A bioceramic microsphere soft tissue injection and a preparation method and application thereof

Abstract

The application discloses a kind of bioceramic microsphere soft tissue injection and its preparation method and application, it is related to the technical field of biomedical materials.It includes the following weight parts raw materials: 100-120 weight parts gel carrier and 50-98 weight parts bioceramic microspheres.The application designs a kind of bioceramic microsphere soft tissue injection, and the soft tissue injection is composed of bioceramic microspheres and gel carrier, and the bioceramic microspheres used in the application reduce the inflammatory response of bioceramic microsphere soft tissue injection while ensuring good cell activity, loading capacity and rheological properties.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical materials, in particular to a bioceramic microsphere soft tissue injection agent and a preparation method and application thereof. BACKGROUND

[0002] Since the latter half of the 20th century, the pursuit of facial rejuvenation has gradually evolved into a popular trend. Injection plastic surgery technology has been widely welcomed due to its simple operation, minimal trauma, high safety, short recovery period, and remarkable effect. This technology not only meets people's pursuit of beauty, but also quickly becomes a popular choice on the market due to its quick-acting characteristics. With the continuous progress of technology and the growing pursuit of beauty, the market demand for injection plastic surgery technology has shown explosive growth, becoming one of the preferred plastic surgery methods for many beauty seekers. In the field of medical cosmetic injection filling, the current mainstream filling materials mainly include non-autologous tissue injections. These materials not only include single-component hyaluronic acid (HA) and collagen, but also composite filling materials that can stimulate collagen and fibroplasia, such as polycaprolactone (PCL), poly-L-lactic acid (PLLA), and calcium hydroxyapatite (CaHA). These advanced materials have begun to be used in clinical applications and gradually occupy a place in the injection micro-plastic cosmetic market. However, hyaluronic acid is easily degraded by enzymes in the body, so it has a short retention time and only provides a temporary volume filling. In order to maintain the desired cosmetic results, regular injections are required. However, excessive injection of hyaluronic acid or its cross-linked form can cause excessive facial swelling and other problems such as puffy face due to high water swelling. Although PLLA or PCL as an injectable filling material has the advantage of stimulating collagen production, it can cause adverse reactions such as local ecchymosis, pain, hematoma, local inflammatory reaction, and edema that can last for 3 to 5 days. Hydroxyapatite, as a naturally occurring material in human tissue, has good biocompatibility and can effectively fill and repair soft tissue, providing appropriate structural support and maintaining the volume and shape after injection.

[0003] Chinese patent CN 110292655A discloses an injection filling preparation containing hydroxyapatite and a preparation method thereof, and evaluates the wrinkle removal effect of hydroxyapatite for soft tissue filling through clinical trials.

[0004] Chinese patent CN 117982735A discloses an antibacterial injectable cosmetic filler and its preparation method, hydroxyapatite microspheres are prepared by oil phase reaction method, and are mixed with collagen paste to be implanted subcutaneously in rabbits to show good filling effect.

[0005] To reduce the foreign body stimulation of the material to the tissue and reduce the occurrence of inflammatory response, hydroxyapatite is usually made into microspheres. Since hydroxyapatite is insoluble in water or organic solvents, it is difficult to prepare microspheres by emulsion polymerization, membrane emulsification, microfluidic method and other methods. It can be made into microspheres by hydrothermal method, spray drying method, sol-gel method, template method, etc. The spray drying method has the advantages of stable microsphere structure, uniform particle size, good result repeatability and easy industrial large-scale production.

[0006] However, due to the relative inertness of hydroxyapatite, the effect of promoting collagen secretion is not ideal. By doping silicon and magnesium elements on the basis of hydroxyapatite, calcium magnesium based silicate bioceramics can be obtained. Compared with hydroxyapatite, it has excellent bioactivity. During the degradation process, it can release Ca 2+ , Si 4+ , Mg 2+ and other ionic products, which can significantly promote vascularization and tissue regeneration, and further produce good collagen neogenesis and long-acting filling effect. Similarly, Chinese patent CN116672496A discloses an injectable tissue filler containing silicate ceramic microspheres and its preparation method and application. Silicate ceramic microspheres are prepared by sol-spray method, which can play a good filling role. However, silicate bioceramics has a high pH, which may cause a high local pH and cause serious inflammatory response, and is not suitable for direct use as a filler.

[0007] Therefore, how to reduce the inflammatory response caused by silicate materials and improve biocompatibility is of great significance to improve the safety of bioceramic materials. SUMMARY

[0008] According to the deficiencies of the prior art, the present application designs a bioceramic microsphere soft tissue injection, which is composed of bioceramic microspheres and gel carriers. The bioceramic microspheres used in the present application reduce the inflammatory response of the bioceramic microsphere soft tissue injection while ensuring good cell activity, loading capacity and rheological properties, thereby solving the technical problems raised in the background art. Specifically, the technical scheme of the present application includes the following contents:

[0009] A bioceramic microsphere soft tissue injection, the bioceramic microsphere soft tissue injection includes the following weight parts of raw materials:

[0010] 100-120 parts by weight of gel carrier and 90-98 parts by weight of bioceramic microspheres.

