CN107625995B - Multilayer coaxial fiber bone repair membrane material and preparation method thereof - Google Patents
Multilayer coaxial fiber bone repair membrane material and preparation method thereof Download PDFInfo
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- CN107625995B CN107625995B CN201710710641.XA CN201710710641A CN107625995B CN 107625995 B CN107625995 B CN 107625995B CN 201710710641 A CN201710710641 A CN 201710710641A CN 107625995 B CN107625995 B CN 107625995B
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Abstract
The invention relates to a multilayer coaxial fibrous bone repair material and a preparation method thereof, belonging to the field of biological materials. The material is prepared by taking biodegradable aliphatic polyester with biocompatibility and natural macromolecules as main raw materials and using four layers of coaxial needles through an electrostatic spinning method. The fiber has a multilayer coaxial structure, and antibacterial and anti-inflammatory drugs and bone-promoting drugs can be added in different layers according to the bone repair process. The material of the invention has excellent biocompatibility and controllable and long-term drug release performance. Meanwhile, along with the progress of the bone repair process, the medicine carried in the fiber is released layer by layer, and corresponding substances required by the process are provided for different stages of the bone repair. The material can realize controllable in vivo degradation according to requirements without taking out the material by a secondary operation.
Description
Technical Field
The invention relates to a preparation method of a multilayer coaxial fiber bone repair membrane material, belongs to the field of biological materials, and particularly relates to a bone repair membrane material which is composed of fibers with a four-layer coaxial structure and is adaptive to each stage of bone repair in a drug release process and a preparation method thereof.
Background
Bone defects are clinically very common wounds, and the process of bone repair is very lengthy, mainly including the hematoma and inflammation phase, the initial callus reaction phase, the chondrogenesis phase, and the bone formation and remodeling phase. In the bone repair process, cells secrete various growth factors to play roles in different time sequences, so that the repair of bone defects is ensured.
Clinically, when a bone defect occurs, it is best to perform bone grafting, mainly including autologous bone grafting, allogeneic bone grafting, non-tissue repair, etc., wherein autologous bone grafting is the best for repairing the bone defect. However, excessive bone access may cause new trauma and complications to the patient; allogeneic bone transplantation can overcome the problems caused by partial autologous bone transplantation, but is limited by donor sources, tissue conditions during transplantation, immunogenicity and the like; non-tissue repair is commonly used in joint replacement surgery, where the main problem is the inability to integrate with surrounding tissue to form a focus of infection. For bone or tissue defects, the ideal solution is to overcome the above three problems. The field of biomaterials offers a potential alternative to this problem.
As an ideal bone repair scaffold material, the following four requirements are required to be met:
(1) has biocompatibility, osteoconductivity and osteoinductivity, and can provide support for normal cell activities;
(2) biodegradability, ability to provide space for the new tissue to grow in and gradually replace with new tissue;
(3) certain mechanical property, can bear the stress in the operation process and the bone growth process;
(4) a coherent porous structure that transports nutrients and waste for the production of new tissue.
The electrostatic spinning technology is a simple and general method for preparing the nano-fiber, and different drugs can be easily loaded into the fiber in the electrostatic spinning process due to the simple and easy drug loading mode, and in addition, the drug can not generate performance change after being loaded into the fiber, can still maintain the performance of the drug, and can be used for preventing postoperative infection or promoting the generation of bones. Therefore, the nanofiber drug-loaded membrane prepared by electrostatic spinning has good clinical application prospect. Meanwhile, the lack or insufficient activity of various growth factors at the bone defect is an important reason for influencing bone regeneration, so that the research on a biological material capable of providing the required active factors for different stages of bone repair is very important.
The Bone defect repair mainly comprises four stages of hematoma and inflammation stage, initial callus reaction stage, cartilage formation stage and Bone repair stage, wherein the side emphasis of each stage of repair is different, and the required growth factors and repair substances are different, until the application date, no Drug loading system capable of providing the growth factors and nutrients for each stage of Bone repair is found, the Materials can regulate the matrix material composition of each layer of membrane material so as to control fiber layer-by-layer degradation, achieve layer-by-layer Drug release, and simultaneously achieve the purpose of multiple and programmed release of different drugs and growth factors required for different stages of Bone formation according to the characteristics of the Bone fracture healing process by the design of four-layer coaxial structure, after loading the Materials into the body, the fiber outermost matrix first starts to degrade while releasing the antibacterial Drug so as to inhibit bacterial infection and inflammation appearing in the blood and inflammation stages, and then the secondary outer matrix starts to release into vascular factors so as to promote the vascularization process at the defect, and finally the inner and innermost matrix once provides the Bone repair with the biological release of the biological dressing, biological, dressing, Bone repair, Bone repair, Bone repair, Bone repair, Bone.
The bone-promoting substances related in the patent mainly refer to hydroxyapatite, graphene oxide, carbon nano tubes, tricalcium phosphate and icariin; the bioglass comprises 45S5, apatite-wollastonite activated glass and machinable bioactive glass; growth factors such as Bone Morphogenetic Proteins (BMPs), Fibroblast Growth Factors (FGFs), transforming growth factors (TGF-BETA), platelet-derived growth factors (PDGFs), Vascular Endothelial Growth Factors (VEGFs) and insulin-like growth factors (IGFs).
Disclosure of Invention
The invention aims to provide a preparation method of a multilayer coaxial fiber bone repair membrane material, which realizes the directional release of antibacterial drugs and bone drugs, can promote the bone defect repair, does not need secondary operation, and can inhibit bacterial infection and inflammation which are easy to occur after the defect occurs.
The multilayer coaxial fiber bone repair membrane material is characterized in that:
(1) the outermost fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural macromolecules as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural macromolecules is 0/100-20/80, and the mass ratio of the antibacterial and anti-inflammatory drugs to the total mass of the mixture of the degradable aliphatic polyester and the degradable natural macromolecules which can be prepared by the outermost matrix material is 1/100-30/100;
(2) the secondary outer layer fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural high polymer as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural high polymer is 20/80-50/50, so that the mass ratio of the bone drug to the total mass of the mixture of degradable aliphatic polyester and degradable natural high polymer which can be obtained by the secondary outer layer matrix material is 1/100-40/100;
(3) the secondary inner layer fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural high polymer as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural high polymer is 50/50-80/20, so that the mass ratio of the bone drug to the total mass of the mixture of degradable aliphatic polyester and degradable natural high polymer which can be obtained by the secondary inner layer matrix material is 1/100-40/100;
(4) the innermost layer fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural high polymer as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural high polymer is 80/20-100/0, so that the mass ratio of the bone drug to the total mass of the mixture of degradable aliphatic polyester and degradable natural high polymer which can be degraded by the innermost layer matrix material is 1/100-40/100;
the degradable aliphatic polyester comprises: one or a mixture of more than two of polylactic acid, polycaprolactone, polylactic acid-glycolic acid copolymer, polylactic acid-caprolactone copolymer and polylactic acid-glycolic acid-caprolactone copolymer; the degradable natural polymer comprises: one or more of type I collagen, gelatin, chitosan, and fibroin.
