Acellular matrix particle and hyaluronic acid-based gel premix material as well as preparation method and application thereof

Abstract

The invention provides an acellular matrix particle and hyaluronic acid-based gel premixed material as well as a preparation method and application thereof, and belongs to the technical field of biomedical materials. The preparation method comprises the following steps: respectively sterilizing the acellular matrix particles and the hyaluronic acid-based gel, and mixing to obtain the premixed material of the acellular matrix particles and the hyaluronic acid-based gel. The method for preparing the acellular matrix particles from the raw material end is provided, the animal-derived raw material is used and subjected to virus inactivation, decellularization, freeze-drying, grinding, sieving and sterilization, the ground acellular matrix particles do not need to be homogenized and subjected to secondary freeze-drying, and the preparation method of the acellular matrix particles is optimized. The prepared premixed material of the acellular matrix particles and the hyaluronic acid-based gel has good stability, is an ideal filling repair material, and has a wide application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical materials, and particularly relates to a premixed material of acellular matrix microparticles and hyaluronic acid-based gel, a preparation method thereof, and an application thereof. Background Technique

[0002] Hyaluronic Acid (HA) is a high-molecular-weight mucopolysaccharide formed by N-acetylglucosamine and D-glucuronic acid linked by β-1,4-glycosidic bonds. The abundant hydroxyl groups and hydrogen bonds in its molecular structure endow it with excellent hydrophilicity. In addition, the sponge-like water absorption characteristics of HA make it a recognized high-efficiency moisturizing agent. Initially, HA was mainly applied in the ophthalmic field. With the introduction of microbial fermentation technology, the extraction cost of HA raw materials has been significantly reduced, and its application scope has gradually expanded to the fields of cosmetics and medical beauty fillers.

[0003] Non-crosslinked or slightly crosslinked HA is mainly used for moisturizing and is suitable for smoothing shallow surface wrinkles such as the eyes and neck; crosslinked HA shows more diverse filling effects according to its own crosslinking degree and single / double-phase properties. The characteristics of low immunogenicity, good support force, and degradability of HA have made it one of the most widely used medical beauty fillers at present.

[0004] With the continuous improvement of the needs of beauty seekers, an ideal filling and repairing material should not only meet the shaping needs, but also have the characteristics of long-term naturalness and low adverse reactions. At present, there are still many problems with HA-related products on the market. HA is prone to the Tyndall phenomenon after injection, and triangular protrusions are likely to appear in some filling areas. Secondly, the maintenance effect of HA products has timeliness, and regular injections are required to maintain the expected beauty effect. Long-term HA injections may lead to changes in the skin tissue structure, including the formation of nodules and hard lumps.

[0005] In order to overcome the limitations of single use of HA, in recent years, researchers have tried to compound HA with other materials. CN 116808290 A discloses an injectable filler and a reconstitution method thereof, which involves mixing hyaluronic acid as a matrix with synthetic polymer microspheres (such as PCL, PLLA, PVA, etc.), and adding an aqueous solvent to form a colloidal gel during clinical use. However, in actual operation, due to the viscosity of HA, the operability of preparing HA into a dry powder for clinical compounding is low, and a uniform gel state can only be obtained through a long-time compounding. Secondly, the hydroxyapatite microspheres currently used as facial fillers are only limited to the tear trough area and the lower orbital margin, and common adverse reactions such as swelling and nodules occur.

[0006] CN 116212112 A discloses an injectable filling material and its preparation and application. The material is prepared by mixing the slurry formed by acellular allogeneic dermal microparticles with an aqueous hyaluronic acid solution and then performing irradiation treatment, which makes up for the defect that the acellular allogeneic dermal filling material is prone to delamination when used alone. From the perspective of raw material sources, the acellular allogeneic dermal microparticles used are autologous, and the acquisition channels are relatively limited, making large-scale production impossible.

[0007] Therefore, there is an urgent need in the art for a filling and repairing material that is convenient for clinical injection and meets the requirements of shaping, naturalness, and low adverse reactions. Summary of the Invention

[0008] The purpose of the present invention is to provide a premixed material of acellular matrix microparticles and hyaluronic acid-based gel, and its preparation method and application.

