Alginate-based facial fillers, methods of making and using the same

Abstract

The application belongs to the technical field of biological medicine and medical cosmetology materials, and discloses a sodium alginate-based facial filler, which comprises 13-26% of sodium alginate-based material, 40-43% of polylactic acid microspheres and 34-44% of lyophilization protectant in percentage by mass; the sodium alginate-based material comprises sodium alginate and oxidized sodium alginate, the G / M ratio of the sodium alginate and the oxidized sodium alginate is 3:1-1:1, and the mass ratio of the oxidized sodium alginate to the sodium alginate is 2:1-1:2. The sodium alginate-based material with biodegradable stimulation characteristics is used, the aldehyde group of the oxidized sodium alginate can react with the amino group on the surface of the tissue to form a Schiff base, thereby helping to anchor the target site, and the sodium alginate-based material can act on cells together with the polylactic acid microspheres to promote collagen secretion, the facial filler also has the functions of timely filling and gradient degradation. The application also provides a preparation method of the sodium alginate-based facial filler.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomedical and medical cosmetology materials, and particularly relates to an alginate-based facial filler and a preparation method and application thereof. BACKGROUND

[0002] With the development of biomaterial science and clinical medical technology, the plastic and cosmetic industry has shifted from surgical beauty to minimally invasive non-surgical beauty, which has promoted the rapid development of minimally invasive injection beauty products and provided some beauty methods with small trauma and fast recovery.

[0003] At present, conventional tissue filling products include hyaluronic acid, collagen, hydroxyapatite, polylactic acid and autologous fat particles, etc. These products have good filling effect and certain biocompatibility. The degradation period of conventional hyaluronic acid and collagen is short, which can be solved by cross-linking or adding high molecular materials with a longer degradation time. However, the degree of cross-linking is difficult to determine. If the degree of cross-linking is small, the effect of extending the cross-linking time cannot be achieved. If the degree of cross-linking is large, the resistance during use will be increased, the hydrophilicity will be poor, and inflammation may also be caused, resulting in induration or granuloma, etc. In addition, there is also a safety risk of residual cross-linking agent. The hydrophilicity of high molecular materials is poor, and the cell affinity is weak, so there is also a risk of causing inflammation. At present, the sodium hyaluronate filler in the dynamic expression area (such as the apple muscle and the command lines) is easily extruded by the muscle, and the filling material is prone to shift or diffuse, resulting in a "steamed bun" appearance, easy to shift, poor stability and short duration.

[0004] The prior art cannot meet the needs of timely filling and continuous stimulation of collagen regeneration to supplement the defect site without falling back. Therefore, it has become an urgent problem to develop a filling material that can be used for medical cosmetology, has the characteristics of gradient degradation, continuous stimulation of collagen regeneration and good compatibility with tissues. SUMMARY

[0005] The present application aims to provide an alginate-based facial filler and a preparation method and application thereof to solve at least one of the above technical problems.

[0006] According to a first aspect of the present application, an alginate-based facial filler is provided, which comprises 13% to 26% alginate-based material, 40% to 43% polylactic acid microspheres and 34% to 44% lyophilization protectant by mass percentage; wherein the alginate-based material comprises sodium alginate and oxidized sodium alginate, the G / M ratio of sodium alginate and oxidized sodium alginate is 3:1 to 1:1, and the mass ratio of oxidized sodium alginate to sodium alginate is 2:1 to 1:2.

[0007] The oxidized sodium alginate is prepared by the following method: sodium alginate is dispersed in anhydrous ethanol to prepare a sodium alginate suspension, then sodium periodate solution is added to the sodium alginate suspension, and the reaction is carried out for 18-32 hours in the dark, and the reaction is terminated by adding ethylene glycol to obtain a reaction mixture, and the reaction mixture is purified and freeze-dried to obtain the oxidized sodium alginate.

[0008] The sodium alginate-based facial filler has good raw material tissue compatibility, the sodium alginate-based material has biodegradable stimulation characteristics, and the sodium alginate-based material and the polylactic acid microspheres have a dual effect on cell stimulation of collagen production. In addition, the oxidized sodium alginate is obtained by oxidizing sodium alginate, part of the hydroxyl groups in the molecule are oxidized to aldehyde groups, the presence of the aldehyde groups makes more active sites appear on the molecular chain, these sites are more easily attacked by external factors such as moisture, oxygen, enzymes, etc., so that the molecular chain is more easily broken, resulting in that the oxidized sodium alginate is more easily degraded than the sodium alginate, and therefore the two together as a facial filler carrier material can realize gradient degradation in the filling site due to different degradation rates.

