CN115671387B

CN115671387B - Bone repair stent for long-segment bone defect and preparation method and application thereof

Info

Publication number: CN115671387B
Application number: CN202211405215.2A
Authority: CN
Inventors: 何志敏; 宋天喜; 崔云; 朱金亮; 胡艳丽; 胡刚; 仇志烨; 崔孟龙; 吴晶晶
Original assignee: Weifang Aojing Health Technology Co ltd; Aojing Medical Technology Co ltd
Current assignee: Weifang Aojing Health Technology Co ltd; Aojing Medical Technology Co ltd
Priority date: 2022-11-10
Filing date: 2022-11-10
Publication date: 2023-12-08
Anticipated expiration: 2042-11-10
Also published as: CN115671387A

Abstract

The invention provides a bone repair stent for long segment bone defect, a preparation method and application thereof, wherein the bone repair stent comprises a stent main body, a stent main body filler filled in the stent main body and an osteoinductive regeneration membrane attached to the surface of the stent main body; the bracket main body is of a three-dimensional reticular frame structure; the osteoinductive regeneration membrane comprises a loose layer and a compact layer; the loose layer is attached to the surface of the bracket main body; the stent main body is prepared from a mixture of biodegradable fibers and polylactic acid; the stent filler comprises biphasic calcium phosphate porous bone powder and biological adhesive; the bioadhesive comprises a bioactive material and a solvent. The bone repair stent provided by the invention has excellent mechanical property, biocompatibility and bone repair capability, can be used as a substitute of autologous bone, and is used for repairing a long-segment bone defect in clinical requirements.

Description

Bone repair stent for long-segment bone defect and preparation method and application thereof

Technical Field

The invention relates to the technical field of bone repair, in particular to a bone repair stent for a long-segment bone defect, and a preparation method and application thereof.

Background

The long-segment bone defect is an orthopedic difficult and complicated disease, and a common treatment method comprises bone grafting, wherein the bone grafting comprises autologous bone grafting, such as common ilium and fibula taking and bone grafting treatment. Because the autologous bone is limited and the demand of the defective bone of the long segment bone is large, the autologous bone transplantation needs multiple operations, which can increase the operation pain of patients; therefore, there is an urgent need for a substitute that can replace autologous bone for repair of a clinically desirable long-segment bone defect.

However, the long-segment bone defect has high requirements on the bone repair material, not only needs to have enough mechanical strength for supporting, but also needs to have good biocompatibility and osteogenesis performance, and the bone repair material prepared by the existing method has single component structure, so that the requirement is difficult to realize; therefore, there is an urgent need for a long-segment bone repair scaffold that can be used in clinical demand to replace autologous bone for repair of long-segment bone defects.

Disclosure of Invention

Aiming at one or more technical problems in the prior art, the invention provides a bone repair bracket for long-segment bone defects, and a preparation method and application thereof.

The present invention provides in a first aspect a bone repair scaffold for a long-segment bone defect, the bone repair scaffold comprising a scaffold body, a scaffold body filler filled inside the scaffold body, and an osteoinductive regeneration membrane conforming to a surface of the scaffold body; the bracket main body is of a three-dimensional reticular frame structure; the osteoinductive regeneration membrane comprises a loose layer and a compact layer; the loose layer is attached to the surface of the bracket main body;

the bracket main body is formed by molding a mixture of biodegradable fibers and polylactic acid; the stent filler comprises biphasic calcium phosphate porous bone powder and a biological adhesive; the bioadhesive comprises a bioactive material and a solvent.

Preferably, the three-dimensional mesh frame structure comprises an outer frame and a mesh structure connected inside the outer frame; the net structure is formed by connecting a plurality of support columns; the support columns are arranged along the length direction of the support main body; the diameter of the support column is 1-3 mm, and the interval between two adjacent support columns is 2-5 mm.

Preferably, screw holes are formed in the support main body, and the compressive strength of the support main body is 80-100 MPa.

Preferably, the compact layer is prepared from mineralized collagen bone powder and collagen solution according to the dosage ratio of (5-8) g to 100 mL; the thickness of the compact layer is 0.5-1.5 mm.

Preferably, the loose layer is prepared from mineralized collagen bone powder and the collagen solution according to the dosage ratio of (10-20) g to 100 mL; the thickness of the loose layer is 0.2-1 mm.

Preferably, the bone repair scaffold further comprises bone nails; the bone nails are connected with the screw holes and used for fixing the bracket main body.

The present invention provides in a second aspect a method for preparing the bone repair scaffold according to the first aspect, the method comprising the steps of:

s1, performing 3D printing by taking a mixture of biodegradable fibers and polylactic acid as a raw material to obtain a bracket main body;

s2, mixing the biphasic calcium phosphate porous bone powder with the biological adhesive to obtain a stent filler; the bioadhesive comprises a bioactive material and a solvent;

s3, performing freeze-drying molding, cross-linking, freeze-drying and rolling on the mixture of the mineralized collagen bone powder and the collagen solution to obtain a compact layer;

s4, brushing the mixture of mineralized collagen bone powder and collagen solution on the surface of the compact layer, and freeze-drying to form a loose layer to obtain the osteoinductive regeneration membrane.

Preferably, after step S4, the method further comprises a step of preparing bone nails by hot melt molding or injection molding using a mixture of biodegradable fibers and polylactic acid as a raw material.

Preferably, in the mixture of the biodegradable fiber and the polylactic acid, the biodegradable fiber accounts for 3 to 10 weight percent; preferably, the molecular weight of the polylactic acid is 10 to 20 ten thousand, and the biodegradable fiber is polyglycolic acid fiber.

Preferably, in step S1, the temperature of the 3D printing is 200-240 ℃, and the printing speed is 0.1-0.2 mm/min.

Preferably, in the step S2, the mass ratio of the biphasic calcium phosphate porous bone powder to the biological adhesive is (20-50): 100; the dosage ratio of the bioactive material to the solvent is (5-8) g to 100mL; preferably, the bioactive material is at least one of medical collagen, chitosan and hyaluronic acid, and the solvent is one of autologous blood, physiological saline and water for injection.

Preferably, the bioadhesive further comprises a growth factor and/or an antibiotic; preferably, the growth factor is one of concentrated growth factor, bone morphogenic protein and transforming growth factor-beta; the antibiotic is one of vancomycin, gentamicin and tobramycin; more preferably, the growth factor accounts for 0.01-5% of the total mass of the biological adhesive; the antibiotics account for 0.01 to 5 percent of the total mass of the biological adhesive.