[0009] The present invention provides a composition, which is composed of acellular matrix microparticles, hyaluronic acid-based gel, and water. Among them, the mass-volume ratio of the acellular matrix microparticles, hyaluronic acid-based gel, and water is (5 - 25) g : (500 - 1500) mL : (100 - 500) mL.

[0010] Further, the mass-volume ratio of the acellular matrix microparticles, hyaluronic acid-based gel, and water is 15 g : 1000 mL : 300 mL; the hyaluronic acid-based gel is a crosslinked or non-crosslinked hyaluronic acid-based gel.

[0011] Further, the above water is sterile water for injection.

[0012] Further, the median particle size of the acellular matrix microparticles is between 50 and 200 μm;

[0013] The hyaluronic acid-based gel refers to a gel mainly composed of hyaluronic acid or its metal salt, and the content of hyaluronic acid or its metal salt in the hyaluronic acid-based gel is 10 - 50 mg / mL.

[0014] Further, the preparation of the acellular matrix microparticles includes the following steps:

[0015] (1) Raw material treatment: Take the acellular matrix raw material, wash it with water to remove fat;

[0016] (2) Virus inactivation: Inactivate the acellular matrix raw material obtained in step (1) with a peracetic acid-ethanol solution;

[0017] (3) Decellularization: Immerse the product obtained in step (2) in a Triton X-100 solution;

[0018] (4) After freeze-drying, grinding, sieving, and irradiation sterilization, the acellular matrix microparticles are obtained.

[0019] Furthermore, in step (1), the acellular matrix raw material is an animal-derived tissue or organ;

[0020] In step (2), the volume percentage concentration of peracetic acid in the peracetic acid-ethanol solution is 0.15-2.2%, the volume percentage concentration of ethanol is 10-35%, and the volume ratio of the peracetic acid-ethanol solution to the acellular matrix raw material is 10-15:1; the inactivation time is 1.5-8 hours, and the temperature is 16-37°C;

[0021] In step (3), the mass percentage of the Triton X-100 solution is 0.15-5.5%, the volume ratio of the Triton X-100 solution to the acellular matrix raw material is (1-10):1, the soaking temperature is 35-45°C, and the soaking time is 1-2 h;

[0022] In step (4), the freeze-drying conditions are: keep warm at -50 to -30°C for 10-18 hours, and then raise the temperature to 30-40°C for freeze-drying and dehydration;

[0023] The temperature inside the equipment for grinding is 0-40°C;

[0024] The dose of the irradiation sterilization is 15-25 kGy.

[0025] Furthermore, in step (2), the volume percentage concentration of peracetic acid in the peracetic acid-ethanol solution is 0.2%, the volume percentage concentration of ethanol is 20%, and the volume ratio of the peracetic acid-ethanol solution to the acellular matrix raw material is 13:1; the inactivation time is 4 hours, and the temperature is 25°C;

[0026] In step (3), the mass percentage of the Triton X-100 solution is 0.25%, the volume ratio of the Triton X-100 solution to the acellular matrix raw material is 6:1, and the soaking temperature is 37°C;

[0027] In step (4), the freeze-drying conditions are: keep warm at -40°C for 12 hours, and then raise the temperature to 35°C for freeze-drying and dehydration;

[0028] The temperature inside the equipment for grinding is 7°C;

[0029] The conditions for sieving are: the median of the particle size distribution for screening is 10-1000 μm, preferably 50-200 μm;

[0030] The irradiation sterilization is Co-60 sterilization or electron beam sterilization, and the irradiation dose is 15 kGy dose.

[0031] Further, the acellular matrix is derived from one or a mixture of pericardium, peritoneum, mucosa, peritoneum, tendon, muscle, bone, cartilage, and skin of animals such as pigs, cows, and sheep.

[0032] Further, the preparation of the non-crosslinked hyaluronic acid-based gel comprises the following steps: dissolving hyaluronic acid or its metal salt in a buffer solution and sterilizing it to obtain the gel.

[0033] Further, the pushing force range of the non-crosslinked hyaluronic acid-based gel is 5 - 25 N.

[0034] Further, the dissolving time is 0.5 - 1.5 h and the temperature is 16 - 37 °C.