[0009] Moreover, the high-G content sodium alginate-based material can have a dual effect on macrophages to promote fibroblasts to produce collagen to promote the repair of damaged tissues, and can be biodegraded into non-toxic and harmless small molecules in the body with metabolism.

[0010] In some embodiments, the mass-volume ratio of sodium alginate to anhydrous ethanol can be 1:4-1:6. Specifically, 4-6 mL of anhydrous ethanol needs to be added for each 1 g of sodium alginate for dispersion.

[0011] In some embodiments, in the sodium periodate solution, the concentration of sodium periodate can be 6-20 wt%, and the molar ratio of ethylene glycol to sodium periodate in the sodium periodate solution can be 1:1.

[0012] In some embodiments, the molar ratio of sodium periodate to sodium alginate monomer units can be 20%-100%. For example, the molar ratio of sodium periodate to sodium alginate monomer units can be 20%, 40%, 60%, 80%, or 100%.

[0013] In some embodiments, the purification and freeze-drying of the reaction mixture can include the following steps: adding the reaction mixture to anhydrous ethanol, precipitating, filtering, and drying the precipitate, then dialyzing the precipitate with water for 24-72 hours, and freeze-drying the liquid in the dialysis bag.

[0014] In some embodiments, the volume ratio of the reaction mixture to anhydrous ethanol can be 1:4-1:6.

[0015] In some embodiments, the drying method of the precipitate can be vacuum drying at 35-45℃.

[0016] In some embodiments, the molecular weight of sodium alginate can be 8-12 kDa.

[0017] In some embodiments, the polylactic acid microspheres can be at least one of poly-L-lactic acid solid smooth microspheres, poly-L-lactic acid porous microspheres, poly-DL-lactic acid solid smooth microspheres, poly-DL-lactic acid porous microspheres, poly-DL-lactic acid-poly-L-lactic acid solid smooth microspheres, and poly-DL-lactic acid-poly-L-lactic acid porous microspheres.

[0018] In some embodiments, the particle size of the polylactic acid microspheres can be in three gradient ranges of 15-25 μm, 25-35 μm, and 35-50 μm, and the microspheres in the three particle size gradient ranges can be in a mass ratio of 3:2:1-2:1:1.

[0019] In some embodiments, the polylactic acid microspheres include first polylactic acid microspheres, second polylactic acid microspheres, and third polylactic acid microspheres, the particle size of the first polylactic acid microspheres can be 15-25 μm, the particle size of the second polylactic acid microspheres can be 25-35 μm, the particle size of the third polylactic acid microspheres can be 35-50 μm, and the mass ratio of the first polylactic acid microspheres, the second polylactic acid microspheres, and the third polylactic acid microspheres can be 3:2:1-2:1:1. In this way, due to the existence of spatial gaps of different sizes, the polylactic acid microspheres of different particle sizes can achieve better filling effect, so that the filling support is better. Moreover, the polylactic acid microspheres of different particle sizes have different degradation rates, and the small polylactic acid microspheres can be preferentially degraded to promote collagen neogenesis, achieving gradient degradation effect.

[0020] In some embodiments, the porosity of the polylactic acid microspheres can be 30%-60%.

[0021] In some embodiments, the lyophilization protectant can be at least one of mannitol, trehalose, and polyethylene glycol.

[0022] According to a second aspect of the present application, a preparation method of a sodium alginate-based facial filler is provided, comprising the following steps:

[0023] S1, adding a sodium alginate-based material into water for injection, stirring uniformly, and then adding a lyophilization protectant to dissolve to obtain a mobile phase;

[0024] S2, adding polylactic acid microspheres as microsphere particles suspended in the mobile phase into the above mobile phase, and after vacuum freeze-drying, packaging and sterilization, the sodium alginate-based facial filler is obtained.

[0025] In some embodiments, the procedure of vacuum freeze-drying is as follows:

[0026]

[0027] In some embodiments, the sterilization treatment method can be irradiation sterilization or sterilization using ethylene oxide.