The medicine loaded on the outermost layer matrix comprises penicillin, cephamycin, tetracycline, chloramphenicol, macrolide, lincomycin, fluoroquinolones, nitroimidazole, polypeptide and quaternary ammonium salt antibacterial medicines, aspirin, indomethacin, naproxen, diclofenac, ibuprofen, nimesulide and celecoxib anti-inflammatory medicines, the medicine loaded on the second outer layer matrix comprises vascular endothelial growth factor VEGF, platelet-derived endothelial growth factor PD-ECGF, heparanase, angiogenin angs, cyclooxygenase COX-2, hypoxia-inducible factor-1, DFO, erythropoietin Epo, β -elemene blood vessel forming medicines, the medicine loaded on the second and innermost layer matrices comprises hydroxyapatite, graphene oxide, carbon nano tube, tricalcium phosphate, icariin, bioglass and/or growth factor, bioglass comprises 45S5, apatite-wollastonite active glass and insulinotropic bioactive glass, and the growth factor is selected from one or more of BMP, TGF, IGF.
The preparation method of the bone repair membrane material comprises the following steps:
(1) dissolving degradable aliphatic polyester in an organic solvent, magnetically stirring at room temperature for 6-12h to obtain a solution A with the mass concentration of the degradable aliphatic polyester being 0.00-0.02g/m L, and obtaining a pure solvent which is the solution A when the content of the degradable aliphatic polyester is 0;
(2) adding degradable natural polymers into the solution A, and magnetically stirring for 6-12h at room temperature to obtain a solution B with the mass concentration of the degradable natural polymers being 0.08-0.10g/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution B is 0/100-20/80;
(3) adding the antibacterial drug 1 into the solution B, and magnetically stirring for 6-12h at room temperature to obtain a solution C with the matrix material concentration of 0.1g/m L, wherein the ratio of the mass of the antibacterial drug 1 in the solution C to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-30/100.
(4) Dissolving the degradable aliphatic polyester in an organic solvent, and magnetically stirring for 6-12h at room temperature to obtain a solution D with the mass concentration of the degradable aliphatic polyester being 0.02-0.05g/m L;
(5) adding degradable natural polymers into the solution D, and magnetically stirring for 6-12h at room temperature to obtain a solution E with the mass concentration of the degradable natural polymers being 0.05-0.08g/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution E is 20/80-50/50;
(6) adding the blood vessel promoting substance 2 into the solution E, and magnetically stirring for 6-12h at room temperature to obtain a solution F with the matrix material mass concentration of 0.10g/m L, wherein the ratio of the mass of the blood vessel promoting substance 2 in the solution F to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-40/100.
(7) Dissolving the degradable aliphatic polyester in an organic solvent, and magnetically stirring for 6-12h at room temperature to obtain a solution G with the mass concentration of the degradable aliphatic polyester being 0.02-0.05G/m L;
(8) adding degradable natural polymers into the solution G, and magnetically stirring for 6-12H at room temperature to obtain a solution H with the mass concentration of the degradable natural polymers being 0.05-0.08G/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution H is 50/50-80/20;
(9) adding bone substance 3 into the solution H, and magnetically stirring at room temperature for 6-12H to obtain solution I with matrix material concentration of 0.1g/m L, wherein the ratio of the mass of the bone substance 3 in the solution I to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-40/100.
(10) Dissolving degradable aliphatic polyester in an organic solvent, and magnetically stirring for 6-12h at room temperature to obtain a solution J with the mass concentration of the degradable aliphatic polyester being 0.08-0.10g/m L;
(11) adding degradable natural polymers into the solution J, magnetically stirring for 6-12h at room temperature to obtain a solution K with the mass concentration of the degradable natural polymers being 0.00-0.02g/m L, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural polymers in the solution K is 80/20-100/0, and when the mass of the degradable natural polymers is 0, the step (11) can be omitted, and the solution K is the solution J substantially;
(12) adding bone substance 4 into the solution K, magnetically stirring at room temperature for 6-12h to obtain solution L with matrix material concentration of 0.1g/m L, wherein the ratio of the mass of the bone substance 4 in the solution L to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-40/100.
(13) Respectively adding the solution C, the solution F, the solution I and the solution L into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 100-600rpm, the flow rate of the innermost layer of the spinning solution is 0.1-1m L/h, the flow rate of the secondary inner layer is 0.5-1.5m L/h, the flow rate of the secondary outer layer is 0.5-1.5m L/h, the flow rate of the outermost layer is 1-3m L/h, the voltage is 15-30kV, the receiving distance is 15-30cm, and the spinning time is 5-30h to obtain an electrospun fiber membrane;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 2-7 days, and then packaged and sterilized.
The invention adopts the electrostatic spinning method to prepare the four-layer coaxial nanofiber bone repair membrane material, but the invention is not limited to the preparation of the bone repair fiber membrane with the structure, and the modification or drug loading and the like on the surface of the bone repair fiber membrane with the structure are all applicable to the invention.
Drawings
FIG. 1 is a schematic illustration of the materials of the present invention;
according to the experimental design, the matrixes of the four layers of coaxial fibers are different, and in the using process, the fibers are degraded layer by layer in the sequence of a first layer, a second layer, a third layer and a fourth layer, and meanwhile, the medicines can be controlled to be released layer by layer in the sequence of antibacterial and anti-inflammatory medicines 1, blood vessel medicines 2, bone medicines 3 and bone medicines 4.
FIG. 2 is an electron microscope picture of the film materials of examples 1 to 4;
as can be seen from the electron microscope pictures, the fibers prepared in examples 1-4 have smooth surfaces without beaded structures, indicating that the drug is well encapsulated in the fibers.
FIG. 3 shows TEM (example 5)
As can be seen from the TEM image of example 5, we successfully produced electrospun nanofibers having a four-layer coaxial structure by using a four-layer coaxial needle.
FIG. 4 is a photograph showing the inhibition of bacteria (Pseudomonas aeruginosa) in examples 6 to 8
The bacteriostatic picture is an experimental picture of a bacteriostatic experiment on the 5 th day, and we can see that the material still has good bacteriostatic performance on the 5 th day. The peak period of inflammation and infection on days 1 to 3 of the occurrence of bone defect, and it was found from the above experiment that the material can help to suppress inflammation and infection in the initial stage of the occurrence of bone defect.
FIG. 5 shows experimental data on cell proliferation of examples 9 to 12
The upper graph is a proliferation experiment picture of MC3T3 cells, and we can see that the proliferation amount of cells on the material is more than 80% compared with that of a blank sample on the 7 th day of proliferation, which indicates that the membrane material can effectively help osteoblasts to adhere and proliferate, and is beneficial to the implementation of an osteogenesis process.
