CN116251236A - Biological breast patch and preparation method thereof - Google Patents
Biological breast patch and preparation method thereof Download PDFInfo
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- CN116251236A CN116251236A CN202310060857.1A CN202310060857A CN116251236A CN 116251236 A CN116251236 A CN 116251236A CN 202310060857 A CN202310060857 A CN 202310060857A CN 116251236 A CN116251236 A CN 116251236A
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- soaking
- breast patch
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- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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Abstract
The invention discloses a biological breast patch and a preparation method thereof. The preparation method of the biological breast patch comprises the following steps: 1) Collecting dermis of mammal, cleaning with water, and filtering; 2) Carrying out virus inactivation treatment on the skin tissue treated in the step 1); 3) Degreasing the skin tissue treated in the step 2); 4) Performing decellularization treatment on the skin tissue treated in the step 3) to obtain a decellularized dermis matrix; 5) Immersing the acellular dermal matrix obtained in the step 4) in an aqueous solution of sodium hyaluronate, and taking out; 6) And 5) freeze-drying the acellular dermal matrix treated in the step 5) to obtain the acellular dermal matrix. The biological breast patch adopts the acellular dermal matrix material, can guide the host autologous cells to grow in, provides a template for reconstructing damaged tissues by cells, can be rapidly vascularized, and gradually degrades along with the construction of the new tissues of the host until the new tissues of the host are completely replaced by the host tissues.
Description
Technical Field
The invention belongs to the technical field of tissue engineering, and particularly relates to a biological breast patch and a preparation method thereof.
Background
The breast reconstruction of breast cancer patients with indications is widely accepted at home and abroad, and the implant breast reconstruction is the most common breast reconstruction mode due to the advantages of short operation time, quick postoperative recovery, no donor area injury, safety, reliability and the like.
The primary condition for breast reconstruction is that the surface of the implanted prosthesis is covered by as much tissue as possible, but mastectomy results in a great amount of muscle loss, and the rest of autologous tissue often cannot achieve the effect of effectively wrapping and supporting the prosthesis, so that the prosthesis can suffer from complications such as sagging and even exposure. To address the problem of prosthetic coverage loss, some biomaterial products (e.g., decellularized dermal matrix, bovine pericardium, etc.) or synthetic materials (e.g., titanium coated polypropylene patches, etc.) are increasingly being used in implant breast reconstruction procedures, with or without associated muscle tissue to cover the prosthetic reconstruction breast. In the patch products for breast reconstruction which are marketed at present, a crosslinking agent is introduced in the preparation process of the patch taking the pericardium as a raw material, so that the natural structure of the material is changed and the degradation of the material in a living body is prevented; the synthetic patch is not degradable, so that the synthetic patch can exist in a patient for a long time, and psychological burden of the patient is increased.
The invention patent (CN 100372511C) discloses a decellularized dermis matrix and a preparation method thereof, and describes the steps of animal body surface skin stripping, cleaning, virus inactivation, immunogen removal, fat removal, forming, packaging and sterilization to obtain medical products with different sizes. The acellular dermal matrix is used as a natural medical material for tissue transplantation, can guide autologous cells to grow in after being implanted into a human body, provides a template for reconstructing damaged tissues by cells, is rapidly vascularized, and gradually degrades along with the construction of new tissues of a host until the new tissues are completely replaced by the host tissues. However, few decellularized dermal matrix materials are reported for use in breast reconstruction at present.
Disclosure of Invention
The invention aims to provide a biological breast patch and a preparation method thereof, wherein the biological breast patch can guide host autologous cells to grow in, provide a template for reconstructing damaged tissues by cells, can be rapidly vascularized, and gradually degrade along with the construction of new tissues of a host until the new tissues of the host are completely replaced by the host tissues.
The invention provides a preparation method of a biological breast patch, which comprises the following steps:
1) Collecting dermis of mammal, cleaning with water, and filtering;
2) Carrying out virus inactivation treatment on the skin tissue treated in the step 1);
3) Degreasing the skin tissue treated in the step 2);
4) Performing decellularization treatment on the skin tissue treated in the step 3) to obtain a decellularized dermis matrix;
5) Immersing the acellular dermal matrix obtained in the step 4) in an aqueous solution of sodium hyaluronate, and taking out;
6) And 5) freeze-drying the acellular dermal matrix treated in the step 5) to obtain the biological breast patch.
In a specific embodiment of the invention, the mammal is a cow; the dermis layer is obtained by peeling off the body surface skin and subcutaneous tissue.