[0028] According to a third aspect of the present application, there is provided use of the above alginate-based facial filler in the preparation of a medical cosmetology product or a cosmetic product, in particular as a tissue damage repair product, a collagen regeneration stimulating product, a facial filler product.

[0029] Compared with the prior art, the present application has the following beneficial effects:

[0030] (1) The alginate-based facial filler of the present application uses an alginate-based material with biodegradable properties as a facial filler carrier raw material, which can be slowly biodegraded into non-toxic and harmless small molecules with metabolism, and can also play a role in stimulating collagen production like polylactic acid microspheres, achieving the purpose of double and gradient stimulation of collagen regeneration;

[0031] (2) In the alginate-based facial filler of the present application, the alginate-based material is composed of oxidized sodium alginate and sodium alginate, and the oxidized sodium alginate is more easily degraded than the sodium alginate. The combination of the two as a facial filling carrier material not only plays a supporting role but also can be degraded in a gradient manner;

[0032] (3) In the alginate-based facial filler of the present application, the aldehyde group of the polyaldehyde-based oxidized sodium alginate obtained by sodium periodate oxidation can react with the amino group on the surface of the tissue to form a Schiff base, producing a chemical bonding effect, making it difficult to produce displacement at the injection filling site, and better anchoring the target site;

[0033] (4) The alginate-based facial filler of the present application has excellent tissue damage repair performance, collagen protein regeneration stimulating performance, timely filling performance, and gradient degradation performance. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 The Fourier transform infrared spectrum of the oxidized sodium alginate prepared in Example 1 of the present application; wherein SA represents sodium alginate, and OSA represents oxidized sodium alginate;

[0035] Figure 2 The microscope pictures of cells at 0h, 12h and 24h in the scratch healing experiment of Experimental Example 1 of the present application. DETAILED DESCRIPTION

[0036] The application will be further described in detail in connection with the embodiments. The examples are only for explanation and do not limit the application in any way. If not otherwise specified, the raw materials and reagents used in the examples are conventional products that can be commercially available; the experimental methods not specified in the examples are usually carried out according to the conventional conditions in the art or according to the conditions recommended by the manufacturers.

[0037] The sodium alginate powder used in the application is commercially available, and the purity of the sodium alginate is 85%, wherein the content of guluronic acid is 50% to 80%. In the following examples, "intensive stirring" refers to the stirring speed of 1200 to 1800 rpm. It should be noted that "G / M" refers to the ratio of guluronic acid (G segment) and mannuronic acid (M segment) in the sodium alginate molecule.

[0038] Example 1

[0039] The present example provides a preparation method of a sodium alginate-based facial filler, comprising the following steps:

[0040] (1) Preparation of oxidized sodium alginate: 10 g of sodium alginate powder (G / M = 3:1) is dispersed in 50 mL of anhydrous ethanol to prepare a sodium alginate suspension; 10 g of sodium periodate is dissolved in 50 mL of water to prepare a sodium periodate solution; the sodium periodate solution is added to the sodium alginate suspension at a molar ratio of 60% of sodium periodate to sodium alginate monomer unit, and the mixture is magnetically stirred at room temperature in the dark for about 24 h, then an equal molar amount of ethylene glycol is added, and the reaction is terminated after about 15 min to obtain a reaction mixture;

[0041] The obtained reaction mixture is poured into anhydrous ethanol under intensive stirring, and the volume ratio of the reaction mixture to anhydrous ethanol is 1:5, and a precipitate is precipitated, then the precipitate is filtered and dried at 40°C under vacuum to obtain a solid powder, the obtained solid powder is dialyzed with distilled water for 24 h to remove unreacted sodium periodate and ethylene glycol and other small molecular impurities, and finally the liquid in the dialysis bag is freeze-dried to obtain oxidized sodium alginate (G / M = 3:1).

[0042] The prepared oxidized sodium alginate is subjected to Fourier transform infrared spectroscopy detection by using a Fourier transform infrared spectrometer (instrument model: Nicolet iS50), and the detection result is shown in Figure 1 From Figure 1 it can be seen that the oxidized sodium alginate (OSA) has a C=O double bond stretching vibration peak at 1735 cm -1 compared with sodium alginate (SA), indicating that the oxidized sodium alginate is successfully prepared.