FIG. 6 shows alkaline phosphatase activity data of examples 13 to 14
The production of alkaline phosphatase is a characteristic which is remarkable in the early stage of osteogenesis, and it is understood from the figure that the content of alkaline phosphatase in the experimental groups corresponding to examples 13 and 14 is high, indicating that the material has a good effect of promoting osteogenesis.
FIG. 7 shows osteopontin contents of examples 15 to 16
Osteopontin is a relatively significant marker in the middle and later stages of osteogenesis, and as can be seen from the above figure, the experimental groups corresponding to example 15 and example 16 have relatively high osteopontin content, which further indicates that the material has a good effect on osteogenesis.
Detailed Description
The present invention will be further illustrated with reference to the following examples, but the present invention is not limited to the following examples.
Example 1
(1) Adding 1g of gelatin into 10m of L trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain a solution A with the gelatin mass concentration of 0.1g/m L;
(2) adding 0.01g of ciprofloxacin into the solution A, and magnetically stirring for 6 hours at room temperature to obtain a solution B, wherein the mass ratio of the ciprofloxacin to the gelatin in the solution B is 1/100;
(3) adding 0.2g of polycaprolactone into 10m L trifluoroethanol solvent, and magnetically stirring at room temperature for 12 hours to obtain solution C with the mass concentration of polycaprolactone of 0.02g/m L;
(4) adding 0.8g of gelatin into the solution C, and magnetically stirring at room temperature for 12 hours to obtain a solution D, wherein the mass ratio of the polycaprolactone to the gelatin in the solution D is 20/80;
(5) adding 0.01g of DFO into the solution D, and magnetically stirring for 6 hours at room temperature to obtain a solution E, wherein the ratio of the mass of DFO in the solution E to the total mass of polycaprolactone and gelatin is 1/100;
(6) adding 0.5g of polycaprolactone into 10m L trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain solution F with the mass concentration of polycaprolactone of 0.05g/m L;
(7) adding 0.5G of gelatin into the solution F, and magnetically stirring for 12 hours at room temperature to obtain a solution G, wherein the mass ratio of the polycaprolactone to the gelatin in the solution G is 50/50;
(8) adding 0.01G of hydroxyapatite into the solution G, and magnetically stirring for 6 hours at room temperature to obtain a solution H, wherein the ratio of the mass of the hydroxyapatite in the solution H to the total mass of the polycaprolactone and the gelatin is 1/100;
(9) adding 1g of polycaprolactone into 10m of L trifluoroethanol solvent, and magnetically stirring for 6 hours at room temperature to obtain a solution I with the mass concentration of 0.1g/m L;
(10) adding 0.01g of tricalcium phosphate into the solution I, and magnetically stirring for 6 hours at room temperature to obtain a solution J, wherein the mass ratio of the tricalcium phosphate to the polycaprolactone in the solution J is 1/100;
(11) respectively adding the solution B, the solution E, the solution H and the solution J into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 0.1m L/H, the flow rate of the secondary inner layer is 1m L/H, the flow rate of the secondary outer layer is 1m L/H, the flow rate of the outermost layer is 2m L/H, the voltage is 24kV, the receiving distance is 20cm, and the spinning time is 16H, so that an electrospun fiber membrane is obtained;
(12) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 3 days, and then packaged and sterilized.
Example 2
(1) Adding 0.2g of polycaprolactone into 10m L trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain solution A with the mass concentration of polycaprolactone of 0.02g/m L;
(2) adding 0.8g of gelatin into the solution A, and magnetically stirring for 12 hours at room temperature to obtain a solution B, wherein the mass ratio of the polycaprolactone to the gelatin in the solution B is 20/80;
(3) adding 0.3g of moxifloxacin into the solution B, and magnetically stirring for 6 hours at room temperature to obtain a solution C, wherein the ratio of the mass of moxifloxacin in the solution C to the total mass of polycaprolactone and gelatin is 30/100;
(4) adding 0.5g of polycaprolactone into 10m L trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain solution D with the mass concentration of polycaprolactone of 0.05g/m L;
(5) adding 0.5g of gelatin into the solution D, and magnetically stirring at room temperature for 12 hours to obtain a solution E, wherein the mass ratio of the polycaprolactone to the gelatin in the solution E is 50/50;
(6) adding 0.10g of VEGF into the solution E, and magnetically stirring for 6 hours at room temperature to obtain a solution F, wherein the ratio of the mass of the VEGF in the solution F to the total mass of the polycaprolactone and the gelatin is 10/100;
(7) adding 0.8G of polycaprolactone into 10m L trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain solution G with the mass concentration of polycaprolactone of 0.08G/m L;
(8) adding 0.2G of gelatin into the solution G, and magnetically stirring at room temperature for 12 hours to obtain a solution H, wherein the mass ratio of the polycaprolactone to the gelatin in the solution H is 80/20;
(9) adding 0.20g of icariin into the solution H, and magnetically stirring for 6 hours at room temperature to obtain a solution I, wherein the ratio of the mass of the icariin in the solution I to the total mass of the polycaprolactone and the gelatin is 20/100;
(10) adding 1g of polycaprolactone into 10m of L trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain a solution J with the mass concentration of 0.1g/m L;
(11) adding 0.01g of tricalcium phosphate into the solution J, and magnetically stirring for 6 hours at room temperature to obtain a solution K, wherein the mass ratio of the tricalcium phosphate to the polycaprolactone in the solution K is 1/100;
(12) respectively adding the solution C, the solution F, the solution I and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 0.1m L/h, the flow rate of the secondary inner layer is 0.5m L/h, the flow rate of the secondary outer layer is 0.5m L/h, the flow rate of the outermost layer is 3m L/h, the voltage is 24kV, the receiving distance is 20cm, and the spinning time is 16h to obtain an electrospun fiber membrane;
(13) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 3 days, and then packaged and sterilized.
