Preparation method of plastic bone grafting material and plastic bone grafting material

Abstract

The invention relates to a preparation method of a plastic bone grafting material and the plastic bone grafting material.The preparation method comprises the following steps that a, phenyl phosphate lithium salt is dissolved in a phosphoric acid buffer salt solution to obtain a mixed solution, and Ge MA is dissolved in the mixed solution to obtain a Ge MA gel solution; b, bone meal and the Ge MA gel solution are subjected to a cross-linking effect, so that the bone meal and the Ge MA gel solution are fully mixed, then light source irradiation is conducted, the bone meal hydrogel bone grafting material is obtained, and the particle size of the bone meal is 0.25-0.5 mm. According to the preparation method of the plastic bone grafting material, provided by the invention, the bone meal and the Ge MA gel solution are subjected to cross-linking and then subjected to light source irradiation, so that the bone meal and the hydrogel are uniformly mixed, the compatibility of an organic phase and a bone meal particle interface is good, and the prepared plastic bone grafting material is relatively high in plasticity and good in osteoinductivity; and the cell adhesion rate is high.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to the technical field of tissue engineering, and more particularly, the present disclosure relates to a preparation method of a plastic bone graft material and the plastic bone graft material. BACKGROUND

[0002] The self-repairing ability of bone defects is usually weak, which brings considerable inconvenience and pain to patients. In clinical treatment, repairing bone defects is a quite challenging problem. Bone powder is a common material for repairing bone defects, but the plasticity of bone powder itself is poor, and the degradation rate in the body is slow, so bone powder is usually combined with other biological materials, such as hydrogel, to improve its mechanical properties and biocompatibility.

[0003] However, the prior art is mostly limited to simple loading of hydrogel and granular bone, and there are problems such as poor interface compatibility between the organic phase and the bone powder particles, and uneven distribution of the bone powder particles in the hydrogel due to the effect of gravity sedimentation, resulting in insufficient bone induction and poor cell adhesion rate. SUMMARY

[0004] Therefore, the present disclosure provides a preparation method of a plastic bone graft material and the plastic bone graft material to solve the technical defects in the prior art.

[0005] To achieve the above-mentioned purpose, the present disclosure adopts the following technical solutions:

[0006] The present disclosure provides a preparation method of a plastic bone graft material, comprising the following steps:

[0007] Step a: Dissolve a lithium phenylphosphate salt in a phosphate buffered saline solution to obtain a mixed solution, and dissolve a Ge l MA in the mixed solution to obtain a Ge l MA gel solution;

[0008] Step b: Cross-link the bone powder and the Ge l MA gel solution to fully mix the bone powder and the Ge l MA gel solution, and then irradiate with a light source to obtain a bone powder hydrogel bone graft material, wherein the particle size of the bone powder is 0.25 mm to 0.5 mm.

[0009] In an embodiment of the present disclosure, the mass of the lithium phenylphosphate salt is 0.05 g, the volume of the phosphate buffered saline solution is 20 ml, and the concentration of the mixed solution is 0.25% w / v.

[0010] In an embodiment of the present disclosure, the concentration of the Ge l MA gel solution is 5% w / v.

[0011] In an embodiment of the present disclosure, the mass ratio of the Ge l MA to the bone powder is 1:1 to 1:4.

[0012] In one embodiment of the present application, in step b, the temperature for cross-linking the bone powder with the Gel MA gel solution is 4°C, and the cross-linking time is 1 min.

[0013] In one embodiment of the present application, in step b, the light source for irradiation is a 405 nm light source, and the irradiation time is 10-30 s.

[0014] In one embodiment of the present application, step b further comprises:

[0015] The bone powder is cross-linked with the Gel MA gel solution, the bone powder and the Gel MA gel solution are fully mixed to obtain a bone powder hydrogel mixed colloid, the bone powder hydrogel mixed colloid is injected into a mold, and then irradiated with a light source to obtain a bone powder hydrogel bone graft material.

