A facial filler and skin texture improvement compound preparation based on decellularized matrix, its preparation method and application

Abstract

This application provides a facial filler and skin texture improvement composite formulation based on acellular matrix, its preparation method, and its application. By weight percentage, the facial filler and skin texture improvement composite formulation comprises 0.3%-20% polysaccharide and / or polysaccharide derivatives, 0.3%-6% osmotic pressure regulator, 0.5%-60% lyophilized decellularized matrix powder, and the balance being water. The polysaccharide is selected from at least one of sodium hyaluronate and carboxymethyl chitosan; the polysaccharide derivative is selected from at least one of PEGDE crosslinked carboxymethyl chitosan and oxidized sodium hyaluronate crosslinked carboxymethyl chitosan; the osmotic pressure regulator is a non-ionic osmotic pressure regulator; and the D50 particle size of the lyophilized decellularized matrix powder is 30-100 μm. This facial filler and skin texture improvement composite formulation based on acellular matrix exhibits excellent injection compliance, good suspension stability, and adjustable rheological properties.

Description

Technical Field

[0001] This application relates to the field of medical aesthetics technology, and in particular to a facial filling and skin texture improvement compound preparation based on decellularized matrix, its preparation method and application. Background Technology

[0002] Facial aging manifests as a decrease in soft tissue volume, sagging ligaments, reduced skin elasticity, and rough skin texture. Common facial rejuvenation treatments include surgical facelifts, laser therapy, and injectable fillers. Among these, injectable fillers have become one of the most popular non-surgical cosmetic procedures due to their minimally invasive nature, rapid results, and high reversibility.

[0003] Currently, commonly used facial filler materials mainly include hyaluronic acid (HA) fillers, poly-L-lactic acid (PLLA), calcium hydroxyapatite (CaHA), and autologous fat. Among them, hyaluronic acid fillers dominate the facial filler market due to their good biocompatibility, immediate filling effect, and biodegradability. However, hyaluronic acid fillers mainly have a locating effect and cannot induce autologous tissue regeneration.

[0004] Decellularized matrix (dECM) is a novel tissue engineering material that removes cellular components from xenogeneic or allogeneic tissues, retaining only the complete three-dimensional network structure of the extracellular matrix, including collagen, elastin, glycosaminoglycans, and various growth factors. As an injectable implantable material, granulated decellularized matrix shows great potential in the field of facial rejuvenation due to its unique tissue regeneration-promoting effects.

[0005] However, existing decellularized matrix (dECM) filler materials have the following technical bottlenecks: (1) High injection pressure: The injection pressure is high after the decellularized matrix is ​​mixed with physiological saline, which can easily clog the needle; (2) Poor suspension stability: It is difficult to achieve uniform suspension of particles with a single medium, and stratification is easy to occur; (3) Rheological properties do not match clinical needs: Different facial areas have different requirements for the rheological parameters of fillers. Summary of the Invention

[0006] To address the aforementioned technical problems, in a first aspect, this application provides a facial filling and skin texture improvement composite formulation based on decellularized matrix, comprising, by weight percentage, 0.3%-20% polysaccharides and / or polysaccharide derivatives, 0.3%-6% osmotic pressure regulators, 0.5%-60% decellularized matrix lyophilized powder, and the balance being water; The polysaccharide is selected from at least one of sodium hyaluronate and carboxymethyl chitosan; the polysaccharide derivative is selected from at least one of PEGDE crosslinked carboxymethyl chitosan and oxidized sodium hyaluronate crosslinked carboxymethyl chitosan; the osmotic pressure regulator is a non-ionic osmotic pressure regulator; The D50 particle size of the decellularized matrix lyophilized powder is 30-100 μm.

[0007] Preferably, the above-mentioned facial filler and skin texture improvement compound preparation based on decellularized matrix comprises, by weight percentage, 0.3%-1.8% polysaccharide and / or polysaccharide derivative, 4%-6% osmotic pressure regulator, 20%-40% decellularized matrix lyophilized powder, and the balance being water.

[0008] Preferably, the osmotic pressure regulator is selected from sorbitol, mannitol, and propylene glycol.