Preferably, the collagen solution is obtained by mixing collagen and purified water in an amount ratio of (3-5) g to 100 mL.

Preferably, in the step S3, the crosslinking is carried out by soaking for 24-28 hours by adopting an ethanol solution of glutaraldehyde with the mass fraction of 0.01%.

The invention provides in a third aspect the use of a bone repair scaffold according to the first aspect in the field of repair of long-segment bone defects.

Compared with the prior art, the invention has at least the following beneficial effects:

(1) The main body of the bone repair stent is formed by the mixture of biodegradable fibers and polylactic acid, and is a degradable material; the support main body is of a three-dimensional reticular frame structure, the support column can improve the mechanical property of the support main body, and the support main body filler filled in the three-dimensional reticular frame structure has excellent biocompatibility and osteogenesis capability; in addition, the three-dimensional net structure of the stent main body also has certain shaping property, so that new tissues can grow along the length direction of the stent main body.

(2) The osteoinductive regeneration membrane in the bone repair stent comprises a compact layer and a loose layer, wherein the loose layer has similar bone components and is attached to the surface of the stent, so that new bone formation can be induced; the compact layer has compact and smooth surface, can well isolate the lesion area from the external environment by utilizing the physical barrier function of the membrane, avoids infection, and ensures that the regeneration function of the tissue to be repaired is exerted to the greatest extent.

(3) The bone repair bracket provided by the invention has excellent mechanical property, biocompatibility and bone repair capability, can be personalized and customized according to the actual bone defect condition, meets different requirements, solves the problem that the clinical long-segment bone defect repair at the present stage adopts autologous bone to carry out multiple transplants, can be used as an autologous bone substitute, and is used for repairing the clinical long-segment bone defect.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic illustration of the structure of a bone repair scaffold for a long segmental bone defect provided by the invention;

FIG. 2 is a schematic illustration of the structure of a mid-osteoinductive regeneration membrane of a bone repair scaffold for long segmental bone defects according to the present invention;

FIG. 3 is a schematic view of a bone repair scaffold according to the present invention using an embedded implant bone defect site;

FIG. 4 is a schematic view of a bone repair scaffold according to the present invention using a shell-type implant to replace a bone defect site;

FIG. 5 is a diagram of liver pathology after 12 weeks of animal experiments (implantation of the bone repair scaffold provided in example 1 for repair of rabbit radius defects);

FIG. 6 is a graph of kidney pathology after 12 weeks of animal experiments (implantation of the bone repair scaffold provided in example 1 for repair of rabbit radius defects);

FIG. 7 is a graph showing lung pathology after 12 weeks of animal experiments (implantation of the bone repair scaffold provided in example 1 for repair of rabbit radius defects);

FIG. 8 is a view of spleen pathology after 12 weeks of animal experiments (implantation of the bone repair scaffold provided in example 1 for repair of rabbit radius defects);

FIG. 9 is an X-ray film 24 weeks after performing an animal experiment (implantation of the bone repair scaffold provided in example 2 for repair of rabbit radius defects);

FIG. 10 is an X-ray film 24 weeks after performing an animal experiment (implantation of the bone repair scaffold provided in comparative example 12 for repair of rabbit radius defects);

in the figure: 1-a stent body; 11-supporting columns; 2-stent filler; 3-osteoinductive regeneration membrane; 31-a loose layer; 32-dense layer; 4-bone nails.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the embodiments described below will be clearly and completely described in conjunction with the technical solutions of the embodiments of the present invention, and it is apparent that the described embodiments are some, but not all, embodiments of the present invention, and all other embodiments obtained by persons of ordinary skill in the art without making any inventive effort based on the embodiments of the present invention are within the scope of protection of the present invention.

1-4, the present invention provides in a first aspect a bone repair scaffold for a long-segment bone defect, the bone repair scaffold comprising a scaffold body, a scaffold body filler filled inside the scaffold body, and an osteoinductive regeneration membrane conforming to a surface of the scaffold body; the bracket main body is of a three-dimensional reticular frame structure; the osteoinductive regeneration membrane comprises a loose layer and a compact layer; the loose layer is attached to the surface of the bracket main body;

The main body of the bone repair stent is formed by the mixture of biodegradable fibers and polylactic acid, and is a degradable material; the support main body is of a three-dimensional reticular frame structure, the support column can improve the mechanical property of the support main body, the three-dimensional reticular structure can ensure that the material can keep good mechanical property in the transverse direction and the longitudinal direction, and the support main body filler (formed by mixing biphasic calcium phosphate porous bone powder and biological adhesive) filled in the three-dimensional reticular frame structure has excellent biocompatibility and osteogenesis capability; in addition, the three-dimensional reticular structure of the bracket main body also has certain shaping property, so that new tissues can be ensured to grow along the length direction of the bracket main body, and the deviation of the repaired damaged part and the self bone can be avoided.

The osteoinductive regeneration membrane of the bone repair stent comprises a compact layer and a loose layer, wherein the loose layer has similar bone components and is attached to the surface of the stent, so that new bone formation can be induced; the compact layer has compact and smooth surface, can well isolate the lesion area from the external environment by utilizing the physical barrier function of the membrane, avoids infection, and ensures that the regeneration function of the tissue to be repaired is exerted to the greatest extent.

The bone repair bracket provided by the invention has excellent mechanical property, biocompatibility and bone repair capability, can be personalized and customized according to the actual bone defect condition, meets different requirements, solves the problem that the clinical long-segment bone defect repair at the present stage adopts autologous bone to carry out multiple transplants, can be used as an autologous bone substitute, and is used for repairing the clinical long-segment bone defect.

According to some preferred embodiments, the three-dimensional mesh frame structure comprises an outer frame and a mesh structure connected inside the outer frame; the net structure is formed by connecting a plurality of support columns; the support columns are arranged along the length direction of the support main body; the diameter of the support column is 1-3 mm (for example, 1mm, 1.2mm, 1.4mm, 1.6mm, 1.8mm, 2mm, 2.2mm, 2.4mm, 2.6mm, 2.8mm or 3 mm), and the interval between two adjacent support columns is 2-5 mm (for example, 2mm, 2.2mm, 2.4mm, 2.6mm, 2.8mm, 3mm, 3.2mm, 3.4mm, 3.6mm, 3.8mm, 4mm, 4.2mm, 4.4mm, 4.6mm, 4.8mm or 5 mm).