[0011] Further, the preparation method of the gel carrier comprises the following steps:

[0012] 110-120 parts by weight of purified water and 2-3 parts by weight of biomacromolecular material are mixed to form a gel carrier by mechanical stirring.

[0013] Further, the purified water can also be replaced by sterile water or deionized water.

[0014] Further, the conditions of the mechanical stirring include stirring temperature 25-30℃, stirring speed 200-300r / min and stirring time 30-40min.

[0015] Further, the biomacromolecular material includes sodium hyaluronate, and can also be sodium carboxymethyl cellulose, collagen or gelatin.

[0016] Further, the preparation method of the bioceramic microsphere comprises the following steps:

[0017] 95-150 parts by weight of calcium phosphate and 95-105 parts by weight of silicate are mixed by ball milling to obtain ceramic powder;

[0018] 190-200 parts by weight of ceramic powder, 4.5-5.0 parts by weight of binder, 0-3.5 parts by weight of wetting agent and 500 parts by weight of purified water are mixed to form a spray slurry;

[0019] The spray slurry is spray granulated at 200-260℃ to form a bioceramic microsphere body;

[0020] The bioceramic microsphere body is sintered at 1100-1400℃ for 1-2h, and then sieved to obtain a bioceramic microsphere.

[0021] Further, the calcium phosphate is selected from any one or both of hydroxyapatite and β-tricalcium phosphate, and the calcium phosphate can also be selected from α-tricalcium phosphate or tetracalcium phosphate.

[0022] Further, the silicate includes white calcium silicate, and can also use silica apatite, bioglass or transparent apatite.

[0023] Further, the conditions of the ball milling include ball milling temperature 25-30℃, ball milling speed 300-400r / min and ball milling time 2-4h.

[0024] Further, the binder includes polyvinyl alcohol or polyvinyl pyrrolidone.

[0025] Further, the wetting agent includes polysorbate 80 or polysorbate 60.

[0026] Further, the particle size range of the screened particles is 20-50 mu m.

[0027] A preparation method of a bioceramic microsphere soft tissue injection, the preparation method comprising the following steps:

[0028] The gel carrier and the bioceramic microspheres are mixed and high-pressure homogenized, then loaded into a syringe and subjected to 121 DEG C high-pressure sterilization for 30 min to obtain the bioceramic microsphere soft tissue injection.

[0029] Further, the high-pressure homogenization conditions include a homogenization pressure of 10-20 MPa, a homogenization rotation speed of 1000-1800 r / min and a homogenization time of 8-10 min.

[0030] An application of a bioceramic microsphere soft tissue injection in the field of collagen regeneration.

[0031] Compared with the prior art, the present application has the following beneficial effects:

[0032] (1) The bioceramic microspheres prepared by the present application are spherical and smooth, the smooth spherical shape makes the bioceramic microspheres more easily pass through the syringe during injection, and the bioceramic microspheres can flow into the soft tissue area to be filled with lower pushing force.

[0033] (2) The bioceramic prepared by the present application has pores on the surface, the pores increase the contact area of the bioceramic microspheres and the gel, increase the friction, and help to maintain the suspended state of the microspheres in the gel. When these microspheres reach the target tissue site, they will provide a growth scaffold for autologous tissue.

[0034] (3) The size of the microspheres prepared by the present application is most ideal between 20-50 mu m, which not only facilitates injection, but also maximally reduces the immune inflammatory reaction caused by the uptake of the microspheres by macrophages.

[0035] (4) The bioceramic microspheres used by the present application belong to calcium phosphate or silicate, contain calcium, phosphorus, silicon, magnesium active elements, and have good biocompatibility and bioactivity, and have low immunogenicity after being injected into soft tissue, and almost no serious inflammatory reaction. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 SEM images of the bioceramic microspheres prepared by the preparation example 5 of the present application, wherein the scale of the (A) figure is 100 mu m, and the scale of the (B) figure is 10 mu m;

[0037] Figure 2 SEM images of the bioceramic microspheres prepared by the preparation example 6 of the present application, wherein the scale of the (A) figure is 100 mu m, and the scale of the (B) figure is 10 mu m;

[0038] Figure 3 SEM images of the bioceramic microspheres prepared in Preparation Example 7 of the present application, wherein the scale of the (A) image is 100 μm and the scale of the (B) image is 10 μm;

[0039] Figure 4 SEM images of the bioceramic microspheres prepared in Preparation Example 13 of the present application, wherein the scale is 20 μm;

