Adiponectin microspheres, adiponectin hydrogel and application of adiponectin microspheres and adiponectin hydrogel in field of bone adhesives

Abstract

The invention provides adiponectin microspheres, adiponectin hydrogel and application of the adiponectin microspheres and the adiponectin hydrogel in the field of adhesives. The adiponectin microsphere comprises adiponectin and a PLGA (poly (lactic-co-glycolic acid)) polymer, the average particle size of the adiponectin microsphere is 40-50 microns, and the adiponectin is wrapped in the PLGA polymer through the porous aperture of the PLGA polymer. The provided adiponectin microspheres can be used for preparing adiponectin hydrogel, and can be used in the field of bone adhesives to promote osteogenic differentiation.

Description

Technical Field

[0001] The invention relates to the field of medical bone adhesives, and in particular to adiponectin microspheres, adiponectin hydrogels and applications thereof in the field of bone adhesives. Background Art

[0002] The current treatment for comminuted fractures is mainly to fix the bone fragments with internal fixators such as steel plates, screws, steel needles, and steel wires, or simply to keep the fracture line stable and let it heal on its own. This often leads to problems such as unstable fixation of bone fragments, malformation, delayed healing or nonunion, and even residual limb dysfunction, and internal fixators are expensive or require a second surgery to remove. Some small pieces of broken bones cannot be fixed and remain free, affecting function or causing inflammation. Mobile bone fragments in key areas can even lead to compression paraplegia. Therefore, the fixation of comminuted fractures is a key problem that needs to be solved urgently in trauma orthopedics.

[0003] Medical bone adhesive is a new type of fracture fixation material that can reposition bone fragments well. It has the advantages of easy shaping, simple operation, stable performance, and few complications. It can solve the problems of unstable fixation of bone fragments, complicated surgery, easy infection, and high cost caused by internal fixation for comminuted fractures. The bonding principle of medical bone adhesive includes intermolecular adsorption, mechanical interlocking, and chemical bond bonding. Due to the low adhesion of medical bone adhesive and poor mechanical properties, biocompatibility, and bone conductivity, it is currently mainly used in the field of orthopedics for joint prosthesis replacement, osteoporosis, bone tumor resection and reconstruction, and percutaneous vertebroplasty for vertebral compression fractures, but is rarely used for fracture fixation. Therefore, it is necessary to find a method to improve medical bone adhesive to improve its adhesion and mechanical properties; at the same time, it can make it physiologically degradable; and locally promote the growth of osteoblasts. Recently, bone tissue engineering has attempted to study its nano-modification through nanotechnology. Summary of the invention

[0004] The present invention aims to solve one of the technical problems in the related art at least to a certain extent. The present invention provides adiponectin microspheres, adiponectin hydrogels and their use in the field of bone adhesives. The provided adiponectin microspheres or adiponectin hydrogels can be used as bone adhesives, or used to prepare bone adhesives to promote osteogenic differentiation.

[0005] Specifically, the present invention provides the following technical solutions:

[0006] In a first aspect of the present invention, adiponectin microspheres are provided, comprising adiponectin and PLGA polymers, wherein the average particle size of the adiponectin microspheres is 40 to 50 microns, and the adiponectin is encapsulated in the PLGA polymer through the porous pores of the PLGA polymer.

[0007] According to an embodiment of the present invention, the drug loading amount of adiponectin in the adiponectin microspheres is at least 10%, preferably at least 20%, and the encapsulation rate of adiponectin is at least 50%, preferably at least 65%.

[0008] According to an embodiment of the present invention, the particle size distribution span of the adiponectin microspheres is 1 to 3 microns.

[0009] In a second aspect of the present invention, a method for preparing adiponectin microspheres is provided, comprising:

[0010] After the PLAG polymer and adiponectin are mixed, self-healing packaging is performed at a predetermined temperature to obtain the adiponectin microspheres.

[0011] The provided method adsorbs and encapsulates adiponectin in microspheres under mild self-healing conditions, avoids contact of adiponectin with organic solvents, and ensures the activity of adiponectin.

[0012] According to an embodiment of the present invention, the mass ratio of the PLGA to the adiponectin microspheres is 1:(300-500).

