CN109331222B - Bone repair material capable of forming 3D porous scaffold in situ and preparation and application thereof - Google Patents

Abstract

The invention discloses a bone repair material, which is prepared by taking calcium sulfate hemihydrate and bioactive ceramic materials as raw materials, uniformly mixing, granulating and then carrying out heat treatment. The bone repair material has injectability, is suitable for minimally invasive surgery, can form a 3D porous support with good mechanical strength in situ after injection, and has excellent performance in the aspects of promoting bone repair and regeneration.

Description

Bone repair material capable of forming 3D porous scaffold in situ and preparation and application thereof

Technical Field

The invention belongs to the technical field of medical materials, and particularly relates to a bone repair material capable of forming a 3D porous scaffold in situ, a preparation method thereof and application thereof in bone repair.

Background

Bone filler materials and bone active implants are currently the most commonly used materials for bone repair and regeneration in surgical procedures. There are approximately over 200 million surgical procedures worldwide each year requiring the use of bone filler material. Among all bone filling materials, autologous bone is most suitable for the bone regeneration conditions of patients, such as bone conduction, bone induction and osteogenesis, and thus is considered as an optimal material, but autologous bone comes from the patients themselves, the source is extremely limited, and bone extraction may increase additional pain to the patients. The use of xenogenic bone also accounts for a large market, such as bovine decalcified bone, but there is a great concern about the use of xenogenic bone due to the uncontrollable source and potential disease infection. Synthetic bone is increasingly popular with doctors and patients due to its lack of source limitation and the absence of potential disease-carrying factors, and its product variety and market are rapidly growing.

There is uncertainty about the topography of the bone defect site, and therefore it is a great challenge how to plasticize bone implants and filler materials. Usually, the bone implant is pre-shaped in vitro according to the wound, and then the wound site is opened and the material is implanted. The entire procedure is complex and the surgery is open. When the repair site is deep and surrounded by soft tissue and body fluids, the difficulty of the operation is high and the patient suffers great pain.

Minimally invasive techniques based on bone repair filling materials have been developed. At present, the bone repair filling material for minimally invasive surgery is generally bone cement which has good fluidity and can be implanted into a bone defect part through injection. However, the existing bone cement used as an implant material has the defects that the molding material is compact and has few pores after being implanted, and the bone regeneration and the blood vessel growth are not facilitated. From the viewpoint of being most suitable for bone repair and regeneration, a porous scaffold active material having a certain pore size (both autologous bone and allogeneic bone are porous materials) is suitable. However, since the bone defect site is usually large, the corresponding large-sized porous scaffold material cannot be implanted by injection. In contrast, researchers have attempted to add adjuvants to bone cement to create holes in situ, but the results have been poor.

Therefore, finding bone repair materials that can be injected and form 3D porous scaffolds is urgently needed in the clinic and market, and has been a great challenge in the field.

Disclosure of Invention

Aiming at the defects and the technical development requirements of the prior art, the invention mainly aims to provide a novel bone repair material which has injectability, is suitable for minimally invasive surgery, can form a 3D porous scaffold with good mechanical strength in situ after injection, and has excellent performance in the aspects of promoting bone repair and regeneration.

The invention also aims to provide a preparation method of the bone repair material and an application of the bone repair material in bone repair.

Still another object of the present invention is to provide the use of the above bone repair material in the preparation of a 3D porous scaffold, the prepared 3D porous scaffold, and the use of the 3D porous scaffold in bone repair.

Compared with the prior art, the invention can obtain excellent technical effects: (1) the bone repair material can be directly injected (namely no liquid medium is added), and a 3D porous scaffold can be formed in situ at a bone defect part after injection; (2) under the matching of the model, the bone repair material can be made into a 3D porous bracket with any shape in advance in vitro so as to adapt to the clinical application requirements under various conditions; (3) the material degradation rate can be regulated and controlled by controlling the material composition and/or the morphology structure, so that the requirement on the degradation rate under different application conditions is met, and the growth of new bones is matched; (4) the preparation process of the bone repair material is simple, and the streamlined production can be carried out by adopting an engineering technology.