[0040] Figure 5 Cell proliferation activity results of the bioceramic microspheres prepared in Preparation Examples 5 to 7 and Preparation Examples 8 to 10 of the present application;

[0041] Figure 6 Rheological properties of the bioceramic microsphere soft tissue injection prepared in Example 3 and Comparative Example 1 of the present application, wherein G' represents storage modulus, G" represents loss modulus, and η represents viscosity;

[0042] Figure 7 Results of H&E staining of the sample at the implantation site after implanting the bioceramic microsphere soft tissue injection prepared in Example 3 and Comparative Example 2 into a rabbit for 2 weeks;

[0043] Figure 8 Results of Sirius red staining of the sample at the implantation site after implanting the bioceramic microsphere soft tissue injection prepared in Example 3 and Comparative Example 2 into a rabbit. DETAILED DESCRIPTION

[0044] The technical solutions of the present application will be described clearly and completely below through the examples of the present application and the accompanying drawings. Obviously, the described examples are only a part of the examples of the present application, but not all the examples. Based on the examples in the present application, all the other examples obtained by those skilled in the art without making creative efforts fall within the scope of protection of the present application.

[0045] Unless otherwise specified, the raw materials and reagents used in the present application below are commercially available or can be prepared by known methods.

[0046] Preparation Example 1

[0047] The preparation method of the gel carrier specifically includes the following processes:

[0048] 0.15 g of sodium hyaluronate was slowly added into 120 g of purified water and mixed, and then placed in a temperature environment of 25°C, and mechanically stirred by a stirring paddle with a rotation speed of 200 r / min for 30 min to complete the preparation of the gel carrier.

[0049] Preparation Example 2

[0050] The preparation method of the gel carrier specifically includes the following steps:

[0051] 4.6g of sodium hyaluronate was slowly added to 110g of purified water and mixed. The mixture was then placed in a 25°C environment and mechanically stirred for 35 minutes using a stirring paddle at 250r / min to complete the preparation of the gel carrier.

[0052] Preparation Example 3

[0053] The preparation method of the gel carrier specifically includes the following steps:

[0054] 3g of sodium hyaluronate was slowly added to 120g of purified water and mixed. The mixture was then placed in a 30°C environment and mechanically stirred for 40 minutes using a stirring paddle at 300r / min to complete the preparation of the gel carrier.

[0055] Preparation Example 4

[0056] The preparation method of the gel carrier specifically includes the following steps:

[0057] 10g of sodium hyaluronate was slowly added to 120g of purified water and mixed. The mixture was then placed in a 30°C environment and mechanically stirred for 40 minutes using a stirring paddle at 300r / min to complete the preparation of the gel carrier.

[0058] Preparation Example 5

[0059] The preparation method of bioceramic microspheres specifically includes the following processes:

[0060] Weigh out 95g of hydroxyapatite and 95g of white calcium silicate, mix them evenly, and put them into a ball mill jar. In an environment of 25℃, ball mill at a speed of 300r / min for 4 hours to obtain ceramic powder.

[0061] Weigh 190g of ceramic powder, 4.5g of polyvinylpyrrolidone and 500g of purified water and mix them evenly to prepare a spray slurry;

[0062] The spray slurry is fed into a spray dryer and sprayed at a temperature of 200°C to obtain bioceramic microsphere preforms;

[0063] The bioceramic microsphere preforms were sintered at 1400℃ for 1 hour. After sintering, they were naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20–50 μm. The bioceramic microspheres were characterized by scanning electron microscopy, and the results are as follows: Figure 1 As shown, bioceramic microspheres have a relatively smooth surface and a certain amount of porosity.

[0064] Preparation Example 6

[0065] A method for preparing bioceramic microspheres, specifically comprising the following processes:

[0066] 65 g of hydroxyapatite, 65 g of beta-tricalcium phosphate, and 100 g of xonotlite are weighed and uniformly mixed, and then are jointly loaded into a ball mill tank, and are ball milled and crushed at a rotation speed of 350 r / min for 3 h in an environment at 25 DEG C to obtain ceramic powder;

[0067] 195 g of the ceramic powder, 4.7 g of polyvinyl alcohol, 3.5 g of polysorbate 80, and 500 g of purified water are uniformly mixed to prepare a spray slurry;

[0068] The spray slurry is fed into a spray dryer, and spray granulation is performed at a temperature of 240 DEG C to obtain bioceramic microsphere blanks;

[0069] The bioceramic microsphere blanks are placed in a high-temperature environment at 1100 DEG C for sintering for 2 h, and after sintering is completed, natural cooling to room temperature is performed and sieving is performed to obtain bioceramic microspheres with a particle size range of 20-50 mu m. The bioceramic microspheres are characterized by scanning electron microscopy, and the results are shown in Figure 2 The bioceramic microspheres have a relatively smooth surface and have certain porosity.

