CN113413462B - Nano material for simulating natural killer cells to efficiently treat intracellular bacteria and preparation method and application thereof - Google Patents
Nano material for simulating natural killer cells to efficiently treat intracellular bacteria and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a nano material for simulating natural killer cells to efficiently treat intracellular bacteria, and a preparation method and application thereof. The nano material for simulating natural killer cells to efficiently treat intracellular bacteria comprises dendritic mesoporous silica which is used as a carrier and doped with disulfide bonds and protease loaded in a pore channel of the carrier; the protease is preferably perforin and/or granzyme. The nano material for simulating the natural killer cells to efficiently treat the intracellular bacteria takes the dendritic mesoporous silica doped with disulfide bonds as a carrier, and the protease is degraded and released in a response manner under a specific oxidation-reduction potential possessed by an intracellular bacteria model, so that the protease not only can avoid killing the normal cells because the protease is hardly degraded in the normal cells, but also can simulate the natural killer cells to efficiently treat the intracellular bacteria through the specific response release so as to realize the sterilization treatment on infected cells, and further cause a spontaneous immune regulation mechanism of organisms to promote efficient antibacterial and anti-infection application.
Description
Technical Field
The invention belongs to the technical field of organic and inorganic hybrid mesoporous molecular materials, and particularly relates to a nano material for simulating natural killer cells to efficiently treat intracellular bacteria, and a preparation method and application thereof.
Background
Perforin and granzyme are glycoproteins present in Cytotoxic T Lymphocytes (CTL) and Natural Killer (NK) cells. It was found that when cells are invaded by virus or transformed into cancer cells, effector cells contact target cells, active channels are formed on target cell membranes through perforin, and granzyme is released into the target cells, and the target cells are induced to be cracked and killed. This immune-activated regulatory pathway was demonstrated (immunological. today 1991,12, (11), 415-. However, many types of cells do not express perforin or form immune synapses, and NK cells activated in vivo immunity release perforin and granzymes in relatively small amounts. In addition, researchers have found that the effect of perforin and granzyme is largely dependent on the environment in which the cells are placed (immunol. today 2000,21, (6), 254-. Therefore, how to safely realize the controllable release of perforin and granzyme so as to lead NK-like cells to effectively kill intracellular bacteria is a major scientific problem to be solved urgently.
Various means of activating the immune system have been devised by this student in an attempt to achieve active targeting by external drug stimulation to activate NK cells to release perforin and granzyme. As shown by researchers in 2007, granzyme H-assisted granzyme B kills cells infected with adenovirus and kills the infected adenovirus cells through the controlled regulation of granzyme H (Trends Immunol.2007,28, (9), 373-375). However, no report about the simulation construction of the artificial natural killer nanomaterial system exists at present.
Disclosure of Invention
The invention aims to provide a nano material for simulating natural killer cells to efficiently treat intracellular bacteria, and a preparation method and application thereof. The nano material for simulating the natural killer cells to efficiently treat the intracellular bacteria takes the dendritic mesoporous silica doped with disulfide bonds as a carrier, and the protease is degraded and released in a response manner under a specific oxidation-reduction potential possessed by an intracellular bacteria model, so that the protease not only can avoid killing the normal cells because the protease is hardly degraded in the normal cells, but also can simulate the natural killer cells to efficiently treat the intracellular bacteria through the specific response release so as to realize the sterilization treatment on infected cells, and further cause a spontaneous immune regulation mechanism of organisms to promote efficient antibacterial and anti-infection application.
In a first aspect, the invention provides a nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria. The nano material for simulating natural killer cells to efficiently treat intracellular bacteria comprises double-sulfur-bond-doped Dendritic Mesoporous silica (DMON) serving as a carrier and protease loaded in a pore channel of the carrier. Preferably, the protease is a glycoprotein. More preferably, the protease is perforin and/or granzyme.
According to the invention, the disulfide bond is doped in the dendritic mesoporous silica framework, and the collapse and degradation of the framework occur under the condition of the surface oxidation-reduction potential specific to intracellular bacteria, so that the loaded nano-protease is efficiently and specifically released, and the treatment effect of killing the intracellular bacteria is achieved. In addition, bacterial fragments generated by the rupture of intracellular bacteria killed by the nano protease are taken as antigens in an organism to stimulate the humoral immunity and the cellular immunity of the organism, the activation of the cellular immunity can further amplify the killing to the intracellular bacteria, and virtuous circle is realized so as to efficiently kill the intracellular bacteria.