In the above method, the virus inactivation treatment in step 2) includes: soaking the skin tissue with an aqueous solution of hydrogen peroxide, and cleaning after the soaking is finished;
the mass concentration of the aqueous solution of hydrogen peroxide can be 0.06-0.1 g/ml, and can be specifically 0.06g/ml;
the soaking time can be 1-1.5 h, and can be 1.5h specifically;
the cleaning may be ultrasonic cleaning.
In the above method, the degreasing treatment in step 3) includes: soaking the skin tissue with a mixed solution of chloroform and ethanol, and cleaning after the soaking is finished;
in the mixed solution, the volume ratio of chloroform to ethanol can be 1: (1-5), specifically 1:3, a step of;
the soaking time can be 30-60 min, and can be 45min specifically;
the cleaning may be ultrasonic cleaning.
In the above method, the decellularizing treatment in step 4) includes: soaking the skin tissue in an aqueous solution of an alkaline reagent, and cleaning after the soaking is finished;
the alkaline reagent may be NaOH or KOH;
the concentration of the alkaline reagent in the aqueous solution of the alkaline reagent may be 0.5 to 5.5mol/L, specifically may be 3mol/L;
the soaking time can be 30-60 min, and can be specifically 30min;
the cleaning may be ultrasonic cleaning;
the number of times of the decellularization treatment may be 3 to 5 times, and specifically may be 3 times.
In the above method, the concentration of the aqueous solution of sodium hyaluronate in step 5) may be 0.01 to 0.1g/ml, the hydration time of the sample after compounding sodium hyaluronate is significantly shortened, and the concentration of the aqueous solution of sodium hyaluronate is preferably 0.03 to 0.1g/ml, more preferably 0.03 to 0.05g/ml or 0.05 to 0.1g/ml, still more preferably 0.03g/ml, 0.05g/ml, 0.1g/ml;
the soaking in the step 5) comprises a first soaking and a second soaking after the first soaking finishes turning over the acellular dermal matrix, wherein the time of the first soaking and the time of the second soaking can be respectively 10-20 min (such as 10 min).
In the above method, the freeze-drying in step 6) may include the steps of:
a) Spreading the sample on a freeze-drying disc, and placing the sample on a partition board in a freeze dryer;
b) Pre-freezing the product: starting a freeze dryer, firstly cooling the box body to-50 ℃, and starting cooling for at least 1 hour;
c) Vacuumizing: after the pre-freezing is finished, starting a vacuum pump, and reducing the vacuum degree of the box body and maintaining the vacuum degree below 20 Pa;
d) And (3) primary drying: the temperature of the box body is increased to-30 ℃, the box body enters a temperature maintaining stage after the temperature is increased, and the total time from the temperature increase is 3 hours;
e) And (5) secondary drying: the temperature of the box body is continuously increased to-15 ℃, the box body enters a temperature maintaining stage after the temperature increase is finished, and the total time from the temperature increase is 1h;
f) And (3) third drying: the temperature of the box body is continuously increased to 0 ℃, the box body enters a temperature maintaining stage after the temperature is increased, and the total time from the temperature increase is 1h;
g) Fourth drying: the temperature of the box body is continuously increased to 25 ℃, the box body enters a temperature maintaining stage after the temperature increase is finished, and the total time from the temperature increase is at least 1h;
h) And (5) after the freeze-drying is finished, taking out the sample.
Further, the method may further comprise, after step 6): cutting the acellular dermal matrix after freeze drying; and, a step of, in the first embodiment,
and (3) punching the acellular dermal matrix after the shearing and cutting.
Still further, the method further comprises the steps of packaging, sterilizing and analyzing sequentially after the punching treatment;
the sterilization is ethylene oxide sterilization or irradiation sterilization.
Preferably, the conditions for ethylene oxide sterilization are as follows: the temperature is 37 ℃ -47 ℃ (42+/-5 ℃), the humidity is 30% -80%, the concentration of ethylene oxide is 500mg/L, and the sterilization time is 3-5 hours (such as 4 hours);
preferably, the temperature of the analysis is 37 ℃ -47 ℃ (i.e. 42 ℃ + -5 ℃), specifically 42 ℃, and the time can be 4-6 days, specifically 6 days in the analysis oven.
Biological breast patches prepared by the method of any of the above are also within the scope of the present invention.