[0070] Preparation Example 7

[0071] A method for preparing bioceramic microspheres, specifically comprising the following processes:

[0072] 150 g of beta-tricalcium phosphate and 105 g of xonotlite are weighed and uniformly mixed, and then are jointly loaded into a ball mill tank, and are ball milled and crushed at a rotation speed of 400 r / min for 2 h in an environment at 30 DEG C to obtain ceramic powder;

[0073] 200 g of the ceramic powder, 5.0 g of polyvinylpyrrolidone, 2 g of polysorbate 60, and 500 g of water are uniformly mixed to prepare a spray slurry;

[0074] The spray slurry is fed into a spray dryer, and spray granulation is performed at a temperature of 260 DEG C to obtain bioceramic microsphere blanks;

[0075] The bioceramic microsphere blanks are placed in a high-temperature environment at 1250 DEG C for sintering for 1 h, and after sintering is completed, natural cooling to room temperature is performed and sieving is performed to obtain bioceramic microspheres with a particle size range of 20-50 mu m. The bioceramic microspheres are characterized by scanning electron microscopy, and the results are shown in Figure 3 The bioceramic microspheres have a relatively smooth surface and have certain porosity.

[0076] Preparation Example 8

[0077] A method for preparing bioceramic microspheres, specifically comprising the following processes:

[0078] Take 200g of hydroxyapatite nano-powder directly as ceramic powder, 5.0g of polyvinyl pyrrolidone, 2g of polysorbate 60 and 500g of water are mixed uniformly to prepare a spray slurry;

[0079] The spray slurry is sent into a spray dryer, and spray granulation is carried out at a temperature of 260℃ to obtain a bioceramic microsphere blank;

[0080] The bioceramic microsphere blank is placed in a high-temperature environment of 1250℃ and sintered for 1h, and then naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20-50μm.

[0081] Preparation Example 9

[0082] The preparation method of the bioceramic microspheres specifically includes the following processes:

[0083] Take 200g of hydroxyapatite nano-powder directly as ceramic powder, 5.0g of polyvinyl pyrrolidone, 2g of polysorbate 60 and 500g of water are mixed uniformly to prepare a spray slurry;

[0084] The spray slurry is sent into a spray dryer, and spray granulation is carried out at a temperature of 260℃ to obtain a bioceramic microsphere blank;

[0085] The bioceramic microsphere blank is placed in a high-temperature environment of 1250℃ and sintered for 1h, and then naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20-50μm.

[0086] Preparation Example 10

[0087] The preparation method of the bioceramic microspheres specifically includes the following processes:

[0088] Take 200g of hydroxyapatite nano-powder directly as ceramic powder, 5.0g of polyvinyl pyrrolidone, 2g of polysorbate 60 and 500g of water are mixed uniformly to prepare a spray slurry;

[0089] The spray slurry is sent into a spray dryer, and spray granulation is carried out at a temperature of 260℃ to obtain a bioceramic microsphere blank;

[0090] The bioceramic microsphere blank is placed in a high-temperature environment of 1250℃ and sintered for 1h, and then naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20-50μm.

[0091] The bioceramic microsphere blank is placed in a high-temperature environment of 1250℃ and sintered for 1h, and then naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20-50μm.

[0092] Preparation Example 11

[0093] A method for preparing bioceramic microspheres, specifically comprising the following processes:

[0094] 50g of beta-tricalcium phosphate and 150g of xonotlite are weighed and uniformly mixed, and then jointly loaded into a ball mill tank, and ball milled at a speed of 400r / min for 2h in an environment of 30°C to obtain ceramic powder;

[0095] 200g of the ceramic powder, 5.0g of polyvinylpyrrolidone, 2g of polysorbate 60, and 500g of water are weighed and uniformly mixed to prepare a spray slurry;

[0096] The spray slurry is sent into a spray dryer, and spray granulation is performed at a temperature of 260°C to obtain bioceramic microsphere blanks;

[0097] The bioceramic microsphere blanks are placed in a high-temperature environment of 1250°C for sintering for 1h, and then naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20-50μm.

[0098] Preparation Example 12

[0099] A method for preparing bioceramic microspheres, specifically comprising the following processes:

[0100] 150g of beta-tricalcium phosphate and 105g of xonotlite are weighed and uniformly mixed, and then jointly loaded into a ball mill tank, and ball milled at a speed of 400r / min for 2h in an environment of 30°C to obtain ceramic powder;

[0101] 200g of the ceramic powder, 5.0g of polyvinylpyrrolidone, 2g of polysorbate 60, and 500g of water are weighed and uniformly mixed to prepare a spray slurry;

[0102] The spray slurry is sent into a spray dryer, and spray granulation is performed at a temperature of 260°C to obtain bioceramic microsphere blanks;

[0103] The bioceramic microsphere blanks are placed in a high-temperature environment of 1000°C for sintering for 1h, and then naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20-50μm.