Detailed Description

Various aspects of the present invention will be described in detail below, with the above objects in mind.

Unless otherwise specified, the abbreviations for some of the compounds used herein are shown in table 1 below.

TABLE 1

Bioactive ceramics	For short	Bioactive ceramics	For short
				CaSO4·0.5H2O	hCS	Ca10(PO4)6(OH)2	HA
MgO-SiO2	MS	Ca9(HPO4)(PO4)5(OH)	CDHA
				α-Ca3(PO4)2	α-TCP	3CaO·SiO2/Ca3SiO5	C3S
β-Ca3(PO4)2	β-TCP	2CaO·SiO2/Ca2SiO4	C2S
				Ca(H2PO4)2·H2O	MCPM	xCaO·SiO2·nH2O	C-S-H
CaHPO4	DCPA	Ca2P2O7	CPP
				CaHPO4·2H2O	DCPD

< bone repair Material >

The bone repair material is prepared by taking calcium sulfate hemihydrate and bioactive ceramic materials as raw materials, uniformly mixing, granulating and then carrying out heat treatment.

The bioactive ceramic material is selected from bioactive magnesium silicate, bioactive calcium phosphate and bioactive calcium silicateAt least one, preferably from MgO-SiO₂、MgSiO₃、α-Ca₃(PO₄)₂、β-Ca₃(PO₄)₂、Ca(H₂PO₄)₂·H₂O、CaHPO₄、CaHPO₄·2H₂O、Ca₁₀(PO₄)₆(OH)₂、Ca₉(HPO₄)(PO₄)₅(OH)、Ca₂P₂O₇、3CaO·SiO₂/Ca₃SiO₅、2CaO·SiO₂/Ca₂SiO₄、xCaO·SiO₂·nH₂At least one of O.

Particularly preferably, the bioactive ceramic-based material is selected from MgSiO₃、MgO-SiO₂、β-Ca₃(PO₄)₂、β-Ca₂P₂O₇And Ca₃SiO₅At least one of (1).

MgO-SiO₂Represented herein by MgO and SiO₂Mixed powder composed of the components according to the molar ratio of 1: 1. Compared with MgSiO which is also a magnesium silicate material₃Using MgO-SiO₂Is beneficial to improving the long-term mechanical strength of the bone repair material and the 3D porous scaffold prepared from the bone repair material.

As raw materials, the calcium sulfate hemihydrate and the bioactive ceramic material are in the form of powder, and the average particle size is preferably less than 5 microns, preferably 100nm to 1 μm.

In the bone repair material of the present invention, the amount of the bioactive ceramic material is 10 to 50%, preferably 20 to 30%, based on the total mass of the calcium sulfate hemihydrate and the bioactive ceramic material.

In addition to the two types of ingredients mentioned above, the raw materials may optionally be (optionally) added to conventional adjuvants known in the art of bone cement applications. Without limitation, based on the purpose of further improving the injection performance of the bone repair material, biopolymers with good biocompatibility such as PEG, PVA, nanocellulose and the like can be added into the raw materials; to further aid in bone repair and regeneration, bioactive substances such as bioactive ions, growth factors, and the like may be added.

The above-mentioned blending and granulation are not particularly limited to obtain the bone repair material of the present invention, and conventional processes in the field of ceramic materials, which are well known to those skilled in the art, may be employed. Without limitation, suitable amounts of agents (e.g., binders, water, etc.) to aid in powder formation can be added during mixing to facilitate particle formation, and/or suitable amounts of pore formers can be added to introduce porosity into the final microparticle product.

In the formation process of the bone repair material, heat treatment is an indispensable key link, and the heat treatment endows the microparticle product with the capability of forming a 3D porous scaffold through in-situ reaction and has an influence on the mechanical strength of the final 3D porous scaffold. The heat treatment temperature is preferably between 100 and 200 ℃, and preferably between 150 and 190 ℃; the treatment time is from 0.5 to 5 hours, preferably from 1 to 3 hours.