Example 3
(1) Adding 0.2g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution A with the mass concentration of the polylactic acid of 0.02g/m of L;
(2) adding 0.8g of chitosan into the solution A, and magnetically stirring for 10 hours at room temperature to obtain a solution B, wherein the mass ratio of polylactic acid to chitosan in the solution B is 20/80;
(3) adding 0.3g of penicillin into the solution B, and magnetically stirring for 6 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the penicillin in the solution C to the total mass of the polylactic acid and the chitosan is 30/100;
(4) adding 0.4g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution D with the mass concentration of the polylactic acid of 0.04g/m of L;
(5) adding 0.6g of chitosan into the solution D, and magnetically stirring for 10 hours at room temperature to obtain a solution E, wherein the mass ratio of the polylactic acid to the chitosan in the solution E is 40/60;
(6) adding 0.40g of PE-ECGF into the solution E, and magnetically stirring for 6 hours at room temperature to obtain a solution F, wherein the ratio of the mass of the PE-ECGF to the total mass of the polylactic acid and the chitosan in the solution F is 40/100;
(7) adding 0.6G of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution G with the mass concentration of the polylactic acid of 0.06G/m of L;
(8) adding 0.4G of chitosan into the solution G, and magnetically stirring for 10 hours at room temperature to obtain a solution H, wherein the mass ratio of polylactic acid to chitosan in the solution H is 60/40;
(9) adding 0.4g of graphene oxide into the solution H, and magnetically stirring for 6 hours at room temperature to obtain a solution I, wherein the ratio of the mass of the graphene oxide in the solution I to the total mass of the polylactic acid and the chitosan is 40/100;
(10) adding 0.8g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution J with the mass concentration of the polylactic acid of 0.08g/m of L;
(11) adding 0.2g of chitosan into the solution J, and magnetically stirring for 10 hours at room temperature to obtain a solution K, wherein the mass ratio of the polylactic acid to the chitosan in the solution K is 80/20;
(12) adding 0.01g of apatite-wollastonite activated glass into the solution K, and magnetically stirring for 6 hours at room temperature to obtain L, wherein the ratio of the mass of the apatite-wollastonite activated glass to the total mass of the polylactic acid and the chitosan in the solution L is 1/100;
(13) respectively adding the solution C, the solution F, the solution I and the solution L into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber to carry out electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 1m L/h, the flow rate of the secondary inner layer is 1m L/h, the flow rate of the secondary outer layer is 1m L/h, the flow rate of the outermost layer is 1m L/h, the voltage is 24kV, the receiving distance is 18cm, and the spinning is carried out for 12 hours to obtain an electrospun fiber membrane;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood for 4 days at room temperature, and then packaged and sterilized.
Example 4
(1) Adding 0.1g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 6 hours at room temperature to obtain a solution A with the mass concentration of the polylactic acid of 0.01g/m of L;
(2) adding 0.9g of chitosan into the solution A, and magnetically stirring for 10 hours at room temperature to obtain a solution B, wherein the mass ratio of polylactic acid to chitosan in the solution B is 10/90;
(3) adding 0.01g of doxycycline into the solution B, and magnetically stirring for 6 hours at room temperature to obtain a solution C, wherein the ratio of the mass of doxycycline in the solution C to the total mass of polylactic acid and chitosan is 1/100;
(4) adding 0.3g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 6 hours at room temperature to obtain a solution D with the mass concentration of the polylactic acid of 0.03g/m of L;
(5) adding 0.7g of chitosan into the solution D, and magnetically stirring for 10 hours at room temperature to obtain a solution E, wherein the mass ratio of the polylactic acid to the chitosan in the solution E is 30/70;
(6) adding 0.40g of heparanase into the solution E, and magnetically stirring for 6 hours at room temperature to obtain a solution F, wherein the ratio of the mass of the heparanase in the solution F to the total mass of the polylactic acid and the chitosan is 40/100;
(7) adding 0.6G of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 6 hours at room temperature to obtain a solution G with the mass concentration of the polylactic acid of 0.06G/m of L;
(8) adding 0.4G of chitosan into the solution G, and magnetically stirring for 10 hours at room temperature to obtain a solution H, wherein the mass ratio of polylactic acid to chitosan in the solution H is 60/40;
(9) adding 0.4g of apatite-wollastonite activated glass into the solution H, and magnetically stirring for 6 hours at room temperature to obtain a solution I, wherein the ratio of the mass of the apatite-wollastonite activated glass to the total mass of the polylactic acid and the chitosan in the solution I is 40/100;
(10) adding 0.8g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 6 hours at room temperature to obtain a solution J with the mass concentration of the polylactic acid of 0.08g/m of L;
(11) adding 0.2g of chitosan into the solution J, and magnetically stirring for 10 hours at room temperature to obtain a solution K, wherein the mass ratio of the polylactic acid to the chitosan in the solution K is 80/20;
(12) adding 0.4g of machinable bioactive glass into the solution K, and magnetically stirring for 6 hours at room temperature to obtain a solution L, wherein the ratio of the mass of the machinable bioactive glass in the solution L to the total mass of the polylactic acid and the chitosan is 40/100;
(13) respectively adding the solution C, the solution F, the solution I and the solution L into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 0.1m L/h, the flow rate of the secondary inner layer is 0.5m L/h, the flow rate of the secondary outer layer is 0.5m L/h, the flow rate of the outermost layer is 3m L/h, the voltage is 24kV, the receiving distance is 18cm, and the spinning time is 18h, so that an electrospun fiber membrane is obtained;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood for 4 days at room temperature, and then packaged and sterilized.
Example 5
(1) Adding 1g of chitosan into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution A with the mass concentration of chitosan of 0.1g/m L;
(2) adding 0.3g of cefixime into the solution A, and magnetically stirring for 6 hours at room temperature to obtain a solution B, wherein the mass ratio of the cefixime in the solution B to the chitosan is 30/100;
(3) adding 0.2g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution C with the mass concentration of the polylactic acid of 0.02g/m of L;
(4) adding 0.8g of chitosan into the solution C, and magnetically stirring for 10 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid to the chitosan in the solution D is 20/80;
(5) adding 0.01g of angiogenin into the solution D, and magnetically stirring for 6 hours at room temperature to obtain a solution E, wherein the ratio of the mass of the angiogenin to the total mass of the polylactic acid and the chitosan in the solution E is 1/100;
(6) adding 0.7g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution F with the mass concentration of the polylactic acid of 0.07g/m of L;
(7) adding 0.3G of chitosan into the solution F, and magnetically stirring for 10 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid to the chitosan in the solution G is 70/30;
(8) adding 0.01G of 45S5 bioglass powder into the solution G, and magnetically stirring for 6 hours at room temperature to obtain a solution H, wherein the ratio of the mass of 45S5 bioglass to the total mass of polylactic acid and chitosan in the solution H is 1/100;
(9) adding 0.9g of polylactic acid into 10m of L N, N-dimethylformamide solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution I with the mass concentration of the polylactic acid of 0.09g/m of L;
(10) adding 0.1g of chitosan into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J, wherein the mass ratio of polylactic acid to chitosan in the solution J is 90/10;
(11) adding 0.01g of TGF-B into the solution J, and magnetically stirring for 6 hours at room temperature to obtain a solution K, wherein the ratio of the mass of TGF-BETA in the solution K to the total mass of polylactic acid and chitosan is 1/100;
(12) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 0.1m L/H, the flow rate of the secondary inner layer is 1.5m L/H, the flow rate of the secondary outer layer is 1.5m L/H, the flow rate of the outermost layer is 2m L/H, the voltage is 24kV, the receiving distance is 18cm, and the spinning time is 5H to obtain an electrospun fiber membrane;
(13) after the electrostatic spinning is finished, the spinning film is placed in a fume hood for 4 days at room temperature, and then packaged and sterilized.