[0016] In one embodiment of the present application, the injection temperature for injecting the bone powder hydrogel mixed colloid into the mold is 37°C.

[0017] In one embodiment of the present application, step a further comprises sterilizing the Gel MA gel solution.

[0018] In one embodiment of the present application, the plastic bone graft material is prepared by the method for preparing a plastic bone graft material as described above.

[0019] The method for preparing a plastic bone graft material provided by the present application first cross-links bone powder with a Gel MA gel solution, and then irradiates with a light source, so that the bone powder and the hydrogel are uniformly mixed, the compatibility of the organic phase and the bone powder particle interface is good, the plastic bone graft material prepared has strong plasticity, good osteoinductivity, and good cell adhesion rate.

[0020] Other features and advantages of the present application will become apparent from the following detailed description of illustrative embodiments thereof, which proceeds with reference to the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0021] The accompanying drawings incorporated in and forming a part of the specification illustrate embodiments of the present application and, together with the description, serve to explain the principles of the application.

[0022] Figure 1 is a process flow chart of the method for preparing a plastic bone graft material provided by one embodiment of the present application;

[0023] Figure 2is a morphology comparison chart of bone powder mixed with Gel MA hydrogel solution without cross-linking and with cross-linking provided by an embodiment of the present disclosure;

[0024] Figure 3 is a schematic diagram of bone powder hydrogel bone grafting material with different concentrations provided by an embodiment of the present disclosure;

[0025] Figure 4 is a schematic diagram of bone powder hydrogel bone grafting material with different shapes provided by an embodiment of the present disclosure;

[0026] Figure 5 is a schematic diagram of observation of adhesion of MSCs on the surface of Gel MA hydrogel and bone powder wrapped by hydrogel provided by an embodiment of the present disclosure;

[0027] Figure 6 is a schematic diagram of evaluation of repair of rabbit skull defects provided by an embodiment of the present disclosure. DETAILED DESCRIPTION

[0028] In order to make the purposes, technical schemes and beneficial technical effects of the present application clearer, the present application will be described in detail below in combination with specific embodiments. It should be understood that the embodiments described in the specification are only for the purpose of explaining the present application, and are not intended to limit the present application.

[0029] For the sake of simplicity, only some numerical ranges are explicitly disclosed herein. However, any lower limit can be combined with any upper limit to form a range not explicitly recited; and any lower limit can be combined with any other lower limit to form a range not explicitly recited, as can any upper limit with any other upper limit to form a range not explicitly recited. Furthermore, although a range of endpoints is recited, each point or individual number within the range is also included in the range. Thus, each point or individual number can be combined with any other point or individual number to form a range not explicitly recited.

[0030] In the description of the present application, it should be noted that, unless otherwise specified, "above", "below" include the number itself, and "several" means two or more.

[0031] The above summary of the application is not intended to describe each disclosed embodiment or implementation of the present application. The following description illustrates the exemplary embodiments in more detail. In many places throughout the application, guidance is provided by a series of embodiments, which can be used in various combinations. In each instance, the list is merely representative of a group, and should not be interpreted as exhaustive.

[0032] The disclosure provides a preparation method of a plastic bone grafting material, comprising the following steps: step a: dissolving a lithium phenylphosphate salt in a phosphate buffered saline solution to obtain a mixed solution, and dissolving a Ge l MA in the mixed solution to obtain a Ge l MA gel solution; and step b: cross-linking the bone powder with the Ge l MA gel solution, fully mixing the bone powder with the Ge l MA gel solution, and then irradiating the bone powder with a light source to obtain a bone powder hydrogel bone grafting material, wherein the particle size of the bone powder is 0.25-0.5 mm.