[0009] Preferably, the polysaccharide and / or polysaccharide derivative is a polysaccharide and polysaccharide derivative in a mass ratio of 3-5:1.

[0010] Preferably, the polysaccharide is sodium hyaluronate, and the polysaccharide derivative is PEGDE crosslinked carboxymethyl chitosan.

[0011] Preferably, the polysaccharide and / or polysaccharide derivative is PEGDE crosslinked carboxymethyl chitosan and sodium oxyhyaluronate crosslinked carboxymethyl chitosan in a mass ratio of 1:3-5.

[0012] Preferably, the preparation method of the PEGDE crosslinked carboxymethyl chitosan includes: dissolving carboxymethyl chitosan in water, adjusting the pH, adding polyethylene glycol diglycidyl ether, stirring at 30-50°C for 10-40 h, and freeze-drying to obtain the PEGDE crosslinked carboxymethyl chitosan.

[0013] Preferably, the mass ratio of the carboxymethyl chitosan to the polyethylene glycol diglycidyl ether is 1-3:1.

[0014] Preferably, the method for preparing sodium oxidized hyaluronic acid crosslinked carboxymethyl chitosan includes the following steps: a: Dissolve sodium hyaluronate in water, add sodium periodate, react, dialyze, and dry to obtain oxidized sodium hyaluronate; b: Dissolve sodium hyaluronate in water to obtain sodium hyaluronate solution, dissolve carboxymethyl chitosan in water to obtain carboxymethyl chitosan solution, mix sodium hyaluronate solution and carboxymethyl chitosan solution, stir, let stand to solidify, freeze dry to obtain sodium hyaluronate crosslinked carboxymethyl chitosan.

[0015] Preferably, in step a, the mass ratio of sodium hyaluronate to sodium periodate is 1.5-2:1.

[0016] Preferably, in step b, the sodium oxidized hyaluronic acid solution has a mass concentration of 1-5 wt%, the carboxymethyl chitosan solution has a mass concentration of 1-5 wt%, and the volume ratio of the sodium oxidized hyaluronic acid solution to the carboxymethyl chitosan solution is 1-1.5:1.

[0017] Preferably, the method for preparing decellularized matrix lyophilized powder includes: taking pig skin, removing the epidermis and subcutaneous fat tissue to obtain the dermis, separating the epidermis using Dispase II, treating dermal cells with Trypsin to remove antigenic components, washing, freeze-drying, pulverizing, and sieving to obtain the decellularized matrix lyophilized powder.

[0018] Preferably, the dosage form of the facial filler and skin texture improvement compound preparation based on decellularized matrix is ​​an injectable.

[0019] Secondly, the present invention provides a method for preparing the above-mentioned facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: dissolving polysaccharides and / or polysaccharide derivatives and osmotic pressure regulators in water to form a carrier solution, adding decellularized matrix lyophilized powder to the carrier solution, homogenizing, filling and sterilizing, to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0020] Preferably, the rotational speed of the homogenization process is 5000-15000 rpm.

[0021] Thirdly, the present invention provides the application of the above-mentioned facial filling and skin texture improvement composite preparation based on decellularized matrix in the preparation of medical devices for deep facial repair, volume filling, contour beautification or skin texture improvement.

[0022] In summary, the present invention has at least one of the following beneficial technical effects: 1. This application provides a facial filling and skin texture improvement composite formulation based on decellularized matrix, which has the following advantages: 1) Excellent injection compliance: Through the synergistic effect of polysaccharide carrier and decellularized matrix particles, the injection pushing force is 3-15N, which is superior to the traditional physiological saline suspension system (>15 N); 2) Good suspension stability: The viscosity of polysaccharides and / or polysaccharide derivatives is moderate, which can maintain the uniform suspension of decellularized matrix particles, and there is no obvious stratification after standing at room temperature for 7 days; 3) Adjustable rheological properties: By adjusting the type / concentration of polysaccharides and the particle size / concentration of decellularized matrix, G' can be precisely controlled in the range of 50-800 Pa; 4) Strong tissue regeneration capacity: Decellularized matrix can induce fibroblast proliferation and angiogenesis; 5) Good biocompatibility: The decellularized matrix has low immunogenicity after decellularization treatment.

