A cosmetic filling material for injection and a method for preparing the same

Abstract

The application discloses an injection cosmetic filling material which is composed of the following components in percentage by mass: 10-40 parts of hydroxyapatite microspheres, 60-90 parts of gel liquid, 2-5 parts of gradient cross-linking modified sodium hyaluronate, 0.5-1 part of vitamin E-tea seed oil-panthenol compound microemulsion and 0.5-1 part of hyaluronic acid-proline graft copolymer. The application further discloses a preparation method of the injection cosmetic filling material, which comprises the following steps: S1, synthesizing hydroxyapatite; S2, preparing hydroxyapatite microspheres; and S3, stirring and mixing. The hydroxyapatite microsphere gel of the application has a good biocompatibility and a good stable filling system, solves the problem of poor compatibility between a thickening agent and a matrix, can be kept in a plastic part for a long time after injection, can provide long-acting filling and support to improve the skin wrinkle effect in the tissue, can provide nourishing repair and deep moisturizing functions at the same time, has the effects of filling and repair, and delays facial aging.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical materials, and particularly relates to a cosmetic filling material for injection and a preparation method thereof. BACKGROUND

[0002] In the field of medical cosmetology, facial soft tissue filling is one of the core means to improve facial aging, and the technical core is to realize facial volume supplement and structure remodeling through the intervention of filling materials. At present, filling materials have evolved from traditional simple physical filling to stimulating tissue regeneration, and regenerative facial soft tissue filling materials have gradually become the mainstream in the market.

[0003] The core advantage of such regenerative materials is that after being injected into the deep layer of the dermis and subcutaneous tissue of the face, they can not only rely on the volume of the material itself to achieve short-term filling, but also continuously stimulate the activity of human fibroblasts and induce the generation of collagen, ultimately forming a volume support structure composed of self-tissue. This mechanism makes the effect more in line with the physiological state, avoids the false face feeling that may occur with traditional materials, and the effect usually lasts for 1-2 years, and some products can even achieve longer duration, significantly reducing the drawbacks of repeated injections required by traditional filling.

[0004] The existing regenerative facial soft tissue filling products on the market mainly use synthetic high molecular materials such as polylactic acid and polycaprolactone as the core component. Although such materials can trigger tissue regeneration through the degradation process, there are natural limitations in biocompatibility with human tissues as artificial synthetic substances. Long-term implantation may cause potential risks such as local foreign body reaction and thickening of the fibrous envelope, and the degradation rate of the material is difficult to accurately match the rhythm of collagen generation, which may cause uneven filling effect and fluctuation in maintenance period in some cases.

[0005] In contrast, hydroxyapatite, as a natural inorganic component of human bone and teeth, has been verified through decades of clinical application for its biocompatibility and safety, and has good affinity with human hard and soft tissues. It is not easy to cause immune rejection after implantation, and can provide a scaffold carrier for fibroblast attachment and proliferation through its unique porous microsphere structure, more efficiently guiding the ordered deposition of collagen. Therefore, it is necessary to propose an injection cosmetic filling material with hydroxyapatite as the core, precise particle size control, excellent dispersibility, and material degradation and tissue regeneration synergy. SUMMARY

[0006] The present application relates to the technical field of biomedical materials, and particularly relates to a cosmetic filling material for injection and a preparation method thereof.

[0007] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0008] A cosmetic filling material for injection, consisting of the following components by mass percentage: 10-40 parts of hydroxyapatite microspheres, 60-90 parts of gel liquid.

[0009] Preferably, the gel liquid is any combination of one or more of cellulose derivatives, chitosan, sodium alginate, polyethylene oxide, polyethylene glycol, which is added to a mixture of injection water and glycerol in proportion, stirred uniformly, and the mixture is placed in a 60℃ oven for 4h to prepare.

[0010] Preferably, it further comprises the following components by mass percentage: 2-5 parts of gradient cross-linked modified sodium hyaluronate, 0.5-1 part of vitamin E-tea seed oil-panthenol complex microemulsion, 0.5-1 part of hyaluronic acid-proline graft copolymer.