[0013] In a third aspect of the present invention, an adiponectin hydrogel mixture is provided, which is obtained by mixing the adiponectin microspheres described in the first aspect with a hydrogel material.

[0014] The fourth aspect of the present invention provides an adiponectin hydrogel, characterized in that it is obtained by photocuring the adiponectin hydrogel mixture described in the third aspect.

[0015] According to an embodiment of the present invention, the adiponectin hydrogel has at least one of the following properties:

[0016] (a) It can solidify within 80 seconds under blue light irradiation without forming crystals or precipitation;

[0017] (b) Promote BMSCs proliferation;

[0018] (c) Promote osteogenic differentiation.

[0019] The fifth aspect of the present invention provides the use of the adiponectin microspheres described in the first aspect, the adiponectin hydrogel mixture described in the third aspect, or the adiponectin hydrogel described in the fourth aspect in the preparation of a drug for treating fractures. According to an embodiment of the present invention, the fracture is a comminuted fracture.

[0020] The sixth aspect of the present invention provides use of the adiponectin microspheres described in the first aspect, the adiponectin hydrogel mixture described in the third aspect, or the adiponectin hydrogel described in the fourth aspect in the field of bone adhesives.

[0021] The beneficial effects achieved by the present invention are at least:

[0022] The adiponectin microspheres provided by the present invention can be used as bone adhesive or bone adhesive component, have good biocompatibility, promote bone growth and fix fractures, can fix bone fragments while rapidly forming bones, and the implant material can also gradually degrade, providing a new idea and method for the repair of fractures, especially comminuted fractures. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the electron microscopic comparison result of the adiponectin microspheres before and after healing provided by the embodiments of the present invention.

[0024] Figure 2 The results of analyzing the pH and temperature stability of adiponectin microspheres using circular dichroism (CD) spectrometer provided in an embodiment of the present invention are as follows.

[0025] Figure 3 This is the result of evaluating biocompatibility with bone cells using the CCK8 method provided by an embodiment of the present invention, wherein ADPN is adiponectin, and ADPN-Gel is adiponectin hydrogel.

[0026] Figure 4 The mechanical characteristics of the adiponectin hydrogel provided by the embodiment of the present invention are characterized. A is the HE staining result; B is the Masion staining result; C is the callus thickness calculated by HE and Masion staining; D is the comparison of callus thickness and cortical bone quality by MicroCT; E is the comparison of the foot contact area of ​​each group of mice by Catwalk; F is the comparison of the callus strength after healing by three-point bending test, where ADPN represents adiponectin and ADPN-Gel is adiponectin hydrogel.

[0027] Figure 5 The results are provided by the embodiments of the present invention for evaluating the proliferation activity of BMSCs using the MTT method.

[0028] Figure 6 The results of promoting the differentiation of BMSCs into bones by the adiponectin hydrogel provided by the embodiments of the present invention, A is the result of calcium nodules shown by Alizarin red staining 21 days after osteogenic induction; B is the result of Oil Red O staining 14 days after adipogenic induction, red is fat cells; C is Alcian blue staining 21 days after chondrogenic induction, blue is chondrocytes; D is ALP staining 11 days after osteogenic induction; E is a comparison of the areas of calcium nodules shown by Alizarin red staining of the three groups after 21 days of osteogenic induction; F is a comparison of the differences in ALP activities of the three groups after 11 days of osteogenic induction; G: PCR comparison of the differences in osteogenic-related genes among the three groups, wherein ADPN represents adiponectin and ADPN-Gel represents adiponectin hydrogel. DETAILED DESCRIPTION

[0029] Embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, but should not be construed as limiting the present invention.