Preferably, the dendritic mesoporous silica has a large pore diameter of between 25 and 30 nm. The aperture breaks through the limitation that the original small-aperture silicon dioxide system can only load small-molecule drugs, and realizes the load of the large-size nanometer protease (the particle size is usually 8-16 nm).
Preferably, the molar ratio of the disulfide bond to the dendritic mesoporous silica is 2-5%, preferably 3-4%. The proportion is controlled within the range, so that the dendritic structure can be constructed and the responsive degradation can be realized.
Preferably, the protease accounts for 2.5-10% by mass of the nano material for simulating natural killer cells to efficiently treat intracellular bacteria, preferably 6-8% by mass of the nano material, and more preferably 4.5-5.5% by mass of the nano material. In this case, intracellular bacteria can be killed by an appropriate drug (protease) concentration while sufficiently utilizing the carrying ability of the carrier.
Preferably, the nano material for simulating natural killer cells to efficiently treat intracellular bacteria is obtained by modifying methoxysilane polyethylene glycol on the surface of the protease-loaded disulfide bond-doped dendritic mesoporous silica. Modification of methoxy Silane polyethylene glycol (mPEG-Silane) can improve the biological safety of the inorganic nano-carrier in vivo, and good biocompatibility and in vivo circulation response are realized while nano-protease is efficiently loaded.
In a second aspect, the present invention provides a method for preparing the nanomaterial for simulating natural killer cells to effectively treat intracellular bacteria. The preparation method comprises the following steps:
(1) synthesizing disulfide bond-doped dendritic mesoporous silica by taking a surfactant as a structure directing agent, alkali as a catalyst and tetraethoxysilane and organic silicon containing disulfide bonds as precursors through a sol-gel reaction;
(2) and (2) taking the disulfide bond-doped dendritic mesoporous silica synthesized in the step (1) as a carrier, and loading protease in a pore channel of the carrier to obtain the nano material for simulating natural killer cells to efficiently treat intracellular bacteria.
The preparation method is simple, convenient and feasible, high in yield, high in efficiency, pollution-free and low in cost, and the obtained nano material for simulating natural killer cells to efficiently treat intracellular bacteria is regular in shape, controllable in pore size, large in pore volume and good in environmental responsiveness, is beneficial to controllable release and efficient action of nano protease on infected parts of the intracellular bacteria, has excellent relieving and treating effects, and is expected to be used for preparing a medicine system for treating intracellular bacteria infection.
Preferably, the surfactant is at least one of a cationic surfactant, an anionic surfactant or a block copolymer surfactant; the alkali catalyst is at least one of sodium hydroxide, ammonia water, triethanolamine, sodium salicylate and sodium carbonate; the organosilicon containing disulfide bonds is bis- [3- (triethoxysilyl) propyl ] -disulfide and/or bis (triethoxysilylpropyl) disulfide.
Preferably, the preparation method further comprises: stirring and mixing the protease-loaded disulfide bond-doped dendritic mesoporous silica and methoxysilane polyethylene glycol in an alcohol solution.
In a third aspect, the present invention provides a use of the nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria in sterilization.
Drawings
FIG. 1 is a flow chart of the preparation of the nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria according to the present invention;
FIG. 2 is TEM images of disulfide bond-doped dendritic mesoporous silica synthesized in example 1 at different scales, (A) is 1 μm, (B) is 100nm, and (C) is 500 nm;
FIG. 3 is an elemental distribution diagram and a spectrum analysis diagram (including a spectrum analysis diagram of O element, Si element and S element) of monodisperse disulfide bond doped dendritic mesoporous silica nanoparticles;
FIG. 4 is TEM images of the responsive degradation of disulfide bond-doped dendritic mesoporous silica in a reduced glutathione environment for 2h, 24h and 48 h;
FIG. 5 is N of disulfide-bond-doped dendritic mesoporous silica2An adsorption-desorption isotherm, the inset is a pore size distribution curve of the disulfide bond doped dendritic mesoporous silica;
fig. 6 is a result of a test of the growth state of bacterial colonies of the control group, experimental group 1, and experimental group 2.
Detailed Description
The present invention is further illustrated by the following examples, which are to be understood as merely illustrative of, and not restrictive on, the present invention. Unless otherwise specified, each percentage means a mass percentage.
The present disclosure provides a nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria, comprising a disulfide bond-doped dendritic mesoporous silica (also referred to as "dendritic organic-inorganic hybrid mesoporous silica") as a carrier, and a protease loaded in a pore channel of the carrier.