[0011] Preferably, the preparation steps of the gradient cross-linked modified sodium hyaluronate are as follows:

[0012] The sodium hyaluronate with a molecular weight of 800-1200kDa is divided into two groups A and B with a mass ratio of 7:3;

[0013] The low cross-linking degree component is prepared by adding 5%-6% glycolaldehyde to group A in an amount of 5%-6% of the mass of group A, and reacting at pH 6.5-7.0 and 35-40℃ for 1.5h, and the high cross-linking degree component is prepared by using a mixed cross-linking agent of glycolaldehyde-adipic acid dihydrazide with a mass ratio of 3:1, and adding a total amount of 7%-8% of the mass of group B, and reacting at pH 7.0-7.2 and 40-45℃ for 2.5h;

[0014] After mixing the two components, ultrasonic dispersion is performed at a power of 300-400W for 10-15min to obtain the gradient cross-linked modified sodium hyaluronate.

[0015] Preferably, the preparation steps of the hyaluronic acid-proline graft copolymer are as follows:

[0016] The hyaluronic acid with a molecular weight of 100-200kDa is mixed with L-proline at a mass ratio of 5:1, and 1-ethyl-carbodiimide hydrochloride solution with a mass concentration of 1%-2% is added in an amount of 3 times the mass of the mixture;

[0017] Stirring and reacting at pH 5.0-5.5 and 30-35℃ for 4-6h, and then purifying by dialysis for 24h with a molecular weight cutoff of 50kDa, and freeze-drying to obtain a white powder of the graft copolymer.

[0018] Preferably, the preparation steps of the vitamin E-tea seed oil-panthenol complex microemulsion are as follows:

[0019] Vitamin E, tea seed oil, and panthenol are mixed in a mass ratio of 3:2:1 as the oil phase;

[0020] Mixing 10%-15% mass concentration of polysorbate 80 and 5%-8% mass concentration of glycerol as the water phase according to a mass ratio of 2:1;

[0021] Slowly adding the oil phase into the water phase, high-speed shearing at 10000-12000 r / min for 15-20 min at 40-45 DEG C, and then ultrasonic emulsification for 5-8 min at 200-300 W to form uniform microemulsion with a particle size of 100-300 nm.

[0022] A preparation method of a cosmetic filling material for injection, comprising the following steps:

[0023] S1, hydroxyapatite synthesis, mixing raw materials calcium nitrate, diammonium hydrogen phosphate and ammonia water into injection water, continuously reacting for 24-30 h under the conditions of 25-40 DEG C, 9.0-10.0 pH value and 200-300 r / min stirring speed, obtaining hydroxyapatite slurry, and then filtering and washing to obtain hydroxyapatite slurry;

[0024] S2, hydroxyapatite microsphere preparation, introducing the slurry into a spray centrifugal dryer to obtain spherical or spherical-like hydroxyapatite powder; calcining the powder microspheres at a high temperature of 800-1000 DEG C to obtain hydroxyapatite microspheres; screening the hydroxyapatite microspheres by ultrasonic vibration to obtain hydroxyapatite microspheres with a particle size range of 5-100 microns; and calcining the screened microspheres at a high temperature of 1100-1300 DEG C to obtain high-purity, biocompatible hydroxyapatite microspheres;

[0025] S3, stirring and mixing, placing the hydroxyapatite microspheres and other components and gel liquid in a stirrer according to a proportion, the stirrer providing a vacuum negative pressure of 50000-100000 Pa, adjusting the stirring paddle speed to 30-100 r / min, stirring for 60-120 min, and simultaneously self-rotating and revolving the barrel.

[0026] Preferably, in the step S1, the molar ratio of calcium nitrate to diammonium hydrogen phosphate is 5:3, and the mass concentration of ammonia water is 25%-28%; and during the reaction process, the dropping speed of ammonia water is controlled to be 1-2 drops / s.

[0027] Preferably, in the step S2, the inlet air temperature of the spray centrifugal dryer is set to 100-300 DEG C, the outlet air temperature is controlled to be 50-150 DEG C, and the centrifugal speed is 10000-30000 r / min.