[0030] The research on adiponectin is mostly focused on metabolic fields such as glucose metabolism, insulin resistance, and atherosclerosis. In recent years, people have begun to discover that there is a continuity between adiponectin and bone repair, but the research is mostly focused on the field of bone metabolism, such as osteoporosis, osteosclerosis, osteoarthritis, bone tumors, etc. Our previous studies have found that adiponectin has the effect of promoting fracture repair. For example, in a mouse fracture model, local application of adiponectin has been found to promote fracture healing, increase bone strength, and improve bone microstructure. However, when adiponectin is used alone, it needs to act for a long time after administration to better promote bone healing. Using polylactic-co-glycolic acid (PLGA) microspheres as carriers, adiponectin can be released in vivo. However, adiponectin is a protein substance. When microspheres are prepared using the classic microsphere preparation method, there is an oil-water interface, which will cause the inactivation of adiponectin protein. During the research process, the inventor creatively designed a self-healing process to prepare adiponectin microspheres, that is, first prepare porous microparticles, and then adsorb and encapsulate adiponectin in the microspheres under mild self-healing conditions, avoiding contact between adiponectin and organic solvents, and ensuring the activity of adiponectin. The hydrogel prepared by using the provided adiponectin microspheres has a long action time, extremely high stability at different pH values, high stability at low and normal temperatures, and the components of the gel are not only non-cytotoxic, but also can promote cell proliferation.

[0031] In a first aspect of the present invention, adiponectin microspheres are provided, comprising adiponectin and PLGA polymers, wherein the average particle size of the adiponectin microspheres is 40 to 50 microns, and the adiponectin is encapsulated in the PLGA polymer through the porous pores of the PLGA polymer.

[0032] The provided adiponectin microspheres can be used as bone adhesive components, and the prepared bone adhesive has good biocompatibility and has the functions of promoting bone growth and fixing fractures.

[0033] According to a specific embodiment, the amount of adiponectin contained in the adiponectin microspheres is at least 10%, and according to a preferred embodiment, it is at least 20%. According to a specific embodiment, the encapsulation rate of adiponectin is at least 50%, for example, about 65%.

[0034] According to an embodiment of the present invention, the particle size distribution span of the adiponectin microspheres is 1 to 3 microns.

[0035] The adiponectin microspheres can be prepared by the following method, including:

[0036] After the PLGA polymer and adiponectin are mixed, self-healing packaging is performed at a predetermined temperature to obtain the adiponectin microspheres.

[0037] During the research, it was found that adiponectin microspheres can be prepared by self-healing with PLGA polymer as a carrier. It was found that the adiponectin self-healing microspheres have ideal encapsulation rate and particle size distribution. Moreover, PLGA and adiponectin can self-heal at room temperature, and the method is simple and easy.

[0038] According to a specific embodiment, the mass ratio of the PLGA to the adiponectin microspheres is 1:(300-500). According to a preferred embodiment, the mass ratio of the PLGA to the adiponectin microspheres is 1:(350-450), for example, 1:400.

[0039] The prepared adiponectin microspheres can be mixed with a hydrogel material to obtain an adiponectin hydrogel mixture, and the mixture can be cured by photocuring to obtain an adiponectin hydrogel.

[0040] Hydrogel material is a kind of skin external dosage form, is a three-dimensional cross-linked network composed of hydrophilic polymers, has a high water content and a structure similar to the extracellular matrix, and is sensitive to the physiological environment, hydrophilic, flexible, etc., and can be used as a carrier material for external administration. According to a specific embodiment, the hydrogel material mentioned includes but is not limited to MeGc (methacrylate glycol chitosan), polyvinyl alcohol hydrogel, hyaluronic acid hydrogel, carbomer hydrogel, etc. According to a preferred embodiment, the hydrogel material is MeGc. Covalently cross-linked hydrogels can be obtained by light curing, and the prepared hydrogel structure is uniform.

[0041] According to a specific embodiment, the adiponectin hydrogel has at least one of the following properties:

[0042] (a) It can solidify within 80 seconds under blue light irradiation without forming crystals or precipitation;

[0043] (b) Promote BMSCs proliferation;

[0044] (c) Promote osteogenic differentiation.

[0045] The adiponectin microspheres or the adiponectin hydrogel mixture or the adiponectin hydrogel described above can be used in the field of bone adhesives to treat fractures, especially comminuted fractures. They have many advantages, including: good biocompatibility; the ability to promote bone growth and fix fractures; rapid osteogenesis and fixation of bone fragments; and the implant material can be gradually degraded.

[0046] The experimental scheme of the present invention is described below by specific examples, which are only used to facilitate the understanding of those skilled in the art and should not be regarded as limiting the scope of protection of the present invention. It should be noted that the reagents used in the examples can be obtained by commercial purchase unless otherwise specified.