[0035] Further, the preparation of the crosslinked hyaluronic acid-based gel comprises the following steps:

[0036] (a) Dissolving hyaluronic acid or its metal salt in water to obtain a solution;

[0037] (b) Adjusting the pH of the solution to 9 - 13, adding a crosslinking agent for reaction, and then adjusting the pH to neutral;

[0038] (c) Adding an alcohol solution to the solution for dialysis, adjusting the osmotic pressure with a buffer solution, and sterilizing it to obtain the gel.

[0039] Further, in step (a), the mass ratio of water to hyaluronic acid or its metal salt is 15:1 - 30:1, the dissolving temperature is 16 - 37 °C; the dissolving time is 0.5 - 1.5 h;

[0040] In step (b), the crosslinking agent is glycidyl ether, diepoxide, dicyclohexylcarbodiimide, divinyl sulfone, or a multifunctional polyethylene glycol-based crosslinking agent, and the mass ratio of hyaluronic acid or its metal salt to the crosslinking agent is 1:(0.2 - 0.3), preferably 1:0.25;

[0041] In step (c), the purity of the alcohol solution is ≥95%, the volume ratio of the alcohol solution to the solution is 20:1 - 50:1; the number of dialysis times is ≥3 times; the osmotic pressure of the buffer solution is 240 - 340 mOsmol / kg; the sterilization is moist heat sterilization, the sterilization temperature is 110 - 115 °C, and the sterilization time is 8 - 15 min.

[0042] Further, the pushing force of the crosslinked hyaluronic acid-based gel (equipped with a 27G injection needle) after sterilization is 5 - 25 N, the median particle size distribution is 100 - 1200 μm, the swelling degree range is 20 - 200, and the elastic modulus at 1 Hz is 100 - 1000 Pa.

[0043] The present invention also provides a method for preparing the above composition, the method comprising the following steps: mixing acellular matrix microparticles, hyaluronic acid-based gel and water to obtain the composition.

[0044] Further, the mixing time is 20 to 60 min.

[0045] Further, the extrusion force range of the composition (equipped with a 27G injection needle) obtained after mixing is 5 to 25 N, the median particle size distribution is 25 - 1200 μm, the swelling degree range is 20 - 200, and the elastic modulus is 100 - 1000 Pa at 1 Hz.

[0046] The present invention also provides the use of the above composition in the preparation of medical beauty products, cosmetics, and wound repair materials.

[0047] Compared with the prior art, the present invention has achieved the following beneficial effects:

[0048] (1) The present invention obtains acellular matrix microparticles with a specific particle size distribution from the raw material end through virus inactivation, decellularization, freeze-drying, grinding, and sieving. The process is simple and can achieve large-scale production.

[0049] (2) The present invention separately sterilizes the acellular matrix microparticles and the hyaluronic acid-based gel, and mixes and fills them during the isolation period to obtain the final premixed material, which can avoid final sterilization of the mixture.

[0050] (3) The premixed material prepared by the present invention avoids the safety risks that may be introduced during the compounding process, and at the same time ensures that the prepared premixed material has good stability and is convenient for direct use.

[0051] (4) The present invention uses hyaluronic acid as the carrier medium of the acellular matrix microparticles to achieve skin volume filling and avoid displacement of the injectable material. The acellular matrix microparticles can stimulate collagen regeneration for a long time, and the two work together to achieve good long-term filling efficacy.

[0052] Obviously, based on the above content of the present invention, according to the common general knowledge and conventional means in the art, without departing from the above basic technical idea of the present invention, various other forms of modifications, substitutions, or changes can be made.

[0053] The above content of the present invention will be further described in detail below through specific embodiments in the form of examples. However, it should not be understood that the scope of the above subject matter of the present invention is limited to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. Detailed Description of the Invention

[0054] The raw materials and equipment used in the present invention are all known products and are obtained by purchasing commercially available products.