Example 6
(1) Adding 0.1g of L-polylactic acid into 10m L hexafluoroisopropanol solvent, and magnetically stirring for 6h at room temperature to obtain solution A with the mass concentration of the L-polylactic acid being 0.01g/m L;
(2) adding 0.9g of type I collagen into the solution C, and magnetically stirring for 10 hours at room temperature to obtain a solution B, wherein the mass ratio of the levorotatory polylactic acid to the type I collagen in the solution B is 10/90;
(3) adding 0.01g of tacrolimus into the solution B, and magnetically stirring for 6 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the tacrolimus in the solution C to the total mass of the L-polylactic acid and the type I collagen is 1/100;
(4) adding 0.3g of L-polylactic acid into 10Ml of hexafluoroisopropanol solvent, and magnetically stirring for 6h at room temperature to obtain a solution D with the mass concentration of the L-polylactic acid being 0.03g/m L;
(5) adding 0.7g of type I collagen into the solution D, and magnetically stirring for 10 hours at room temperature to obtain a solution E, wherein the mass ratio of the levorotatory polylactic acid to the type I collagen in the solution E is 30/70;
(6) adding 0.40g of IGF into the solution E, and magnetically stirring for 6h at room temperature to obtain a solution F, wherein the ratio of the mass of the IGF in the solution F to the total mass of the L-polylactic acid and the type I collagen is 40/100;
(7) adding 0.6G of L-polylactic acid into 10m L hexafluoroisopropanol solvent, and magnetically stirring for 6h at room temperature to obtain solution G with the mass concentration of the L-polylactic acid being 0.06G/m L;
(8) adding 0.4G of type I collagen into the solution G, and magnetically stirring for 10 hours at room temperature to obtain a solution H, wherein the mass ratio of the levorotatory polylactic acid to the type I collagen in the solution H is 60/40;
(9) adding 0.4g of bone drug bioglass 45S5 into the solution H, and magnetically stirring for 6 hours at room temperature to obtain a solution I, wherein the ratio of the mass of the bioglass 45S5 in the solution I to the total mass of the L-polylactic acid and the type I collagen is 40/100;
(10) adding 0.8g of L-polylactic acid into 10m L hexafluoroisopropanol, and magnetically stirring for 6h at room temperature to obtain a solution J with the mass concentration of the L-polylactic acid being 0.08g/m L;
(11) adding 0.2g of type I collagen into the solution J, and magnetically stirring for 10 hours at room temperature to obtain a solution K, wherein the mass ratio of the levorotatory polylactic acid to the type I collagen in the solution K is 80/20;
(12) dissolving 0.01g of BMP-2 and 20mg of BSA in 500 mu L of deionized water to obtain a solution L;
(13) adding the solution L into the solution K, magnetically stirring for 6h at room temperature, and performing ultrasonic treatment for 5min to obtain a solution M, wherein the ratio of the mass of BMP-2 in the solution M to the total mass of the L-polylactic acid and the type I collagen is 1/100;
(14) respectively adding the solution C, the solution F, the solution I and the solution M into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 0.1M L/h, the flow rate of the secondary inner layer is 0.5M L/h, the flow rate of the secondary outer layer is 0.5M L/h, the flow rate of the outermost layer is 3M L/h, the voltage is 24kV, the receiving distance is 18cm, and the spinning time is 18h, so that an electrospun fiber membrane is obtained;
(15) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 2 days, and then packaged and sterilized.
Example 7
(1) Adding 1.0g of chitosan into 10ml of trifluoroethanol solvent, and magnetically stirring at room temperature for 12 hours to obtain a solution A with the mass concentration of chitosan being 0.10 g/ml;
(2) adding 0.02g of metronidazole into the solution A, and magnetically stirring at room temperature for 12 hours to obtain a solution B, wherein the mass ratio of the metronidazole to the chitosan in the solution B is 2/100;
(3) adding 0.25g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution C with the polylactic acid mass concentration of 0.025 g/ml;
(4) adding 0.75g of chitosan into the solution C, and magnetically stirring for 6 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid to the chitosan in the solution D is 25/75;
(5) adding 0.01g of angiogenin into the solution D, and magnetically stirring for 6 hours at room temperature to obtain a solution E, wherein the mass ratio of the angiogenin to the total mass of the polylactic acid and the chitosan in the solution E is 1/100;
(6) adding 0.5g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution F with the mass concentration of the polylactic acid of 0.05 g/ml;
(7) adding 0.5G of chitosan into the solution F, and magnetically stirring for 6 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid to the chitosan in the solution G is 50/50;
(8) adding 0.05G of TGF- α into the solution G, and uniformly stirring to obtain a solution H with the total polymer mass concentration of 0.10G/ml, wherein the ratio of the mass of TGF- α in the solution H to the total mass of polylactic acid and chitosan is 5/100;
(9) adding 0.75g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring at room temperature for 12 hours to obtain a solution I with the polylactic acid mass concentration of 0.075 g/ml;
(10) adding 0.25g of chitosan into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J;
(11) adding 0.4g of FGF into the solution J, and uniformly stirring to obtain a solution K with the total mass concentration of the high polymers being 0.10g/ml, wherein the ratio of the mass of the FGF in the solution K to the total mass of the polylactic acid and the chitosan is 40/100;
(12) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 600rpm, the flow rate of the innermost layer of the spinning solution is 1m L/H, the flow rate of the secondary inner layer is 1.5m L/H, the flow rate of the secondary outer layer is 1.5m L/H, the flow rate of the outermost layer is 3m L/H, the voltage is 30kV, the receiving distance is 15cm, and the spinning time is 5H, so that an electrospun fiber membrane is obtained;
(13) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 5 days, and then packaged and sterilized.
Example 8
(1) Adding 0.1g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 6h at room temperature to obtain a solution A with the mass concentration of the polylactic acid of 0.01 g/ml;
(2) adding 0.9g of fibroin into the solution A, and magnetically stirring for 6 hours at room temperature to obtain a solution B, wherein the mass ratio of the polylactic acid to the fibroin in the solution B is 10/90;
(3) adding 0.2g of quaternary ammonium salt into the solution B, and magnetically stirring for 6 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the quaternary ammonium salt in the solution C to the total mass of the polylactic acid and the fibroin is 20/100;
(4) adding 0.4g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring at room temperature for 12 hours to obtain a solution C with the polylactic acid mass concentration of 0.04 g/ml;
(5) adding 0.6g of fibroin into the solution C, and magnetically stirring for 12 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid to the fibroin in the solution D is 40/60;
(6) adding β -elemene 0.3g into the solution D, and magnetically stirring at room temperature for 12h to obtain a solution E, wherein the ratio of the mass of the β -elemene in the solution E to the total mass of the polylactic acid and the fibroin is 30/100;
(7) adding 0.6g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution F with the mass concentration of the polylactic acid of 0.06 g/ml;
(8) adding 0.4G of fibroin into the solution F, and magnetically stirring for 9 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid to the fibroin in the solution G is 60/40;
(9) adding 0.2G of graphene oxide into the solution G, and uniformly stirring to obtain a solution H with the total mass concentration of high molecules of 0.10G/ml, wherein the total mass ratio of the graphene oxide to the polylactic acid and the fibroin in the solution H is 60/40;
(10) adding 0.8g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 9h at room temperature to obtain solution I with the mass concentration of the polylactic acid of 0.08g/ml
(11) Adding 0.2g of fibroin into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J, wherein the mass ratio of the polylactic acid to the fibroin in the solution J is 80/20;
(12) adding 0.2g of carbon nano tubes into the solution J, and uniformly stirring to obtain a solution K, wherein the mass ratio of the carbon nano tubes in the solution K to the total mass of the polylactic acid and the fibroin is 20/100;
(13) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 200rpm, the flow rate of the innermost layer of the spinning solution is 0.1m L/H, the flow rate of the secondary inner layer is 1m L/H, the flow rate of the secondary outer layer is 1m L/H, the flow rate of the outermost layer is 2m L/H, the voltage is 28kV, the receiving distance is 15cm, and the spinning time is 15H, so that an electrospun fiber membrane is obtained;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 6 days, and then packaged and sterilized.