[0104] Preparation Example 13

[0105] A method for preparing bioceramic microspheres, specifically comprising the following processes:

[0106] 150g of beta-tricalcium phosphate and 105g of xonotlite are weighed and uniformly mixed, and then jointly loaded into a ball mill tank, and ball milled at a speed of 400r / min for 2h in an environment of 30°C to obtain ceramic powder;

[0107] Weigh 200g of ceramic powder, 5.0g of polyvinylpyrrolidone, 2g of polysorbate 60 and 500g of water and mix them evenly to prepare a spray slurry;

[0108] The spray slurry is fed into a spray dryer and sprayed at a temperature of 260°C to obtain bioceramic microsphere preforms;

[0109] The bioceramic microsphere preforms were sintered at 1500℃ for 2 hours. After sintering, they were naturally cooled to room temperature and sieved to obtain bioceramic microspheres with a particle size range of 20–50 μm. The bioceramic microspheres were characterized by scanning electron microscopy, and the results are as follows: Figure 4 As shown, the microspheres melted and adhered severely.

[0110] (1) Verification of the cell activity of the bioceramic microspheres obtained in Examples 5-13:

[0111] The cell viability of the bioceramic microspheres prepared in Examples 5-13 was evaluated using the CCK-8 assay. Human fibroblasts (MRC5) were seeded onto the surface of the bioceramic microspheres prepared in Examples 5-13, and cell proliferation rate was measured by absorbance on days 1, 3, 5, and 7. The results are as follows: Figure 5 As shown.

[0112] pass Figure 5 It can be observed that, compared with preparations 8-13, the bioceramic microspheres prepared in preparations 5-7 exhibit significantly higher cell proliferation rates, indicating that the bioceramic microspheres obtained in preparations 5-7 have better biocompatibility. Compared with bioceramic microspheres prepared using only hydroxyapatite, magnesium feldspar, or calcium silicate, the bioceramic microspheres obtained in preparations 5-7 through the combined action of calcium phosphate and silicate may have a richer and more balanced ion release capacity containing calcium, phosphorus, silicon, and magnesium, which is conducive to cell adhesion, migration, and ingrowth of new tissue for local repair, thus resulting in a much higher cell proliferation rate than the bioceramic microspheres obtained in preparations 8-10. Through preparation examples 11 and 7, it was found that when the amount of β-tricalcium phosphate was reduced and the amount of leucosite was increased, the cell proliferation rate of the bioceramic microspheres prepared in preparation example 11 was significantly worse. A comparison of preparation examples 12 and 13 with preparation example 7 revealed that the cell proliferation rates of the bioceramic microspheres prepared in preparation examples 12 and 13 were relatively poor. This may be because the raw materials used in this system have certain requirements for calcination temperature; exceeding a certain range may result in insufficient or excessive calcination, affecting the activity of the bioceramic microspheres. This indicates that the raw materials, proportions, and calcination conditions have a significant impact on the bioceramic microspheres prepared according to this invention, and not all proportions can produce bioceramic microspheres with good cell activity according to the system of this invention.

[0113] (2) Test the maximum loading capacity, pH and push force of the bioceramic microspheres prepared in Preparation Examples 5-13:

[0114] Prepare six homogenizer mixing tanks numbered ①-⑨ respectively, and add 113.52 g of the gel carrier prepared in Preparation Example 3 to each of the nine mixing tanks;

[0115] Then, first add 1 g of the bioceramic microspheres prepared in Preparation Example 5 to the ① tank, and then put it into the homogenizer, control the homogenization pressure to be 20 MPa and the homogenization speed to be 1800 r / min, and perform high-pressure homogenization treatment, then add 1 g of the bioceramic microspheres prepared in Preparation Example 5 in small amounts and multiple times during the homogenization process until the gel in the ① tank appears a hard particle block, and record the cumulative addition amount of the bioceramic microspheres prepared in Preparation Example 5;

[0116] Similarly, Preparation Examples 6-13 are respectively dispersed into the ②-⑨ mixing tanks according to the above method, and the cumulative addition amount of the bioceramic microspheres prepared in Preparation Examples 6-13 is recorded respectively;

[0117] Finally, calculate the maximum loading capacity of the bioceramic microspheres prepared in Preparation Examples 5-13, and respectively test the pH using a pH meter, and test the push force using a universal testing machine at a speed of 30 mm / min, and the test results are shown in Table 1.

[0118] Table 1 Comparison of maximum loading capacity, pH and push force

[0119]

[0120]

[0121] *Note: In Table 1, " / " represents that it cannot be extruded or the needle is broken during the test, so the data cannot be measured.