The bone repair material of the present invention may be microparticles of any shape, and preferably are round or cylindrical from the viewpoint of easy injection. The average particle size of the microparticles ranges from a few tens of microns to a few millimeters, preferably between 250 and 1000 microns.

Microparticles may be dense or may have a porous structure, depending on a combination of cost and performance considerations. The pore size in the microparticles may range from nanometers to sub-centimeters, such as from 10 nanometers to 300 microns, preferably from 50 nanometers to 100 microns. The introduction of the porous structure in the preparation of the ceramic powder is a well-known technique that is very easily known to those skilled in the art, such as adding pore-forming agent to the raw materials, adjusting the solid-to-liquid ratio when mixing the raw materials, etc., and will not be described in detail herein.

The bone repair material microparticle product has good mechanical strength, and the compressive strength of the bone repair material microparticle product is usually 2-20MPa, preferably 4-10 MPa.

< preparation of bone repair Material >

Corresponding to the bone repair material, the invention also relates to a preparation method of the bone repair material, which comprises the following steps: the calcium sulfate hemihydrate, the bioactive ceramic material and other optional components are weighed according to the proportion, evenly mixed, granulated and then subjected to heat treatment.

In theory, any process that meets the above synthetic concepts is suitable for synthesizing the bone repair material of the present invention, such as a crushing method, a template method, an engineering method, and the like.

Illustratively, the pulverization method includes: premixing calcium sulfate hemihydrate and bioactive ceramic micropowder, adding a proper amount of water or a water solution containing a biopolymer, and uniformly mixing to form a paste; and (3) after the paste is cured and dried in a room-temperature or low-temperature oven, grinding and crushing the dried product and sieving to obtain particles with the required particle size, and then carrying out heat treatment on the obtained particles to obtain the final granular bone repair material.

Illustratively, the template method includes: premixing calcium sulfate hemihydrate and bioactive ceramic micropowder, adding a proper amount of water or a water solution containing a biopolymer, and uniformly mixing to form a paste; and (3) coating the paste into a porous template with a set pore size, curing and drying in a room-temperature or low-temperature oven, demolding to obtain particles, and then carrying out heat treatment on the particles to obtain the final granular bone repair material. The template used may be a polymer material such as Teflon, PLA, polyester, ABS, etc., among others.

Illustratively, the engineering method comprises: premixing calcium sulfate hemihydrate and bioactive ceramic micropowder, adding a small amount of liquid medium or solution containing a biopolymer, uniformly mixing to form dough or putty, performing extrusion molding to obtain fibers or short rods with a set diameter, forming cylindrical or spherical particles by using a spheroidization device, and performing heat treatment on the obtained particles to obtain the final granular bone repair material.

< use of bone repair Material >

The bone repair material has injectability, and can form a 3D porous scaffold in situ after injection, thereby playing the roles of repairing bone defects and inducing bone regeneration. Any situation where a bone defect is present may be suitable, including but not limited to: bone defects caused by trauma (such as trauma, car accident and fracture), bone tumor, osteoporosis, osteonecrosis and the like, alveolar bone repair and the like. The novel material can be shaped spontaneously according to the shape of a defect part to form a 3D porous scaffold with a corresponding structure, and simultaneously plays a supporting role. Therefore, the invention also relates to the application of the bone repair material in bone repair.

When the bone repair material of the present invention is injected, the injection method is not particularly limited, and any conventional method known in the art for injecting bone cement can be applied. The bone repair material can be directly injected by dry powder, and also can be dispersed in a proper amount of liquid medium for injection. The type of liquid medium suitable for use is readily determinable by one skilled in the art and is generally a biocompatible liquid (e.g., water), and more preferably a biocompatible biopolymer soluble in the liquid to ensure dispersion homogenization of the system and stable injection of the bone repair material microparticles.