[0047] Example First, adiponectin microspheres were prepared and biomechanical properties and biocompatibility were analyzed. Then, adiponectin hydrogels were prepared and their cell proliferation activity, osteogenic differentiation, intracellular distribution and toxicity were tested.

[0048] Example 1

[0049] Example 1 successfully prepared adiponectin microspheres, comprising the following steps:

[0050] First, prepare a PLGA porous microsphere suspension (dissolve about 320 mg of dried PLGA porous microspheres in water) and add it to a centrifuge tube. Remove the supernatant after centrifugation. Then add 4 ml of a 0.2 mg / ml adiponectin aqueous solution to the centrifuge tube, place it in a 4°C shaker, and incubate it for 12 hours. Then transfer it to a water bath shaker and incubate it for 18 hours at 37°C and 100 rpm to prepare adiponectin microspheres.

[0051] Observation under electron microscope showed that the pores on the surface of PLGA microspheres were significantly reduced, and the prepared adiponectin microspheres healed well. Figure 1 shown. Figure 1 The microspheres before healing refer to the PLGA porous microsphere material itself, and the microspheres after healing refer to the prepared adiponectin microspheres.

[0052] Then the drug loading and encapsulation efficiency of the prepared adiponectin microspheres were analyzed. The prepared adiponectin microspheres were centrifuged at 4000g for 2 minutes in a high-speed centrifuge, and the supernatant was taken and the amount of free adiponectin was measured at 562nm using a micro BCA protein kit.

[0053] Drug loading % = amount of adiponectin loaded in microspheres / weight of microspheres after freeze-drying * 100%

[0054] Encapsulation rate % = adiponectin loading / total adiponectin dosage * 100%

[0055] The experimental results showed that the drug loading and encapsulation efficiency of adiponectin microspheres were 12% and 57.8%, respectively.

[0056] Example 2

[0057] Example 2 The particle size of the adiponectin microspheres prepared in Example 1 was investigated using a dry micron particle size analyzer, and the particle size was determined using a laser diffraction method. Parameter settings: the instrument dispersion pressure was 3 bar, and the injection concentration was >2%. The D10, D50, D90 and Span values ​​of the microspheres were recorded to determine the particle size and distribution. The span of the size distribution of the microspheres was calculated by the following method:

[0058] Span = (D90-D10) / D50 (Span is the dispersion coefficient, D10, D50, and D90 are the microsphere diameters measured at 10%, 50%, and 90% of the particle size, respectively)

[0059] The average particle size of adiponectin microspheres measured experimentally was 44.51 μm, and the particle size distribution span was 1.28 μm.

[0060] Then the pH and temperature stability of the prepared adiponectin microspheres were investigated. The experiment investigated the stability of adiponectin microspheres at different pH and temperatures, and circular dichroism (CD) was used to detect the secondary structure of adiponectin under different temperature and pH conditions.

[0061] The results show Figure 2 As shown in the figure, the experimental results show that adiponectin has good stability at pH=3 and pH=5; it has good stability at 37°C, and when the temperature is greater than 45°C, the secondary structure of adiponectin will be destroyed. Therefore, as mentioned in Example 1, the self-healing condition of 37°C will not affect the secondary structure of adiponectin in adiponectin microspheres.

[0062] Example 3

[0063] Example 3 Adiponectin hydrogel was prepared by the following method, comprising:

[0064] The MeGc hydrogel material was added to the ep tube, and then the adiponectin hydrogel and initiator were added, and PBS was added. After thorough vortex mixing, the bubbles were removed by simple centrifugation to obtain the adiponectin hydrogel mixture. The adiponectin hydrogel mixture was then added to the bone defect and irradiated with a blue light curing lamp (Careet LY-A180) for 80 seconds for curing.

[0065] The mass ratio of adiponectin hydrogel and MeGC hydrogel material is 1:3, and the initiator used is riboflavin.