Because the disulfide bond is doped in the framework of the dendritic mesoporous silica, the doping of the disulfide bond does not influence the physical parameters of the dendritic organic-inorganic hybrid mesoporous silica. The disulfide bond doped dendritic mesoporous silica maintains the morphology of dendritic mesoporous silica nanoparticles, and is suitable for serving as a good protease carrier. The disulfide bonds are uniformly distributed on the framework of the dendritic mesoporous silica, and can be degraded in a better response manner after reaching the intracellular bacteria infected part, so that the controllable release of the protease is realized. In some embodiments, the dendritic organic-inorganic hybrid mesoporous silica nanoparticles have a spherical morphology with dendritic pores. Wherein the diameter of the dendritic pores may be in the range of 10 to 30nm, preferably with a large pore diameter between 25 to 30 nm.
In addition, the particle diameter of the dendritic organic-inorganic hybrid mesoporous silica nano-particle can be 220-280nm, and the specific surface area can be 300-550m2Per g, pore volume of 0.6-1.8m2/g。
The invention selects dendritic organic-inorganic hybrid mesoporous silica nanoparticles as carriers: firstly, the special design of the dendritic carrier breaks through the application limitation of the original small-aperture silicon dioxide system for loading large analysis drugs. Specifically, the conventional small molecule drug can be loaded by using small-aperture silica, but the large-molecular structure of the nano-protease enables the aperture required by the loading to be far larger than that of other mesoporous silica, so that the dendritic organic-inorganic hybrid mesoporous silica nanoparticle with the dendritic framework can provide the large aperture for loading the nano-protease. Secondly, the disulfide bond is doped into the framework of the dendritic mesoporous silica, so that the framework can collapse and degrade under the action of intracellular bacteria high redox potential, and the defect that the degradation of a carrier framework material by a surface modification disulfide bond is limited is overcome. The dendritic organic-inorganic hybrid mesoporous silica disclosed by the invention has the characteristics of specific response, large aperture, high specific surface area, uniform appearance and safety in biodegradation, and is beneficial to construction of a nano material for simulating natural killer cells to efficiently treat intracellular bacteria.
The nanometer protease is loaded in the pore canal of the dendritic mesoporous silica doped with the disulfide bond. The aperture of the dendritic mesoporous silica doped with the disulfide bond is larger than the size of the nano proteaseThe protease can be loaded by physical adsorption. Intracellular bacteria, a specific disease model, exist in biological microenvironments with high redox potentials. The response is preferably GSH and H2O2A double response. The disulfide bond doped in the dendritic mesoporous silica framework is H2O2The mesoporous silica nano particle is broken to form-SOOH bonds under the action of oxidation and is broken to form-SH bonds under the action of GSH (glutathione) reduction, so that the mesoporous silica nano particle is broken due to broken bonds of disulfide bonds in a framework, and the nano protease positioned in a pore channel is released out of the carrier material. When the loaded protease is perforin and granzyme, the perforin perforates the surface of a cell membrane to mediate granzyme to effectively kill intracellular bacteria, and the important problem that the body autoimmune system or conventional antibacterial drugs cannot identify and kill bacteria (intracellular bacteria) hidden in cells is solved. And the nano protease carrier material can effectively cause the immune regulation of organisms in the process of realizing the killing of intracellular bacteria so as to accelerate the killing of pathogenic bacteria and the healing of inflammatory infection parts.
The nano material for simulating natural killer cells to efficiently treat intracellular bacteria can also modify methoxysilane polyethylene glycol on the surface of the protease-loaded disulfide bond-doped dendritic mesoporous silica. The main function of the modified methoxy silane polyethylene glycol is to improve the biocompatibility of the nano material. Meanwhile, the methoxysilane group and the silicon dioxide system have better consistency, and the introduction of other groups can be reduced.
The following description is given to an exemplary method for preparing the nanomaterial for simulating natural killer cells to effectively treat intracellular bacteria according to the present invention with reference to fig. 1. The preparation method disclosed herein is convenient and easy to implement, and the operation conditions are precise and controllable.