When in use, the orthopedic syringes with different inner diameters are selected according to the size of the bone repair material microparticles. After being injected into a bone defect part, the microparticles are naturally accumulated into a shape corresponding to a defect area, and under the action of body fluid or blood, the surfaces of the microparticles are hydrated, so that chemical bonding is formed among the microparticles through self-curing reaction, and a 3D porous scaffold is further formed in situ.

The 3D porous scaffold formed after injection has a porous structure that facilitates the ingrowth of blood vessels, migration of bone cells, and the generation of new bone. The pore size is determined primarily by the particle size of the microparticles and illustratively may be in the range of 10 to 500 microns.

The formed 3D porous scaffold has good mechanical strength, the compressive strength is generally 0.5-4MPa, and sufficient supporting capability can be provided.

In addition, compared with the prior art, the 3D porous scaffold formed by applying the bone repair material has advantages in degradation rate. In general, the larger the surface area, the faster the degradation rate. The purpose of regulating and controlling the degradation rate of the scaffold can be achieved by controlling the size, the internal pore diameter and the porosity of the microparticles, so that the scaffold meets the requirements on the degradation rate under different application conditions and is matched with the growth of new bones. Meanwhile, due to the different biological solubility of different materials, the composition of the microparticles is also an important factor for regulating the degradation rate, for example, the introduction of bioactive ceramics with lower solubility can reduce the degradation rate, and vice versa. Tests have shown that the degradation rate of the 3D porous scaffolds of the present invention is typically 10-30 wt% weight loss in one month.

<3D porous scaffold >

Besides the application mode of injecting the bone repair material microparticles, the bone repair material can be prepared into a 3D porous scaffold in vitro in advance and then used in bone repair, so that more options are provided for clinical application. Correspondingly, the invention also relates to application of the bone repair material in preparation of the 3D porous scaffold, the prepared 3D porous scaffold and application of the 3D porous scaffold in bone repair.

Representatively, the 3D porous scaffold may be prepared by using a mold, and the preparation process may include: the desired mold is printed out using 3D printing techniques, and the microparticles are filled or injected into the mold, which is then provided with a suitable environment to form the 3D porous scaffold in situ.

The mold may be of any shape and size, such as cylindrical, spherical, polygonal, ribbon, Y-shaped, star-shaped, and the like. After the microparticles are injected into the mold, the microparticles may be formed into a 3D porous scaffold in situ, without limitation, by:

(1) injecting deionized water or an aqueous solution (such as a phosphate solution, a biocompatible polymer solution and the like) into the mold, standing for at least 5 minutes, and demolding to obtain a 3D porous scaffold;

(2) and (3) placing the whole mould in an environment with 100% humidity for at least half an hour, and demoulding to obtain the 3D porous scaffold.

From the same bone repair material microparticles and similar molding conditions, the in vitro formed 3D porous scaffold has similar performance to the porous scaffold formed in situ after microparticle injection, the compressive strength is in the range of 0.5-4MPa, and the degradation rate is 10-30 wt% of weight loss in one month.

The 3D porous scaffold prepared by utilizing the bone repair material can be directly used clinically, the specific use method is easy to determine for a person skilled in the art, and the method can refer to the existing various bone repair implantation technologies.

Drawings

Fig. 1 is an XRD pattern of the bone repair materials 1-3. Wherein fig. 1A is an XRD pattern of the bone repair material 1, and fig. 1B is an XRD pattern of the bone repair material 2; fig. 1C is an XRD pattern of the bone repair material 3.

Fig. 2 is an optical photograph of the appearance of the bone repair material and the 3D porous scaffold prepared from the same. Wherein, fig. 2A is an optical photograph of the bone repair material 1; fig. 2B shows the injectability of the bone repair material 1; fig. 2C is a circular ring-shaped porous scaffold made of the bone repair material 1; fig. 2D is a semi-circular ring-shaped porous scaffold made of bone repair material 1; fig. 2E is a cylindrical porous scaffold made of bone repair material 1; fig. 2F is a human bone-shaped porous scaffold made of the bone repair material 2; fig. 2G is a spherical porous scaffold made of bone repair material 3.