[0033] The preparation method of the plastic bone grafting material provided by the disclosure first cross-links the bone powder with the Ge l MA gel solution and then irradiates the bone powder with a light source, so that the bone powder and the hydrogel are uniformly mixed, the compatibility of the organic phase and the bone powder particle interface is good, the prepared bone grafting material has strong plasticity, good osteoinduction, and good cell adhesion rate.

[0034] As shown in Figure 1 The preparation method of the plastic bone grafting material provided by the disclosure comprises the following steps:

[0035] Step a: dissolving a lithium phenylphosphate salt in a phosphate buffered saline solution to obtain a mixed solution, and dissolving a Ge l MA in the mixed solution to obtain a Ge l MA gel solution.

[0036] Specifically, as shown in Figures 2-3 The lithium phenylphosphate salt is dissolved in the phosphate buffered saline solution to obtain a mixed solution, wherein the lithium phenylphosphate salt is a photoinitiator with good water solubility and biocompatibility, and a lithium phenyl(2,4,6-trimethylbenzoyl) phosphate salt is specifically selected as the photoinitiator in the embodiment. After being irradiated by ultraviolet light, the lithium phenyl(2,4,6-trimethylbenzoyl) phosphate salt can absorb the energy of light, split into two active free radicals, initiate the chain polymerization of the photosensitive resin and the active diluent, cross-link and cure the adhesive, and the phosphate buffered saline solution can adjust the pH value of the solution and maintain the stability of the solution.

[0037] GelMA is dissolved in a mixed solution to obtain a GelMA gel solution, wherein GelMA is methacrylated gelatin. GelMA is prepared from methacrylic anhydride (MA) and gelatin and is a photosensitive bio-hydrogel material. GelMA is biocompatible and can be excited by ultraviolet or visible light to form a three-dimensional structure with a certain strength suitable for cell growth and differentiation. In GelMA, the cross-linking reaction is photoinduced in the photo-initiated free radical polymerization reaction. When exposed to a specific wavelength, the photoinitiator splits into two free radicals, which immediately activates the cross-linking reaction. The free radicals are unstable and contain unpaired electrons, which can gain electrons from MA (the double bond of GelMA), converting GelMA into a new free radical called a monomer free radical. This free radical can donate electrons to another MA, leading to a chain growth reaction. It can also combine with another monomer free radical in the coupling reaction, terminating the reaction and ultimately forming a polymer molecular network.

[0038] Step b: Cross-link the bone powder with the Ge l MA gel solution to ensure thorough mixing, and then irradiate with a light source to obtain a bone powder hydrogel bone graft material, wherein the particle size of the bone powder is 0.25 mm to 0.5 mm.

[0039] Specifically, bone powder is cross-linked with GelMA gel solution. In this embodiment, the bone powder used is Geri Natural Calcined Bone Repair Material / Bone Powder, model CBB-G1-C, with a particle size of 0.25 mm to 0.5 mm. Due to the relatively large particle size of the bone powder, gravity sedimentation occurs after mixing with GelMA gel solution, resulting in uneven distribution of the bone powder in the GelMA gel solution.

[0040] like Figure 2 As shown in (a), when bone powder is mixed with GelMA hydrogel solution and directly crosslinked by light, the bone powder will separate into layers under gravity, making it impossible for the GelMA hydrogel solution to encapsulate the bone powder, resulting in uneven distribution of the bone powder in the GelMA hydrogel solution; as Figure 2 As shown in (b), crosslinking at 4°C for 1 min allows the GelMA hydrogel solution to undergo reversible gelation before being uniformly mixed with bone powder. Subsequent photocrosslinking enables uniform distribution of bone powder within the GelMA hydrogel solution. Therefore, first mixing bone powder with the GelMA gel solution for crosslinking ensures thorough and uniform mixing. Further irradiation after mixing reinforces the crosslinking process, yielding a bone powder hydrogel bone graft material.

[0041] In one embodiment of the present disclosure, the mass of the lithium phenylphosphate salt is 0.05 g, the volume of the phosphate buffered saline solution is 20 ml, and the concentration of the mixed solution is 0.25% w / v.