[0023] 2. This application provides a method for preparing the above-mentioned facial filling and skin texture improvement compound preparation based on decellularized matrix. The preparation method is simple and easy to produce.

[0024] 3. This invention achieves precise control of rheological properties through the synergistic design of polysaccharide carriers and decellularized matrix. It provides differentiated formulation parameters for four indications: deep repair, volume filling, contour enhancement, and skin texture improvement. Experimental data validates key conclusions such as the feasibility of using carboxymethyl chitosan as a substitute for hyaluronic acid and the feasibility of a suitable polysaccharide concentration range. This formulation exhibits advantages such as good injection compliance, high suspension stability, good biocompatibility, and the ability to induce tissue regeneration. Detailed Implementation

[0025] The present invention will be further described below with reference to specific embodiments. However, the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified, specific conditions in the following embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the methods used are conventional methods known in the art, and the consumables and reagents used are commercially available. Unless otherwise stated, the technical and scientific terms used herein have the same meaning as those familiar with the art. Furthermore, any methods or materials similar to or equivalent to those described herein may also be applied to the present invention. The present application will be further described in detail below with reference to specific embodiments.

[0026] Carboxymethyl chitosan, degree of deacetylation (DD) ≥60%, degree of substitution ≥80%.

[0027] Sodium hyaluronate was purchased from Bloomage Biotechnology Co., Ltd., with a molecular weight ≥700,000 Daltons.

[0028] Preparation Example 1 The preparation method of PEGDE crosslinked carboxymethyl chitosan includes the following steps: dissolve 3 g of carboxymethyl chitosan (CMCS) in 100 mL of deionized water, adjust the pH to 8-9, stir until completely dissolved, add 1.5 g of PEGDE (polyethylene glycol diepoxyglycerol ether), stir at 45°C for 24 h, freeze dry to obtain PEGDE crosslinked carboxymethyl chitosan.

[0029] Preparation Example 2 The preparation method of oxidized sodium hyaluronate crosslinked carboxymethyl chitosan (OHA / CMCS) includes the following steps: a: Dissolve 5g of sodium hyaluronate in 250mL of water, add 3g of sodium periodate, stir for 10h in the dark at 30°C and pH 4.5-4.9, add ethylene glycol to terminate the reaction, dialyze the reaction solution and dry to obtain oxidized sodium hyaluronate (OHA). b: Dissolve sodium oxyhyaluronate in deionized water to prepare a 3wt% sodium oxyhyaluronate solution. Dissolve carboxymethyl chitosan in deionized water to prepare a 3wt% carboxymethyl chitosan solution. Mix the 3wt% sodium oxyhyaluronate solution and the 3wt% carboxymethyl chitosan solution at a volume ratio of 5:4. Stir at 25°C for 1 min, let stand to solidify for 12 h, freeze, and dry to obtain sodium oxyhyaluronate crosslinked carboxymethyl chitosan (OHA / CMCS).

[0030] Preparation Example 3 The method for preparing decellularized matrix (dECM) lyophilized powder includes the following steps: 1): Take fresh pig skin, remove the epidermis and subcutaneous fat tissue, retain the dermal papillary layer, rinse 3 times with phosphate-buffered saline (PBS) to remove residual blood and impurities, and obtain the dermal layer; 2): Immerse the dermis in a mixed solvent of chloroform and methanol (chloroform to methanol volume ratio of 2:1) at 4°C for 6 hours each time, for a total of 3 treatments to remove fat. After treatment, allow it to air dry overnight. 3): Immerse the defatted dermis in Dispase II solution (1.2 U / mL, PBS) and treat at 4°C for 12 h. After treatment, gently scrape off the epidermis, leaving the dermis intact, and rinse with PBS. Treat with Trypsin-EDTA solution (0.25%) at 37°C for 2 h, then terminate the reaction with fetal bovine serum. 4) The hypertonic salt SDS method was used, with an SDS concentration of 0.5% and a sodium chloride concentration of 0.5M. Treatment was carried out at room temperature for 48 hours, during which a horizontal shaker was used to agitate the solution at 100 rpm. After treatment, the solution was repeatedly washed with ultrapure water until no foam was produced. 5): Freeze-dry, sterilize, and test: DNA residue <50 ng / mg, SDS residue <0.1%, to obtain decellularized matrix (dECM) lyophilized powder.