FIG. 3 is a micro-CT image of the cylindrical 3D porous scaffold shown in FIG. 2E described above.

Fig. 4 is a graph of data from a 3D porous scaffold degradation performance test. Wherein, fig. 4A is weight loss data of porous scaffold samples in PBS; FIGS. 4B and 4C show changes in the calcium ion concentration and the magnesium ion concentration in PBS, respectively; FIG. 4D shows the change in pH of the solution in the degradation experiment.

FIG. 5 is an optical photograph showing the result of ALP staining of cells in vitro. Wherein, group A represents cells of a control group, and group B represents cells of an experimental group.

FIG. 6 is an optical photograph showing the result of alizarin red staining in vitro. Wherein, group A represents cells of a control group, and group B represents cells of an experimental group.

FIG. 7 is a micro-CT image of bone repair in animal experiments. Wherein, fig. 7A is the blank control composition bone condition, fig. 7B is the experimental composition bone condition, and fig. 7C is the original bone defect area morphology.

Detailed Description

The present invention is further illustrated by the following specific examples, which should not be construed as limiting the scope of the invention.

1. Preparation of bone repair material

1.1 preparation of example 1

Mixing 800g of calcium sulfate hemihydrate (average particle size about 0.75 microns), 200g of magnesium oxide (average particle size about 0.35 microns) + silicon oxide (average particle size about 0.08 microns) ((R))MgO and SiO₂The molar ratio of (1: 1) was put into a centrifugal tube (falcon tube), and the mixture was mixed by shaking for 30 seconds using a shaker (model SA8Vortex Mixer) to obtain a mixed powder. 300g of deionized water was added to the mixed powder, and the mixture was mixed uniformly to form a paste. The paste was applied to a porous template having a pore size of 1mm and a thickness of 1mm, dried at room temperature for 3 hours, and then demolded to obtain primary particles. The initial granules were treated at 150 ℃ for 3 hours to give the final granular bone repair material, designated bone repair material 1.

1.2 preparation of example 2

700g of calcium sulfate hemihydrate (average particle size about 0.75 microns), 270g of beta-Ca₃(PO₄)₂(average particle size about 0.3 μm) and 30g of beta-Ca₂P₂O₇The powder (average particle size about 0.3 μm) was put into a centrifugal tube, and mixed by shaking with a shaker for 60 seconds to obtain a mixed powder. To the mixed powder, 200g of additive polyethylene glycol (PEG-300) was added and mixed uniformly, and then the mixture was fed into an extrusion molding machine (model Caleva Multi Lab-extruder, extrusion pore size 1000 μm) to obtain an extrudate at an extrusion speed of 30rpm, and then fed into a spheroidizing machine (model Caleva Multi Lab-spherizer) to operate at 1500rpm for 3 minutes to obtain pellets. The granules were sieved (mesh size 0.5mm) and then treated at 170 ℃ for 2 hours to give the final granular bone repair material, designated bone repair material 2.

1.3 preparation of example 3

750g of calcium sulfate hemihydrate (average particle size about 0.8 μm), 250g of Ca₃SiO₅The powder (average particle size about 0.7 μm) was put into a centrifuge tube, and mixed by shaking with a shaker for 45 seconds to obtain a mixed powder. 400g of deionized water was added to the mixed powder, and the mixture was mixed uniformly to form a paste. The paste was cured and dried in a low temperature oven (40 ℃) for 10 hours to give a block. The blocks were ground and crushed and then sieved (mesh size 800 microns) and the particles obtained after sieving were treated at 190 ℃ for 1.5 hours to give the final granular bone repair material, designated bone repair material 3.

Preparation of 3D porous scaffolds

The bone repair material 1-3 is used as a raw material, a 3D printing technology is matched, and a mold is used for preparing the 3D porous support.