Example 9
(1) Taking 0.05g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 7 hours at room temperature to obtain a solution A with the mass concentration of the polycaprolactone of 0.005 g/ml;
(2) adding 0.95g of chitosan into the solution A, and magnetically stirring for 7 hours at room temperature to obtain a solution B, wherein the mass ratio of polycaprolactone to chitosan in the solution B is 5/95;
(3) adding 0.02g of aspirin into the solution B, and magnetically stirring for 7 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the aspirin to the total mass of the polycaprolactone and the chitosan in the solution C is 2/100;
(4) taking 0.3g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution C with the mass concentration of the polycaprolactone of 0.03 g/ml;
(5) adding 0.7g of chitosan into the solution C, and magnetically stirring for 9 hours at room temperature to obtain a solution D, wherein the mass ratio of polycaprolactone to chitosan in the solution D is 30/70;
(6) adding 0.15g of DFO into the solution D, and magnetically stirring for 9 hours at room temperature to obtain a solution E, wherein the ratio of the mass of DFO in the solution E to the total mass of polycaprolactone and chitosan is 15/100;
(7) taking 0.7g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 11 hours at room temperature to obtain a solution F with the mass concentration of the polycaprolactone of 0.07 g/ml;
(8) adding 0.3G of chitosan into the solution F, and magnetically stirring for 11 hours at room temperature to obtain a solution G, wherein the mass ratio of polycaprolactone to chitosan in the solution G is 70/30;
(9) adding 0.11G of BMP-2 into the solution G, and uniformly stirring to obtain a solution H, wherein the ratio of the mass of the BMP-2 in the solution H to the total mass of the polycaprolactone and the chitosan is 11/100;
(10) adding 0.95g of polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring at room temperature for 9 hours to obtain a solution I with the polycaprolactone mass concentration of 0.095g/ml
(11) Adding 0.05g of chitosan into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J;
(12) adding 0.24g of IGF into the solution J, and uniformly stirring to obtain a solution K, wherein the ratio of the mass of the IGF in the solution K to the total mass of the polycaprolactone and the chitosan is 24/100;
(13) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 400rpm, the flow rate of the innermost layer of the spinning solution is 0.3m L/H, the flow rate of the secondary inner layer is 0.6m L/H, the flow rate of the secondary outer layer is 0.9m L/H, the flow rate of the outermost layer is 1.2m L/H, the voltage is 27kV, the receiving distance is 26cm, and the spinning time is 10 hours to obtain an electrospun fiber membrane;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 2 days, and then packaged and sterilized.
Example 10
(1) Taking 0.05g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 7 hours at room temperature to obtain a solution A with the mass concentration of the polycaprolactone of 0.01 g/ml;
(2) adding 0.95g of fibroin into the solution A, and magnetically stirring for 7 hours at room temperature to obtain a solution B, wherein the mass ratio of polycaprolactone to fibroin in the solution B is 5/95;
(3) adding 0.1g of naproxen into the solution B, and magnetically stirring for 7 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the naproxen to the total mass of the polycaprolactone and the fibroin in the solution C is 10/100;
(4) taking 0.3g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution C with the mass concentration of the polycaprolactone of 0.03 g/ml;
(5) adding 0.7g of fibroin into the solution C, and magnetically stirring for 9 hours at room temperature to obtain a solution D, wherein the mass ratio of polycaprolactone to fibroin in the solution D is 30/70;
(6) adding 0.36g of DFO into the solution D, and magnetically stirring for 9 hours at room temperature to obtain a solution E, wherein the ratio of the mass of DFO in the solution E to the total mass of polycaprolactone and fibroin is 36/100;
(7) taking 0.7g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 11 hours at room temperature to obtain a solution F with the mass concentration of the polycaprolactone of 0.07 g/ml;
(8) adding 0.3G of fibroin into the solution F, and magnetically stirring for 11 hours at room temperature to obtain a solution G, wherein the mass ratio of polycaprolactone to fibroin in the solution G is 70/30;
(9) adding 0.08G of apatite-wollastonite activated glass into the solution G, and uniformly stirring to obtain a solution H, wherein the mass ratio of the apatite-wollastonite activated glass to the total mass of the polycaprolactone and the fibroin in the solution H is 8/100;
(10) adding 0.95g of polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring at room temperature for 9 hours to obtain a solution I with the polycaprolactone mass concentration of 0.095g/ml
(11) Adding 0.05g of fibroin into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J;
(12) adding 0.19g of hydroxyapatite into the solution J, and uniformly stirring to obtain a solution K, wherein the ratio of the mass of the hydroxyapatite in the solution K to the total mass of the polycaprolactone and the fibroin is 19/100;
(13) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 500rpm, the flow rate of the innermost layer of the spinning solution is 0.4m L/H, the flow rate of the secondary inner layer is 0.8m L/H, the flow rate of the secondary outer layer is 0.8m L/H, the flow rate of the outermost layer is 2m L/H, the voltage is 30kV, the receiving distance is 22cm, and the spinning time is 14H to obtain an electrospun fiber membrane;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood for 4 days at room temperature, and then packaged and sterilized.
