A recombinant collagen type iii complex cosmetic injection filler and a preparation method thereof
By combining genipin crosslinking with bacterial cellulose and catechin-oxidized sodium alginate microspheres, a stable collagen hydrogel is formed, solving the durability and safety issues of recombinant type III collagen injectable fillers and achieving excellent cosmetic repair effects.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- ANHUI KEMEN BIOTECHNOLOGY CO LTD
- Filing Date
- 2023-07-31
- Publication Date
- 2026-06-19
AI Technical Summary
Existing recombinant type III collagen injectable fillers are difficult to provide long-lasting filling and repair effects, and their safety and compatibility need to be improved.
Genipin was used as a crosslinking agent to form a hydrogel, and bacterial cellulose and catechin-oxidized sodium alginate microspheres were added to form a stable composite material. Through intertwining and slow release of catechins, the stability and durability of the collagen network were enhanced.
It achieves excellent filling and repair effects and can maintain the filling effect for a long time, improving safety and compatibility, making it a highly efficient cosmetic repair material.
Smart Images
Figure BDA0004368565300000081
Abstract
Description
Technical Field
[0001] This invention relates to the field of cosmetic injection filling technology, and in particular to a recombinant type III collagen composite cosmetic injection filler and its preparation method. Background Technology
[0002] In recent years, the increasing awareness of beauty has driven the rapid development of the medical aesthetics industry. Injectable fillers are a non-surgical cosmetic procedure that involves injecting biological or synthetic biocompatible materials into the dermis or subcutaneous layer. This method can fill depressions or eliminate wrinkles, thereby achieving a more youthful appearance. Currently, common biological materials used in injectable fillers include hyaluronic acid and collagen. Compared to synthetic materials, they have higher compatibility with human tissues, are less prone to complications such as hardening and edema, and offer greater safety. Recombinant type III collagen has a flexible triple helix structure that can form a dense and well-organized collagen fiber network, providing excellent skin repair and regeneration functions. However, how to provide recombinant type III collagen injectable fillers with excellent filling and repair effects that last for a long time remains a challenge that requires further improvement. Summary of the Invention
[0003] Based on the technical problems existing in the background technology, the present invention proposes a recombinant type III collagen composite cosmetic injectable filler and its preparation method.
[0004] The present invention proposes a recombinant type III collagen composite cosmetic injectable filler, comprising the following raw materials in parts by weight: 0.5-1 parts sodium hyaluronate, 0.1-0.2 parts recombinant type III humanized collagen, 0.02-0.05 parts genipin, 0.03-0.08 parts catechin-oxidized sodium alginate microspheres, 0.01-0.03 parts bacterial cellulose, and 100 parts physiological saline.
[0005] Preferably, the preparation method of the catechin-oxidized sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to an aqueous solution of oxidized sodium alginate, dispersed by high-speed stirring, then calcium chloride solution is added and stirred at high speed to react, and then freeze-dried to obtain the final product.
[0006] Preferably, in the preparation method of the catechin-oxidized sodium alginate microspheres, the high-speed stirring dispersion speed is 5000-10000 r / min and the time is 20-40 min, and the high-speed stirring reaction speed is 5000-8000 r / min and the time is 1-3 h.
[0007] Preferably, in the preparation method of the catechin-oxidized sodium alginate microspheres, the ratio of catechin, Span80, liquid paraffin and anhydrous ethanol is 1g:(0.1-0.3)g:(5-10)mL:(20-30)mL, the volume ratio of oil phase to aqueous solution of oxidized sodium alginate and calcium chloride solution is 1:(3-5):(3-5), the mass concentration of aqueous solution of oxidized sodium alginate is 1-2%, and the mass concentration of calcium chloride solution is 1-2%.
[0008] Oxidized sodium alginate is formed by oxidizing the cis-diol structure of sodium alginate molecules with oxidants such as sodium periodate and potassium permanganate to produce a dialdehyde structure. The preparation method can employ conventional methods in the field, for example: adding 10-20% (by mass) of sodium periodate to a 1-3% sodium alginate aqueous solution, reacting in the dark for 18-24 hours, then terminating the reaction with an appropriate amount of ethylene glycol, adding anhydrous ethanol to precipitate the product, dissolving it in deionized water, and repeating the precipitation-precipitation step 2-3 times. The resulting product is then vacuum dried to obtain the final product.