[0066] Example 4

[0067] Example 4 The biocompatibility of the adiponectin hydrogel prepared in Example 3 was tested. The adiponectin hydrogel was solidified on the bottom of a 96-well plate according to the above method, and a monolayer of adherent MC3T3E1 cells in the logarithmic growth phase was taken and incubated at 5×10 3 The cells were inoculated at a density of 1 / well in a 96-well plate, with 200 μL per well. After culturing for 1, 4, and 7 days, 10 μL of CCK-8 solution was added to each well. The cells were placed in a cell culture incubator and cultured for 2 h. The absorbance was measured with an enzyme-labeled instrument (detection wavelength: 450 nm, reference wavelength: 650 nm).

[0068] The results are as follows Figure 3 The results showed that compared with the control group, both adiponectin and adiponectin hydrogel had no toxicity to MC3T3E1 cells and promoted cell growth.

[0069] At the same time, the safety of adiponectin hydrogel implantation in animals was evaluated. Under sterile conditions, the muscles of the left femur of rats were separated to form a muscle cavity, and the gel was implanted as the experimental group; the right side was used as a blank control group. The animals' eating, drinking, and activities within one week after implantation, as well as the incision reaction and healing were observed. No abnormalities or toxicity were found.

[0070] An ideal bone adhesive can fix fractures, especially those in weight-bearing areas, to achieve stability and replace the role of traditional internal fixation. This is the fundamental property of adiponectin hydrogel for fracture treatment. The material mechanics of the prepared adiponectin hydrogel was investigated below, and the in vivo and in vitro mechanics of the prepared adiponectin hydrogel after fracture in mice were studied.

[0071] Example 5

[0072] First, the material mechanical properties of the adiponectin hydrogel prepared in Example 3 were investigated.

[0073] Under the clean bench, dispense the uncured hydrogel into different ep tubes, add different concentrations of adiponectin microspheres and initiators, fill the volume with PBS, and simply centrifuge to remove bubbles. The gel will not solidify and aggregate into small lumps. Take a suitable volume and add it to the bone defect, and cure it after 80 seconds of irradiation with a blue light curing lamp.

[0074] Because the gel is easy to solidify, there is no need to consider cellular uptake in the treatment of fractures. Therefore, it is placed at the bottom of the cell culture dish and co-cultured with the cells during cell experiments, so there is no need for special cellular uptake studies.

[0075] The biomechanical properties of adiponectin hydrogel were then characterized.

[0076] The experimental mice were divided into control, adiponectin (ADPN), and adiponectin hydrogel (ADPN-Gel) groups. Catwalk tests were performed 4 weeks after tibial fracture surgery. The mice were then killed, the complete tibia was obtained, and the muscles, fascia and other soft tissues attached to the tibia were cleaned. The tibia specimens fixed with 4% paraformaldehyde for 3 days were rinsed with clean water for 15 minutes, and then placed in a pre-prepared neutral 10% disodium EDTA decalcification solution, placed on an oscillator for decalcification, and the solution was changed every other day. After 6 weeks of decalcification, HE and MS staining were performed. After Micro-CT examination of the other side of the tibia, a three-point bending test was performed to record the maximum mechanical strength when the bone was broken.

[0077] The results are as follows Figure 4 The bone healing of adiponectin and adiponectin hydrogel groups was significantly earlier, the thickness of the bone cortex was better, the bone microstructure of the bone cortex was better, the Catwalk sole contact area was larger, and the three-point bending test showed that its strength and toughness were better.

[0078] Through the above research, it was found that the mechanical advantages of adiponectin hydrogel are: adiponectin hydrogel can solidify within 80 seconds under blue light irradiation; it will not form crystals and is evenly distributed, and will not solidify and aggregate into small precipitates in the gel; there are no crystalline particles, so there is no need to perform intracellular distribution detection; and it can form gel quickly without affecting the release of adiponectin protein.

[0079] The cell proliferation activity, osteogenic differentiation effect and mechanism of action of adiponectin hydrogel were explored below.

[0080] Example 6

[0081] Example 6 first studied the effect of adiponectin hydrogel on the proliferation activity of BMSCs.

[0082] 200 μL of BMSCs suspension was added at a rate of 2×10 3 The cells were inoculated into a 96-well plate at a concentration of 100 μg / well and cultured in a constant temperature cell culture incubator for 1, 4, and 7 days. After that, 20 μL of 5 mg / mL MTT solution was added to each well under light-proof conditions. The cells were incubated in a constant temperature cell culture incubator for 4 hours. 150 μL of DMSO solution was added and the absorbance of each well at a wavelength of 490 nm was measured using an automatic microplate reader. The results are shown in Figure 2. Figure 5 As shown, the results showed that adiponectin and adiponectin hydrogel had a significant effect in promoting the proliferation of BMSCs.