[0122] As can be seen from Table 1, Preparation Examples 5-7 achieve higher microsphere loading capacity while maintaining lower push force. As can be seen from the table, with the change of the composition of the microspheres, the pH also changes significantly, the pH of Preparation Examples 5-7 is below 8, which is neutral or weakly alkaline; the pH of Preparation Examples 8-12 is above 8, which is alkaline. Sodium hyaluronate will completely ionize in an alkaline environment with high pH, resulting in an increase in the ionic strength of the solution, thereby reducing the flexibility of the molecular chain and the viscosity of the solution. This change makes the microspheres more likely to settle and difficult to form a stable suspended state in the gel carrier; although Preparation Example 13 has a pH close to neutral, the sintering temperature is too high, resulting in microsphere melting and serious adhesion, which can be seen from the fact that the needle is broken during the test. Preparation Example 13 has a pH close to neutral, but the sintering temperature is too high, resulting in microsphere melting and serious adhesion, which can be seen from the fact that the needle is broken during the test. Figure 4SEM images show that, therefore, it is extremely difficult to extrude. This can be due to the fact that in the present system, there are certain selective requirements for the raw materials, raw material ratio and sintering temperature for the preparation of bioceramic microspheres. Low pushing force and high microsphere load improve the convenience of the injection in clinical operation, and help to prolong the durability of collagen regeneration and repair effect.

[0123] Example 1

[0124] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0125] Take 50g of the bioceramic microspheres obtained in Preparation Example 5 and slowly add them to 120g of the gel carrier obtained in Preparation Example 1 for mixing, and then put them into a homogenizer, control the homogenization pressure at 10MPa and the homogenization speed at 1000r / min, and high-pressure homogenization treatment for 8min, so that the bioceramic microspheres are fully dispersed in the gel carrier; after the homogenization treatment is completed, the obtained composite material is loaded into a 1.5mL syringe, and then placed in a high-pressure sterilizer at 121℃ for sterilization for 30min, to obtain a bioceramic microsphere soft tissue injection, and test its pushing force.

[0126] Example 2

[0127] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0128] Take 98g of the bioceramic microspheres obtained in Preparation Example 6 and slowly add them to 100g of the gel carrier obtained in Preparation Example 2 for mixing, and then put them into a homogenizer, control the homogenization pressure at 15MPa and the homogenization speed at 1300r / min, and high-pressure homogenization treatment for 9min, so that the bioceramic microspheres are fully dispersed in the gel carrier; after the homogenization treatment is completed, the obtained composite material is loaded into a 1.5mL syringe, and then placed in a high-pressure sterilizer at 121℃ for sterilization for 30min, to obtain a bioceramic microsphere soft tissue injection, and test its pushing force.

[0129] Example 3

[0130] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0131] Take 98g of the bioceramic microspheres obtained in Preparation Example 7 and slowly add them to 120g of the gel carrier obtained in Preparation Example 3 for mixing, and then put them into a homogenizer, control the homogenization pressure at 20MPa and the homogenization speed at 1800r / min, and high-pressure homogenization treatment for 10min, so that the bioceramic microspheres are fully dispersed in the gel carrier; after the homogenization treatment is completed, the obtained composite material is loaded into a 1.5mL syringe, and then placed in a high-pressure sterilizer at 121℃ for sterilization for 30min, to obtain a bioceramic microsphere soft tissue injection, and test its pushing force.

[0132] Comparative Example 1

[0133] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0134] Take 98 g of the bioceramic microspheres obtained in Preparation Example 7 and slowly add them to 120 g of the gel carrier obtained in Preparation Example 4 for mixing, and then put them into a homogenizer, control the homogenization pressure at 20 MPa and the homogenization speed at 1800 r / min, and high-pressure homogenize for 10 min, so that the bioceramic microspheres are fully dispersed in the gel carrier; after the homogenization is completed, the obtained composite material is loaded into a 1.5 mL syringe, and then placed in a high-pressure sterilizer at 121 ℃ for sterilization for 30 min, to obtain a bioceramic microsphere soft tissue injection, and test the pushing force thereof.

[0135] Comparative Example 2

[0136] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0137] Take 30 g of the bioceramic microspheres obtained in Preparation Example 7 and slowly add them to 125 g of the gel carrier obtained in Preparation Example 3 for mixing, and then put them into a homogenizer, control the homogenization pressure at 20 MPa and the homogenization speed at 1800 r / min, and high-pressure homogenize for 10 min, so that the bioceramic microspheres are fully dispersed in the gel carrier; after the homogenization is completed, the obtained composite material is loaded into a 1.5 mL syringe, and then placed in a high-pressure sterilizer at 121 ℃ for sterilization for 30 min, to obtain a bioceramic microsphere soft tissue injection, and test the pushing force thereof.