The diameter and the interval of the support columns are controlled in the range, so that the compressive strength of the support main body can be ensured, and enough space can be provided for the growth of new tissues; if the diameter is too large and the interval is too small, the gap in the stent body is small, and enough space cannot be provided for the growth of new tissues; the degradation speed of the bracket main body is low, and the bracket main body cannot be matched with the growth of new tissue cells, so that the repair of bone defects is not facilitated; if the diameter is too small and the interval is too large, the compressive strength of the stent main body is low, and the repair of the bone defect of a longer section cannot be satisfied; the design and selection can be made within the above range according to the actual situation, if the defect is smaller, the diameter and spacing of the support columns cannot be too large, if the defect itself is larger in size, such as the legs of an adult male, the diameter and spacing of the support columns can be selected to be a little larger.

According to some preferred embodiments, the bracket main body is provided with screw holes, and the compressive strength of the bracket main body is 80-100 MPa; the screw holes are arranged on the bracket main body and are used for being connected with the bone nails, so that the bracket main body is fixed; the strength of the bracket main body is 80-100 MPa (for example, 80MPa, 82MPa, 84MPa, 86MPa, 88MPa, 90MPa, 92MPa, 94MPa, 96MPa, 98MPa or 100 MPa) and can meet the requirement of repairing the defect of the long-segment bone.

In some specific embodiments of the present invention, screw holes are provided at both ends of the bracket body, and the screw holes are symmetrically provided along the center line of the bracket body.

According to some preferred embodiments, the dense layer is made from mineralized collagen bone meal and collagen solution in a dose ratio of (5-8) g:100mL (e.g., can be 5g:100mL, 6g:100mL, 7g:100mL, or 8g:100 mL); the dense layer has a thickness of 0.5 to 1.5mm (e.g., may be 0.5mm, 0.6mm, 0.7mm, 0.8mm, 0.9mm, 1mm, 1.1mm, 1.2mm, 1.3mm, 1.4mm, or 1.5 mm).

According to some preferred embodiments, the porous layer is made from mineralized collagen bone powder and the collagen solution in a dose ratio of (10-20) g:100mL, which may be, for example, 10g:100mL, 11g:100mL, 12g:100mL, 13g:100mL, 14g:100mL, 15g:100mL, 16g:100mL, 17g:100mL, 18g:100mL, 19g:100mL or 20g:100 mL); the thickness of the porous layer is 0.2 to 1mm (e.g., may be 0.2mm, 0.3mm, 0.4mm, 0.5mm, 0.6mm, 0.7mm, 0.8mm, 0.9mm, or 1 mm).

The ratio of the mineralized collagen bone powder to the collagen solution in the compact layer to the loose layer is controlled in the range, so that the loose layer with similar components to the autologous bone and the compact layer with smooth and compact surface can be obtained, the osteoinductive regeneration film can be ensured to have excellent osteogenesis inducing capability, and the regeneration film can be better isolated from the external environment before the growth of new tissues of the bone defect part, thereby avoiding infection and enabling the regeneration function of the tissues to be repaired to be exerted to the greatest extent. The thickness of the compact layer is controlled in the range, so that a better isolation effect with the external environment can be ensured; the thickness of the porous layer is controlled within the above range, and excellent osteogenesis inducing ability can be ensured.

According to some preferred embodiments, the bone repair scaffold further comprises bone nails; the bone nails are connected with the screw holes and used for fixing the bracket main body; the bone nail is used for fixing the bracket, can effectively prevent the bracket from slipping, and solves the problem that the bone defect repairing bracket is easy to slip after being implanted into the surface of the broken end of the bone defect.

The invention is to say that the biphasic calcium phosphate porous bone powder adopted by the invention is prepared by referring to the preparation method of Chinese patent CN 108553691A; the method comprises the following steps: the preparation components of the biphasic calcium phosphate porous bone powder comprise a first calcium salt, polyvinyl alcohol and PMMA microspheres, wherein the mass ratio of the first calcium salt to the PMMA microspheres is (0.1-0.5) 3:2, and the preparation method comprises the following steps of: (a) Preparing polyvinyl alcohol solution with the concentration of 0.2-0.5 g/mL; (b) preparing a biphasic phosphate suspension; adding hydroxyapatite and beta-calcium phosphate into PBS solution to form suspension; (c) Adding the polyvinyl alcohol solution into the biphase phosphate suspension, stirring for 0.5-1 hour, adding PMMA microspheres, and stirring for 0.5-1 hour to obtain a sintering base solution; (d) Placing the sintering base solution into sintering equipment for sintering to obtain a sintering material; the sintering comprises the following stages: the first stage: the temperature rising rate is 5-10 ℃/min, the target temperature is 400-800 ℃, and the constant temperature time is 300-350 min; and a second stage: the temperature rising rate is 5-10 ℃/min, the target temperature is 1000-1200 ℃, and the constant temperature time is 180-200 min; and a third stage: stopping heating the sintering equipment, and naturally cooling to room temperature; (e) pulverizing the sintered material; and (f) screening to obtain the granules with the particle size of 1-2 mm.

It should be noted that the mineralized collagen bone powder adopted by the invention is prepared by referring to the preparation method of Chinese patent CN 108421088A; the method comprises the following steps: s1, dissolving collagen in acetic acid to prepare an acid solution of the collagen, wherein the concentration of the collagen is 1mg/mL; s2, continuously stirring the solution obtained in the step S1, and slowly dropwise adding a solution containing calcium ions, wherein the adding amount of the calcium ions is 0.05mol of the calcium ions added per gram of collagen; s3, continuously stirring the solution obtained in the step S2, slowly dropwise adding a solution containing phosphate ions, wherein the molar ratio of the adding amount of the phosphate ions to the adding amount of the calcium ions in the step S2 is Ca/P=1.65-1.82; s4, continuously stirring the solution obtained in the step S3, slowly dropwise adding a NaOH solution to the mixed system with the pH value of 6-8, wherein when the pH value is 5-6, precipitation of the mixed system is started, and when the pH value is 7, a white suspension is formed in the mixed system; s5, standing the mixed system obtained in the step 1.4 for 24-48 hours, separating out precipitate, washing away impurity ions, then freeze-drying, and grinding to obtain mineralized collagen bone powder.

The invention does not limit the size of the bracket main body specifically, can design and select printing parameters according to the actual bone defect condition, obtain bracket main bodies with different sizes, can realize personalized customization and meets different requirements.