[0083] Then the effect of adiponectin hydrogel on the osteogenic differentiation of BMSCs was investigated.

[0084] BMSCs of passage 3 were taken and 5×10 were added to each well. 3The cells were inoculated in a gelatin-coated 12-well plate (control, adiponectin, adiponectin self-healing microsphere hydrogel) at a density of 10 cells, and osteogenic induction medium (containing sodium β-glycerophosphate, vitamin C and dexamethasone) was added. After 11 days of induction, 1 mL of 4% paraformaldehyde was used for fixation for 15 minutes, and the sample was added according to the instructions of the alkaline phosphatase activity detection kit. The OD value at 405 nm was measured using a microplate reader, and the number of DEA enzyme activity units of ALP was calculated. After 21 days of induction, after alizarin red staining, obvious red-stained round calcium nodules could be seen.

[0085] Real-time PCR detection was performed at the same time: after 11 days of induction, the culture medium of each group was discarded, and the total RNA of each group of cells was extracted according to the instructions of Cell Total RNA Isolation Kit. The ratio of A260nm / A280nm and RNA concentration were measured to determine its purity. TM RT reagent Kit with gDNA Eraser Description Reverse transcription to synthesize cDNA, TB Premix Ex Taq TM II kit was used to prepare 20 μL reaction system, and the PCR reaction was completed in the qRT-PCR instrument. The osteogenesis-related genes detected included ABMP2, osteoprotegerin (OPG), and Runx-related transcription factor 2 (Runx2).

[0086] The results are as follows Figure 6 As shown, the results showed that compared with the control group, the osteogenic effect of adiponectin and adiponectin hydrogel was significantly stronger, the ALP activity was increased, and the expression of osteogenic-related genes was significantly enhanced.

[0087] Based on the above results, it can be seen that adiponectin can promote the proliferation of BMSCs and their osteogenic differentiation, and the promoting effect of adiponectin hydrogel is more obvious than that of adiponectin alone.

[0088] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "specific implementation", or "some implementations" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0089] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. Adiponectin microsphere, characterized in that: The invention comprises adiponectin and PLGA polymer. The average particle size of the adiponectin microsphere is 40-50 microns. The adiponectin is wrapped into the PLGA polymer through the porous pores of the PLGA polymer.

2. The adiponectin microspheres according to claim 1, characterized in that The drug loading amount of the adiponectin in the adiponectin microsphere is at least 10%, and the encapsulation rate of the adiponectin is at least 50%.

3. The adiponectin microspheres according to claim 1, characterized in that The particle size distribution span of the adiponectin microspheres is 1 to 3 microns.

4. A method for preparing adiponectin microspheres, characterized in that: include: After the PLAG polymer and adiponectin are mixed, self-healing packaging is performed at a predetermined temperature to obtain the adiponectin microspheres.

5. The method according to claim 4, characterized in that The mass ratio of the PLGA to the adiponectin microspheres is 1:(300-500).

6. The method according to claim 4, characterized in that The predetermined temperature is 20 to 37 degrees Celsius.

7. An adiponectin hydrogel mixture, characterized in that: The adiponectin hydrogel mixture is obtained by mixing the adiponectin microspheres according to any one of claims 1 to 3 with a hydrogel material; Optionally, the hydrogel material is MeGc.

8. An adiponectin hydrogel, characterized in that: The method is obtained by photocuring the adiponectin hydrogel mixture according to claim 7.

9. Use of the adiponectin microspheres according to any one of claims 1 to 3, the adiponectin hydrogel mixture according to claim 7, or the adiponectin hydrogel according to claim 8 or 9 in the preparation of a drug for treating bone fractures.

10. Use of the adiponectin microspheres according to any one of claims 1 to 3, the adiponectin hydrogel mixture according to claim 7, or the adiponectin hydrogel according to claim 8 or 9 in the field of bone adhesives.

Images