[0138] Comparative Example 3

[0139] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0140] Take 120 g of the bioceramic microspheres obtained in Preparation Example 7 and slowly add them to 80 g of the gel carrier obtained in Preparation Example 3 for mixing, and then put them into a homogenizer, control the homogenization pressure at 20 MPa and the homogenization speed at 1800 r / min, and high-pressure homogenize for 10 min, so that the bioceramic microspheres are fully dispersed in the gel carrier; after the homogenization is completed, the obtained composite material is loaded into a 1.5 mL syringe, and then placed in a high-pressure sterilizer at 121 ℃ for sterilization for 30 min, to obtain a bioceramic microsphere soft tissue injection, and test the pushing force thereof.

[0141] Comparative Example 4

[0142] A preparation method of a bioceramic microsphere soft tissue injection, specifically comprising the following processes:

[0143] 63.23 g of the bioceramic microspheres obtained in Preparation Example 8 were slowly added into 154.81 g of the gel carrier obtained in Preparation Example 3, mixed, and then placed into a homogenizer, and the homogenization pressure was controlled at 20 MPa, the homogenization speed was controlled at 1800 r / min, and the bioceramic microspheres were fully dispersed in the gel carrier by high-pressure homogenization for 10 min; after the homogenization, the obtained composite material was loaded into a 1.5 mL syringe, and then placed into a high-pressure sterilizer at 121 ℃ for sterilization for 30 min, to obtain a bioceramic microsphere soft tissue injection, and the push force thereof was tested.

[0144] The push forces of the soft tissue injections obtained in Examples 1 to 3 and Comparative Examples 1 to 4 are shown in Table 2 below.

[0145] Table 2: Comparison of push forces

[0146] Material source Pushing force (N) Example 1 20.34 Example 2 37.20 Example 3 30.41 Comparative Example 1 129.12 Comparative Example 2 9.03 Comparative Example 3 109.59 Comparative Example 4 103.26

[0147] As can be seen from the comparison of the push forces in Table 2, in the present system, as the content of the bioceramic microspheres increases, the push force of the injection increases, that is, the higher the loading amount, the more difficult the injection is to be extruded. The push forces of Examples 1 to 3 are small, and the extrusion is smooth; the material of Comparative Example 1 is hydroxyapatite, and the bioceramic microspheres prepared by the method are difficult to disperse in the gel, which may be due to the fact that the amount of the biological macromolecule in the gel carrier is too high, so that the filler prepared is not uniform, and it is difficult to extrude; Comparative Example 2 contains less bioceramic microspheres, so the extrusion is smooth, but reducing the amount of the bioceramic microspheres is not conducive to the function of the injection; Comparative Example 3 contains more bioceramic microspheres than the ratio provided in the present application, and the push force is high, which is difficult to extrude. Comparative Example 4 uses hydroxyapatite nano-powder as the ceramic powder, and the bioceramic microspheres prepared therefrom have a large push force, which is not suitable for injection, which may be due to the fact that in the present system, the raw materials and the ratio for preparing the bioceramic microspheres have certain selectivity requirements. It is indicated that the ratio of the bioceramic microspheres and the gel carrier and the selection of the formula raw materials are changed at will, which can result in poor loading amount of the bioceramic microspheres of the injection, and can easily increase the viscosity of the injection, which is not conducive to use.

[0148] (3) Evaluation of the rheological properties of the soft tissue injection

[0149] The rheological properties of the bioceramic microsphere soft tissue injections obtained in Example 3 and Comparative Example 1 were evaluated using a parallel plate rheometer (Haake Mars 40, Germany). Tests involved measuring rheological properties under applied stress, recording the deformation behavior of the material under constant stress and its recovery under zero stress, and determining the composite modulus by applying oscillating stress within the viscoelastic range of the soft tissue injection. Viscosity, storage modulus, and loss modulus were measured at a frequency of 1 Hz and an oscillation force of 100 Pa. Results are shown below. Figure 6 In this context, (A) represents the rheological properties of Example 3, and (B) represents the rheological properties of Comparative Example 1.

[0150] pass Figure 6 It can be observed that the bioceramic microsphere soft tissue injection prepared in Example 3 exhibits good shear-thinning behavior, meaning that the viscosity of the injection significantly decreases with increasing shear rate. This characteristic not only helps improve the flowability of the material during injection but also reduces the applied force, making the operation more convenient. Meanwhile, G' and G" also reflect the good elasticity and deformation resistance of the bioceramic microsphere soft tissue injection prepared in Example 3, indicating that it can partially recover its initial structure after shear stress relief, thus better maintaining the stability of the filling morphology after injection, contributing to improved injection durability and aesthetic effects. In contrast, the bioceramic microsphere soft tissue injection prepared in Comparative Example 1 exhibits weakened shear-thinning ability and high viscosity. Although it has stronger load-bearing capacity, it is not conducive to its extrusion from the syringe. This indicates that, with a fixed loading of bioceramic microspheres, the ratio of the gel carrier has a significant impact on the final prepared bioceramic microsphere soft tissue injection; not all ratios can achieve good rheological properties.