[0028] The present application has the following beneficial effects:

[0029] 1. This invention uses hydroxyapatite microspheres as the core to form a microsphere gel. Compared with traditional filler materials with synthetic polymers such as polylactic acid and polycaprolactone as the core, it has better biocompatibility and no cytotoxicity. After injection, the microspheres are fixed by fibrous tissue and are not easy to move. The gel components gradually degrade and are replaced by human tissue. The microspheres degrade slowly and can be retained for a long time. It can not only improve skin wrinkles through stable support to achieve long-lasting filling, but also simultaneously play a role in nourishing and repairing and deep moisturizing, achieving the dual effect of filling and anti-aging repair, thus delaying the facial aging process.

[0030] 2. This invention uses a gradient crosslinking process to prepare sodium hyaluronate. By combining components with different degrees of crosslinking, it avoids the defects of traditional single crosslinking products that degrade too quickly or have excessively high mechanical strength. The elastic modulus of the prepared filler material is significantly higher than that of single crosslinking products. This not only prolongs the duration of the filling effect but also ensures the material's flexibility and natural feel, reducing the risk of hardening and foreign body reactions. At the same time, relying on the hyaluronic acid-proline graft copolymer to regulate skin cell activity, it breaks through the limitation of traditional materials that can only fill. By promoting collagen regeneration, it improves aging problems such as collagen loss and decreased elasticity in the dermis, achieving a synergistic effect of filling and anti-aging.

[0031] 3. This invention utilizes composite microemulsion carrier technology to stabilize active ingredients such as vitamin E and panthenol through high-speed shearing and ultrasonic emulsification, preventing their oxidation and failure or clogging of the needle, and ensuring the effective performance of their antioxidant, repair, and other biological functions. At the same time, it combines vacuum negative pressure and barrel rotation-revolution synergistic stirring process to solve the problem of poor compatibility between thickener and matrix, prevent phase separation, and achieve an instability coefficient within 16 hours that is much lower than that of atmospheric pressure stirring. This ensures smooth injection, reduces accelerated material degradation and tissue irritation risks, and improves the stability and safety of the filling system. Attached Figure Description

[0032] Figure 1 This is a flowchart of the hydroxyapatite slurry synthesis process proposed in this invention;

[0033] Figure 2 This is a flowchart of the hydroxyapatite microsphere preparation process proposed in this invention;

[0034] Figure 3 This is an overall appearance diagram of the hydroxyapatite microspheres proposed in this invention;

[0035] Figure 4 This is an enlarged view of the hydroxyapatite microspheres proposed in this invention;

[0036] Figure 5 This is a crystallinity analysis diagram of the hydroxyapatite microspheres proposed in this invention;

[0037] Figure 6 The images show the viscoelasticity analysis of the filler materials in the embodiments and comparative examples proposed in this invention at 25°C and 0.1Hz. DETAILED DESCRIPTION

[0038] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments of the present application.

[0039] The cosmetic filling material for injection is composed of the following components in percentage by mass: 10-40 parts of hydroxyapatite microspheres, and 60-90 parts of gel liquid.

[0040] The gel liquid is one or more of any combination of cellulose derivatives, chitosan, sodium alginate, polyethylene oxide, and polyethylene glycol, which is added to a mixed solution of injection water and glycerol in proportion, stirred uniformly, and placed in a 60℃ oven for 4h to obtain.

[0041] The following components are further included: 2-5 parts of gradient crosslinking modified sodium hyaluronate, 0.5-1 part of vitamin E-tea seed oil-panthenol composite microemulsion, and 0.5-1 part of hyaluronic acid-proline graft copolymer.

[0042] The preparation steps of the gradient crosslinking modified sodium hyaluronate are as follows:

[0043] The sodium hyaluronate with a molecular weight of 800-1200kDa is divided into two groups A and B with a mass ratio of 7:3;

[0044] The low crosslinking degree component is prepared by using 5%-6% glycolaldehyde with an addition amount of 5%-6% of the mass of group A, at pH 6.5-7.0 and 35-40℃ for 1.5h, and the high crosslinking degree component is prepared by using a mixed crosslinking agent of glycolaldehyde and adipic acid dihydrazide with a mass ratio of 3:1, with a total addition amount of 7%-8% of the mass of group B, at pH 7.0-7.2 and 40-45℃ for 2.5h;

[0045] After mixing the two components, ultrasonic dispersion is performed at a power of 300-400W for 10-15min to obtain the gradient crosslinking modified sodium hyaluronate.