[0009] Bacterial cellulose (BC) is a porous, network-like, nanoscale biopolymer material synthesized by microorganisms, such as Acetobacter xylinum, through fermentation. Preferably, the length of the bacterial cellulose is 20-200 nm.
[0010] A method for preparing the recombinant type III collagen composite cosmetic injectable filler includes the following steps:
[0011] S1. Dissolve sodium hyaluronate and recombinant type III humanized collagen in an aqueous sodium hydroxide solution, then add genipin to carry out a cross-linking reaction. Wash the resulting gel with water, freeze-dry it, and grind it into powder to obtain gel powder.
[0012] S2. Add bacterial cellulose to physiological saline, disperse it evenly by ultrasonication, and then add gel powder and catechin-oxidized sodium alginate microspheres and mix evenly to obtain the final product.
[0013] Preferably, the concentration of the sodium hydroxide solution is 0.05-0.1 mol / L.
[0014] Preferably, the crosslinking reaction is carried out at a temperature of 30-60°C for 3-8 hours.
[0015] The beneficial effects of this invention are as follows:
[0016] This invention uses genipin as a crosslinking agent to moderately crosslink recombinant type III humanized collagen and sodium hyaluronate to form a hydrogel. Furthermore, it adds porous, network-like nanoscale biopolymer bacterial cellulose, whose structure is similar to collagen, providing synergistic support. Oxidized sodium alginate microspheres loaded with catechins, an effective collagen-promoting component, serve two purposes: firstly, the microspheres act as crosslinking points, allowing bacterial cellulose and collagen to intertwine more tightly, forming a more stable composite hydrogel that is less prone to degradation in vivo, thus maintaining a long-lasting filling effect; secondly, the slow release of catechins from within the oxidized sodium alginate microspheres allows newly formed collagen to cover the bacterial cellulose and collagen network structure, further enhancing the filler's durability. Therefore, this composite cosmetic injectable filler not only offers excellent filling and repair effects but also maintains its effect for a long time, making it a highly promising cosmetic repair material. Detailed Implementation
[0017] The technical solution of the present invention will now be described in detail through specific embodiments.
[0018] Example 1
[0019] A recombinant type III collagen composite cosmetic injectable filler comprises the following raw materials in parts by weight: 0.8 parts sodium hyaluronate, 0.15 parts recombinant type III humanized collagen, 0.03 parts genipin, 0.05 parts catechin-oxidized sodium alginate microspheres, 0.02 parts bacterial cellulose, and 100 parts physiological saline.
[0020] The preparation method of catechin-oxidized sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin, and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to a 1.5% (w / w) aqueous solution of oxidized sodium alginate, and the mixture is dispersed by high-speed stirring at 8000 r / min for 30 min; then a 1.5% (w / w) calcium chloride solution is added, and the mixture is stirred at 6000 r / min for 2 h; finally, the mixture is freeze-dried to obtain the final product. The ratio of catechin, Span80, liquid paraffin, and anhydrous ethanol is 1 g:0.2 g:6 mL:25 mL, and the volume ratio of the oil phase to the aqueous solution of oxidized sodium alginate and the calcium chloride solution is 1:4:4.
[0021] The preparation method of oxidized sodium alginate is as follows: Add sodium periodate equivalent to 15% of the mass of sodium alginate to a 2% sodium alginate aqueous solution, react in the dark for 20 hours, then add an appropriate amount of ethylene glycol to terminate the reaction, add anhydrous ethanol to precipitate the precipitate, and then dissolve it in deionized water. Repeat the precipitation-precipitation step 3 times, and then vacuum dry the obtained product to obtain the product.
[0022] The preparation method of the above-mentioned recombinant type III collagen composite cosmetic injectable filler is as follows:
[0023] S1. Sodium hyaluronate and recombinant type III humanized collagen were dissolved in a 0.05 mol / L sodium hydroxide aqueous solution, and then genipin was added. The cross-linking reaction was carried out at 50°C for 5 hours. The resulting gel was washed with water, freeze-dried, and then ground into powder to obtain gel powder.
[0024] S2. Add bacterial cellulose to physiological saline, disperse it evenly by ultrasonication, and then add gel powder and catechin-oxidized sodium alginate microspheres and mix evenly to obtain the final product.
[0025] Example 2
[0026] A recombinant type III collagen composite cosmetic injectable filler comprises the following raw materials in parts by weight: 0.5 parts sodium hyaluronate, 0.1 parts recombinant type III humanized collagen, 0.02 parts genipin, 0.03 parts catechin-oxidized sodium alginate microspheres, 0.01 parts bacterial cellulose, and 100 parts physiological saline.