The invention firstly adopts a mixture of biodegradable fibers and polylactic acid to print a stent main body through 3D, then mixes biphasic calcium phosphate porous bone powder and biological adhesive, fills the mixture into the stent main body, prepares the osteoinductive regeneration membrane with a compact layer and a loose layer by taking mineralized collagen bone powder and collagen solution as raw materials, and attaches one surface of the loose layer of the osteoinductive regeneration membrane to the surface of the stent main body to obtain the bone repair stent capable of being used for long-segment bone defects.

The preparation method of the bone repair stent is simple in process, and the prepared bone repair stent has excellent mechanical property, biocompatibility and bone repair capability, can be personalized and customized according to the actual bone defect condition, meets different requirements, solves the problem that the autologous bone needs to be transplanted for multiple times in the clinical long-segment bone defect repair at the present stage, and can be used as an autologous bone substitute for repairing the clinical long-segment bone defect.

The main body of the bone repair stent prepared by the invention is prepared by adopting a mixture of biodegradable fibers and polylactic acid through 3D printing, and is a degradable material; the biphasic calcium phosphate porous bone powder and biological adhesive mixture filled in the stent main body has excellent biocompatibility and osteogenesis capability.

In some embodiments, the method of preparing a bone repair scaffold comprises: s1, taking a mixture of biodegradable fibers and polylactic acid as a raw material (the biodegradable fibers account for 3-10 wt%) and obtaining a bracket main body through a 3D printing fusion sedimentation technology under the conditions that the printing temperature is 200-240 ℃ and the printing speed is 0.1-0.2 mm/min; s2, mixing the biphasic calcium phosphate porous bone powder and the biological adhesive according to the mass ratio of (20-50): 100 to obtain a stent filler; s3, mixing the mineralized collagen bone powder and the collagen solution according to the dosage ratio (5-8) g to 100mL, stirring for 4-6 hours at the rotating speed of 100-150 r/min, freeze-drying and forming, soaking and cross-linking for 24-28 hours by using a cross-linking agent, washing for 24-48 hours by using purified water, freeze-drying, and finally rolling on a roll squeezer to obtain the compact layer; the cross-linking agent is ethanol solution of glutaraldehyde with the mass fraction of 0.01%; the rolling pressure is 10-15 MPa, and the rolling speed is 5-10 mm/min; s4, mixing mineralized collagen bone powder and a collagen solution according to the dosage ratio (10-20) g to 100mL, stirring for 4-6 h at the rotating speed of 100-150 r/min, then coating the mixture on the surface of the compact layer, and forming a loose layer through freeze drying to obtain the osteoinductive regeneration membrane.

In some specific embodiments of the invention, the freeze-drying and freeze-drying forming comprises a pre-freezing stage, a first stage and a second stage, wherein the process conditions of each stage are as follows:

pre-freezing: keeping the temperature at-20 ℃ for 120-160 min, keeping the temperature at-10 ℃ for 240-280 min (vacuumizing, aerating for 100+/-10 Pa),

first stage (maintained at 0 ℃): at 0 ℃, the constant temperature time is 480-500 min (vacuuming, aerating at 100+ -10 Pa);

second stage (sublimation): firstly, keeping the constant temperature at 10 ℃ for 120-150 min; then, the constant temperature is kept at 20 ℃ for 120-150 min; finally, the constant temperature is kept at 30 ℃ for 120-150 min.

In the process of preparing the compact layer, after freeze-drying and forming, ethanol solution with mass fraction of glutaraldehyde of 0.01% is adopted for soaking and crosslinking, so that a more stable three-dimensional network structure can be formed between molecules, and the performances of strength and the like of the compact layer can be improved; washing with purified water to remove residual glutaraldehyde in ethanol; the rolling pressure and speed are controlled in the range, so that the smooth and compact surface of the compact layer can be ensured, the lesion area is better isolated from the external environment, the infection is avoided, the regeneration function of specific tissues is exerted to the greatest extent, and the mechanical property of the compact layer can be further improved; if the rolling step is not performed, the density of the obtained compact layer is insufficient, and the lesion area cannot be effectively isolated from the external environment.

According to some preferred embodiments, after step S4, the method further comprises a step of preparing bone nails by hot melt molding or injection molding using a mixture of biodegradable fibers and polylactic acid as a raw material.

The main body of the bracket and the bone nail are both prepared from a mixture of biodegradable fibers and polylactic acid serving as raw materials and are degradable materials.

According to some preferred embodiments, the biodegradable fiber comprises 3 to 10wt% (e.g., may be 3wt%, 3.5wt%, 4wt%, 4.5wt%, 5wt%, 5.5wt%, 6wt%, 6.5wt%, 7wt%, 7.5wt%, 8wt%, 8.5wt%, 9wt%, 9.5wt%, or 10 wt%) in the mixture of the biodegradable fiber and polylactic acid; preferably, the molecular weight of the polylactic acid is 10 to 20 ten thousand (for example, 10 ten thousand, 11 ten thousand, 12 ten thousand, 13 ten thousand, 14 ten thousand, 15 ten thousand, 16 ten thousand, 17 ten thousand, 18 ten thousand, 19 ten thousand or 20 ten thousand) and the biodegradable fiber is polyglycolic acid fiber.

The stent main body is prepared from a mixture of biodegradable fibers and polylactic acid through 3D printing, and is a degradable material; the inventor finds that when the mass ratio of the biodegradable fiber is in the range, the compressive strength of the bracket main body can be ensured to be 80-100 MPa, and the bracket can be used for repairing the defect of the long-segment bone; the content of biodegradable fibers is too small to meet the compressive strength required for repair of long segmental bone defects.

According to some preferred embodiments, in step S1, the 3D printing is performed at a temperature of 200-240 ℃ (e.g., may be 200 ℃, 205 ℃, 210 ℃, 215 ℃, 220 ℃, 225 ℃, 230 ℃, 235 ℃, or 240 ℃), and at a printing speed of 0.1-0.2 mm/min (e.g., may be 0.1mm/min, 0.12mm/min, 0.14mm/min, 0.16mm/min, 0.18mm/min, or 0.2 mm/min).

The temperature control of the 3D printing can ensure the fluidity of the mixture of the biodegradable fiber and the polylactic acid in the 3D printing process, and is convenient for printing; the temperature is too low, the mixture of the biodegradable fiber and the polylactic acid has poor fluidity, and the caking phenomenon is easy to occur; the invention controls the temperature and the printing speed of 3D printing in the above range so as to obtain the bracket main body with uniform and stable performance.