2.1 mold fabrication

A mold file was made using Solidworks, and then a mold was made by 3D printing. The mould outward appearance is the rectangle, and inside is the cavity that has different structures, includes: spherical (diameter 5mm), cylindric (diameter 3mm, height 10mm), ring (internal diameter 5mm, external diameter 10mm, height 3mm), semicircle ring (internal diameter 5mm, external diameter 10mm, height 3mm), human bone shape (length 20mm, thickness 3 mm).

2.23D preparation of porous scaffolds

The bone repair material was filled into the cavity of the mold using an orthopedic syringe with an inner diameter of 1.5mm, followed by standing in a 100% humidity environment for 5 hours and demolding to obtain a 3D porous scaffold.

3. Characterization of structure and physical and chemical properties

The structures and partial physical and chemical properties of the prepared bone repair material and the 3D porous scaffold are characterized, and the physical and chemical properties mainly comprise phase composition, morphology, mechanical strength and degradation performance.

3.1 phase composition of bone repair Material

The phase composition of the samples was determined by XRD analysis (model Siemens diffractometer 5000).

Fig. 1A is an XRD pattern of the bone repair material 1, the crystalline phase of which is composed mainly of calcium sulfate hemihydrate, with a small amount of calcium sulfate dihydrate, while the other components are present in amorphous form.

FIG. 1B is an XRD pattern of the bone repair material 2 with its crystalline phases predominantly present as calcium sulfate hemihydrate and β -Ca₃(PO₄)₂。

FIG. 1C is an XRD pattern of the bone repair material 3 showing that its crystalline phase is composed primarily of calcium sulfate hemihydrate and tricalcium silicate, i.e., Ca₃SiO₅And (4) forming.

As noted above, heat treatment is an indispensable key in the formation of bone repair materials, which gives the microparticles the ability to react in situ to form a 3D porous scaffold. The reason for this is that the raw material calcium sulfate hemihydrate is easily hydrated to form calcium sulfate dihydrate during the processing, thereby losing or losing the in-situ reaction function. The heat treatment can again convert the calcium sulfate dihydrate to calcium sulfate hemihydrate, thereby ensuring and enhancing its ability to form a 3D porous scaffold in situ.

3.2 morphology

(1) Appearance and appearance

Fig. 2 is an optical photograph of the appearance of the bone repair material and the 3D porous scaffold prepared from the same. Wherein 2A is an optical photograph of the bone repair material 1; 2B demonstrates the injectability of the bone repair material 1; 2C is a circular porous scaffold made of the bone repair material 1; 2D is a semicircular annular porous scaffold made of bone repair material 1; 2E is a cylindrical porous scaffold made of bone repair material 1; 2F is a human bone-shaped porous scaffold made of the bone repair material 2; 2G is a spherical porous scaffold made of bone repair material 3.

As can be seen from fig. 2C-2G, the bone repair material can not only form a good filling in the simulated bone defect site (i.e., the mold), but also form various 3D porous scaffold structures corresponding to the mold structure through in situ surface reaction. The formed 3D porous scaffold is in a particle assembly form, and is stable in structure and not loose.

(2) micro-CT image analysis

The morphology and cross-section of the 3D porous scaffold samples were analyzed by micro-CT analysis (Skyscan 1172, Bruker Corporation, Germany).

FIG. 3 is a micro-CT image of the cylindrical 3D porous scaffold shown in FIG. 2E described above. As shown, the entire scaffold is composed of microparticles and has a porous structure inside.

3.3 mechanical Strength

The resulting bone repair materials 1-3 and 3D porous scaffolds were tested for compressive strength with reference to ISO 5833:2002 and ASTM F451-99a standards.

(1) Bone repair material microparticles

TABLE 2

The results in table 2 show that the bone repair materials 1-3 of the present invention have satisfactory compressive strength and the strength of the microparticles in a liquid environment is greatly improved, which provides support for the use of the materials in vivo.

(2)3D porous scaffold

The compressive strength of the cylindrical porous scaffold shown in the above FIG. 2E was tested, and the result showed that the compressive strength was 1.196MPa, which showed a higher compressive strength.