[0042] It is considered that the concentration of the GeIMA gel can affect the physical properties of the constructed hydrogel network, such as mechanical strength, porosity, degradation rate, etc. Different concentrations of GeIMA gel solution are suitable for different tissue engineering needs, can simulate the mechanical properties of different tissues, control the release rate of drugs, or adjust the behavior and differentiation of cells in a three-dimensional culture environment. Therefore, in one embodiment of the present disclosure, the concentration of the GeIMA gel solution is 5% w / v to meet the requirements of the prepared plastic bone graft material for mechanical strength, porosity, and degradation rate, etc. In this embodiment, the concentration of the GeIMA gel solution can also be adjusted according to the speed of bone formation, which is adjustable, so that the concentration of the GeIMA gel solution matches the degradation rate of the bone graft material.

[0043] It is considered that when the proportion of bone powder is too high, the degradation rate of the entire composite material will be significantly reduced, which will delay the complete replacement process of new bone tissue and affect the repair effect. And excessive bone powder will increase the brittleness of the material, reduce its elasticity and toughness, affect the stability and bearing capacity of the material in the early stage of implantation, and cannot provide sufficient mechanical support, which is not conducive to the initial healing and reconstruction of bone tissue. If the proportion of bone powder is too small, the material will be too weak to be shaped or maintain the predetermined structure. Therefore, the proportion of GeIMA and bone powder needs to be adjusted to an appropriate range.

[0044] In one embodiment of the present disclosure, the mass ratio of GeIMA to bone powder is 1:1 to 1:4. As shown in Figure 3 different mass ratios (1:1, 1:2, 1:3, 1:4) of GeIMA and bone powder are mixed, the hydrogel can completely wrap the bone powder, and the hardness of the plastic bone graft material increases with the increase of the mass of the bone powder. In addition, the degradation rate of GeIMA can be adjusted by changing its cross-linking density, and after adding bone powder, the degradation rate of the entire composite material can be effectively slowed down due to the slow degradation rate of the bone powder itself. By adjusting the proportion of GeIMA and bone powder, the degradation rate of the material can be controlled to match the speed of tissue repair or regeneration while maintaining the stability of the structure, avoiding the adverse effects caused by too fast or too slow degradation. Controlling the mass ratio of GeIMA and bone powder in the range of 1:1 to 1:4 can ensure that the final product has good forming performance and faster degradation rate, and can also improve the surface properties of the material, promote cell adhesion and proliferation.

[0045] In one embodiment of the present disclosure, in step b, the temperature for cross-linking the bone powder with the Ge l MA gel solution is 4°C, and the cross-linking time is 1 min.

[0046] Specifically, the Ge l MA gel solution is a kind of temperature-sensitive bio-hydrogel material. The cross-linking reaction of Ge l MA usually requires photo-initiation or chemical initiator. In the case of photo-initiation, although the main driving force is light, proper temperature control is still crucial. Within a certain temperature range, the Ge l MA solution will transform from a liquid state to a gel state with viscoelasticity. This transition point, i.e., the lower critical solution temperature (LCST), can be fine-tuned by adjusting the concentration and cross-linking degree of Ge l MA. Maintaining the temperature at a level slightly higher than the LCST before or during mixing with the bone powder can ensure that the Ge l MA rapidly and uniformly transforms into a gel state, thereby better wrapping and fixing the bone powder particles and forming a stable composite structure.

[0047] At a suitable temperature, the viscosity of the Ge l MA gel increases, which helps to evenly distribute the bone powder particles in the gel network and reduce aggregation. Uniformly distributed bone powder not only improves the mechanical properties of the composite material but also ensures uniform transmission of biological signals (such as osteoinduction) within the material, which is beneficial for cell growth and differentiation.