According to some preferred embodiments, in step S2, the mass ratio of the biphasic calcium phosphate porous bone meal and the biological adhesive is (20-50): 100 (e.g. may be 20:100, 25:100, 30:100, 35:100, 40:100, 45:100 or 50:100); the biphasic calcium phosphate porous bone powder and the biological adhesive are controlled in the range, so that the mixed stent filler is ensured to be in a dough shape (paste shape) and fully filled into the stent main body, and the degradation speed of the stent filler can be better matched with the growth speed of new tissues in the bone repair process; if the content of the biphasic calcium phosphate porous bone powder is too high, the biphasic calcium phosphate porous bone powder is insufficiently mixed with the biological adhesive, the filler is dried and diverged, the filling effect is poor, the degradation speed of the filler is too slow, and the filler cannot be matched with the growth speed of new tissues, so that the bone repair is not facilitated; if the content of the biphasic calcium phosphate porous bone powder is too low, the biphasic calcium phosphate porous bone powder is not agglomerated after being mixed with the biological adhesive, is easy to overflow, has poor filling effect, and has too high degradation speed of the filler, so that new tissues can not grow as soon as possible, and the bone repair is not facilitated.

According to some preferred embodiments, the ratio of the amount of the bioactive material to the solvent is (5-8) g:100mL (e.g., may be 5g:100mL, 6g:100mL, 7g:100mL, or 8g:100 mL); preferably, the bioactive material is at least one of medical collagen, chitosan and hyaluronic acid, and the solvent is one of autologous blood, physiological saline and water for injection; the dosage of the active material and the solvent is controlled in the range, so that the active material can be ensured to be uniformly dispersed, the mixture has good fluidity and good plasticity, and the mixture is convenient to be mixed with the biphasic calcium phosphate porous bone powder and then filled in the meshes of the main body bracket net structure.

According to some preferred embodiments, the bioadhesive further comprises a growth factor and/or an antibiotic; preferably, the growth factor is one of concentrated growth factor, bone morphogenic protein and transforming growth factor-beta; the antibiotic is one of vancomycin, gentamicin and tobramycin; more preferably, the growth factor accounts for 0.01-5% of the total mass of the biological adhesive, for example, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% or 5%); the antibiotic may be, for example, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% or 5% by weight of the total mass of the bioadhesive.

The bioadhesives of the present invention may also contain growth factors and/or antibiotics; selecting according to the specific situation of the bone defect; when the bone forming capability of the patient with the bone defect is poor, the growth factors can be added according to the actual situation to promote the generation of new tissues; when a patient has bone tumor or inflammation, a proper amount of antibiotics can be added according to the actual situation so as to avoid infection and obstruct the bone repair process; when the patient has poor bone forming capability and bone tumor or inflammation, the growth factors and antibiotics can be added in proper amounts according to the actual situation.

According to some preferred embodiments, in step S3, the crosslinking is carried out by soaking for 24-28 hours (for example, 24 hours, 25 hours, 26 hours, 27 hours, or 28 hours) with an ethanol solution of glutaraldehyde with a mass fraction of 0.01%.

According to some preferred embodiments, the collagen solution is obtained by mixing collagen and purified water in a dosage ratio of (3-5) g:100mL (e.g., may be 3g:100mL, 3.5g:100mL, 4g:100mL, 4.5g:100mL, or 5g:100 mL); the collagen of the invention is type I collagen, which is similar to the collagen component in bone tissue and has better biocompatibility with bone tissue.

In some specific embodiments, the collagen solution is prepared by: according to the dosage ratio of the type I collagen to the purified water of 3-5g to 100mL, dissolving the collagen in the purified water, and stirring for 24-36 h under the condition of cooling water (ensuring the temperature is lower than 50 ℃) at the rotating speed of 150-200 r/min to obtain the collagen solution.

When the bone repair stent is applied to long-segment bone repair, after the stent is implanted into a bone defect part, the stent is fixed through the bone nails and screw holes arranged on a stent main body, and then the bone defect part is covered by the bone induction regeneration film, so that the bone induction regeneration film is attached to the surface of the stent, the length of the bone induction regeneration film is 5-10 mm longer than the distance between two broken ends of the bone defect, the bone induction regeneration film is ensured to better wrap the part to be repaired, the part to be repaired is isolated from the external environment, infection is avoided, and the regeneration function of the tissue to be repaired is exerted to the greatest extent.

The bracket of the invention can be embedded (as shown in figure 3) and sleeved (as shown in figure 4) when being implanted into the bone defect part; when the embedded type bone fracture device is adopted, the bracket is inserted into the marrow cavities of two broken ends at the bone fracture position; when the external sleeving type is adopted, holes with the depth of 5-10 mm are reserved at the two ends of the bracket, the sizes of the holes are set and selected according to the size condition of the actual defect part, and the requirement that the autologous bone is just embedded in the holes can be met.

In order to more clearly illustrate the technical scheme and advantages of the present invention, the present invention will be further described below with reference to examples.

It should be noted that the materials and reagents in the invention can be obtained directly or by self-synthesis in the market, and the specific model is not limited.

The performance test methods of the bone repair scaffolds prepared in the examples and comparative examples of the present invention refer to the following methods:

compressive strength test: the axial loading was measured using a cylindrical sample (bone repair scaffold prepared in examples and comparative examples) with a height h and diameter corresponding to a dimensional ratio of 1.5.ltoreq.h/d.ltoreq.2.0, with reference to the method specified in GB23101.1-20084.4, and the sample (bone repair scaffold prepared in examples and comparative examples) was placed in a material mechanical tester for compression test with a compression rate of 0.5mm/min, calculated by measuring the average of the loading forces recorded at the moment the stress began to drop.

Cytotoxicity test: the bone repair scaffolds of the example and the comparative example were subjected to irradiation sterilization, the bone repair scaffolds of the example and the comparative example were leached in a ratio of 0.2g/mL using a cell culture medium containing 10% calf serum as a leaching medium, and inoculated into a 96-well culture plate after cell counting, and placed in CO ₂ Culturing in incubator at 37 deg.c for 24 hr, and eliminating culture liquid; adding fresh cell culture solution into blank control group, respectively adding leaching solutions of the bone repair stent of the example and the comparative example into experimental group, and placing into CO ₂ Culturing in an incubator for 24 hours, observing cell morphology by using a microscope, adding 20 mu L of MTT solution with mass concentration of 5g/L into each hole, continuously culturing for 4 hours, discarding liquid in the hole, adding 200 mu L of LDMSO, measuring absorbance at 570nm and 650nm wavelength of an enzyme label instrument, and calculating relative increment rate, wherein the relative increment rate is = (the average value of absorbance of an experimental group/the average value of absorbance of a blank control group) multiplied by 100%. The reference standard of the grade of the relative increment rate is shown in the table below, and the lower the grade is, the lower the biotoxicity is.