3.4 degradation Properties

The 3D porous scaffold sample to be tested was placed in phosphate buffered saline at pH 7.4 for degradation experiments. During the experiment, the ion concentration in the solution was analyzed by ICP-OES (Avio 200, PerkinElmer).

The test samples are represented by the cylindrical porous scaffold shown in FIG. 2E above; simultaneously referring to the manufacturing method of the bracket, the initial raw materials of calcium sulfate hemihydrate and MgO-SiO₂The proportion of the two is adjusted from 80 percent to 20 percent to 90 percent to 10 percent and 50 percent to obtain the other two porous scaffolds with similar structures.

Figure 4A shows the weight loss data for three porous scaffold samples in phosphate buffered solution. The data in the figure show that the weight loss of the scaffold in the solution gradually increases with time, and can reach 21-31% after 36 days.

FIGS. 4B and 4C show the changes in the calcium ion concentration and the magnesium ion concentration in PBS, respectively. It can be seen that the concentrations of calcium and magnesium ions in the solution reached the highest at 7-11 days and then began to decrease. The increase in concentration results from the gradual release of calcium and magnesium ions from the scaffold, and the subsequent decrease is mainly due to the biological activity of the scaffold, which induces the formation of hydroxyapatite, thereby depleting the solution of inorganic ions.

FIG. 4D shows the change in pH of the solution in the degradation experiment. The pH was 7-8 in PBS for 1 day and around 6.5-7.7 after 36 days. In this interval, no significant effect on bone cells is observed.

4. Bone repair applications

4.1 in vitro experiments

Culturing to the third generation by using the separated human bone marrow mesenchymal stem cells. The bone repair material 1 (experimental group) shown in fig. 2A was placed in a cell culture medium, placed in a cell culture chamber at 37 ℃ and left to stand for 24 hours, and the mixed solution was filtered to obtain an experimental group cell culture medium. The control group culture medium is a common cell culture medium.

Dividing cells into two groups, respectively inoculating the two groups into 6-pore plates, culturing a control group by using a common cell culture medium, and changing the liquid every three days; the experimental group was cultured using the medium after the filtration of the standing solution, and the solution was changed every three days. After the cells were cultured for 14 days, two sets of cells were subjected to ALP and alizarin red staining, respectively.

FIG. 5 is an optical photograph showing the result of cell ALP staining, wherein group A represents the cells of the control group and group B represents the cells of the experimental group. The more blue, the stronger the osteogenic capacity.

FIG. 6 is an optical photograph showing the result of alizarin red staining, wherein group A represents the cells of the control group and group B represents the cells of the experimental group. More red spots indicate a stronger osteogenic capacity.

The results of fig. 5 and 6 both show that the experimental group has stronger osteogenesis ability of the static solution cultured cells, which is significantly better than the control group.

4.2 animal experiments/osteogenesis experiments

In the experiment, 3-3.5kg of New Zealand white rabbits are selected, anesthetized by phenobarbital through ear marginal veins, fixed to an operating table, the knee joints on the left side are selected for disinfection and drape treatment, the skin and fascia are cut layer by layer to expose the femoral condyles, holes with the Kirschner wire are drilled to form the hole channels with the length of 1cm and the diameter of 0.5cm, and the appearance of the bone defect area is shown as the figure 7C.

Animals were divided into two groups: one group was an experimental group, and the bone repair material 1 shown in fig. 2A was injected to the bone defect; one group was a blank control group with no treatment after drilling (no particle injection).

After 4 weeks, the femurs are taken out respectively, and micro-CT scanning is carried out to detect the osteogenesis condition of the experimental group and the blank control group. The experimental results show that the experimental group has obvious bone ingrowth at the bone defect (figure 7B) and has obvious advantages in the aspects of bone defect repair and regeneration induction compared with the blank control group (figure 7A).

Claims (22)

1. A bone repair material characterized by: calcium sulfate hemihydrate and bioactive ceramic materials are used as raw materials, evenly mixed, granulated and then subjected to heat treatment to obtain the calcium sulfate hemihydrate and bioactive ceramic materials;

the heat treatment is to convert calcium sulfate dihydrate which is easily hydrated and formed in the processing process of the raw material calcium sulfate hemihydrate into calcium sulfate hemihydrate;

the bone repair material is microparticles, and the average particle size of the microparticles is 250-1000 microns.