[0055] Example 1: Preparation of acellular matrix microparticles

[0056] (1) Raw material treatment: Take the acellular matrix raw material, repeatedly rinse it with clear water until no obvious foreign objects are visible to the naked eye, and use a scraper or scraping equipment to remove excess fat;

[0057] (2) Virus inactivation: Treat the above-obtained product with peracetic acid-ethanol solution, where the volume percentage concentration of peracetic acid is 0.2% and the volume percentage concentration of ethanol is 20%. The volume ratio of peracetic acid-ethanol solution to the acellular matrix raw material is 13:1, the inactivation time is 4 hours, and the temperature is 25°C;

[0058] (3) Decellularization: Immerse the above-obtained product in a 0.25% (mass percentage) Triton X-100 solution for 1.5 h. The volume ratio of Triton X-100 solution to the acellular matrix raw material is 6:1, and the temperature is 37°C;

[0059] (4) Freeze-drying: Put the obtained acellular matrix material into a vacuum freeze-dryer. The freezing temperature is -40°C, keep it at this temperature for 12 hours, and then gradually raise the temperature to 35°C for freeze-drying dehydration;

[0060] (5) Grinding: Grind the above-obtained product, and the temperature inside the equipment where the material is located during this process is 7°C;

[0061] (6) Sieving: Sieve the above-obtained product through a 200-μm sieve;

[0062] (7) Sterilization: Spread the above-obtained product on a covered glass dish for irradiation sterilization, and the irradiation dose is 15 kGy.

[0063] Among them, the median particle size of the acellular matrix microparticles prepared according to the above steps is 50 - 200 μm, and the purity of the acellular matrix microparticles is 80% - 90%.

[0064] Example 2: Preparation of non-crosslinked sodium hyaluronate gel

[0065] Dissolve sodium hyaluronate with a molecular weight of 866 KDa in phosphate buffer solution. Place it in a water bath at 37°C and stir for 1.5 h, and then perform moist heat sterilization (sterilize at 115°C for 15 min) to finally obtain a non-crosslinked sodium hyaluronate gel with a sodium hyaluronate content of 20 mg / mL.

[0066] Example 3: Preparation of non-crosslinked sodium hyaluronate gel

[0067] Refer to the method of Example 2, the difference is only that sodium hyaluronate with a molecular weight of 866 KDa is replaced by sodium hyaluronate with a molecular weight of 1500 KDa, and finally a non-crosslinked sodium hyaluronate gel with a sodium hyaluronate content of 20 mg / mL is obtained.

[0068] Example 4: Preparation of Non-crosslinked Sodium Hyaluronate Gel

[0069] Referring to the method of Example 2, the difference is only that the sodium hyaluronate with a molecular weight of 866KDa is replaced by sodium hyaluronate with a molecular weight of 2860KDa, and finally a non-crosslinked sodium hyaluronate gel with a sodium hyaluronate content of 20mg / mL is obtained.

[0070] Example 5: Preparation of Crosslinked Sodium Hyaluronate Gel

[0071] (1) Dissolve sodium hyaluronate with a molecular weight of 866KDa in water for injection, and the mass ratio of water for injection to sodium hyaluronate is 20:1. Place it in a water bath at 37°C and stir for 1.5h.

[0072] (2) Adjust the pH of the solution to 11.5 with 0.1% sodium hydroxide, add 1,4-butanediol diglycidyl ether (BDDE) to the mixed solution for crosslinking reaction, sodium hyaluronate:BDDE (mass ratio) = 1:0.25. After the crosslinking reaction is completed, add hydrochloric acid to neutralize the pH to neutral.

[0073] (3) Add ethanol solution for dialysis 3 times, and the mass ratio of ethanol to the material for each dialysis is 2:1. After dialysis, add phosphate buffer solution to adjust the osmotic pressure to 330mOsmol / kg, and sterilize by moist heat (sterilize at 115°C for 15min). Finally, a crosslinked sodium hyaluronate gel with a sodium hyaluronate content of 20mg / mL is obtained.

[0074] Example 6: Preparation of Crosslinked Sodium Hyaluronate Gel

[0075] Referring to the method of Example 5, the difference is only that the sodium hyaluronate with a molecular weight of 866KDa is replaced by sodium hyaluronate with a molecular weight of 1500KDa, and finally a crosslinked sodium hyaluronate gel with a sodium hyaluronate content of 20mg / mL is obtained.

[0076] Example 7: Preparation of Crosslinked Sodium Hyaluronate Gel

[0077] Referring to the method of Example 5, the difference is only that the sodium hyaluronate with a molecular weight of 866KDa is replaced by sodium hyaluronate with a molecular weight of 2860KDa, and finally a crosslinked sodium hyaluronate gel with a sodium hyaluronate content of 20mg / mL is obtained.