[0043] (2) Preparation of alginate-based facial filler: 13.5% alginate-based material, 42.6% poly-L-lactic acid solid smooth microspheres (particle size 30 μm, porosity 40%) and 43.9% mannitol, a freeze-drying protective agent, were prepared in mass percentage, wherein the alginate-based material included oxidized sodium alginate (G / M = 3:1) and sodium alginate (G / M = 3:1). The above-mentioned oxidized sodium alginate (G / M = 3:1) and sodium alginate (G / M = 3:1) were added to water for injection in a mass ratio of 2:1 and stirred uniformly, and then the freeze-drying protective agent was added and dissolved to obtain a mobile phase;

[0044] Finally, the poly-L-lactic acid solid smooth microspheres were added to the above-mentioned mobile phase as microsphere particles suspended in the mobile phase, and after vacuum freeze-drying according to the program in Table 1, packaging and sterilization, an alginate-based facial filler was obtained.

[0045] Table 1. Program of vacuum freeze-drying

[0046]

[0047] Example 2

[0048] The present embodiment provides a preparation method of an alginate-based facial filler, comprising the following steps:

[0049] (1) Preparation of oxidized sodium alginate: 10 g of sodium alginate powder (G / M = 3:1) was dispersed in 50 mL of anhydrous ethanol to prepare a sodium alginate suspension; 10 g of sodium periodate was dissolved in 50 mL of water to prepare a sodium periodate solution; the sodium periodate solution was added to the sodium alginate suspension in a molar ratio of 60% of sodium periodate to sodium alginate monomer units, and the mixture was stirred magnetically at room temperature for about 24 h in the dark, then an equal molar amount of ethylene glycol was added, and the reaction was terminated after about 15 min to obtain a reaction mixture;

[0050] The obtained reaction mixture was poured into vigorously stirred anhydrous ethanol, and the volume ratio of the reaction mixture to anhydrous ethanol was 1:5, and a precipitate was precipitated, then the precipitate was filtered and vacuum dried at 40°C to obtain a solid powder, the obtained solid powder was dialyzed against distilled water for 24 h to remove unreacted sodium periodate and ethylene glycol and other small molecular impurities, and finally the liquid in the dialysis bag was freeze-dried to obtain oxidized sodium alginate (G / M = 3:1).

[0051] (2) Preparation of alginate-based facial filler: 13.5% alginate-based material, 42.6% poly-L-lactic acid porous microspheres (particle size 30 μm, porosity 50%) and 43.9% mannitol, a freeze-drying protective agent, were prepared according to the mass percentage. The alginate-based material includes oxidized sodium alginate (G / M=3:1) and sodium alginate (G / M=3:1). The above-mentioned oxidized sodium alginate (G / M=3:1) and sodium alginate (G / M=3:1) were added to water for injection in a mass ratio of 2:1 and stirred uniformly, and then a freeze-drying protective agent was added to obtain a mobile phase;

[0052] Finally, the poly-L-lactic acid porous microspheres were added to the above-mentioned mobile phase as microsphere particles suspended in the mobile phase, and vacuum freeze-drying was carried out according to the procedure in Table 1, and then the alginate-based facial filler was packaged, sterilized and obtained.

[0053] Example 3

[0054] The present embodiment provides a preparation method of an alginate-based facial filler, comprising the following steps:

[0055] (1) Preparation of oxidized sodium alginate: 10 g of sodium alginate powder (G / M=3:1) was dispersed in 50 mL of anhydrous ethanol to prepare a sodium alginate suspension; 10 g of sodium periodate was dissolved in 50 mL of water to prepare a sodium periodate solution; the sodium periodate solution was added to the sodium alginate suspension in a molar ratio of 60% of sodium periodate to sodium alginate monomer units, and the mixture was stirred magnetically at room temperature for about 24 h in the dark, then an equal molar amount of ethylene glycol was added, and the reaction was terminated after about 15 min to obtain a reaction mixture;

[0056] The obtained reaction mixture was poured into vigorously stirred anhydrous ethanol, and the volume ratio of the reaction mixture to anhydrous ethanol was 1:5, and a precipitate was precipitated, then the precipitate was filtered and vacuum dried at 40°C to obtain a solid powder. The obtained solid powder was dialyzed with distilled water for 24 h to remove unreacted sodium periodate and ethylene glycol and other small molecular impurities, and finally the liquid in the dialysis bag was freeze-dried to obtain oxidized sodium alginate (G / M=3:1).