Semi-quantitative analytical scoring criteria for bone repair healing are seen in the following table;

score value	Description of bone repair situation
		0	Repair of bone is not seen
1	<10% of the defective portions formed bone tissue or bone-like tissue
		2	10 to 25 percent of defect part forms bone tissue or bone-like tissue
3	25% -50% of the defect part forms bone tissue or bone-like tissue
		4	50% -75% of the defect part forms bone tissue or bone-like tissue
5	>75% of the defect portions form bone tissue or bone-like tissue

Example 1:

s1, performing 3D printing (the printing temperature is 220 ℃ and the printing speed is 0.15 mm/min) by adopting a mixture of polyglycolic acid fibers and polylactic acid (the polyglycolic acid fibers account for 6%) to obtain a bracket main body; screw holes are formed in the bracket main body; the bracket main body is a three-dimensional reticular frame structure with a plurality of supporting columns (the diameter is 2mm; the interval between adjacent supporting columns is 4 mm) arranged inside;

S2, uniformly mixing the biphasic calcium phosphate porous bone powder and the biological adhesive in a mass ratio of 40:100, and filling the mixture into the bracket main body; the biological adhesive comprises a biologically active material (medical collagen) and a solvent (autologous blood) in an amount ratio of 6g to 100 mL;

s3, adding mineralized collagen bone powder into a collagen solution (the dosage ratio of the mineralized collagen bone powder to the collagen solution is 7g:100 mL), stirring for 6 hours under the condition of the rotating speed of 120r/min, uniformly mixing, freeze-drying and forming, soaking and crosslinking for 28 hours by adopting an ethanol solution of glutaraldehyde with the mass fraction of 0.01%, washing for 36 hours by using purified water to remove a residual crosslinking agent, freeze-drying, and rolling (the rolling pressure is 12MPa and the rolling speed is 8 mm/min) to obtain a compact layer;

s4, adding mineralized collagen bone powder into a collagen solution (the dosage ratio of the mineralized collagen bone powder to the collagen solution is 15g:100 mL), stirring for 5 hours under the condition of the rotating speed of 150r/min, brushing the mixture on the surface of a compact layer after uniform mixing, and forming a loose layer on the surface of the compact layer through freeze drying to obtain an osteoinduction regeneration film;

s5, adopting a mixture of biodegradable fibers and polylactic acid (polyglycolic acid fibers account for 3 percent), and obtaining the bone nail through hot melting and pressing at 220 ℃ and connecting the bone nail with screw holes on the stent main body.

S6, attaching a loose layer of the osteoinductive regeneration membrane to the surface of the bracket main body to obtain the bone repair bracket.

Example 2:

s1, performing 3D printing (the printing temperature is 200 ℃ and the printing speed is 0.1 mm/min) by adopting a mixture of polyglycolic acid fibers and polylactic acid (the polyglycolic acid fibers account for 3%) to obtain a bracket main body; screw holes are formed in the bracket main body; the bracket main body is a three-dimensional reticular frame structure with a plurality of supporting columns (the diameter is 1mm; the interval between adjacent supporting columns is 2 mm) arranged inside;

s2, uniformly mixing the biphasic calcium phosphate porous bone powder and the biological adhesive in a mass ratio of 20:100, and filling the mixture into the bracket main body; the biological adhesive comprises a biologically active material (chitosan) and a solvent (physiological saline) in the dosage ratio of 5g to 100 mL;

s3, adding mineralized collagen bone powder into a collagen solution (the dosage ratio of the mineralized collagen bone powder to the collagen solution is 5g:100 mL), stirring for 4 hours at the rotating speed of 150r/min, uniformly mixing, freeze-drying, forming, soaking and crosslinking for 28 hours by adopting an ethanol solution of glutaraldehyde with the mass fraction of 0.01%, washing for 24 hours by using purified water to remove a residual crosslinking agent, freeze-drying, and rolling (the rolling pressure is 10MPa and the rolling speed is 5 mm/min) to obtain a compact layer;

S4, adding mineralized collagen bone powder into a collagen solution (the dosage ratio of the mineralized collagen bone powder to the collagen solution is 10g:100 mL), stirring for 6 hours under the condition of the rotating speed of 100r/min, brushing the mixture on the surface of a compact layer after uniform mixing, and forming a loose layer on the surface of the compact layer through freeze drying to obtain an osteoinduction regeneration film;

s5, preparing the bone nail by adopting a mixture of biodegradable fibers and polylactic acid (polyglycolic acid fibers account for 3%) through hot melt molding at 200 ℃, and connecting the bone nail with screw holes on the stent main body.

Example 3:

s1, performing 3D printing (the printing temperature is 240 ℃ and the printing speed is 0.2 mm/min) by adopting a mixture of polyglycolic acid fibers and polylactic acid (the polyglycolic acid fibers account for 10%) to obtain a bracket main body; screw holes are formed in the bracket main body; the bracket main body is a three-dimensional reticular frame structure with a plurality of supporting columns (the diameter is 3mm; the interval between adjacent supporting columns is 5 mm) arranged inside;

s2, uniformly mixing the biphasic calcium phosphate porous bone powder and the biological adhesive in a mass ratio of 50:100, and filling the mixture into the bracket main body; the biological adhesive comprises a biologically active material (hyaluronic acid) and a solvent (water for injection) in an amount ratio of 8g to 100 ml;

S3, adding mineralized collagen bone powder into a collagen solution (the dosage ratio of the mineralized collagen bone powder to the collagen solution is 8g:100 mL), stirring for 6 hours at the rotating speed of 100r/min, uniformly mixing, freeze-drying, forming, soaking and crosslinking for 28 hours by adopting an ethanol solution of glutaraldehyde with the mass fraction of 0.01%, washing for 48 hours by using purified water to remove a residual crosslinking agent, freeze-drying, and rolling (the rolling pressure is 15MPa and the rolling speed is 10 mm/min) to obtain a compact layer;

s4, adding mineralized collagen bone powder into a collagen solution (the dosage ratio of the mineralized collagen bone powder to the collagen solution is 20g:100 mL), stirring for 6 hours under the condition of the rotating speed of 150r/min, brushing the mixture on the surface of a compact layer after uniform mixing, and forming a loose layer on the surface of the compact layer through freeze drying to obtain an osteoinduction regeneration film;

s5, preparing the bone nail by injection molding at 240 ℃ by adopting a mixture of biodegradable fibers and polylactic acid (polyglycolic acid fibers account for 3%), and connecting the bone nail with screw holes in the stent main body.