[0046] The preparation steps of the hyaluronic acid-proline graft copolymer are as follows:

[0047] The hyaluronic acid with a molecular weight of 100-200kDa is mixed with L-proline at a mass ratio of 5:1, and 1-ethyl-carbodiimide hydrochloride solution with a mass concentration of 1%-2% is added in an amount of 3 times the mass of the mixture;

[0048] Stirring reaction is performed at pH 5.0-5.5 and 30-35℃ for 4-6h, and after reaction, dialysis purification is performed for 24h with a molecular weight cutoff of 50kDa, and white powder graft copolymer is obtained after freeze-drying.

[0049] The preparation steps of the vitamin E-tea seed oil-panthenol complex microemulsion are as follows:

[0050] Vitamin E, tea seed oil and panthenol are mixed in a mass ratio of 3:2:1 as the oil phase;

[0051] Polysorbate 80 with a mass concentration of 10%-15% and glycerol with a mass concentration of 5%-8% are mixed in a mass ratio of 2:1 as the water phase;

[0052] The oil phase is slowly added to the water phase, and high-speed shearing is carried out at 10,000-12,000 r / min for 15-20 min at 40-45°C, and then ultrasonic emulsification is carried out for 5-8 min, to form a uniform microemulsion with a particle size of 100-300 nm.

[0053] A preparation method of an injection cosmetic filling material, comprising the following steps:

[0054] S1, hydroxyapatite synthesis, raw materials calcium nitrate, diammonium hydrogen phosphate and ammonia are mixed into injection water, the molar ratio of calcium nitrate to diammonium hydrogen phosphate is 5:3, the mass concentration of ammonia is 25%-28%, during the reaction process, the dropwise adding speed of ammonia is controlled to be 1-2 drops per second, the reaction is continuously carried out at 25-40°C, 9.0-10.0 pH value and 200-300 r / min stirring speed for 24-30 h, to obtain hydroxyapatite slurry, and then the hydroxyapatite slurry is filtered and washed to obtain hydroxyapatite slurry; the synthesis process flow of the hydroxyapatite slurry is as shown in Figure 1 , wherein raw materials A, B and C represent calcium nitrate, diammonium hydrogen phosphate and ammonia respectively;

[0055] S2, hydroxyapatite microsphere preparation, the specific process is as shown in Figure 2 , the slurry is introduced into a spray centrifugal dryer, the inlet air temperature of the spray centrifugal dryer is set to be 180-220°C, the outlet air temperature is controlled to be 80-100°C, and the centrifugal speed is 8,000-10,000 r / min, to obtain spherical or spherical-like hydroxyapatite powder; the powder microsphere is calcined at a high temperature of 800-1,000°C, to obtain a hydroxyapatite microsphere; the hydroxyapatite microsphere is sieved by ultrasonic vibration, to obtain a hydroxyapatite microsphere with a suitable particle size range of 5-100 microns; and the sieved microsphere is calcined at a high temperature of 1,100-1,300°C, to obtain a hydroxyapatite microsphere with high purity and good biocompatibility; the prepared microsphere data are as follows: the morphology, the surface is relatively smooth, and the microsphere is composed of a solid sphere; the crystallinity, 90-100%; the particle size, 5-100 microns, and the average particle size, 25-45 microns; the specific shape appearance of the microsphere is as shown in Figure 3 and Figure 4 , and the crystallinity is as shown in Figure 5 .

[0056] S3, stirring mixing, the hydroxyapatite microspheres and other components with the gel liquid in proportion in the blender, the blender provides 50000-100000 Pa vacuum negative pressure, the stirring paddle speed is adjusted to 30-100 r / min, stirring for 60-120 min, the temperature is 25-30℃, the barrel rotates simultaneously, the rotation makes the components stir uniformly, the revolution generates centrifugal force to make the bubbles separate, and the cosmetic filling material mainly formed by the microsphere gel is formed.

[0057] I. Example design Example 1

[0058] A cosmetic filling material for injection, consisting of the following components by mass percentage: 20g of hydroxyapatite microspheres per 100g, 80g of gel liquid.