Firstly, preparing the dendritic organic-inorganic hybrid mesoporous silica. For example, the dendritic organic-inorganic hybrid mesoporous silica is synthesized by a sol-gel method by using a surfactant as a structure directing agent, using an alkali catalyst, and using Tetraethoxysilane (TEOS) and bis- [3- (triethoxysilyl) propyl ] -disulfide (BTEE) containing a disulfide bond as precursors. At this time, the disulfide bond is uniformly doped in the framework of the dendritic mesoporous silica. The sol-gel reaction temperature can be 75-85 ℃, and the reaction time can be 9-15 h. The process can improve the existing preparation process of the mesoporous silica with small aperture (less than 5nm) so as to prepare the dendritic organic-inorganic hybrid mesoporous silica with the aperture between 25 and 30 nm.
The surfactant may be a cationic surfactant, an anionic surfactant, or a block copolymer surfactant, including but not limited to cetyltrimethylammonium chloride (CTAC) and/or cetyltrimethylammonium bromide (CTAB). The base catalyst includes, but is not limited to, at least one of sodium hydroxide, ammonia, Triethanolamine (TEA), sodium salicylate (NaSal), and sodium carbonate.
In some embodiments, TEA is dissolved in water and mixed with stirring at 80 ℃ for 0.5h, then CTAB and NaSal are added and stirred at 80 ℃ for 1h, then TEOS and BTEE are added to the solution and the reaction is continued overnight, cooling after completion of the reaction. The product was collected by high speed centrifugation and washed with ethanol to remove residual reactants. And (3) extracting the collected product at 60 ℃ by using an ethanol solution of ammonium nitrate to remove the surfactant, and performing vacuum drying overnight at room temperature to obtain the dendritic organic-inorganic hybrid mesoporous silica.
During the reaction, the volume ratio of TEOS to BTEE is 3: 2-4: 5. the proper silicon source proportion is beneficial to preparing the dendritic silicon dioxide doped with disulfide bonds with regular shapes, otherwise, framework collapse or disulfide bond doping failure can be caused. In some examples, the volume ratio of TEOS to BTEE is 6: 5,7: 6, optimally 1: 1. further, the mass ratio of CTAB, TEA and NaSal may be 5: 1: 2-7: 1: 4.
and (3) loading protease on the dendritic organic-inorganic hybrid mesoporous silica. The kind of protease may be changed adaptively as required, and a nanoscale protease is preferable. More preferably, perforin and/or granular enzyme are supported on the dendritic organic-inorganic hybrid mesoporous silica. The mass ratio of perforin to granzyme is preferably 4: 3-4: 5. the nano material obtained at the moment can simulate the treatment concept of natural killer cells for efficiently treating intracellular bacteria (CAR-NK), the dendritic mesoporous silica doped with disulfide bonds is specifically degraded at the intracellular bacteria infection part of an organism to release perforin and granzyme (the NK cells are simulated to release perforin and granzyme), the perforin punches a cell membrane so as to mediate the granzyme to enter the cells to kill the intracellular bacteria, and the problem that the conventional antibiotics cannot kill the intracellular bacteria is solved. Moreover, the nano material can cause the spontaneous immune regulation mechanism of the organism to play a role in high-efficiency synergistic antibacterial and anti-infection effects in the process of effectively realizing killing of intracellular bacteria.
The method for loading the protease is not the innovation point of the invention, and the loading method commonly used in the field can be adopted. In some embodiments, the dendritic organic-inorganic hybrid mesoporous silica and the protease are stirred in a polar solvent. The polar solvent is a polar solvent commonly used in the art, and is preferably water. The stirring time can be 12-24 h.
In some embodiments, the dendritic organic-inorganic hybrid mesoporous silica is dispersed in a protease solution and stirred. By way of example, the concentration of perforin and granzyme in the protease solution may be 20-40U/mL. The stirring temperature can be room temperature or a low-temperature ice-water bath.
Methoxy Silane polyethylene glycol (mPEG-Silane) can also be modified on the surface of the dendritic organic-inorganic hybrid mesoporous silica (disulfide bond doped dendritic mesoporous silica) loaded with protease. The modification method can be that the disulfide bond doped dendritic mesoporous silica loaded with protease and methoxysilane polyethylene glycol (mPEG-Silane) are stirred in an alcohol solution. Thus, the surface of the protease-loaded disulfide bond doped dendritic mesoporous silica is modified by the mPEG-Silane through covalent bond combination. The molecular weight of the methoxy silane polyethylene glycol can be 1000-10000. In some embodiments, the mass ratio of the dendritic organic-inorganic hybrid mesoporous silica to the mPEG-Silane can be 2: (5-11), for example, 2: 7. 2: 9, etc. In this case, a good surface modification effect can be obtained. The stirring temperature may be 20-30 deg.C, preferably 25 deg.C. The stirring time may be 15 to 48 hours, preferably 30 hours.