[0151] (4) Evaluation of collagen regeneration performance of bioceramic microsphere soft tissue injection

[0152] To verify the collagen regeneration performance of the bioceramic microsphere soft tissue injection prepared in this invention, subcutaneous implantation experiments were conducted on animals using the bioceramic microsphere soft tissue injections prepared in Example 3 (experimental group) and Comparative Example 2 (control group). The local inflammatory response and collagen regeneration were observed in rabbits after implantation of the injection.

[0153] Experimental methods

[0154] 1) General anesthesia was administered via intravenous injection of 10% chloral hydrate. The rabbit's back was shaved to prepare the injection area. The prepared experimental animal was placed in a prone position on the operating table.

[0155] 2) Bioceramic microsphere soft tissue injection was implanted into the subcutaneous tissue of the rabbit's back using a 27G injection needle;

[0156] 3) After the procedure, each animal was housed in a separate cage.

[0157] Experimental observation

[0158] 1) Observe the condition of the subcutaneous filler and local tissues at specific time points after the filler procedure;

[0159] 2) Specimens were taken from the implantation site, fixed with 10% formaldehyde, and after routine decalcification, dehydration, paraffin embedding, and serial sectioning, hematoxylin-eosin (H&E) staining and Sirius red staining were performed.

[0160] 3) Observe the inflammatory response and collagen regeneration around the implantation site under an optical microscope, and observe the type of collagen under a polarized light microscope.

[0161] Result evaluation: such as Figure 7 , Figure 8 As shown, the bioceramic microsphere soft tissue injection in the experimental group (Example 3) exhibited good biocompatibility. No obvious inflammatory reaction was observed around the material, with only a small number of macrophages surrounding it, and fibroblast formation was visible, forming a thin and flat capsule around the microspheres. Furthermore, the fibroblasts participated in collagen regeneration and showed a significant collagen regeneration effect. Polarized light microscopy observation results showed that within 2 weeks after injection, it mainly promoted the production of type III collagen; by 2 months, type I collagen began to be produced; and by 6 months, a large amount of type I collagen was produced, thereby forming a collagen network structure with good support in the skin. In contrast, the bioceramic microsphere soft tissue injection in the control group (Comparative Example 2) also had good biocompatibility, but the collagen regeneration effect was significantly weaker than that in the experimental group. It can be seen that although reducing the number of bioceramic microspheres can improve the pushing effect of the prepared bioceramic microsphere soft tissue injection, the corresponding therapeutic effect is reduced, which is not conducive to use.

[0162] The results above show that the bioceramic microsphere soft tissue injection prepared according to the preparation method provided by the present invention has good biocompatibility and collagen regeneration effect.

[0163] The embodiments described above, along with the accompanying drawings, have provided a detailed explanation of the technical solutions and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Various changes and modifications can be made to the present invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed.

Claims

1. A bioceramic microsphere soft tissue injectable characterized in that, The bioceramic microsphere soft tissue injection includes the following raw materials by weight: 100-120 parts by weight of a gel carrier and 50-98 parts by weight of bioceramic microspheres; The preparation method of the gel carrier includes the following steps: 110-120 parts by weight of purified water and 0.15-4.6 parts by weight of a biomacromolecular material are mixed to form a gel carrier by mechanical stirring; The biomacromolecular material is sodium hyaluronate; The preparation method of the bioceramic microspheres includes the following steps: 95-150 parts by weight of calcium phosphate and 95-105 parts by weight of silicate are mixed by ball milling to obtain ceramic powder; 190-200 parts by weight of the ceramic powder, 4.5-5.0 parts by weight of a binder, 0-3.5 parts by weight of a wetting agent, and 500 parts by weight of purified water are mixed to form a spray slurry; The spray slurry is spray granulated at 200-260°C to form a bioceramic microsphere blank; The bioceramic microsphere blank is sintered at 1100-1400°C for 1-2 hours, and then sieved to obtain bioceramic microspheres; The calcium phosphate is selected from any one or both of hydroxyapatite and β-tricalcium phosphate; The silicate is white calcio-silicate; The sieved particle size ranges from 20 μm to 50 μm.

2. The bioceramic microsphere soft tissue injectable of claim 1, wherein, The binder includes polyvinyl alcohol or polyvinylpyrrolidone.

3. The bioceramic microsphere soft tissue injectable of claim 1, wherein, The wetting agent includes polysorbate 80 or polysorbate 60.

4. The method for preparing the soft tissue injection of the bioceramic microspheres according to any one of claims 1-3, characterized in that, The preparation method includes the following steps: The gel carrier and the bioceramic microspheres are mixed by high-pressure homogenization, then loaded into a syringe, and subjected to high-pressure sterilization at 121°C for 30 minutes to obtain the bioceramic microsphere soft tissue injection.

5. Use of the bioceramic microsphere soft tissue injection of any one of claims 1-3 or prepared by the preparation method of claim 4 in the field of bioceramic materials.

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