[0057] (2) Preparation of alginate-based facial filler: 13.5% alginate-based material, 42.6% poly-L-lactic acid porous microspheres (particle size 30 μm, porosity 50%) and 43.9% mannitol, a freeze-drying protective agent, were prepared according to the mass percentage. The alginate-based material includes oxidized sodium alginate (G / M=3:1) and sodium alginate (G / M=3:1). The above-mentioned oxidized sodium alginate (G / M=3:1) and sodium alginate (G / M=3:1) were added to water for injection in a mass ratio of 2:1 and stirred uniformly, and then a freeze-drying protective agent was added to obtain a mobile phase;

[0058] Finally, the poly-DL-lactic acid solid smooth microspheres are added into the above-mentioned mobile phase as microsphere particles suspended in the mobile phase, and the vacuum freeze-drying is carried out according to the procedure in Table 1, and then the alginic acid-based facial filler is obtained after packaging and sterilization.

[0059] Example 4

[0060] The present example provides a preparation method of an alginic acid-based facial filler, comprising the following steps:

[0061] (1) Preparation of oxidized sodium alginate: 10 g of sodium alginate powder (G / M = 3:1) is dispersed in 50 mL of anhydrous ethanol to prepare a sodium alginate suspension; 10 g of sodium periodate is dissolved in 50 mL of water to prepare a sodium periodate solution; the sodium periodate solution is added to the sodium alginate suspension at a molar ratio of 60% of sodium periodate to sodium alginate monomer units, and the mixture is magnetically stirred at room temperature in the dark for about 24 h, then an equal molar amount of ethylene glycol is added, and the reaction is terminated after about 15 min to obtain a reaction mixture;

[0062] The obtained reaction mixture is poured into anhydrous ethanol under vigorous stirring, and the volume ratio of the reaction mixture to anhydrous ethanol is 1:5, and a precipitate is precipitated, then the precipitate is filtered and vacuum dried at 40°C to obtain a solid powder, and the obtained solid powder is dialyzed with distilled water for 24 h to remove unreacted sodium periodate and ethylene glycol and other small molecular impurities, and finally the liquid in the dialysis bag is freeze-dried to obtain oxidized sodium alginate (G / M = 3:1).

[0063] (2) Preparation of the alginic acid-based facial filler: according to the mass percentage, 13.5% of the alginic acid-based material, 42.6% of the poly-DL-lactic acid porous microspheres (particle size of 30 μm, porosity of 50%), and 43.9% of the freeze-drying protective agent mannitol are prepared, wherein the alginic acid-based material includes oxidized sodium alginate (G / M = 3:1) and sodium alginate (G / M = 3:1). The above-mentioned oxidized sodium alginate (G / M = 3:1) and sodium alginate (G / M = 3:1) are added to water for injection at a mass ratio of 2:1 and stirred uniformly, and then the freeze-drying protective agent is added and dissolved to obtain a mobile phase;

[0064] Finally, the poly-DL-lactic acid porous microspheres are added into the above-mentioned mobile phase as microsphere particles suspended in the mobile phase, and the vacuum freeze-drying is carried out according to the procedure in Table 1, and then the alginic acid-based facial filler is obtained after packaging and sterilization.

[0065] Comparative Example 1

[0066] The present comparative example provides a preparation method of an alginic acid-based facial filler, comprising the following steps:

[0067] (1) Preparation of oxidized sodium alginate: 10 g of sodium alginate powder (G / M=1:2) was dispersed in 50 mL of anhydrous ethanol to prepare a sodium alginate suspension; 10 g of sodium periodate was dissolved in 50 mL of water to prepare a sodium periodate solution; the sodium periodate solution was added to the sodium alginate suspension at a molar ratio of sodium periodate to sodium alginate monomer unit of 60%, and the mixture was stirred magnetically at room temperature for about 24 h in the dark, then an equal molar amount of ethylene glycol was added, and the reaction was terminated after about 15 min to obtain a reaction mixture;

[0068] The obtained reaction mixture was poured into anhydrous ethanol stirred vigorously, and the volume ratio of the reaction mixture to anhydrous ethanol was 1:5, and a precipitate was separated out, then the precipitate was filtered and dried at 40°C under vacuum to obtain a solid powder, and the obtained solid powder was dialyzed against distilled water for 24 h to remove unreacted sodium periodate and ethylene glycol and other small molecular impurities, and finally the liquid in the dialysis bag was freeze-dried to obtain oxidized sodium alginate (G / M=1:2).