The nano material for simulating natural killer cells to efficiently treat intracellular bacteria can provide a new way for efficiently and controllably treating bacterial infection, inflammation and tissue healing by regulating perforin and granzyme released by NK cells. The nano material for simulating natural killer cells to efficiently treat intracellular bacteria can simultaneously load perforin and granular enzyme, and can perform specific response degradation under the high reduction potential environment of the intracellular bacteria to release the perforin and the granular enzyme, so that the perforation of the cell membrane surface and the killing treatment of bacteria in the cells are realized. Moreover, the nano material for simulating natural killer cells to efficiently treat intracellular bacteria can effectively kill the intracellular bacteria to realize infection resistance, simultaneously stimulate the spontaneous immunity of organisms so as to greatly improve the treatment effect, and the immunity memory enables the resistance of the organisms to be greatly improved. The nano material for simulating natural killer cells to efficiently treat intracellular bacteria has excellent and wide application prospect in CAR-NK therapy for killing intracellular bacteria to resist infection and application in the aspect of cooperative immunity.
The present invention will be described in detail by way of examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art may be made in light of the above teachings. The specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and are not limited to the specific values exemplified below.
Example 1
Step (1) is to prepare the disulfide bond doped dendritic mesoporous silica. 70mg TEA, 400mg CTAB and 200mg NaSal were dissolved in 25mL deionized water, mixed well and stirred in a water bath at 80 ℃ for 1.5 hours, then 2mL TEOS and 2mL BTEE were added to the solution and the reaction was continued for 12 hours, followed by cooling and centrifugation at 12000r/min for 30min to collect the crude product. The crude product was washed 2 times each by centrifugation using water and ethanol, the washed crude product was dispersed in an ethanol solution of ammonium nitrate of 8mg/mL, refluxed overnight, centrifuged, and the product was collected and washed with ethanol. To adequately remove the CTAB surfactant, the ethanol reflux and washing steps of ammonium nitrate were repeated 3 times each.
And (2) loading perforin and granzyme. Dispersing 10mg of disulfide bond-doped dendritic mesoporous silica nanoparticles into 3mL of nano protease aqueous solution. The concentrations of perforin and granzyme in the aqueous solution were both 30U/mL. Stirring the solution at low temperature (2-5 ℃) for 6 hours, then centrifugally washing, and collecting a product to obtain the protease-loaded disulfide bond-doped dendritic mesoporous silica.
And (3) modifying the surface of PEG. And (3) adding 10mL of the product obtained in the step (2), namely the nano-protease-loaded disulfide bond-doped dendritic mesoporous silica into 100mL of ethanol, then adding 50mg of methoxysilane polyethylene glycol, stirring at normal temperature for 3h, and then carrying out centrifugal washing to obtain the final product.
Fig. 2 is a TEM distribution plot of disulfide-bond doped dendritic mesoporous silica nanoparticles on copper mesh carbon film after sonication. The disulfide bond doped mesoporous silica has good dispersibility, a spherical dendritic structure, uniform size and regular appearance.
Fig. 3 is an elemental distribution diagram and an energy spectrum analysis diagram of monodisperse disulfide bond doped dendritic mesoporous silica nanoparticles. The sulfur element is successfully doped in the framework of the dendritic mesoporous silica.
And (3) testing degradability: adding the disulfide bond doped dendritic mesoporous silica nanoparticles into a 10mM GSH solution, stirring for 2h, 12h, 24h and 48h at the temperature of 20-30 ℃, and observing the change of the morphology structure of the obtained degradation product under a TEM. Fig. 4 shows that the dendritic framework structure collapses and the pore channels of the framework carrier are opened along with the extension of the degradation time, so that the good release effect of the nano protease can be realized. This shows that the disulfide bond doped dendritic mesoporous silica has excellent time-dependent degradability and specific response degradability. The time-dependent degradability means that the disulfide bond-doped dendritic mesoporous silica gradually becomes a broken structure with increasing collapse degree with the time. The specific response degradability is to use GSH to simulate the specific microenvironment of intracellular bacteria in an organism, namely to respond to the degradation of the microenvironment of the intracellular bacteria, and basically not to degrade in other microenvironments. The nano material for simulating natural killer cells to efficiently treat intracellular bacteria can simulate the specificity of selective killing bacteria of NK cells, can controllably release nano protease along with the lapse of time, has the effect of sustained release of medicaments, and can realize a good antibacterial effect through the release of the nano protease.