[0059] The preparation process of hydroxyapatite microspheres in this example strictly follows the steps S1 and S2 in the technical scheme, the vacuum stirring parameters are 80000 Pa negative pressure, 100 r / min speed, stirring for 120 min, and the temperature is 30℃. The gel liquid is prepared by adding chitosan and polyethylene glycol to the mixed solution of injection water and glycerol in proportion and stirring uniformly, and the mixture is placed in a 60℃ oven for 4h. Example 2

[0060] A cosmetic filling material for injection, consisting of the following components by mass percentage: 20g of hydroxyapatite microspheres per 100g, 75.5g of gel liquid, 3g of gradient cross-linked modified sodium hyaluronate, 0.8g of vitamin E-tea seed oil-panthenol composite microemulsion, and 0.7g of hyaluronic acid-proline graft copolymer.

[0061] In addition to the basic steps, in the preparation of the filling material, the gradient cross-linked modified sodium hyaluronate, the composite microemulsion, and the graft copolymer are added to the blender at the same time as the hydroxyapatite microspheres and the gel liquid, and the vacuum stirring parameters are 80000 Pa negative pressure, 80 r / min speed, stirring for 55 min, and the temperature is 27℃. The gel liquid is prepared by adding chitosan and polyethylene glycol to the mixed solution of injection water and glycerol in proportion and stirring uniformly, and the mixture is placed in a 60℃ oven for 4h. Example 3

[0062] A cosmetic filling material for injection, the components of this embodiment are the same as those of Example 2, only the content of each component is adjusted, and the specific weight of each component is as follows: 18 g of hydroxyapatite microspheres, 78.5 g of gel liquid, 2.5 g of gradient cross-linking modified sodium hyaluronate, 0.5 g of vitamin E-tea seed oil-panthenol composite microemulsion, and 0.5 g of hyaluronic acid-proline graft copolymer per 100 g of filling material. The gel liquid is prepared by adding chitosan and polyethylene glycol to a mixture of injection water and glycerol in a proportion, stirring uniformly, and placing the mixture in an oven at 60°C for 4 h.

[0063] The preparation process of this embodiment is the same as that of Example 2, and the vacuum stirring parameters are 80000 Pa negative pressure, 80 r / min rotation speed, 55 min stirring time, and 27°C temperature.

[0064] II. Design of Comparative Examples

[0065] Comparative Example 1

[0066] A cosmetic filling material for injection, which uses a common regenerative facial soft tissue filling product on the market as the core ingredient, mainly composed of synthetic polymer materials such as polylactic acid and polycaprolactone.

[0067] Comparative Example 2

[0068] A cosmetic filling material for injection, the components of this embodiment are the same as those of Example 2, only the content of each component is adjusted, and the specific weight of each component is as follows: 18 g of hydroxyapatite microspheres, 78.5 g of gel liquid, 2.5 g of gradient cross-linking modified sodium hyaluronate, 0.5 g of vitamin E-tea seed oil-panthenol composite microemulsion, and 0.5 g of hyaluronic acid-proline graft copolymer per 100 g of filling material. The gel liquid is prepared by adding chitosan and polyethylene glycol to a mixture of injection water and glycerol in a proportion, stirring uniformly, and placing the mixture in an oven at 60°C for 4 h.

[0069] The preparation process of this embodiment is the same as that of Example 2, and the vacuum stirring parameters are 80000 Pa negative pressure, 80 r / min rotation speed, 55 min stirring time, and 27°C temperature.

[0070] Comparative Example 3

[0071] A cosmetic filling material for injection, the components of this embodiment are the same as those of Example 2, only the content of each component is adjusted, and the specific weight of each component is as follows: 18 g of hydroxyapatite microspheres, 78.5 g of gel liquid, 2.5 g of gradient cross-linking modified sodium hyaluronate, 0.5 g of vitamin E-tea seed oil-panthenol composite microemulsion, and 0.5 g of hyaluronic acid-proline graft copolymer per 100 g of filling material. The gel liquid is prepared by adding chitosan and polyethylene glycol to a mixture of injection water and glycerol in a proportion, stirring uniformly, and placing the mixture in an oven at 60°C for 4 h.