[0078] Example 8: Preparation of Premixed Material of Decellularized Matrix Particles and Non-crosslinked Hyaluronic Acid-based Gel

[0079] Mix 15 g of the acellular matrix microparticles prepared in Example 1, 1000 mL of the non-crosslinked sodium hyaluronate gel solution prepared in Example 2, and 300 mL of sterilized injection water in an isolator for 20 minutes. Then fill the mixed gel into a prefilled syringe to obtain a premixed material of acellular matrix microparticles and non-crosslinked sodium hyaluronate gel.

[0080] Example 9: Preparation of a premixed material of acellular matrix microparticles and non-crosslinked hyaluronic acid-based gel

[0081] Refer to the method of Example 8, with the only difference being that the non-crosslinked sodium hyaluronate gel solution in Example 2 is replaced with the non-crosslinked sodium hyaluronate gel solution in Example 3 to obtain the product.

[0082] Example 10: Preparation of a premixed material of acellular matrix microparticles and non-crosslinked hyaluronic acid-based gel

[0083] Refer to the method of Example 8, with the only difference being that the non-crosslinked sodium hyaluronate gel solution in Example 2 is replaced with the non-crosslinked sodium hyaluronate gel solution in Example 4 to obtain the product.

[0084] Example 11: Preparation of a premixed material of acellular matrix microparticles and crosslinked hyaluronic acid-based gel

[0085] Refer to the method of Example 8, with the only difference being that the non-crosslinked sodium hyaluronate gel solution in Example 2 is replaced with the crosslinked sodium hyaluronate gel solution in Example 5 to obtain the product.

[0086] Example 12: Preparation of a premixed material of acellular matrix microparticles and crosslinked hyaluronic acid-based gel

[0087] Refer to the method of Example 8, with the only difference being that the non-crosslinked sodium hyaluronate gel solution in Example 2 is replaced with the crosslinked sodium hyaluronate gel solution in Example 6 to obtain the product.

[0088] Example 13: Preparation of a premixed material of acellular matrix microparticles and crosslinked hyaluronic acid-based gel

[0089] Refer to the method of Example 8, with the only difference being that the non-crosslinked sodium hyaluronate gel solution in Example 2 is replaced with the crosslinked sodium hyaluronate gel solution in Example 7 to obtain the product.

[0090] The beneficial effects of the present invention are demonstrated by the following experimental examples.

[0091] Experimental Example 1: Testing the particle size distribution of acellular matrix microparticles

[0092] 1. Experimental method

[0093] Prepare acellular matrix microparticles (Table 1: Samples 1-6) respectively according to the method of Example 1. Disperse the obtained acellular matrix microparticles in purified water, and measure the particle size distribution by a laser particle size analyzer equipped with a wet dispersion instrument.

[0094] 2. Experimental results

[0095] Table 1 Test results of particle size distribution of acellular matrix microparticles

[0096]

[0097] The median particle size distribution results of the acellular matrix microparticles meet the requirements between 10 and 1000 μm. The results in Table 1 show that the median particle size of the acellular matrix microparticles of the present invention is between 50 and 200 μm, meeting the requirements. Experimental Example 2. Physicochemical property tests of non-crosslinked sodium hyaluronate gel and crosslinked sodium hyaluronate gel

[0098] 1. Experimental method

[0099] (1) Pushing force test: Use a universal material testing machine to push the sodium hyaluronate gel and crosslinked sodium hyaluronate gel out of a 27G injection needle respectively at a pushing rate of 30 mm / min, and record the average value of the required force.

[0100] (2) Particle size distribution test: Disperse the crosslinked sodium hyaluronate gel prepared in Examples 5-7 in a physiological saline solution and measure it by a laser particle size analyzer equipped with a wet dispersion instrument.

[0101] (3) Swelling degree test: Place the crosslinked sodium hyaluronate gel on a 500-mesh sieve, put it in a petri dish, add 0.9% sodium chloride solution to completely soak the sample. After the sample swells sufficiently, take out the sieve and blot the liquid at the bottom and around the sieve with filter paper, then weigh it and dry it to a constant weight at 80 °C.

[0102] (4) Elastic modulus test: Use a rheometer to test. At 25 ± 2 °C, use a plate diameter of 40 mm at a gap height of 1 mm, and perform the test at a strain of 0.8% and a frequency of 1 Hz each time.