Example 4:

example 4 is substantially the same as example 1, except that: the bioadhesive also contains a growth factor (bone morphogenic protein) in an amount of 1% of the total bioadhesive.

Example 5:

example 5 is substantially the same as example 1, except that: the biological adhesive also contains antibiotics, and the dosage of the antibiotics is 0.01 percent of the total weight of the biological adhesive.

Example 6:

example 6 is substantially the same as example 1, except that: the biological adhesive also comprises a growth factor and an antibiotic, wherein the dosage of the growth factor is 1 percent of the total amount of the biological adhesive, and the dosage of the antibiotic is 0.01 percent of the total amount of the biological adhesive.

Comparative example 1:

comparative example 1 is substantially the same as example 1 except that: in step S1, the bracket main body is not provided with a support column.

Comparative example 2:

comparative example 2 is substantially the same as example 1 except that: in the step S2, the mass ratio of the biphasic calcium phosphate porous bone powder to the biological adhesive is 5:100.

Comparative example 3:

comparative example 3 is substantially the same as example 1 except that: in the step S2, the mass ratio of the biphasic calcium phosphate porous bone powder to the biological adhesive is 70:100.

Comparative example 4:

comparative example 4 is substantially the same as example 1 except that: the bioadhesive comprises a bioactive material (medical collagen) and a solvent (autologous blood) in an amount ratio of 2g to 100 ml.

Comparative example 5:

comparative example 5 is substantially the same as example 1 except that: the bioadhesive comprises a bioactive material (medical collagen) and a solvent (autologous blood) in an amount ratio of 10g to 100 ml.

Comparative example 6:

comparative example 6 is substantially the same as example 1 except that: in step S3, the ratio of mineralized collagen bone powder to collagen solution was 2 g/100 mL.

Comparative example 7:

comparative example 7 is substantially the same as example 1 except that: in step S3, the ratio of mineralized collagen bone powder to collagen solution was 15 g/100 mL.

Comparative example 8:

comparative example 8 is substantially the same as example 1 except that: in step S3, the step of immersing and crosslinking for 28 hours with an ethanol solution of glutaraldehyde with a mass fraction of 0.01% is not included.

Comparative example 9:

comparative example 9 is substantially the same as example 1 except that: in step S4, the ratio of mineralized collagen bone powder to collagen solution is 2 g/100 mL.

Comparative example 10:

comparative example 10 is substantially the same as example 1 except that: in step S4, the usage ratio of mineralized collagen bone powder to collagen solution is 30 g/100 mL.

Comparative example 11:

comparative example 11 is substantially the same as example 1 except that: there are no steps S3 and S4.

Comparative example 12:

comparative example 12 is substantially the same as example 2 except that: there is no step S2.

Comparative example 13:

comparative example 13 is substantially the same as example 1 except that: and step S4 is omitted, and the compact layer is directly attached to the surface of the bracket main body in step S5.

Comparative example 14:

comparative example 14 is substantially the same as example 1 except that: and step S3 is omitted, and the loose layer is directly attached to the surface of the bracket main body in step S5.

Comparative example 15:

comparative example 15 is substantially the same as example 1 except that: in step S3, the step of rolling is not included.

The performance data of the bone repair scaffolds provided by the examples and comparative examples of the present invention are shown in table 1.

TABLE 1

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As can be seen from table 1, the stent body of comparative example 1 is a net-shaped frame structure without a stent body, the compressive strength of the bone repair stent is significantly reduced, and the mechanical support required for the long-segment bone defect indication cannot be satisfied; in comparative example 2, the content of the biphasic calcium phosphate porous bone powder is too low, the content of the biological adhesive is too high, so that the mixture is not agglomerated, the mixture is easy to flow, has overflow risk, has poor filling effect, and is fast in filler degradation and mismatched with the osteogenesis rate; in comparative example 3, the content of the biphasic calcium phosphate porous bone powder is too high, the content of the biological adhesive is too low, the biphasic calcium phosphate porous bone powder is insufficiently infiltrated, is unevenly mixed, and is dried and diverged, so that a doughy form cannot be formed, the porous bone powder is easy to scatter during filling, the filling effect is poor, the degradation rate of the filler is reduced, and the porous bone powder is not matched with the osteogenesis rate; comparative example 4 has too low bioactive material content, too high solvent content, insufficient viscosity of the bioadhesive, and material divergence after mixing with the biphasic calcium phosphate porous bone powder, inconvenience in filling, poor filling effect, poor osteoinductive performance and reduced osteogenic capacity; comparative example 5 the bioactive material (medical collagen) content was too high and the solvent content was too low, resulting in insufficient dispersion of the bioactive material (medical collagen), occurrence of caking or floc phenomenon, uneven dispersion, poor filling effect, poor osteoinductive performance, and reduced osteogenic capacity; the collagen content in the compact layer of comparative example 6 is too high, the degradation rate is high, the osteogenic performance is slightly reduced, and the problems of solubility absorption, early membrane cracks and the like are easy to occur, so that the bone healing condition is influenced; the compact layer of comparative example 7 has too low collagen content and insufficient toughness, and is easy to crack, so that the filler of the stent main body overflows, the osteoinductive performance and the osteogenesis capability are reduced, and the bone repair is not facilitated; comparative example 8 did not undergo cross-linking treatment, the dense layer was low in strength, was prone to fracture during encapsulation, and was fast in degradation rate, and not matched with the osteogenesis rate, not conducive to bone repair; comparative example 9 the content of mineralized collagen bone powder in the porous layer was too low, the osteogenic performance was reduced, the degradation rate was faster, the rate was not matched with the osteogenic rate, early degradation of the membrane occurred, and bone healing was not favored; the loose layer of comparative example 10 had too high content of mineralized collagen powder, and could not be completely impregnated when mixed with collagen solution, and could not be smoothly coated to form a loose layer; comparative example 11 does not include an osteoinductive regeneration membrane, and the osteogenic performance is reduced while the physical barrier function is not sufficiently achieved, and infection is easily caused in the later stage, affecting bone healing; comparative example 12 lacks a stent body filler having osteoinductive properties, resulting in an internal cavity, bone tissue does not grow well into the interior, and osteogenic capacity is lowered; comparative example 13 bone repair scaffold without loose layer, induction performance was reduced; the comparative example 14 bone repair scaffold does not contain a compact layer, the strength of the regeneration induction membrane is insufficient, the regeneration induction membrane is easy to break at early stage, the effect of a physical barrier cannot be fully achieved, infection is easy to cause at later stage, and bone healing is affected; comparative example 15 the dense layer was not roll-treated during preparation, the strength of the regeneration-inducing membrane was reduced, it was easily broken, the dense layer had a large porosity, it was unable to act as a physical barrier, it was easy to cause infection in the latter stage, and bone healing was affected.