[0072] The comparative example is prepared by a process different from that of Example 3, in that step S5 microsphere gel preparation uses normal pressure stirring instead of vacuum stirring, and the stirring speed is only 20 r / min, the stirring time is 45 min, and the barrel rotation and revolution are not coordinated.

[0073] III. Comparative Test Analysis

[0074] Comparative Test One: Gel System Stability Analysis

[0075] The volume concentration, particle size, particle shape, and particle migration of the dispersion system can all cause changes in the light transmittance signal of the sample. For a filler with uniform and stable dispersion, the particle sedimentation rate is small and stable, and there is no rapid particle aggregation or system flocculation, so the change value and rate of the light transmittance are small. Therefore, under a specific excitation wavelength, the change in the light transmittance recorded by the analyzer can indicate the change in the state of the dispersion system, and thus evaluate the stability and dispersion uniformity of the system. The sample light transmittance was monitored in real time for 16 h, and the instability coefficient was calculated. The higher the instability coefficient, the more unstable the sample. The specific recorded data are shown in the following table:

[0076] Table 1: Instability Coefficient Comparative Data Record Table

[0077]

[0078] Based on the above table, the instability index of the samples of Example 1, Example 2, and Example 3 remained around 0 within 16 h, while the instability index of the sample of Comparative Example 1 gradually increased within the first 6 hours and then stabilized at a level much higher than that of the three example samples after 6 hours. The comparison between Example 2 and Comparative Example 2 shows that the use of gradient cross-linking modification of sodium hyaluronate gives the gel a more stable microstructure. The comparison between Example 3 and Comparative Example 3 shows that vacuum negative pressure stirring combined with self-rotation and revolution stirring can remove the bubbles in the cosmetic filler, make the components uniformly dispersed, and improve the stability of the dispersion system.

[0079] Comparative Test Two: Pushing Force Test

[0080] The cosmetic fillers prepared in Example 1 and Example 2 were used for testing. A 1-milliliter syringe was used to extract the cosmetic filler after standing, a 27G needle was added, and a digital push-pull force testing machine was used to detect the average pushing force required for the cosmetic filler to be pushed out through the injection needle. The specific recorded data are shown in the following table:

[0081] Table 2: Average Pushing Force (N) Comparative Data Record Table

[0082]

[0083] In combination with the above table, the average pushing force of Example 1, Example 2 and Example 3 is basically consistent at about 18N, while the average pushing force of the comparative sample is higher than 10N, which shows that the gel system of the examples has good injectability. The comparison results of Example 1 and Example 2 show that in the case of more solutes in Example 2, the gradient cross-linked sodium hyaluronate, the composite microemulsion and the graft copolymer can improve the injection performance of the material.

[0084] Comparative Test 3: Viscoelasticity Test

[0085] The viscoelasticity test of the filling materials of Examples 1-3 and Comparative Examples 1-3 was carried out at 25°C and 0.1Hz, and the results were as follows: Figure 6 The elastic modulus G' of each group was greater than G", which proved that the material had good deformation resistance and could effectively restore its original shape after stress was applied. The specific recorded data are shown in the following table:

[0086] Table 3 Comparison data record table of elastic modulus G' (Pa) and storage modulus G" (Pa)

[0087]

[0088] In combination with the above table, the comparison of Example 1 and Example 2, Example 2 and Example 3, Example 1 and Comparative Example 1, Example 2 and Comparative Example 2, and Example 3 and Comparative Example 3 was made respectively.

[0089] The comparison results of Example 1 and Comparative Example 1 show that the elastic modulus of Example 1 is at a higher level, higher than that of Comparative Example 1, which shows that the strength of the filling material prepared in Example 1 is higher, which proves that the material has good deformation resistance and can effectively restore its original shape after stress is applied, which is better than the existing commercial products, and has better effect on filling skin wrinkles and can promote rapid repair of the skin sebum membrane layer.