Example 11
(1) Adding 1g of type I collagen into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution A with the mass concentration of the type I collagen of 0.1 g/ml;
(2) adding 0.02g of nimesulide into the solution A, and magnetically stirring for 3 hours at room temperature to obtain a solution B with the concentration of a base material of 0.1g/ml, wherein the mass ratio of nimesulide to type I collagen in the solution B is 2/100;
(3) taking 0.75g of polylactic acid-glycolic acid-caprolactone copolymer, adding into 10ml of trifluoroethanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution C with the mass concentration of the polylactic acid-glycolic acid-caprolactone copolymer of 0.75 g/ml;
(4) adding 0.25g of type I collagen into the solution C, and magnetically stirring for 9 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid-glycolic acid-caprolactone copolymer to the type I collagen in the solution D is 75/25;
(5) adding 0.02g of angiogenin into the solution D, and magnetically stirring for 6 hours at room temperature to obtain a solution E, wherein the ratio of the mass of the angiogenin in the solution E to the total mass of the polylactic acid-glycolic acid-caprolactone copolymer and the type I collagen is 2/100;
(6) adding 0.5g of polylactic acid-glycolic acid-caprolactone copolymer into 9.5ml of trifluoroethanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution F;
(7) adding 0.5G of type I collagen into the solution F, and magnetically stirring for 9 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid-glycolic acid-caprolactone copolymer to the type I collagen in the solution G is 50/50;
(8) adding 0.46ml of IGF and BSA aqueous solution (IGF: 0.02 g; BSA: 80mg) to the solution H, adding 40. mu.l of Span80, and magnetically stirring at room temperature for 10min to obtain a solution I, wherein the ratio of the mass of the IGF in the solution I to the total mass of the polylactic acid-glycolic acid-caprolactone copolymer and the type I collagen is 2/100;
(9) adding 1g of polylactic acid-glycolic acid-caprolactone copolymer into 10ml of trifluoroethanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution J with the mass concentration of the polylactic acid-glycolic acid-caprolactone copolymer of 0.10 g/ml;
(10) adding 0.4g of FGF into the solution J, and performing ultrasonic treatment for 20min to obtain a solution K, wherein the mass ratio of the FGF in the solution K to the polylactic acid-glycolic acid-caprolactone copolymer is 40/100;
(11) respectively adding the solution B, the solution E, the solution I and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber to carry out electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 300rpm, the flow rate of the innermost layer of the spinning solution is 0.1ml/h, the flow rate of the secondary inner layer is 0.5ml/h, the flow rate of the secondary outer layer is 0.8ml/h, the flow rate of the outermost layer is 1.7ml/h, the voltage is 27kV, the receiving distance is 30cm, and the spinning time is 26h to obtain an electrospun fiber membrane;
(12) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 5 days, and then packaged and sterilized.
Example 12
(1) Adding 1g of type I collagen into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution A with the mass concentration of the type I collagen of 0.1 g/ml;
(2) adding 0.3g of pentamycin into the solution A, and magnetically stirring for 3 hours at room temperature to obtain a solution B with the concentration of a matrix material of 0.1g/ml, wherein the mass ratio of the pentamycin to the I-type collagen in the solution B is 30/100;
(3) taking 0.5g of polylactic acid-glycolic acid-caprolactone copolymer, adding into 10ml of trifluoroethanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution C with the mass concentration of the polylactic acid-glycolic acid-caprolactone copolymer of 0.05 g/ml;
(4) adding 0.5g of type I collagen into the solution C, and magnetically stirring for 9 hours at room temperature to obtain a solution D with the total mass concentration of high molecules of 0.1g/ml, wherein the mass ratio of the polylactic acid-glycolic acid-caprolactone copolymer to the type I collagen in the solution D is 50/50;
(5) adding 0.4g of Desferrioxamine (DFO) into the solution D, and magnetically stirring for 6h at room temperature to obtain a solution E with the total polymer mass concentration of 0.10g/ml, wherein the ratio of the mass of the Desferrioxamine (DFO) in the solution E to the total mass of the polylactic acid-glycolic acid-caprolactone copolymer and the type I collagen is 40/100;
(6) adding 0.75g of polylactic acid-glycolic acid-caprolactone copolymer into 9.5ml of trifluoroethanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution F;
(7) adding 0.25G of type I collagen into the solution F, and magnetically stirring for 9 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid-glycolic acid-caprolactone copolymer to the type I collagen in the solution G is 75/25;
(8) adding 0.46ml of TGF-B and BSA aqueous solution (TGF-B: 0.35 g; BSA: 80mg) into the solution H, adding 40 mul of Span80, and magnetically stirring at room temperature for 10min to obtain a solution I, wherein the ratio of the mass of the TGF-B in the solution I to the total mass of the polylactic acid-glycolic acid-caprolactone copolymer and the type I collagen is 35/100;
(9) adding 1g of polylactic acid-glycolic acid-caprolactone copolymer into 10ml of trifluoroethanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution J with the mass concentration of the polylactic acid-glycolic acid-caprolactone copolymer of 0.10 g/ml;
(10) adding 0.07g of tricalcium phosphate into the solution J, and performing ultrasonic treatment for 20min to obtain a solution K, wherein the mass ratio of the tricalcium phosphate to the polylactic acid-glycolic acid-caprolactone copolymer in the solution K is 7/100;
(11) respectively adding the solution B, the solution E, the solution I and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber to carry out electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 300rpm, the flow rate of the innermost layer of the spinning solution is 0.1ml/h, the flow rate of the secondary inner layer is 0.5ml/h, the flow rate of the secondary outer layer is 1.0ml/h, the flow rate of the outermost layer is 3ml/h, the voltage is 28kV, the receiving distance is 20cm, and the spinning time is 18h to obtain an electrospun fiber membrane;
(12) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 3 days, and then packaged and sterilized.
Example 13
(1) Taking 0.2g of polylactic acid-caprolactone copolymer, adding into 10ml of hexafluoroisopropanol solvent, and magnetically stirring for 8 hours at room temperature to obtain a solution A with the mass concentration of the polylactic acid-caprolactone copolymer of 0.04 g/ml;
(2) adding 0.8g of chitosan into the solution A, and magnetically stirring for 8 hours at room temperature to obtain a solution B, wherein the mass ratio of the polylactic acid-caprolactone copolymer to the chitosan in the solution B is 20/80;
(3) adding 0.3g of ibuprofen into the solution B, and magnetically stirring for 10 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the ibuprofen in the solution C to the total mass of the polylactic acid-caprolactone copolymer and the chitosan is 30/100;
(4) taking 0.35g of polylactic acid-caprolactone copolymer, adding into 10ml of hexafluoroisopropanol solvent, and magnetically stirring for 12 hours at room temperature to obtain a solution C with the mass concentration of the polylactic acid-caprolactone copolymer being 0.035 g/ml;
(5) adding 0.65g of chitosan into the solution C, and magnetically stirring for 9 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid-caprolactone copolymer to the chitosan in the solution D is 35/65;
(6) adding 0.31g of epoxidase into the solution D, and magnetically stirring for 8 hours at room temperature to obtain a solution E, wherein the ratio of the mass of the epoxidase in the solution E to the total mass of the polylactic acid-caprolactone copolymer and the chitosan is 31/100;
(7) taking 0.65g of polylactic acid-caprolactone copolymer, adding the polylactic acid-caprolactone copolymer into 10ml of hexafluoroisopropanol solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution F with the mass concentration of the polylactic acid-caprolactone copolymer of 0.65 g/ml;
(8) adding 0.35G of chitosan into the solution F, and magnetically stirring for 10 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid-caprolactone copolymer to the chitosan in the solution G is 65/35;
(9) adding 0.37G of TGF-B into the solution G, and uniformly stirring to obtain a solution H, wherein the ratio of the mass of the TGF-B in the solution H to the total mass of the polylactic acid-caprolactone copolymer and the chitosan is 37/100;
(10) taking 0.8g of polylactic acid-caprolactone copolymer, adding into 10ml of hexafluoroisopropanol solvent, magnetically stirring for 9h at room temperature to obtain solution I with the mass concentration of the polylactic acid-caprolactone copolymer being 0.08g/ml
(11) Adding 0.2g of chitosan into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J, wherein the mass ratio of the polylactic acid-caprolactone copolymer to the chitosan in the solution J is 80/20;
(12) adding 0.18g of tricalcium phosphate into the solution J, and uniformly stirring to obtain a solution K, wherein the mass ratio of the tricalcium phosphate to the total mass of the polylactic acid-caprolactone copolymer and the chitosan is 18/100;
(13) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 500rpm, the flow rate of the innermost layer of the spinning solution is 0.2m L/H, the flow rate of the secondary inner layer is 0.8m L/H, the flow rate of the secondary outer layer is 0.8m L/H, the flow rate of the outermost layer is 1m L/H, the voltage is 21kV, the receiving distance is 24cm, and the spinning time is 23H to obtain an electrospun fiber membrane;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 3 days, and then packaged and sterilized.