2. The bone repair material of claim 1, wherein: the bioactive ceramic material is at least one of magnesium silicate, calcium phosphate and calcium silicate bioactive materials.

3. Bone repair material according to claim 1 or 2, characterized in that: the bioactive ceramic material is selected from MgO-SiO₂、MgSiO₃、α-Ca₃(PO₄)₂、β-Ca₃(PO₄)₂、Ca(H₂PO₄)₂·H₂O、CaHPO₄、CaHPO₄·2H₂O、Ca₁₀(PO₄)₆(OH)₂、Ca₉(HPO₄)(PO₄)₅(OH)、Ca₂P₂O₇、Ca₃SiO₅、Ca₂SiO₄、xCaO·SiO₂·nH₂At least one of O.

4. Bone repair material according to claim 1 or 2, characterized in that: the bioactive ceramic material is selected from MgSiO₃、MgO-SiO₂、β-Ca₃(PO₄)₂、β-Ca₂P₂O₇And Ca₃SiO₅At least one of (1).

5. The bone repair material of claim 1, wherein: the calcium sulfate hemihydrate and bioactive ceramic-based materials have an average particle size of less than 5 microns.

6. The bone repair material of claim 1, wherein: the calcium sulfate hemihydrate and the bioactive ceramic-based material have an average particle size of 100nm to 1 μm.

7. The bone repair material of claim 1, wherein: the dosage of the biological active ceramic material is 10-50% of the total mass of the calcium sulfate hemihydrate and the biological active ceramic material.

8. The bone repair material of claim 1, wherein: the dosage of the biological active ceramic material is 20-30% of the total mass of the calcium sulfate hemihydrate and the biological active ceramic material.

9. The bone repair material of claim 1, wherein: the raw material also contains biopolymer and/or bioactive substances.

10. The bone repair material of claim 1, wherein: the heat treatment temperature is between 100 ℃ and 200 ℃; the treatment time is 0.5-5 hours.

11. The bone repair material of claim 10, wherein: the heat treatment temperature is 150-190 ℃.

12. The bone repair material of claim 10, wherein: the treatment time is 1-3 hours.

13. A method of preparing a bone repair material according to any one of claims 1 to 12 comprising: the calcium sulfate hemihydrate, the bioactive ceramic material and other optional components are weighed according to the proportion, evenly mixed, granulated and then subjected to heat treatment.

14. Use of a bone repair material according to any one of claims 1 to 12 in the preparation of a bone repair material.

15. Use according to claim 14, characterized in that: can be used for repairing bone defect caused by wound, bone tumor, osteoporosis, osteonecrosis, or alveolar bone.

16. Use according to claim 14 or 15, characterized in that: the bone repair material is directly injected in dry powder or dispersed in a proper amount of liquid medium for injection.

17. Use of the bone repair material according to any one of claims 1 to 12 for the preparation of a 3D porous scaffold.

18. A method of making a 3D porous scaffold comprising:

providing a mould;

filling or injecting the bone repair material microparticles of any one of claims 1 to 12 into a mold;

providing a proper environment to enable the bone repair material microparticles to form the 3D porous scaffold in situ.

19. The method of claim 18, wherein: the mold is prepared by a 3D printing technology.

20. The method of claim 18, wherein one of the following operations is selected to form the microparticles into a 3D porous scaffold in situ:

(1) injecting deionized water or aqueous solution into the mold, standing for at least 5 minutes, and demolding to obtain a 3D porous support;

(2) and (3) placing the whole mould in an environment with 100% humidity for at least half an hour, and demoulding to obtain the 3D porous scaffold.

21. A 3D porous scaffold prepared by the method of any one of claims 18-20.

22. Use of the 3D porous scaffold of claim 21 in the preparation of a bone repair material.

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