[0090] The comparison results of Example 1 and Comparative Example 2 show that the elastic modulus of Example 2 is at a high level, higher than that of Example 1, which shows that the gradient cross-linked modified sodium hyaluronate can form a stable elastic support network;

[0091] The comparison results of Example 2 and Comparative Example 3 show that the filling material with higher solute content has higher elastic modulus;

[0092] The comparison results of Example 2 and Comparative Example 2 show that the elastic modulus of Example 2 is higher than that of Comparative Example 2, which proves that the gradient cross-linked sodium hyaluronate can form a synergistic effect with the graft copolymer to provide a stable microenvironment for collagen synthesis, and has better filling effect, and a single cross-linked structure cannot achieve the same effect;

[0093] The comparison results of example 3 and comparative example 3 show that the elastic modulus of example 3 is significantly higher than that of comparative example 3, because the material of comparative example 3 has a large number of bubble components mixed unevenly, which destroys the structural stability and leads to poor deformation resistance.

[0094] Test four: cytotoxicity test

[0095] The cosmetic filling material prepared in example 1 was taken for testing. The cells in vigorous growth were digested and configured into a cell suspension of 1x10 5 The 96-well plate was taken, 100 μL of the cell suspension was added to each well, and the plate was cultured at 37°C in a 5% CO2concentration for 24 h; after 24 h, the original culture solution was discarded, and 100 μL of each concentration of sample extract (0.2 g / ml extraction ratio, 37°C extraction for 24 h), negative control and positive control were added, respectively. At least 6 replicates were made for each group, and the plate was cultured at 37°C in a 5% CO2concentration for 24 h. Then, cell morphology observation was performed. All the liquid in the 96-well plate was discarded, 50 μL of MTT solution was added, and the plate was incubated at 37°C for 2 h. The MTT solution was removed, 100 μL of isopropyl alcohol was added to each well, the plate was shaken, and the absorbance was detected by an enzyme-labeled instrument and the survival rate was calculated. The test results of example 1 show that the filling material has good biocompatibility and does not produce cytotoxicity.

[0096] Table 4 survival rate record table

[0097]

[0098] The present application is even only a formula containing basic components, and the biocompatibility, filling stability, injectability and deformation resistance are significantly better than those of conventional commercially available synthetic high molecular filling materials, which embodies the core advantages of natural-based materials.

[0099] The addition of three functional components of gradient cross-linking modified sodium hyaluronate, composite microemulsion and graft copolymer, and the vacuum self-rotation and revolution stirring process can significantly improve the performance of the filling material in all dimensions, which is the core of the technical breakthrough of the present application. The stirring process proposed in the present application, i.e., vacuum negative pressure and self-rotation and revolution cooperation, is the key production link to ensure the performance of the material. Even if the optimized full-function formula is used, if the process has the defects of not being defoamed and being inhomogeneous, the filling stability, safety, repair ability and long-acting property of the material will still be significantly worse than those of the process optimization group, and the optimal potential of the formula cannot be realized.

[0100] The above is only a preferred specific embodiment of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can make equivalent replacement or change within the technical scope disclosed in the present application according to the technical solution and inventive concept of the present application, which should be covered within the protection scope of the present application.

Claims

1. A cosmetic filling material for injection, characterized by, 10-40 parts of hydroxyapatite microspheres, 60-90 parts of gel liquid, 2-5 parts of gradient crosslinking modified sodium hyaluronate, 0.5-1 part of vitamin E-tea seed oil-panthenol compound microemulsion, 0.5-1 part of hyaluronic acid-proline graft copolymer, by mass percentage; The preparation steps of the gradient crosslinking modified sodium hyaluronate are as follows: The sodium hyaluronate with a molecular weight of 800-1200 kDa is divided into two groups A and B with a mass ratio of 7:3; The A group is reacted with 5%-6% mass of glycolaldehyde, and the addition amount is 5%-6% of the mass of the A group, to prepare a low crosslinking degree component under the conditions of pH 6.5-7.0 and 35-40 DEG C for 1.5 h; the B group is reacted with a glycolaldehyde-adipic acid dihydrazide mixed crosslinking agent with a mass ratio of glycolaldehyde to adipic acid dihydrazide of 3:1, and the total addition amount is 7%-8% of the mass of the B group, to prepare a high crosslinking degree component under the conditions of pH 7.0-7.2 and 40-45 DEG C for 2.5 h; The two components are mixed and ultrasonically dispersed at a power of 300-400 W for 10-15 min to obtain the gradient crosslinking modified sodium hyaluronate; The preparation steps of the vitamin E-tea seed oil-panthenol compound microemulsion are as follows: Vitamin E, tea seed oil and panthenol are mixed in a mass ratio of 3:2:1 as an oil phase; Polysorbate 80 with a mass concentration of 10%-15% and glycerol with a mass concentration of 5%-8% are mixed in a mass ratio of 2:1 as an aqueous phase; The oil phase is slowly added into the aqueous phase, and high-speed shearing is carried out at 10000-12000 r / min for 15-20 min at 40-45 DEG C, and then ultrasonic emulsification is carried out at 200-300 W for 5-8 min to form a uniform microemulsion with a particle size of 100-300 nm.

