and closed positions and regulate the inflow or inflow of gas. 3. A piston pump for displacing a volume of gas, comprising: a chamber configured to hold a volume of gas and comprising a piston plate having one or more lumens for gas to enter the chamber, wherein one or more, or each, of the lumens includes a valve movable between an open and a closed position, and controlling the inflow or inflow of gas and including a base containing one or more threaded or non-threaded holes containing a module for containing the cell culture medium, or for monitoring the conditions in the chamber, or for sampling the gas, or for characterizing the gas; the first channel for receiving gas containing the first valve for adjusting the flow of gas passing through the first channel, and the specified first valve is movable between open and closed positions; and a second passage capable of venting gas when contained in the chamber and containing a second valve for adjusting the flow of gas passing through the second passage, said valve being movable between open and closed positions. 4. A gas displacement pump, comprising: a chamber configured to hold a volume of gas, said chamber comprising a base and one or more modules for holding a cell culture medium, or for monitoring conditions in the chamber, or for sampling a gas, or for characterizing a gas; and a channel configured to receive and release gas. 5. A connection structure configured to connect at least two pumps for transferring gas between them, the connection structure comprising a hollow channel and one or more threaded or non-threaded holes in the walls of the connection structure, the holes containing a module for containing a cell culture medium , or to monitor conditions in a chamber, or to take gas samples, or to characterize a gas. 6. A system for determining the interaction between the test atmosphere and the airway model, containing the pump according to claim 2 or 3 and the pump according to claim 4. 7. A method for modeling the interaction between the test atmosphere and the model of the respiratory tract, including the use of the system according to claim 1 or 6. 8. Use of the system according to claim 1 or 6 to simulate the interaction between the test atmosphere and the airway model. 9. A method for determining the effect of the test atmosphere on a cell culture contained in a model respiratory tract, including the use of a system according to claim 1 or 6. 10. Use of the system according to claim 1 or 6 to determine the effect of the test atmosphere on the cell culture contained in the airway model. 11. A method for determining the effect of the test atmosphere on a cell culture contained in a model of the respiratory tract, including: providing a system according to claim 1 or 6, wherein the system contains a cell culture in one or more modules; and comparison of the cell culture before and/or after exposure to the test atmosphere, wherein the difference between the cell culture before and/or after exposure of the cells to the test atmosphere is an indication that the test atmosphere affects the cell culture. 12. A method for modeling the interaction between the test atmosphere and the airway model in the system according to claim 1 or 6, including: with the first valve of the first pump open and the second valve of the first pump closed, providing a gas containing the test atmosphere to the first pump through the first passage; closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; actuating the second pump to draw the test atmosphere into the connecting structure and blowing the chamber of the first pump and the connecting structure with ambient air; opening the first valve of the first pump towards the ambient air and creating a tight connection between the first channel and the second channel of the first pump; and after a certain period of time, the use of the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump. 13. A method for determining the effect of the test atmosphere on the airway model in the system according to claim 1 or 6, including: with the first valve of the first pump open and the second valve of the first pump closed, providing a gas containing the test atmosphere to the first pump through the first passage; closing the first valve and opening the second valve of the first pump and closing the valves on the piston plate of the first pump; actuating the second pump to draw the test atmosphere through the connecting structure and blowing the chamber of the first pump and the connecting structure with ambient air; opening the first valve of the first pump towards the ambient air and creating a tight connection between the first channel and the second channel of the first pump; and after a certain period of time, using the second pump to move the test atmosphere through the connecting structure and through the first valve of the first pump; in this case, the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump, or the connecting structure, or the second pump, or a combination of two or more of these options, and this method includes an additional step of determining the effect of the test atmosphere on cell culture, with the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture. 14. A method for modeling the interaction between the test atmosphere and the model of the respiratory tract, including: providing a test atmosphere in the chamber of the first pump; diverting the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; blowing the first pump and at least part of the connecting structure with ambient air; maintaining the test atmosphere in the second pump and connecting structure for a certain period of time; displacement of the test atmosphere into the connecting structure and the first pump by means of the second pump; and performing one or more cycles of pumping ambient air in the second pump; while the test atmosphere comes into contact with the cell culture located in the first pump, or the connecting structure, or the second pump, or a combination of two or more of these options. 15. A method for determining the impact of the test atmosphere on the airway model, including: providing a test atmosphere in the chamber of the first pump; diverting the test atmosphere from the first pump to a connection structure connecting the first pump to the second pump; blowing the first pump and at least part of the connecting structure with ambient air; maintaining the test atmosphere in the second pump and connecting structure for a certain period of time; displacement of the test atmosphere through the connecting structure and the first pump with the help of the second pump; and performing one or more cycles of pumping ambient air in the second pump; in this case, the test atmosphere comes into contact with the cell culture located in one or more modules located in the first pump, or the connecting structure, or the second pump, or a combination of two or more of these options, and this method includes an additional step of determining the effect of the test atmosphere on cell culture, with the difference in cell culture before and/or after exposure to the test atmosphere is an indication that the test atmosphere affects the cell culture. 16. An aerosol generating device capable of or adapted to carry out the method of claim 14 or 15.

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