In the biological breast patch described above, the biological breast patch may be rectangular or crescent;
the diameter of the through holes in the biological breast patch is 0.5-2.5 mm, and the distance between two adjacent through holes can be 1-2 cm; the through hole can be a round hole;
the diameter of the through holes in the biological breast patch can be 0.5-2.5 mm (such as 2 mm), and the distance between two adjacent through holes can be 1-2 cm (such as 2 cm);
the biological breast patch is crescent-shaped, and the through holes are distributed at the middle position of the biological breast patch.
The breast patch provided by the invention is an acellular dermal matrix material, and has the following advantages:
(1) The breast patch provided by the invention effectively carries out virus inactivation and antigen removal treatment, eliminates the risk of zoonosis transmission, has excellent biocompatibility and small rejection reaction after implantation into a human body;
(2) According to the breast patch provided by the invention, a fixing agent is not used in the treatment process, and sodium hyaluronate is introduced, so that the hydration speed and effect of the material are increased by the composite sodium hyaluronate, and the proliferation and differentiation of host cells in acellular dermal matrixes can be promoted, so that tissue regeneration is facilitated;
(3) The breast patch provided by the invention is processed by a vacuum freeze drying technology, so that the natural loose porous structure of the acellular dermal matrix is maintained to the greatest extent, and the cell ingrowth and tissue regeneration can be effectively guided;
(4) According to the breast patch provided by the invention, the surface of the product is perforated, and the crescent specification is designed in combination with clinical requirements, so that complications such as seroma and the like caused by operation effusion can be avoided, and the pruning flow of a clinician is greatly reduced;
(5) The breast patch provided by the invention adopts ethylene oxide sterilization, greatly reduces the damage of radiation sterilization to the collagen fiber bundles of the material, increases the mechanical strength of the patch, prolongs the degradation time, and is more suitable for implant breast reconstruction.
Drawings
Fig. 1 is a schematic diagram of punching a breast patch in example 1 of the present invention, the patch shapes are rectangular (a) and crescent (B), respectively.
Fig. 2 is a photograph of wet (a) and dry (B) breast patches of example 1 of the present invention.
Fig. 3 is a scanning electron microscope image of the smooth surface (a), rough surface (B) and cross section (C) of the breast patch in example 1 of the present invention.
Fig. 4 is a HE staining chart of the breast patch in example 1 of the present invention.
FIG. 5 is a graph showing the results of HE staining of 12-week old materials from the implantation of a breast patch in New Zealand white rabbits according to example 1 of the present invention.
Fig. 6 is a graph comparing the measured data of tensile strength and tearing force of the breast patches prepared in example 1 and example 2 of the present invention.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods used in the following examples are all conventional methods unless otherwise specified; materials, reagents and the like used are commercially available unless otherwise specified.
Example 1 preparation of a biological Breast Patch by ethylene oxide Sterilization
1. And (3) raw material treatment: thawing cow leather, peeling off the skin and subcutaneous tissues, retaining only dermis, washing skin with purified water and draining off water;
2. virus inactivation: soaking skin tissue in 0.06g/ml hydrogen peroxide solution for 1.5 hr, and cleaning in an ultrasonic cleaner;
3. degreasing: the chloroform was used: ethanol is 1:3 (v/v) soaking skin tissue for 45min, and cleaning in an ultrasonic cleaner after soaking;
4. cell removal treatment: soaking skin tissue in 3mol/L NaOH solution for 30min, cleaning in an ultrasonic cleaner after soaking, and repeating the steps for 3 times to obtain acellular dermal matrix;
5. active material compounding: soaking the acellular dermal matrix in 0.03g/ml sodium hyaluronate solution for 10min, turning over the acellular dermal matrix after soaking, and continuously soaking for 10min to compound the acellular dermal matrix;
6. and (3) freeze drying: spreading the acellular dermal matrix obtained in the step (5) in a stainless steel freeze-drying plate, and then placing the stainless steel freeze-drying plate in a vacuum freeze-drying machine, and freeze-drying according to a preset program to obtain a freeze-dried acellular dermal matrix material;
the lyophilization procedure was:
a) Spreading the sample on a freeze-drying disc, and placing the sample on a partition board in a freeze dryer;
b) Pre-freezing the product: starting a freeze dryer, firstly cooling the box body to-50 ℃, and starting cooling for at least 1 hour;
c) Vacuumizing: after the pre-freezing is finished, starting a vacuum pump, and reducing the vacuum degree of the box body and maintaining the vacuum degree below 20 Pa;
d) And (3) primary drying: the temperature of the box body is increased to-30 ℃, the box body enters a temperature maintaining stage after the temperature is increased, and the total time from the temperature increase is 3 hours;
e) And (5) secondary drying: the temperature of the box body is continuously increased to-15 ℃, the box body enters a temperature maintaining stage after the temperature increase is finished, and the total time from the temperature increase is 1h;
f) And (3) third drying: the temperature of the box body is continuously increased to 0 ℃, the box body enters a temperature maintaining stage after the temperature is increased, and the total time from the temperature increase is 1h;
g) Fourth drying: the temperature of the box body is continuously increased to 25 ℃, the box body enters a temperature maintaining stage after the temperature increase is finished, and the total time from the temperature increase is at least 1h;
h) And (5) after the freeze-drying is finished, taking out the sample.