[0027] The preparation method of catechin-oxidized sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin, and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to a 1% (w / w) aqueous solution of oxidized sodium alginate, and the mixture is dispersed by high-speed stirring at 5000 r / min for 20 min; then a 1% (w / w) calcium chloride solution is added, and the mixture is stirred at 5000 r / min for 1 h; finally, the mixture is freeze-dried to obtain the final product. The ratio of catechin, Span80, liquid paraffin, and anhydrous ethanol is 1 g: 0.1 g: 5 mL: 20 mL, and the volume ratio of the oil phase to the aqueous solution of oxidized sodium alginate and the calcium chloride solution is 1:3:3.
[0028] The preparation method of oxidized sodium alginate is as follows: Add sodium periodate equivalent to 10% of the mass of sodium alginate to a 1% sodium alginate aqueous solution, react in the dark for 18 hours, then add an appropriate amount of ethylene glycol to terminate the reaction, add anhydrous ethanol to precipitate the precipitate, and then dissolve it in deionized water. Repeat the precipitation-precipitation step 3 times, and then vacuum dry the obtained product to obtain the product.
[0029] The preparation method of the above-mentioned recombinant type III collagen composite cosmetic injectable filler is as follows:
[0030] S1. Sodium hyaluronate and recombinant type III humanized collagen were dissolved in a sodium hydroxide aqueous solution with a concentration of 0.05 mol / L. Then, genipin was added, and the cross-linking reaction was carried out at 30°C for 3 hours. The resulting gel was washed with water, freeze-dried, and then ground into powder to obtain gel powder.
[0031] S2. Add bacterial cellulose to physiological saline, disperse it evenly by ultrasonication, and then add gel powder and catechin-oxidized sodium alginate microspheres and mix evenly to obtain the final product.
[0032] Example 3
[0033] A recombinant type III collagen composite cosmetic injectable filler comprises the following raw materials in parts by weight: 1 part sodium hyaluronate, 0.2 parts recombinant type III humanized collagen, 0.05 parts genipin, 0.08 parts catechin-oxidized sodium alginate microspheres, 0.03 parts bacterial cellulose, and 100 parts physiological saline.
[0034] The preparation method of catechin-oxidized sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin, and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to a 2% (w / w) aqueous solution of oxidized sodium alginate, and dispersed by high-speed stirring at 10000 r / min for 40 min; then a 2% (w / w) calcium chloride solution is added, and the mixture is stirred at 8000 r / min for 3 h; finally, it is freeze-dried to obtain the final product. The ratio of catechin, Span80, liquid paraffin, and anhydrous ethanol is 1 g:0.3 g:10 mL:30 mL, and the volume ratio of the oil phase to the aqueous solution of oxidized sodium alginate and the calcium chloride solution is 1:5:5.
[0035] The preparation method of oxidized sodium alginate is as follows: Add sodium periodate equivalent to 20% of the mass of sodium alginate to a 3% sodium alginate aqueous solution, react in the dark for 24 hours, then add an appropriate amount of ethylene glycol to terminate the reaction, add anhydrous ethanol to precipitate the precipitate, dissolve it in deionized water, repeat the precipitation-precipitation step twice, and then vacuum dry the obtained product to obtain the product.
[0036] The preparation method of the above-mentioned recombinant type III collagen composite cosmetic injectable filler is as follows:
[0037] S1. Sodium hyaluronate and recombinant type III humanized collagen were dissolved in a 0.1 mol / L sodium hydroxide aqueous solution, and then genipin was added. The cross-linking reaction was carried out at 60°C for 8 hours. The resulting gel was washed with water, freeze-dried, and then ground into powder to obtain gel powder.
[0038] S2. Add bacterial cellulose to physiological saline, disperse it evenly by ultrasonication, and then add gel powder and catechin-oxidized sodium alginate microspheres and mix evenly to obtain the final product.
[0039] Comparative Example 1
[0040] A recombinant type III collagen composite cosmetic injectable filler comprises the following raw materials in parts by weight: 0.8 parts sodium hyaluronate, 0.15 parts recombinant type III humanized collagen, 0.03 parts genipin, 0.05 parts catechin-oxidized sodium alginate microspheres, and 100 parts physiological saline.