[0103] 2. Experimental results

[0104] Table 2 Performance test of non-crosslinked sodium hyaluronate gel

[0105] Molecular weight (KDa) Ejection force (N) Example 2 866 7.25 Example 3 1500 8.35 Example 4 2860 9.32

[0106] The results in Table 2 show that the pushing forces of the non-crosslinked sodium hyaluronate gels prepared in Examples 2-4 are within the set range (5-25 N).

[0107] Table 3 Performance test of crosslinked sodium hyaluronate gel

[0108]

[0109]

[0110] Combining the results in Tables 2 and 3, it can be seen that the extrusion force of the crosslinked sodium hyaluronate gels prepared in Examples 5-7 is higher than that of the non-crosslinked sodium hyaluronate gel but is also within the set range (5-25 N), which can meet the appropriate feel of clinicians. The elastic modulus and swelling degree also meet the requirements and can meet the expected use for facial filling.

[0111] Experimental Example 3. Performance Test of Acellular Matrix Particle Premixed Non-Crosslinked Hyaluronic Acid-Based Gel

[0112] 1. Experimental Method

[0113] As described in Experimental Example 2.

[0114] 2. Experimental Results

[0115] Table 4. Performance Test of Acellular Matrix Particle Premixed Non-Crosslinked Hyaluronic Acid-Based Gel

[0116]

[0117] The results in Table 4 show that there is not much change in the extrusion force of the premixed material of the acellular matrix particle premixed non-crosslinked sodium hyaluronate gel compared with that before mixing (Table 2), and the median of the particle size distribution after mixing meets the requirements.

[0118] Experimental Example 4. Performance Test of Acellular Matrix Particle Premixed Crosslinked Hyaluronic Acid-Based Gel

[0119] 1. Experimental Method

[0120] As described in Experimental Example 2.

[0121] 2. Experimental Results

[0122] Table 5. Performance Test of Acellular Matrix Particle Premixed Crosslinked Hyaluronic Acid-Based Gel

[0123]

[0124] The results in Table 5 show that there is not much change in each property of the premixed material of the acellular matrix particle premixed crosslinked sodium hyaluronate gel compared with that before mixing (Table 3), and each index after mixing still meets the requirements.

[0125] Experimental Example 5. Long-Term Storage Experiment of Premixed Material of Acellular Matrix Particle and Crosslinked Hyaluronic Acid-Based Gel

[0126] 1. Experimental Method

[0127] Select the premixed material prepared in Example 12 (i.e., the sample obtained by mixing acellular matrix microparticles with a median particle size distribution in the range of 50 - 200 μm and cross-linked sodium hyaluronate gel with a raw material molecular weight of 1500 KDa), place it under laboratory conditions (temperature 20 - 25 °C, humidity 50% - 70% RH), and conduct long-term placement tests, push-out force, particle size distribution, and swelling degree tests (the test methods for push-out force, particle size distribution, and swelling degree are as described in Experimental Example 2).

[0128] 2. Experimental Results

[0129] Table 6 Test Results of Long-Term Placement Performance of Premixed Material of Acellular Matrix Microparticles and Cross-Linked Hyaluronic Acid-Based Gel

[0130]

[0131] The results in Table 6 show that the appearance of the premix samples at each time point was stable, no delamination was observed, and there were no significant differences in the push-out force, median particle size distribution, and swelling degree, indicating that the premixed material of acellular matrix microparticles and hyaluronic acid-based gel prepared in the present invention has good stability.

[0132] In summary, the present invention provides a premixed material of acellular matrix microparticles and hyaluronic acid-based gel, its preparation method and application. In the present invention, the acellular matrix microparticles and hyaluronic acid-based gel are sterilized separately and then mixed to obtain the premixed material of acellular matrix microparticles and hyaluronic acid-based gel. The present invention provides a method for preparing acellular matrix microparticles from the raw material end. Using animal-derived raw materials, through virus inactivation, decellularization, freeze-drying, grinding and sieving, and sterilization, the ground acellular matrix microparticles do not require homogenization and secondary freeze-drying, optimizing the preparation method of acellular matrix microparticles. The premixed material of acellular matrix microparticles and hyaluronic acid-based gel prepared in the present invention has good stability, is an ideal filling and repair material, and has broad application prospects.