7. Cutting and shearing: and (5) shearing the freeze-dried acellular dermal matrix material according to the size and specification of the product. The invention particularly designs a crescent-shaped product in combination with clinical use requirements of breast reconstruction, as shown in fig. 1. The crescent product is used in the process that the upper short arc is sutured with the pectoral large muscle of the patient, and the lower long arc is sutured with the chest wall of the patient, so that the operation time of a clinician is greatly reduced, and the infection risk caused by patch trimming is reduced;
8. punching and packaging: the breast patch which is cut by the special puncher is punched by parameters of 2mm diameter of round hole and 2cm spacing, and the punching effect is shown in fig. 1 (A) and 1 (B). Crescent product, the through-hole mainly distributes in the intermediate position that easily accumulates the hydrops, and does not do the perforation to the upper and lower end of the patch that needs to carry out the sewing operation, guarantees that patch mechanical properties is not influenced, and its advantage lies in can showing the reduction doctor's in the art operation volume. After the punching is finished, packaging the breast patch; packaging is performed using blister & te Wei Jianggai material, which requires sterile handling and manipulation.
9. And (3) sterilization analysis: the material is sterilized and analyzed by ethylene oxide, and the sterilization conditions are as follows: the temperature is 42+/-5 ℃, the humidity is 30% -80%, the concentration of ethylene oxide is 500mg/L, and the sterilization time is 4 hours; the analysis process comprises the following steps: in a special analytical oven, the temperature was controlled at 42℃for 6 days to obtain a breast patch.
Example 2 preparation of a biological Breast Patch by radiation Sterilization
1. And (3) raw material treatment: the same as in example 1, 1.
2. Virus inactivation: the same as in example 1, 2.
3. Degreasing: the same as in 3 of example 1.
4. Cell removal treatment: the same as in example 1, 4.
5. Active material compounding: the same as in 5 of example 1.
6. And (3) freeze drying: the same as in example 1, 6.
7. Cutting and shearing: the same as in example 1, 7.
8. Punching and packaging: the same as in 8 of example 1.
9. And (3) sterilization: the product is sterilized by electronic book radiation after punching and packaging, and the radiation dose is 15KGy.
Example 3 preparation of biological Breast Patch compounded with active substances at different concentrations
1. And (3) raw material treatment: the same as in example 1, 1.
2. Virus inactivation: the same as in example 1, 2.
3. Degreasing: the same as in 3 of example 1.
4. Cell removal treatment: the same as in example 1, 4.
5. Active material compounding: soaking the acellular dermal matrix in sodium hyaluronate solution of 0, 0.01, 0.03, 0.05 and 0.1g/ml for 10min, turning over the acellular dermal matrix after soaking, and continuously soaking for 10min to compound the acellular dermal matrix;
6. and (3) freeze drying: the same as in example 1, 6.
7. Cutting and shearing: the same as in example 1, 7.
8. Punching and packaging: the same as in 8 of example 1.
9. And (3) sterilization analysis: the same as in 9 of example 1.
Example 4 detection of biological Breast patches with different Sterilization modes, biological Breast patches compounded with different concentration of active substances
1. Detection of ethylene oxide sterilized biological breast patches
A schematic of the breast patch of example 1 before and after hydration is shown in fig. 2. The breast patch treated by the vacuum freeze drying technology removes most of free water in the material, greatly reduces the risk of bacteria contamination and is more beneficial to long-time storage.