[0031] Example 1

[0032] Example 1 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.5g sodium hyaluronate, 5g sorbitol, 30g decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0033] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain decellularized matrix (dECM) lyophilized powder with a D50 particle size of 100 μm.

[0034] Step 2): Dissolve 1.5g of sodium hyaluronate (HA) and 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 particle size of 100μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0035] The facial filler and skin texture improvement compound preparation based on decellularized matrix provided in this embodiment can be used for deep repair and lifting.

[0036] Example 2

[0037] Example 2 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.5g sodium hyaluronate, 5g mannitol, 30g decellularized matrix lyophilized powder (D50=80μm), and water for injection to 100g.

[0038] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 80 μm.

[0039] Step 2): Dissolve 1.5g of sodium hyaluronate (HA) and 5g of osmotic pressure regulator (mannitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 80μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0040] The facial filler and skin texture improvement compound preparation based on decellularized matrix provided in this embodiment can be used for volume filling.

[0041] Example 3

[0042] Example 3 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.2g sodium hyaluronate, 0.3g PEGDE crosslinked carboxymethyl chitosan prepared by the method of Preparation Example 1, 5g sorbitol, 30g decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0043] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0044] Step 2): Dissolve 1.2g of sodium hyaluronate (HA), 0.3g of PEGDE cross-linked carboxymethyl chitosan prepared by the method of Preparation Example 1, and 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm to the carrier solution, homogenize at 10000rpm for 5min, add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0045] The facial filler and skin texture improvement compound preparation based on decellularized matrix provided in this embodiment can be used for contour enhancement.

[0046] Example 4

[0047] Example 4 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.2g of sodium oxyhyaluronate cross-linked carboxymethyl chitosan prepared by the preparation method of Preparation Example 2, 0.3g of PEGDE cross-linked carboxymethyl chitosan prepared by the preparation method of Preparation Example 1, 5g of sorbitol, 30g of decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0048] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0049] Step 2): 1.2g of sodium oxyhyaluronate cross-linked carboxymethyl chitosan prepared by the preparation method of Preparation Example 2, 0.3g of PEGDE cross-linked carboxymethyl chitosan prepared by the preparation method of Preparation Example 1, and 5g of osmotic pressure regulator (sorbitol) were dissolved in 60g of water for injection to form a carrier solution. Then, 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm was added to the carrier solution, homogenized at 10000rpm for 5min, and water for injection was added to 100g. The mixture was mixed, filled and sterilized to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0050] The facial filler and skin texture improvement compound preparation based on decellularized matrix provided in this embodiment can be used for contour enhancement.

[0051] Example 5

[0052] Example 5 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.5g of PEGDE crosslinked carboxymethyl chitosan prepared by the method of Preparation Example 1, 5g of sorbitol, 30g of decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0053] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0054] Step 2): Dissolve 1.5g of PEGDE cross-linked carboxymethyl chitosan prepared by the method of Preparation Example 1 and 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with D50 of 100μm to the carrier solution, homogenize at 10000rpm for 5min, add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0055] Example 6

[0056] Example 6 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.5g of sodium oxyhyaluronate cross-linked carboxymethyl chitosan prepared by the preparation method of Example 2, 5g of sorbitol, 30g of decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0057] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0058] Step 2): Dissolve 1.5g of sodium oxyhyaluronate cross-linked carboxymethyl chitosan prepared by the preparation method of Preparation Example 2 and 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with D50 of 100μm to the carrier solution, homogenize at 10000rpm for 5min, add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0059] Example 7

[0060] Example 7 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1g carboxymethyl chitosan, 5g propylene glycol, 25g decellularized matrix lyophilized powder (D50=30μm), and water for injection to 100g.