[0048] In addition, the cross-linking time is closely related to the temperature. Under suitable temperature conditions, the cross-linking reaction rate increases, which can shorten the molding time and improve the production efficiency. However, too high a temperature will affect the biocompatibility and cell affinity of the material. Therefore, it is crucial to choose a temperature range that can ensure rapid and effective cross-linking without compromising the biological performance of the material. Therefore, when the bone powder is cross-linked with the Ge l MA gel solution, the temperature is controlled at 4°C, and the cross-linking time is 1 min. At this temperature, the Ge l MA gel solution exhibits a viscous gel state, which can better mix with the bone powder.

[0049] In one embodiment of the present disclosure, in step b, the light source for irradiation is a 405 nm light source, and the irradiation time is 10-30 s.

[0050] Specifically, photocrosslinking typically relies on light of a specific wavelength to activate the photoinitiator. Therefore, the light source must emit a wavelength that matches the absorption spectrum of the photoinitiator used to ensure an effective photoinitiation reaction. The intensity of the light source determines the speed of the photoinitiation reaction; a high-intensity light source can accelerate the curing process and improve production efficiency. Simultaneously, the uniformity of the light source is crucial for ensuring uniform curing of all parts of the material and avoiding localized over-curing or under-curing. The irradiation time directly affects the degree of curing of the material. Too short an irradiation time may lead to incomplete curing and poor mechanical properties of the material; too long an irradiation time may cause over-curing, affecting the material's flexibility or generating stress. By adjusting the irradiation time, the crosslinking density of the material can be controlled to a certain extent, thereby affecting the material's hardness, durability, and other physical properties. Reasonable control of the irradiation time can balance production efficiency and product quality; too long a curing time will reduce the production rate, while too short a time may require rework of defective products. Therefore, in this embodiment, a 405nm light source is selected, and the irradiation time is controlled within the range of 10s to 30s to ensure both the degree of curing and production efficiency.

[0051] In one embodiment of this disclosure, step b further includes: cross-linking bone powder with Ge 1 MA gel solution to obtain a bone powder hydrogel mixture after the bone powder and Ge 1 MA gel solution are fully mixed; injecting the bone powder hydrogel mixture into a mold; and then irradiating it with a light source to obtain a bone powder hydrogel bone graft material.

[0052] Since different shapes of bone graft materials are required under different circumstances, the bone powder hydrogel mixture is injected into a mold. Taking advantage of its plasticity, it is solidified by the mold and then irradiated with a light source to finally obtain bone powder hydrogel bone graft materials of different shapes.

[0053] like Figure 4 As shown, in the material prepared by mixing bone powder and Ge l MA hydrogel at a ratio of 1:4, the bone powder is uniformly distributed inside the material and is completely encapsulated by Ge l MA hydrogel; after photocrosslinking, it can be prepared into different shapes and is not easily deformed, exhibiting good plasticity and compressive strength.

[0054] Considering the temperature-sensitive nature of Ge l MA itself, in one embodiment of this disclosure, the injection temperature for injecting the bone meal hydrogel mixture into the mold is 37°C.

[0055] To reduce the risk of viral infection in medical bio-derived tissue repair materials and ensure the safety of the materials, in one embodiment of this disclosure, step a further includes: sterilizing the Ge l MA gel solution.

[0056] This disclosure also provides a malleable bone graft material, which is prepared by the method described above. The appropriate surface properties and porosity of the malleable bone graft material provided in this embodiment can simulate the composition, structure, and properties of biological bone tissue, and the specific pore size and porosity also facilitate material exchange. Finally, it also possesses osteoinductive and osteoconductive properties, inducing adjacent mesenchymal cells to differentiate into osteoblasts or facilitating the migration of adjacent bone tissue, thus promoting bone regeneration.