In example 1, the microstructure of the breast patch is shown in a scanning electron microscope image of fig. 3, and as can be seen from fig. 3, the breast patch prepared by the method has a smooth surface (fig. 3A), a rough surface (fig. 3B) and a cross section (fig. 3C), the collagen fiber bundles on the smooth surface are arranged in a relatively ordered and dense manner, the collagen fiber bundles on the rough surface are arranged in a relatively sparse manner, and the collagen fibers are crisscrossed, loose and porous, so that the breast patch is more suitable for cell crawling and ingrowth.
The effect of removing cells of the breast patch in example 1 is shown in fig. 4, and as can be seen from fig. 4, the breast patch prepared by the invention completely removes various cells in skin tissues, greatly reduces immunogenicity of materials, and ensures that the breast patch does not cause immune response of a host after being implanted into a patient.
In example 1, the breast patch was implanted in leg muscle of New Zealand white rabbit, and after 12 weeks of implantation, the material was taken and subjected to hematoxylin-eosin (HE) staining treatment, and the result is shown in FIG. 5. As can be seen from FIG. 5, the material is relatively complete, the collagen fibers of the under-the-mirror material are completely visible in structure, fibroblasts and fibroblasts are visible among the collagen fibers, and the inflammatory reaction degree is extremely low. The phenomenon shows that the breast patch prepared by the invention can be kept in vivo for a long time, plays a long-term mechanical supporting role, and can effectively induce surrounding host cells and tissues to grow into the breast patch to integrate materials and tissues.
2. Biological breast patch mechanical property detection with different sterilization modes
The breast patches prepared in the different sterilization modes in the example 1 and the example 2 are subjected to mechanical performance detection such as tensile strength, tearing force and the like. Determination of tensile Properties of plastics according to GB/T1040.3-2006 section 3: the test conditions of the films and sheets were examined by the method specified in the test conditions of the films and sheets, cutting the test specimen into a dumbbell shape having a width of about 10mm and a length of about 30mm, hydrating the purified water, and testing with a universal material tester at a speed of 25mm/min, and separately detecting the tensile strength. The testing machine speed was set at 100mm/min and the tearing force was measured separately, and the measurement results are shown in Table 1 and FIG. 6.
TABLE 1 comparison of mechanical Properties of Breast Patches with different Sterilization modes
| Sterilization mode | Tensile strength/MPa | Tear force/N |
| Irradiation sterilization | 25.09±6.25 | 16.50±2.39 |
| Sterilization of ethylene oxide | 24.92±7.59 | 28.07±3.56 |
As can be seen from Table 1 and FIG. 6, the tensile strengths of the breast patches sterilized by ethylene oxide and irradiation were 24.92.+ -. 7.59, 25.09.+ -. 6.25, respectively, without significant differences; however, the tearing force of the breast patch sterilized by ethylene oxide can reach 28.07+/-3.56, and the tearing force of the breast patch sterilized by irradiation is only 16.50+/-2.39. The reason for this result may be that irradiation sterilization breaks hydrogen bonds between parts of collagen molecules that maintain the collagen structure order, destroys the collagen structure order, and the most affected tearing force is significantly reduced, whereas ethylene oxide sterilization does not have any effect on collagen fibers, so that the tearing force can be maintained at a higher level. Thus, the present invention selects ethylene oxide sterilization as the final sterilization means for the breast patch. In conclusion, the breast patch prepared by the invention has better mechanical strength, can play a role in suspending and bearing the prosthesis and surrounding tissues after being implanted into a human body, and ensures the aesthetic feeling of the reconstructed breast patch.
3. Hydration time detection of biological breast patches compounded by active substances with different concentrations
The biological breast patches prepared in example 3 and compounded by active substances with different concentrations are placed in purified water, the time is counted from the instant of soaking until the product is completely hydrated, and the difference of hydration time of the breast patches soaked by sodium hyaluronate with different concentrations is compared, and the result is shown in Table 2.
TABLE 2 influence of sodium hyaluronate concentration on hydration time
As is clear from Table 2, when sodium hyaluronate is not compounded, the sample hydration time is 5.5s, and the sample hydration time is remarkably shortened after sodium hyaluronate is compounded, but the shortening of the sample hydration time is not remarkable as the concentration is increased to 0.05g/ml or even 0.1 g/ml. The parameters for determining the concentration of the breast patch composite sodium hyaluronate were determined to be 0.03g/ml in combination with hydration effect and cost analysis.
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced in a wide variety of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains.