FIG. 5 is a process for N-doping a disulfide-bond-doped dendritic mesoporous silica2Adsorption-desorption isotherms and pore size profiles. The disulfide bond doped dendritic mesoporous silica has larger aperture, the particle size of dendritic holes is 25-30nm, the space condition of loading nano protease (the size is about 10nm) is met, and the disulfide bond doped dendritic mesoporous silica is suitable for being used as a carrier for efficiently loading enzyme substances.
And (3) antibacterial experiment: gram-positive S.aureus and gram-negative E.coli were grown overnight on sheep blood agar plates (5-7% by volume sheep blood) at 37 ℃. Individual colonies of the strain were picked and cultured overnight at 37 ℃ in 4mL tryptone soy broth. The suspension of Staphylococcus aureus and Escherichia coli was serially diluted 10-fold to obtain a concentration of 1X 10 in trypticase Soytone broth6CFUs mL-1The bacterial liquid of (4). Staphylococcus aureus was diluted to a concentration of 1X 10 with PBS7CFU mL-1. The antibacterial activity test was performed by the extension plate colony counting method. The experiment group 2 is that the nano material for simulating natural killer cells to efficiently treat intracellular bacteria is added into the solution with the concentration of 1 multiplied by 106CFU mL-1The bacterial liquid is cultured for 12-48h, and the concentration of the nano material for simulating natural killer cells to efficiently treat the intracellular bacteria is 10-200 mu g mL-1. The experiment group 1 replaces the nano material for simulating natural killer cells to efficiently treat intracellular bacteria with protease, and the addition amount of the protease is completely consistent with the type and the content of the protease loaded in the nano material of the experiment group 2. The control group is used as a blank sample, and compared with the experimental group 2, the difference is that the nano material for efficiently treating the intracellular bacteria is not added with the simulated natural killer cells. The antibacterial test result shows that the nano material for simulating natural killer cells to efficiently treat intracellular bacteria can effectively kill bacterial colonies.
Claims (5)
1. The nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria is characterized by comprising dendritic mesoporous silica serving as a carrier and doped with disulfide bonds and protease loaded in pore channels of the carrier; the protease is perforin and/or granzyme;
the dendritic mesoporous silica has a large aperture with the aperture between 25 and 30 nm;
the disulfide bond-doped dendritic mesoporous silica generates collapse of a mesoporous silica framework under the specific oxidation-reduction potential of intracellular bacteria so as to release the loaded protease;
the mol ratio of disulfide bonds in the dendritic mesoporous silica is 2-5%;
the protease accounts for 2.5-10% of the nano material for simulating natural killer cells to efficiently treat intracellular bacteria by mass percent;
the nano material for simulating natural killer cells to efficiently treat intracellular bacteria is modified by methoxysilane polyethylene glycol on the surface of the protease-loaded disulfide bond-doped dendritic mesoporous silica.
2. The method for preparing a nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria according to claim 1, wherein the method comprises the following steps:
(1) synthesizing disulfide bond doped dendritic mesoporous silica by using a surfactant as a structure directing agent, alkali as a catalyst and tetraethoxysilane and organic silicon containing disulfide bonds as precursors through a sol-gel reaction; the organosilicon containing disulfide bonds is bis- [3- (triethoxysilyl) propyl ] -disulfide and/or bis (triethoxysilylpropyl) disulfide, and the volume ratio of ethyl orthosilicate to organosilicon containing disulfide bonds is 3: 2-4: 5;
(2) taking the disulfide bond doped dendritic mesoporous silica synthesized in the step (1) as a carrier, and loading protease in a pore channel of the carrier to obtain the nano material for simulating natural killer cells to efficiently treat intracellular bacteria.
3. The production method according to claim 2, characterized in that the surfactant is at least one of a cationic surfactant, an anionic surfactant, or a block copolymer surfactant; the alkali catalyst is at least one of sodium hydroxide, ammonia water, triethanolamine, sodium salicylate and sodium carbonate.
4. The production method according to claim 2 or 3, characterized by further comprising: stirring and mixing the protease-loaded disulfide bond-doped dendritic mesoporous silica and methoxysilane polyethylene glycol in an alcohol solution.
5. The use of the nanomaterial for simulating natural killer cells to efficiently treat intracellular bacteria according to claim 1 in the preparation of products for killing gram-positive staphylococcus aureus and gram-negative escherichia coli.
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