2. The cosmetic filler material for injection according to claim 1, characterized in that, The gel liquid is prepared by adding one or more of cellulose derivatives, chitosan, sodium alginate, polyethylene oxide and polyethylene glycol in any combination into a mixed solution of injection water and glycerol, stirring uniformly, and placing the mixture in an oven at 60 DEG C for 4 h.

3. The cosmetic filler material for injection according to claim 1, wherein The preparation steps of the hyaluronic acid-proline graft copolymer are as follows: Hyaluronic acid with a molecular weight of 100-200 kDa is mixed with L-proline in a mass ratio of 5:1, and 1-ethyl-carbodiimide hydrochloride solution with a mass concentration of 1%-2% is added in an amount of 3 times the mass of the mixture; Stirring is carried out at pH 5.0-5.5 and 30-35 DEG C for 4-6 h, and then the reaction product is purified by dialysis for 24 h with a molecular weight cutoff of 50 kDa, and then freeze-drying is carried out to obtain a white powder of the graft copolymer.

4. A method of preparing a cosmetic filling material for injection according to any one of claims 1 to 3, characterized in that, The following steps are included: S1, hydroxyapatite synthesis, raw materials calcium nitrate, diammonium hydrogen phosphate and ammonia are mixed into injection water, and the reaction is carried out at 25-40 DEG C, pH 9.0-10.0 and a stirring speed of 200-300 r / min for 24-30 h to obtain a hydroxyapatite slurry, which is then filtered and washed to obtain a hydroxyapatite slurry; S2, hydroxyapatite microspheres are prepared, the slurry is introduced into a spray centrifugal dryer to obtain spherical or spherical-like hydroxyapatite powder; the powder microspheres are calcined at a high temperature of 800-1000 DEG C to obtain hydroxyapatite microspheres; the hydroxyapatite microspheres are sieved by ultrasonic vibration to obtain hydroxyapatite microspheres with a particle size range of 5-100 microns; and the sieved microspheres are calcined at a high temperature of 1100-1300 DEG C to obtain high-purity hydroxyapatite microspheres with good biocompatibility; S3, stirring and mixing, the hydroxyapatite microspheres and other components are proportionally placed in a stirring machine with a gel liquid, the stirring machine provides a vacuum negative pressure of 50000-100000 Pa, the stirring paddle rotation speed is adjusted to 30-100 r / min, stirring is performed for 60-120 min, the temperature is 25-30 DEG C, and the barrel is simultaneously rotated and revolved for stirring.

5. The method of claim 4, wherein the cosmetic filler material for injection is prepared by mixing the hyaluronic acid and the cross-linking agent in a ratio of 1:0.01 to 1:0.1 (w / w) and then cross-linking the mixture. In the step S1, the molar ratio of calcium nitrate to diammonium hydrogen phosphate is 5:3, the mass concentration of ammonia water is 25%-28%, and the ammonia water is added at a speed of 1-2 drops per second during the reaction.

6. The method of claim 4, wherein the cosmetic filler material for injection is prepared by mixing the hyaluronic acid and the cross-linking agent in a ratio of 1:0.01 to 1:0.

1. In the step S2, the air inlet temperature of the spray centrifugal dryer is set to 100-300 DEG C, the air outlet temperature is controlled to 50-150 DEG C, and the centrifugal rotation speed is 10000-30000 r / min.

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