After the bone repair stent of example 1 was implanted for repairing rabbit radius defects for 12 weeks, the liver, kidney, lung and spleen histopathological analysis of the experimental rabbits were performed; as shown in fig. 5, liver tissue sections show that liver cells and liver plaques are normal, aligned, and central veins are not abnormal; no pathological changes such as cyst, fibrosis, hepatocyte loss, necrosis or lymphocyte aggregation of liver tissues caused by inflammatory reaction are found, and no obvious difference exists in morphology; as shown in FIG. 6, the kidney tissue section shows that the tissue has no pathological changes such as the loss of collecting tube cells, the loss of glomeruli, cyst, fibrosis, necrosis or lymphocyte aggregation and nodule. In addition, glomeruli are free of inflammatory cell infiltration, inflammation or edema; as shown by the lung tissue section in FIG. 7, the implanted material does not induce lung inflammation, does not cause the destruction of alveolar structure, and does not have the phenomena of fibrous tissue hyperplasia, small abscess formation and the like; from the spleen tissue section shown in fig. 8, the histological analysis showed that the spleen tissue was free of pathological changes such as morphological changes of spleen tissue, lymphocyte loss, cyst, fibrosis, epithelial cell and radial cell loss, necrosis or large lymphocyte aggregation, and no inflammatory changes. In summary, kidney, liver, lung and spleen were stained with H & E at week 12, and no organ damage, structural deformity or necrosis was found; furthermore, no abnormal inflammatory cell infiltration and no abnormal increase in macrophages were observed in any organ.

As can be seen from X-ray films after the bone repair stent was implanted for 24 weeks in FIGS. 9 to 10, the bone healing condition after the bone repair stent of example 2 of the present invention was implanted was significantly better than that of comparative example 12, and the bone repair stent of the present invention had excellent bone repair performance.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A bone repair scaffold for a long segmental bone defect, characterized by:

the bone repair stent comprises a stent main body, a stent main body filler filled in the stent main body and an osteoinductive regeneration membrane attached to the surface of the stent main body; the bracket main body is of a three-dimensional reticular frame structure; the osteoinductive regeneration membrane comprises a loose layer and a compact layer; the loose layer is attached to the surface of the bracket main body; the compact layer is prepared from mineralized collagen bone powder and collagen solution according to the dosage ratio of (5-8) g to 100 mL; the loose layer is prepared from mineralized collagen bone powder and collagen solution according to the dosage ratio of (10-20) g to 100 mL;

The bracket main body is formed by molding a mixture of biodegradable fibers and polylactic acid; the stent filler comprises biphasic calcium phosphate porous bone powder and a biological adhesive; the mass ratio of the biphasic calcium phosphate porous bone powder to the biological adhesive is (20-50): 100; the biological adhesive comprises a bioactive material and a solvent; the dosage ratio of the bioactive material to the solvent is (5-8) g/100 mL.

2. A bone repair stent as claimed in claim 1, wherein:

the three-dimensional mesh frame structure comprises an outer frame and a mesh structure connected inside the outer frame; the net structure is formed by connecting a plurality of support columns;

the support columns are arranged along the length direction of the support main body; the diameter of the support column is 1-3 mm, and the interval between two adjacent support columns is 2-5 mm.

3. A bone repair stent as claimed in claim 1, wherein:

screw holes are formed in the support main body, and the compressive strength of the support main body is 80-100 MPa;

the thickness of the compact layer is 0.5-1.5 mm; and/or

The thickness of the loose layer is 0.2-1 mm.

4. A bone repair stent according to claim 3, wherein:

The bone repair scaffold further comprises bone nails; the bone nails are connected with the screw holes and used for fixing the bracket main body.

5. A method of preparing a bone repair scaffold according to any one of claims 1-4, comprising the steps of:

6. The method of manufacturing according to claim 5, wherein:

after the step S4, the method further comprises a step of preparing bone nails by hot melt molding or injection molding by using a mixture of biodegradable fibers and polylactic acid as a raw material.

7. The method of manufacturing according to claim 6, wherein:

In the mixture of the biodegradable fiber and the polylactic acid, the biodegradable fiber accounts for 3 to 10 weight percent.

8. The method of manufacturing according to claim 7, wherein:

the molecular weight of the polylactic acid is 10-20 ten thousand, and the biodegradable fiber is polyglycolic acid fiber.

9. The method of manufacturing according to claim 5, wherein:

in the step S1, the temperature of the 3D printing is 200-240 ℃, and the printing speed is 0.1-0.2 mm/min;

in step S2, the bioactive material is at least one of medical collagen, chitosan and hyaluronic acid, and the solvent is one of autologous blood, physiological saline and water for injection; and/or

In the step S3, the crosslinking is carried out by soaking for 24-28 h by adopting 0.01% glutaraldehyde ethanol solution by mass fraction.

10. The method of manufacturing according to claim 5, wherein:

the bioadhesive further comprises a growth factor and/or an antibiotic; and/or

The collagen solution is obtained by mixing collagen and purified water according to the dosage ratio of (3-5) g to 100 mL.

11. The method of manufacturing according to claim 10, wherein:

the growth factor is one of concentrated growth factor, bone morphogenetic protein and transforming growth factor-beta; the antibiotic is one of vancomycin, gentamicin and tobramycin.

12. The method of manufacturing according to claim 11, wherein:

the growth factor accounts for 0.01-5% of the total mass of the biological adhesive; the antibiotics account for 0.01 to 5 percent of the total mass of the biological adhesive.