[0041] The preparation method of catechin-oxidized sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin, and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to a 1.5% (w / w) aqueous solution of oxidized sodium alginate, and the mixture is dispersed by high-speed stirring at 8000 r / min for 30 min; then a 1.5% (w / w) calcium chloride solution is added, and the mixture is stirred at 6000 r / min for 2 h; finally, the mixture is freeze-dried to obtain the final product. The ratio of catechin, Span80, liquid paraffin, and anhydrous ethanol is 1 g:0.2 g:6 mL:25 mL, and the volume ratio of the oil phase to the aqueous solution of oxidized sodium alginate and the calcium chloride solution is 1:4:4.
[0042] The preparation method of oxidized sodium alginate is as follows: Add sodium periodate equivalent to 15% of the mass of sodium alginate to a 2% sodium alginate aqueous solution, react in the dark for 20 hours, then add an appropriate amount of ethylene glycol to terminate the reaction, add anhydrous ethanol to precipitate the precipitate, and then dissolve it in deionized water. Repeat the precipitation-precipitation step 3 times, and then vacuum dry the obtained product to obtain the product.
[0043] The preparation method of the above-mentioned recombinant type III collagen composite cosmetic injectable filler is as follows:
[0044] S1. Sodium hyaluronate and recombinant type III humanized collagen were dissolved in a 0.05 mol / L sodium hydroxide aqueous solution, and then genipin was added. The cross-linking reaction was carried out at 50°C for 5 hours. The resulting gel was washed with water, freeze-dried, and then ground into powder to obtain gel powder.
[0045] S2. Add the gel powder and catechin-oxidized sodium alginate microspheres to physiological saline and mix evenly to obtain the final product.
[0046] Comparative Example 2
[0047] A recombinant type III collagen composite cosmetic injectable filler comprises the following raw materials in parts by weight: 0.8 parts sodium hyaluronate, 0.15 parts recombinant type III humanized collagen, 0.03 parts genipin, 0.05 parts catechin-sodium alginate microspheres, 0.02 parts bacterial cellulose, and 100 parts physiological saline.
[0048] The preparation method of catechin-sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin, and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to a 1.5% sodium alginate aqueous solution and dispersed at 8000 r / min for 30 min by high-speed stirring; then a 1.5% calcium chloride solution is added and reacted at 6000 r / min for 2 h by high-speed stirring; finally, the mixture is freeze-dried to obtain the final product. The ratio of catechin, Span80, liquid paraffin, and anhydrous ethanol is 1 g: 0.2 g: 6 mL: 25 mL, and the volume ratio of the oil phase to the sodium alginate aqueous solution and the calcium chloride solution is 1:4:4.
[0049] The preparation method of the above-mentioned recombinant type III collagen composite cosmetic injectable filler is as follows:
[0050] S1. Sodium hyaluronate and recombinant type III humanized collagen were dissolved in a 0.05 mol / L sodium hydroxide aqueous solution, and then genipin was added. The cross-linking reaction was carried out at 50°C for 5 hours. The resulting gel was washed with water, freeze-dried, and then ground into powder to obtain gel powder.
[0051] S2. Add bacterial cellulose to physiological saline, disperse it evenly by ultrasonication, and then add gel powder and catechin-oxidized sodium alginate microspheres and mix evenly to obtain the final product.
[0052] Comparative Example 3
[0053] A recombinant type III collagen composite cosmetic injectable filler comprises the following raw materials in parts by weight: 0.8 parts sodium hyaluronate, 0.15 parts recombinant type III humanized collagen, 0.03 parts genipin, 0.05 parts catechin-sodium alginate microspheres, and 100 parts physiological saline.
[0054] The preparation method of catechin-sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin, and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to a 1.5% sodium alginate aqueous solution and dispersed at 8000 r / min for 30 min by high-speed stirring; then a 1.5% calcium chloride solution is added and reacted at 6000 r / min for 2 h by high-speed stirring; finally, the mixture is freeze-dried to obtain the final product. The ratio of catechin, Span80, liquid paraffin, and anhydrous ethanol is 1 g: 0.2 g: 6 mL: 25 mL, and the volume ratio of the oil phase to the sodium alginate aqueous solution and the calcium chloride solution is 1:4:4.