[0057] In bone defect repair engineering, cells come into contact with bone graft particles, endocytose the particles, and degrade them in lysosomes within the cells, thus completing the entire bone grafting process. Therefore, it is necessary to observe the cell adhesion of the bone graft material. In this embodiment, cell adhesion is observed using scanning electron microscopy. The specific operating steps are as follows:

[0058] (1) Prepare a sterile mixture of Ge l MA and bone powder in a ratio of 1:4 according to the above method, add 100 μL to each well of a 24-well plate, and irradiate with a 405 nm light source for 10-30 s to enhance gelation.

[0059] (2) Then, BMSCs (bone marrow mesenchymal stem cells) are cultured until the confluence reaches about 80%, digested and centrifuged, and seeded with cells 10 4 Individual cells / well, incubated overnight;

[0060] (3) Fix with 2.5% glutaraldehyde overnight at 4°C. Dehydrate with gradient ethanol (50%, 70%, 85% and 95%) for 15 min each, dehydrate with anhydrous ethanol 3 times for 15 min each time, and then dry with supercritical fluid;

[0061] (4) Sputter gold onto the sample for 130 seconds using an ion sputtering instrument (current 10mA, voltage 150V), and observe cell adhesion using a scanning electron microscope.

[0062] like Figure 5 As shown, when BMSCs (bone marrow mesenchymal stem cells) are seeded on the surface of a malleable bone graft material, the cells can adhere to the surface of the malleable bone graft material, which is conducive to cell proliferation.

[0063] The following section provides a further explanation of this embodiment using specific experiments.

[0064] To further evaluate the bone regeneration capacity of the malleable bone graft material in vivo, five male New Zealand white rabbits (average age 12 months) were selected as a rabbit skull defect model. The specific procedures are as follows:

[0065] New Zealand white rabbits were anesthetized by intramuscular injection of levofloxacin according to the standard dosage of 0.2 mL / kg. The skull was prepared and disinfected to expose the skull, and the periosteum was removed. Circular skull defects with a diameter of 10 mm were created on both sides of the top of the rabbit skull using an electric drill. Bone powder and malleable bone graft material were transplanted to the defects on the left and right sides of the rabbit skull, respectively, with 3 rabbits per group. The mass ratio of bone powder to GelMA in the malleable bone graft material was 1:4. Simultaneously, bone powder was transplanted to the right side of the rabbit skull, while the left side served as a control, with 2 rabbits per group. The material surface was made flush with the skull surface, the wound was sutured, and disinfected. The rabbits were injected with penicillin for 5 consecutive days postoperatively to prevent wound infection.

[0066] After constructing the rabbit skull defect model, Micro-CT and tissue section staining analysis were performed.

[0067] Six and twelve weeks after implantation of the material into the bone defect, rabbits were euthanized using the air embolization method, and skull samples were collected and fixed in 4% paraformaldehyde for approximately 48 hours. First, the samples were replaced with physiological saline three times. After simple cleaning of surrounding muscles and other tissues, the samples were scanned using Micro-CT (microcomputed tomography). Global threshold conversion was used to distinguish between the implanted material and newly formed bone tissue, and the scan data were reconstructed into three-dimensional images using analysis software. Then, the samples were fixed in 4% paraformaldehyde solution for another 7 days, decalcified with EDTA (ethylenediaminetetraacetic acid) for 2 months, dehydrated with graded alcohols, and embedded in paraffin. The embedded samples were then sectioned into approximately 7 μm thick sections using a microtome and stained with hematoxylin and eosin (HE) and Masson's trichrome to observe bone regeneration.

[0068] like Figure 6 As shown in (a), from left to right, the blank control group, the bone defect repair group treated with bone powder alone, and the repair group treated with malleable bone graft material are shown. After creating a 1 cm diameter defect in the rabbit skull, the rabbits were treated with bone powder and malleable bone graft material respectively. It was observed that the malleable bone graft material could completely adhere to the wound, while the bone powder particles were easily dispersed.

[0069] Postoperative observation via Micro-CT at 12 weeks, such as Figure 6 As shown in (b), yellow represents bone tissue and blue represents undegraded bone powder. It can be seen that, compared with the blank group and the bone powder group, the plastic bone graft material group has more bone tissue formation and less bone powder residue.