[0069] (2) Preparation of the alginate-based facial filler: 13.5% alginate-based material, 42.6% poly-L-lactic acid solid smooth microspheres (particle size 30 μm, porosity 50%) and 43.9% mannitol, a freeze-drying protective agent, were mixed to prepare the alginate-based facial filler, wherein the alginate-based material included oxidized sodium alginate (G / M=1:2) and sodium alginate (G / M=1:2). The above-mentioned oxidized sodium alginate (G / M=1:2) and sodium alginate (G / M=1:2) were added to water for injection at a mass ratio of 2:1 and stirred until uniform, and then the freeze-drying protective agent was dissolved to obtain a mobile phase;

[0070] Finally, the poly-L-lactic acid solid smooth microspheres were added to the above-mentioned mobile phase as microsphere particles suspended in the mobile phase, and vacuum freeze-drying was performed according to the procedure in Table 1, and then the alginate-based facial filler was packaged and sterilized.

[0071] Experimental Example 1

[0072] To verify whether the alginate-based facial filler of the present application achieves the desired effect, the alginate-based facial fillers of Examples 1-4 and Comparative Example 1 were subjected to a macrophage and fibroblast co-culture test to verify the collagen secretion promoting effect.

[0073] 1. Preparation of conditioned medium

[0074] The conditioned medium was the supernatant of the culture medium produced by co-culturing M0, M1 and M2 macrophages with the scaffold. The specific preparation method of the supernatant of the culture medium was as follows: macrophages were inoculated in a 48-well plate coated with the material at a density of 2×10 4 cells / well, and the supernatant of the culture medium (CM) was collected after co-culturing for 48 h, centrifuged and sterilized by filtering through a 0.22 μm sterile microporous filter, and then stored for use.

[0075] 2. Co-culture group

[0076] (1) Add 5 mL of physiological saline to the alginate-based facial filler, and after complete dissolution, mix the above mixture with the culture medium at a mass ratio of 1:10 to obtain a mixed culture medium. M0, M1 and M2 macrophages are inoculated into the Transwell at a density of 1 x 10 5 cells / well, and after 24 h of culture, the mixed culture medium is added; at the same time of inoculating the macrophages, fibroblasts are inoculated into the 24-well plate at a density of 1 x 10 5 cells / well, and after 24 h of culture, the Transwell is placed into the 24-well plate for co-culture, and after 3 d of culture, PCR detection is performed.

[0077] (2) Wound healing: the co-cultured fibroblasts are inoculated into a 6-well plate at a density of 1 x 10 4 cells / well, and incubated overnight until the cells reach a 90% confluence; then a sterile 200 μL pipette tip is used to draw a scratch on the bottom of the well plate to form a cell-free strip in the center of the monolayer; the original culture medium is discarded, and washed with PBS solution to remove unadhered cells and cell debris. The conditioned medium of M1 cells is added, and then incubated in the cell culture box, and the scratch is photographed using a microscope at 0 h, 12 h and 24 h.

[0078] 3. Experimental results

[0079] The PCR detection results of M0, M1 and M2 macrophages co-cultured with fibroblasts for 3 d are shown in Tables 2 and 3. As can be seen from Table 2, after the alginate-based facial filler of the comparative example is co-cultured with macrophages and fibroblasts for 3 d, the expression values of collagen type I in M0 and M2 macrophages are higher than those of Examples 1-4. As can be seen from Table 3, the expression values of collagen type III in M0, M1 and M2 macrophages obtained by co-culturing Examples 1-4 are higher than those of Comparative Example 1, indicating that in the case of co-culturing macrophages and fibroblasts, the use of oxidized sodium alginate and sodium alginate with high G content as raw materials has excellent performance in promoting macrophages to secrete collagen type III, and thus helps to repair tissue damage.