[0055] The preparation method of the above-mentioned recombinant type III collagen composite cosmetic injectable filler is as follows:
[0056] S1. Sodium hyaluronate and recombinant type III humanized collagen were dissolved in a 0.05 mol / L sodium hydroxide aqueous solution, and then genipin was added. The cross-linking reaction was carried out at 50°C for 5 hours. The resulting gel was washed with water, freeze-dried, and then ground into powder to obtain gel powder.
[0057] S2. Add the gel powder and catechin-sodium alginate microspheres to physiological saline and mix evenly to obtain the final product.
[0058] Test case
[0059] Rats weighing 180±20g were selected as experimental animals. Fifty eligible rats were randomly divided into 5 groups, and 0.1mL of the filler from Example 1, Comparative Examples 1-3, and physiological saline were injected subcutaneously into their backs, respectively. The initial volume V0 of the swollen area was measured after injection. The volume V1 of the swollen area was measured again 60 days after injection. The 60-day residual rate was calculated as V1 / V0 × 100%. The experimental results are shown in Table 1.
[0060] Table 1
[0061]
[0062] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.
Claims
1. A recombinant collagen type III composite cosmetic injectable filler, characterized in that, The raw materials include the following parts by weight: 0.5-1 parts sodium hyaluronate, 0.1-0.2 parts recombinant type III humanized collagen, 0.02-0.05 parts genipin, 0.03-0.08 parts catechin-oxidized sodium alginate microspheres, 0.01-0.03 parts bacterial cellulose, and 100 parts physiological saline. The preparation method of the recombinant type III collagen composite cosmetic injectable filler includes the following steps: S1. Dissolve sodium hyaluronate and recombinant type III humanized collagen in an aqueous sodium hydroxide solution, then add genipin to carry out a cross-linking reaction. Wash the resulting gel with water, freeze-dry it, and grind it into powder to obtain gel powder. S2. Add bacterial cellulose to physiological saline, disperse it evenly by ultrasonication, and then add gel powder and catechin-oxidized sodium alginate microspheres and mix evenly to obtain the final product.
2. The recombinant type III collagen composite cosmetic injectable filler according to claim 1, characterized in that, The preparation method of the catechin-oxidized sodium alginate microspheres is as follows: catechin, Span80, liquid paraffin and anhydrous ethanol are mixed evenly to obtain an oil phase; the oil phase is added dropwise to an aqueous solution of oxidized sodium alginate, and the mixture is dispersed by high-speed stirring; then calcium chloride solution is added and stirred at high speed to react; and finally, the mixture is freeze-dried to obtain the final product.
3. The recombinant type III collagen composite cosmetic injectable filler according to claim 2, characterized in that, In the preparation method of the catechin-oxidized sodium alginate microspheres, the high-speed stirring dispersion speed is 5000-10000 r / min and the time is 20-40 min, and the high-speed stirring reaction speed is 5000-8000 r / min and the time is 1-3 h.
4. The recombinant type III collagen composite cosmetic injectable filler according to claim 2, characterized in that, In the preparation method of the catechin-oxidized sodium alginate microspheres, the ratio of catechin, Span80, liquid paraffin and anhydrous ethanol is 1g:(0.1-0.3)g:(5-10)mL:(20-30)mL, the volume ratio of oil phase to aqueous solution of oxidized sodium alginate and calcium chloride solution is 1:(3-5):(3-5), the mass concentration of aqueous solution of oxidized sodium alginate is 1-2%, and the mass concentration of calcium chloride solution is 1-2%.
5. The recombinant type III collagen composite cosmetic injectable filler according to claim 2, characterized in that, The method for preparing the oxidized sodium alginate is as follows: add sodium periodate equivalent to 10-20% of the mass of sodium alginate to an aqueous solution of sodium alginate with a mass concentration of 1-3%, react in the dark for 18-24 hours, then add an appropriate amount of ethylene glycol to terminate the reaction, add anhydrous ethanol to precipitate the precipitate, then dissolve it in deionized water, repeat the precipitation-precipitation step 2-3 times, and then vacuum dry the obtained product to obtain the product.
6. The recombinant type III collagen composite cosmetic injectable filler according to claim 1, characterized in that, The bacterial cellulose has a length of 20-200 nm.
7. The recombinant type III collagen composite cosmetic injectable filler according to claim 1, characterized in that, The concentration of the sodium hydroxide solution is 0.05-0.1 mol / L.
8. The recombinant type III collagen composite cosmetic injectable filler according to claim 1, characterized in that, The cross-linking reaction is carried out at a temperature of 30-60℃ for 3-8 hours.