Claims (10)
1. A method for preparing a biological breast patch, comprising the following steps:
1) Collecting dermis of mammal, cleaning with water, and filtering;
2) Carrying out virus inactivation treatment on the skin tissue treated in the step 1);
3) Degreasing the skin tissue treated in the step 2);
4) Performing decellularization treatment on the skin tissue treated in the step 3) to obtain a decellularized dermis matrix;
5) Immersing the acellular dermal matrix obtained in the step 4) in an aqueous solution of sodium hyaluronate, and taking out;
6) And 5) freeze-drying the acellular dermal matrix treated in the step 5) to obtain the biological breast patch.
2. The method according to claim 1, characterized in that: the virus inactivation treatment in step 2) comprises: soaking the skin tissue with an aqueous solution of hydrogen peroxide, and cleaning after the soaking is finished;
the mass concentration of the aqueous solution of the hydrogen peroxide is 0.06-0.1 g/ml;
the soaking time is 1 to 1.5 hours;
the cleaning is ultrasonic cleaning.
3. The method according to claim 1 or 2, characterized in that: the degreasing treatment in step 3) includes: soaking the skin tissue with a mixed solution of chloroform and ethanol, and cleaning after the soaking is finished;
in the mixed solution, the volume ratio of chloroform to ethanol is 1: (1-5);
the soaking time is 30-60 min;
the cleaning is ultrasonic cleaning.
4. A method according to any one of claims 1-3, characterized in that: the decellularization treatment in step 4) includes: soaking the skin tissue in an aqueous solution of an alkaline reagent, and cleaning after the soaking is finished;
the alkaline reagent is NaOH or KOH;
the concentration of the alkaline reagent in the aqueous solution of the alkaline reagent is 0.5-5.5 mol/L;
the soaking time is 30-60 min;
the cleaning is ultrasonic cleaning;
the times of the decellularization treatment are 3-5 times.
5. The method according to any one of claims 1-4, wherein: the concentration of the aqueous solution of sodium hyaluronate in the step 5) is 0.01-0.1 g/ml;
the soaking in the step 5) comprises a first soaking and a second soaking after the first soaking finishes turning over the acellular dermal matrix, wherein the time of the first soaking and the time of the second soaking are respectively 10-20 min.
6. The method according to any one of claims 1-5, wherein: the method further comprises the following steps of 6): cutting the acellular dermal matrix after freeze drying; and, a step of, in the first embodiment,
and (3) punching the acellular dermal matrix after the shearing and cutting.
7. The method according to claim 6, wherein: the method further comprises the steps of packaging, sterilizing and analyzing sequentially after the punching treatment;
the sterilization is ethylene oxide sterilization or irradiation sterilization.
8. The method according to claim 7, wherein: the conditions for sterilizing the ethylene oxide are as follows: the temperature is 37-47 ℃, the humidity is 30-80%, the concentration of ethylene oxide is 500mg/L, and the sterilization time is 3-5 hours;
the analysis is carried out in an analysis oven at 37-47 ℃ for 4-6 days.
9. A biologic breast patch prepared by the method of any one of claims 1-8.
10. The biologic breast patch of claim 9, wherein: the biological breast patch is rectangular or crescent;
the diameter of the through holes in the biological breast patch is 0.5-2.5 mm, and the distance between two adjacent through holes is 1-2 cm;
the biological breast patch is crescent-shaped, and the through holes are distributed at the middle position of the biological breast patch.
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| CN105903080A (en) * | 2016-05-23 | 2016-08-31 | 苏州恒瑞迪生医疗科技有限公司 | Breast patch and preparation method thereof |
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| CN112618796A (en) * | 2020-11-16 | 2021-04-09 | 北京奥精医疗器械有限责任公司 | Acellular dermal matrix material and preparation method and application thereof |
| WO2022014769A1 (en) * | 2020-07-13 | 2022-01-20 | 주식회사 엘앤씨바이오 | Hydrated acellular skin substitute and manufacturing method therefor |
| CN114699565A (en) * | 2022-03-31 | 2022-07-05 | 杭州恒正医源生物科技有限公司 | Biological patch and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105903080A (en) * | 2016-05-23 | 2016-08-31 | 苏州恒瑞迪生医疗科技有限公司 | Breast patch and preparation method thereof |
| CN107890586A (en) * | 2017-10-27 | 2018-04-10 | 百澳瑞派（天津）生物科技有限公司 | A kind of preparation method of allogeneic biological sticking patch |
| WO2022014769A1 (en) * | 2020-07-13 | 2022-01-20 | 주식회사 엘앤씨바이오 | Hydrated acellular skin substitute and manufacturing method therefor |
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