[0061] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 30 μm.

[0062] Step 2): Dissolve 1g of carboxymethyl chitosan and 5g of osmotic pressure regulator (propylene glycol) in 60g of water for injection to form a carrier solution. Then add 25g of decellularized matrix (dECM) lyophilized powder with a D50 of 30μm to the carrier solution. Homogenize at 5000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0063] The facial filler and skin texture improvement compound preparation based on decellularized matrix provided in this embodiment can be used for skin texture improvement.

[0064] Example 8

[0065] Example 8 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 1.5g carboxymethyl chitosan, 5g sorbitol, 30g decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0066] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0067] Step 2): Dissolve 1.5g of carboxymethyl chitosan and 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0068] Example 9

[0069] Example 9 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 0.75g sodium hyaluronate, 0.75g carboxymethyl chitosan, 5g sorbitol, 30g decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0070] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0071] Step 2): Dissolve 0.75g of sodium hyaluronate, 0.75g of carboxymethyl chitosan, and 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm to the carrier solution. Homogenize at 10000rpm for 5min, add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0072] The facial filler and skin texture improvement compound preparation based on decellularized matrix provided in this embodiment can be used for contour enhancement.

[0073] Comparative Example 1

[0074] Comparative Example 1 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 5g sorbitol, 30g decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0075] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0076] Step 2): Dissolve 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0077] Comparative Example 2

[0078] Comparative Example 2 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 5g sorbitol, 30g decellularized matrix lyophilized powder (D50=80μm), and water for injection to 100g.

[0079] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 80 μm.

[0080] Step 2): Dissolve 5g of osmotic pressure regulator (sorbitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 80μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0081] Comparative Example 3

[0082] Comparative Example 3 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 5g mannitol, 30g decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100mL.

[0083] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0084] Step 2): Dissolve 5g of osmotic pressure regulator (mannitol) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0085] Comparative Example 4

[0086] Comparative Example 4 provides a facial filling and skin texture improvement compound preparation based on decellularized matrix, comprising: 0.63g sodium chloride, 30g of decellularized matrix lyophilized powder (D50=100μm), and water for injection to 100g.

[0087] The preparation method of a facial filler and skin texture improvement compound based on decellularized matrix includes: Step 1): The decellularized matrix (dECM) lyophilized powder obtained in Preparation Example 3 was pulverized and sieved to obtain a decellularized matrix (dECM) lyophilized powder with a D50 of 100 μm.

[0088] Step 2): Dissolve 0.63g of osmotic pressure regulator (sodium chloride) in 60g of water for injection to form a carrier solution. Then add 30g of decellularized matrix (dECM) lyophilized powder with a D50 of 100μm to the carrier solution. Homogenize at 10000rpm for 5min. Add water for injection to 100g, mix well, fill and sterilize to obtain the facial filling and skin texture improvement compound preparation based on decellularized matrix.

[0089] test: Rheological parameters were measured using a rotational rheometer (25°C, 1Hz); injection extrusion force was measured using a universal testing machine (27G needle, 30mm / min); among them, the data for G' (storage modulus), G'' (loss modulus), viscosity, and cohesion are mean ± standard deviation (n=3). Suspension stability: refers to whether the suspension layer separates after standing at 25°C. "Stable" means no separation after standing at 25°C for 7 days, and "unstable" means separation within 24 hours after standing at 25°C.

[0090] Layering rate: The facial filler and skin texture improvement composite formulations based on decellularized matrix obtained in the examples and comparative examples were loaded into 25 mL stoppered graduated cylinders and left to stand at 37°C for 28 days. The layering interface height was recorded. Layering rate = H 上清 / H 总 , where H 上清 H is the distance between the stratified interface and the liquid surface. 总 The total height of the facial filler and skin-improving compound formulation based on decellularized matrix filled into a graduated cylinder. The stratification rate is the average of three parallel measurements.

[0091] The test results are shown in Table 1.