[0080] Table 2 Co-culture group COL-1 (collagen type I expression)

[0081]

[0082] Table 3 Co-culture group COL-3 (collagen type III expression)

[0083]

[0084] Microscopic images of cells at 0h, 12h, and 24h in the scratch healing assay are shown below. Figure 2 As shown. From Figure 2 It can be seen that the scratch width observed at 24h was significantly lower than that observed at 0h and 12h, indicating that the alginate-based facial filler of the present invention can promote the enhanced migration ability of fibroblasts in a scratch healing experiment lasting 24 hours, which helps surrounding tissues to gather towards the damaged site and ultimately achieve tissue repair.

[0085] The above experimental results show that the alginate-based facial filler of the present invention has excellent collagen-promoting properties and the potential to promote tissue repair.

[0086] The above descriptions are merely some embodiments of the present invention. Those skilled in the art can make various modifications and improvements without departing from the inventive concept of the present invention, and these all fall within the scope of protection of the present invention.

Claims

1. An alginate-based facial filler, characterized in that, The product, by weight percentage, is prepared from 13%–26% alginate-based material, 40%–43% polylactic acid microspheres, and 34%–44% lyophilization protectant via the following method: The alginate-based material is added to water for injection and stirred, then the lyophilization protectant is added to dissolve it and obtain a mobile phase; the polylactic acid microspheres are added as microsphere particles suspended in the mobile phase, and after vacuum freeze-drying, packaged and sterilized, the product is obtained; the vacuum freeze-drying procedure is as follows: The alginate-based material comprises sodium alginate and sodium oxidized alginate, wherein the G / M ratio of sodium alginate and sodium oxidized alginate is 3:1 to 1:1, and the mass ratio of sodium oxidized alginate to sodium alginate is 2:1 to 1:2; the polylactic acid microspheres have particle sizes in three gradient ranges: 15μm to 25μm, 25μm to 35μm, and 35μm to 50μm, and the mass ratio of microspheres in the three particle size gradient ranges is 3:2:1 to 2:1:1, and the porosity of the polylactic acid microspheres is 30% to 60%; The oxidized sodium alginate was prepared by the following method: sodium alginate was dispersed in anhydrous ethanol to prepare a sodium alginate suspension, then sodium periodate solution was added to the sodium alginate suspension, and the reaction was carried out under light-protected conditions for 18-32 h. Ethylene glycol was added to terminate the reaction and the reaction mixture was obtained. The reaction mixture was purified and freeze-dried to obtain the final product.

2. The alginate-based facial filler according to claim 1, characterized in that, The concentration of sodium periodate in the sodium periodate solution is 6-20 wt%, the molar ratio of ethylene glycol to sodium periodate in the sodium periodate solution is 1:1, and the molar ratio of sodium periodate to sodium alginate monomer unit is 20%-100%.

3. The alginate-based facial filler according to claim 2, characterized in that, The mass-to-volume ratio of sodium alginate to anhydrous ethanol is 1:4 to 1:

6.

4. The alginate-based facial filler according to claim 2, characterized in that, The purification and freeze-drying of the reaction mixture includes the following steps: adding the reaction mixture to anhydrous ethanol, precipitating the precipitate, filtering, drying the precipitate, dialyzing with water for 24-72 hours, and freeze-drying the liquid in the dialysis bag.

5. The alginate-based facial filler according to any one of claims 1-4, characterized in that, The freeze-drying protectant is selected from at least one of mannitol, trehalose, and polyethylene glycol.

6. The method for preparing the alginate-based facial filler according to any one of claims 1 to 5, characterized in that, Includes the following steps: S1. The alginate-based material is added to water for injection and stirred, and then a lyophilization protectant is added to dissolve it to obtain a mobile phase; S2. Polylactic acid microspheres are added to the above mobile phase as microsphere particles suspended in the mobile phase. After vacuum freeze-drying, they are packaged and sterilized to obtain the final product.

7. The method for preparing the alginate-based facial filler according to claim 6, characterized in that, The vacuum freeze-drying procedure is as follows: 。 8. The use of the alginate-based facial filler according to any one of claims 1 to 5 in the preparation of medical aesthetic products or cosmetics.

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