Claims

1. A composition, characterized in that The invention is composed of decellularized matrix particles, hyaluronic acid-based gel and water, wherein the mass volume ratio of the decellularized matrix particles, hyaluronic acid-based gel and water is (5-25) g: (500-1500) mL: (100-500) mL.

2. The composition according to claim 1, characterized in that The mass volume ratio of the decellularized matrix particles, hyaluronic acid-based gel and water is 15g:1000mL:300mL; the hyaluronic acid-based gel is a cross-linked or non-cross-linked hyaluronic acid-based gel.

3. The composition according to claim 1, characterized in that The median particle size of the acellular matrix particles is between 50 and 200 μm; The hyaluronic acid-based gel refers to a gel with hyaluronic acid or its metal salt as the main component, and the content of hyaluronic acid or its metal salt in the hyaluronic acid-based gel is 10 to 50 mg / mL.

4. The composition according to claim 1, characterized in that The preparation of the decellularized matrix microparticles comprises the following steps: (1) Raw material processing: Take the decellularized matrix raw material, wash it with water, and remove fat; (2) Virus inactivation: The decellularized matrix material obtained in step (1) is inactivated using a peracetic acid-ethanol solution; (3) Decellularization: Soak the product obtained in step (2) in a Triton X-100 solution; (4) After freeze-drying, grinding, sieving, and irradiation sterilization, the decellularized matrix microparticles are obtained.

5. The composition according to claim 4, characterized in that In step (2), the volume percentage concentration of peracetic acid in the peracetic acid-ethanol solution is 0.15-2.2%, the volume percentage concentration of ethanol is 10-35%, and the volume ratio of the peracetic acid-ethanol solution to the decellularized matrix raw material is 10-15:1; the inactivation time is 1.5-8 hours, and the temperature is 16-37° C.; In step (3), the mass percentage of the Triton X-100 solution is 0.15-5.5%, the volume ratio of the Triton X-100 solution to the decellularized matrix material is (1-10):1, the immersion temperature is 35-45° C., and the immersion time is 1-2 h; In step (4), the freeze-drying conditions are: keeping warm at -50 to -30°C for 10 to 18 hours, then heating to 30 to 40°C for freeze-drying and dehydration; The temperature inside the grinding equipment is 0-40°C; The radiation sterilization dosage is 15 to 25 kGy.

6. The composition according to claim 2, characterized in that The preparation of the non-cross-linked hyaluronic acid-based gel comprises the following steps: dissolving hyaluronic acid or its metal salt in a buffer solution and sterilizing the solution.

7. The composition according to claim 2, characterized in that The preparation of the cross-linked hyaluronic acid-based gel comprises the following steps: (a) dissolving hyaluronic acid or its metal salt in water to obtain a solution; (b) adjusting the pH of the solution to 9-13, adding a cross-linking agent for reaction, and then adjusting the pH to neutral; (c) Add alcohol solution to the solution for dialysis, add buffer to adjust the osmotic pressure, and sterilize to obtain the product.

8. The composition according to claim 7, characterized in that In step (a), the mass ratio of water to hyaluronic acid or its metal salt is 15:1 to 30:1, the dissolution temperature is 16 to 37° C., and the dissolution time is 0.5 to 1.5 h; In step (b), the cross-linking agent is glycidyl ether, diepoxide, biscarbodiimide, divinyl sulfone or multifunctional polyethylene glycol-based cross-linking agent, and the mass ratio of the hyaluronic acid or its metal salt to the cross-linking agent is 1:(0.2-0.3), preferably 1:0.25; In step (c), the purity of the alcohol solution is ≥95%, the volume ratio of the alcohol solution to the solution is 20:1 to 50:1; the number of dialysis is ≥3 times; the osmotic pressure of the buffer solution is 240 to 340 mOsmol / kg; the sterilization is wet heat sterilization, the sterilization temperature is 110 to 115° C., and the sterilization time is 8 to 15 min.

9. A method for preparing the composition according to any one of claims 1 to 8, characterized in that: The method comprises the following steps: mixing decellularized matrix microparticles, hyaluronic acid-based gel and water to obtain the product.

10. Use of the composition according to any one of claims 1 to 8 in the preparation of medical cosmetology, cosmetics, and wound repair materials.

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