[0070] like Figure 6As shown in (c)-(d) in FIG. 6, more bone tissue was observed in the skull defect treated with the bone powder hydrogel bone grafting material by HE staining, and less undegraded bone powder was observed; the Masson staining results showed that more collagen was deposited in the skull defect treated with the plastic bone grafting material, so the material can significantly promote bone defect repair and bone tissue regeneration.

[0071] The preparation method of the plastic bone grafting material provided by the present disclosure first cross-links the bone powder and the gel MA gel solution, and then irradiates it with a light source, so that the bone powder and the hydrogel are uniformly mixed, the compatibility of the organic phase and the bone powder particle interface is good, the plastic bone grafting material prepared has strong plasticity, good osteoinduction, and good cell adhesion rate. The plastic bone grafting material is used in bone grafting engineering, begins to degrade in the body, provides support during the degradation process, acts as a scaffold for new bone formation, and is eventually replaced by the body bone after degradation. And the appropriate surface properties and porosity of the plastic bone grafting material can simulate the composition, structure and performance of biological bone tissue, and the specific pore size and porosity are also beneficial to material exchange. Finally, it also has bone induction and bone conduction properties, induces adjacent mesenchymal cells to differentiate into osteoblasts or facilitates adjacent bone tissue to crawl, which helps to promote bone regeneration.

[0072] The above has described various embodiments of the present disclosure, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes are obvious to those skilled in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles, practical applications, or technical improvements in the market of the embodiments, or to enable other ordinary skilled persons in the art to understand the embodiments disclosed herein. The scope of the present disclosure is defined by the appended claims.

Claims

1. A method for producing a plastic bone graft material, characterized by, The method comprises the following steps: Step a: dissolving a lithium phenylphosphate salt in a phosphate buffered saline solution to obtain a mixed solution, dissolving GelMA in the mixed solution to obtain a GelMA gel solution; Step b: cross-linking bone powder with the GelMA gel solution, fully mixing the bone powder and the GelMA gel solution, and then irradiating a light source to obtain a bone powder hydrogel bone graft material, wherein the particle size of the bone powder is 0.25-0.5 mm.

2. The method of producing a plastic bone graft material according to claim 1, characterized by, The mass of the lithium phenylphosphate salt is 0.05 g, the volume of the phosphate buffered saline solution is 20 ml, and the concentration of the mixed solution is 0.25% w / v.

3. The method of preparing a malleable bone graft material according to claim 1, wherein, The concentration of the GelMA gel solution is 5% w / v.

4. The method of preparing a malleable bone graft material according to claim 1, wherein, The mass ratio of the GelMA to the bone powder is 1:1-1:

4.

5. The method of preparing a malleable bone graft material according to claim 1, wherein, In step b, the cross-linking temperature of the bone powder with the GelMA gel solution is 4°C, and the cross-linking time is 1 min.

6. The method of preparing a malleable bone graft material of claim 1, wherein, In step b, the light source for irradiation is a 405 nm light source, and the irradiation time is 10-30 s.

7. The method of preparing a malleable bone graft material according to claim 1, wherein Step b further comprises: Cross-linking the bone powder with the GelMA gel solution, fully mixing the bone powder and the GelMA gel solution to obtain a bone powder hydrogel mixed colloid, injecting the bone powder hydrogel mixed colloid into a mold, and then irradiating a light source to obtain a bone powder hydrogel bone graft material.

8. The method of producing a plastic bone graft material according to claim 7, characterized by, The injection temperature of the bone powder hydrogel mixed colloid into the mold is 37°C.

9. The method of preparing a malleable bone graft material of claim 1, wherein, Step a further comprises sterilizing the GelMA gel solution.

10. A malleable bone grafting material, characterized in that, The plastic bone graft material is prepared by the method of any one of claims 1-9.

Images