[0092] Table 1. Performance test results of the facial filler and skin texture improvement composite formulations based on decellularized matrix obtained from the examples and comparative examples.

[0093] The DNA residue of the facial filling and skin improvement compound preparations based on decellularized matrix obtained in the examples and comparative examples was <50 ng / mg.

[0094] As shown in Table 1, the addition of the polysaccharide carrier significantly reduced the injection pushing force (down to 13.3 N) and improved injection compliance and suspension stability. As can be seen from the comparison of Examples 1-8 and Table 1, by adjusting the type of polysaccharide, particle size and concentration of decellularized matrix, G' can be precisely controlled within the range of 50-800 Pa to meet the needs of different indications.

[0095] As shown in Table 1, when carboxymethyl chitosan was used to replace sodium hyaluronate by the same amount, G', viscosity, and extrusion force all changed, indicating that the type of polysaccharide has a significant impact on rheological properties. As shown in Table 1, the combination of HA and CMCS in Example 9 can achieve intermediate value control of rheological properties, providing more options for formulation optimization. Furthermore, the combination of HA and CMCS has a synergistic effect on improving the stability of the formulation.

[0096] The above are all preferred embodiments of this application, and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, shape and principle of this application should be covered within the scope of protection of this application.

Claims

1. A facial filler and skin texture improvement compound preparation based on decellularized matrix, characterized in that, By weight percentage, it includes 0.3%-20% polysaccharides and polysaccharide derivatives, 0.3%-6% osmotic pressure regulators, 0.5%-60% decellularized matrix lyophilized powder, and the balance being water; The polysaccharide is selected from at least one of sodium hyaluronate and carboxymethyl chitosan; the polysaccharide derivative is sodium oxyhyaluronate crosslinked with carboxymethyl chitosan; the osmotic pressure regulator is a nonionic osmotic pressure regulator; The D50 particle size of the decellularized matrix lyophilized powder is 30-100 μm; The polysaccharides and polysaccharide derivatives are polysaccharides and polysaccharide derivatives with a mass ratio of 3-5:

1.

2. The facial filling and skin texture improvement compound preparation based on decellularized matrix according to claim 1, characterized in that, By weight percentage, it includes 0.3%-1.8% polysaccharides and polysaccharide derivatives, 4%-6% osmotic pressure regulators, 20%-40% decellularized matrix lyophilized powder, and the balance being water.

3. The facial filling and skin texture improvement compound preparation based on decellularized matrix according to claim 1, characterized in that, The osmotic pressure regulator is selected from sorbitol, mannitol, and propylene glycol.

4. The facial filling and skin texture improvement compound preparation based on decellularized matrix according to claim 1, characterized in that, The method for preparing the decellularized matrix freeze-dried powder includes: taking pig skin, removing the epidermis and subcutaneous fat tissue to obtain the dermis, removing fat, separating the epidermis using Dispase II, treating dermal cells with Trypsin to remove antigen components, washing, freeze-drying, pulverizing, and sieving to obtain the decellularized matrix freeze-dried powder.

5. The facial filling and skin texture improvement compound preparation based on decellularized matrix according to claim 1, characterized in that, The dosage form of the facial filling and skin texture improvement compound preparation based on decellularized matrix is ​​an injectable.

6. A method for preparing a facial filler and skin texture improvement composite formulation based on a decellularized matrix as described in any one of claims 1-5, characterized in that, include: The polysaccharide and its derivatives, along with the osmotic pressure regulator, are dissolved in water to form a carrier solution. The decellularized matrix lyophilized powder is then added to the carrier solution, homogenized, filled, and sterilized to obtain the facial filling and skin texture improvement composite formulation based on the decellularized matrix.

7. The method for preparing the facial filler and skin texture improvement composite formulation based on decellularized matrix according to claim 6, characterized in that, The homogenization process is carried out at a rotation speed of 5000-15000 rpm.

8. The use of the facial filler and skin texture improvement composite formulation based on decellularized matrix as described in any one of claims 1-5 in the preparation of implantable medical devices for deep facial repair, volume filling, contour enhancement or skin texture improvement.

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