Example 14
(1) Taking 0.2g of polylactic acid-glycolic acid copolymer, adding into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 8 hours at room temperature to obtain a solution A with the mass concentration of the polylactic acid-glycolic acid copolymer being 0.02 g/ml;
(2) adding 0.8g of fibroin into the solution A, and magnetically stirring for 8 hours at room temperature to obtain a solution B, wherein the mass ratio of the polylactic acid-glycolic acid copolymer to the fibroin in the solution B is 20/80;
(3) adding 0.3g of diclofenac into the solution B, and magnetically stirring for 10 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the diclofenac in the solution C to the total mass of the polylactic acid-glycolic acid copolymer and the fibroin is 30/100;
(4) taking 0.35g of polylactic acid-glycolic acid copolymer, adding into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 12 hours at room temperature to obtain a solution C with the mass concentration of the polylactic acid-glycolic acid copolymer being 0.035 g/ml;
(5) adding 0.65g of fibroin into the solution C, and magnetically stirring for 9 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid-glycolic acid copolymer to the fibroin in the solution D is 35/65;
(6) adding 0.4g of heparanase into the solution D, and magnetically stirring for 8 hours at room temperature to obtain a solution E, wherein the ratio of the mass of the heparanase in the solution E to the total mass of the polylactic acid-glycolic acid copolymer and the fibroin is 40/100;
(7) taking 0.65g of polylactic acid-glycolic acid copolymer, adding into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 10 hours at room temperature to obtain a solution F with the mass concentration of the polylactic acid-glycolic acid copolymer being 0.65 g/ml;
(8) adding 0.35G of fibroin into the solution F, and magnetically stirring for 10 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid-glycolic acid copolymer to the fibroin in the solution G is 65/35;
(9) adding 0.4G of TGF-B into the solution G, and uniformly stirring to obtain a solution H, wherein the ratio of the mass of the TGF-B in the solution H to the total mass of the polylactic acid-glycolic acid copolymer and the fibroin is 40/100;
(10) taking 0.8g of polylactic acid-glycolic acid copolymer, adding into 10ml of N, N-dimethyl amide solvent, magnetically stirring for 9h at room temperature to obtain solution I with the mass concentration of the polylactic acid-glycolic acid copolymer being 0.08g/ml
(11) Adding 0.2g of fibroin into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J, wherein the mass ratio of the polylactic acid-glycolic acid copolymer to the fibroin in the solution J is 80/20;
(12) adding 0.4g of icariin into the solution J, and uniformly stirring to obtain a solution K, wherein the ratio of the mass of the icariin in the solution K to the total mass of the polylactic acid-glycolic acid copolymer and the fibroin is 40/100;
(13) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 500rpm, the flow rate of the innermost layer of the spinning solution is 0.2m L/H, the flow rate of the secondary inner layer is 0.6m L/H, the flow rate of the secondary outer layer is 0.8m L/H, the flow rate of the outermost layer is 2m L/H, the voltage is 28kV, the receiving distance is 23cm, and the spinning time is 28H, so that an electrospun fiber membrane is obtained;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood for 4 days at room temperature, and then packaged and sterilized.
Example 15
(1) Taking 0.1g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 6 hours at room temperature to obtain a solution A with the mass concentration of the polycaprolactone of 0.01 g/ml;
(2) adding 0.9g of type I collagen into the solution A, and magnetically stirring for 6 hours at room temperature to obtain a solution B, wherein the mass ratio of the polycaprolactone to the type I collagen in the solution B is 10/90;
(3) adding 0.01g of indomethacin into the solution B, and magnetically stirring for 6 hours at room temperature to obtain a solution C, wherein the ratio of the mass of the indomethacin in the solution C to the total mass of the polycaprolactone and the type I collagen is 1/100;
(4) taking 0.4g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 12 hours at room temperature to obtain a solution C with the mass concentration of the polycaprolactone of 0.04 g/ml;
(5) adding 0.6g of type I collagen into the solution C, and magnetically stirring for 12 hours at room temperature to obtain a solution D, wherein the mass ratio of the polycaprolactone to the type I collagen in the solution D is 40/60;
(6) adding 0.01g of hypoxia inducible factor-1 into the solution D, and magnetically stirring for 12 hours at room temperature to obtain a solution E, wherein the ratio of the mass of the hypoxia inducible factor-1 in the solution E to the total mass of the polycaprolactone and the type I collagen is 1/100;
(7) taking 0.6g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution F with the mass concentration of the polycaprolactone of 0.06 g/ml;
(8) adding 0.4G of type I collagen into the solution F, and magnetically stirring for 9 hours at room temperature to obtain a solution G, wherein the mass ratio of the polycaprolactone to the type I collagen in the solution G is 60/40;
(9) adding 0.01G of carbon nano tubes into the solution G, and uniformly stirring to obtain a solution H with the total mass concentration of the high polymer of 0.10G/ml, wherein the ratio of the mass of the carbon nano tubes in the solution H to the total mass of the polycaprolactone and the type I collagen is 1/100;
(10) taking 0.95g of polycaprolactone, adding the polycaprolactone into 10ml of N, N-dimethyl amide solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution I with the mass concentration of the polycaprolactone of 0.95 g/ml;
(11) adding 0.05g of type I collagen into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J, wherein the mass ratio of the polycaprolactone to the type I collagen in the solution J is 95/5;
(12) adding 0.01g of machinable bioactive glass into the solution J, and uniformly stirring to obtain a solution K, wherein the ratio of the mass of the machinable bioactive glass in the solution K to the total mass of the polycaprolactone and the type I collagen is 1/100;
(13) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 200rpm, the flow rate of the innermost layer of the spinning solution is 0.2m L/H, the flow rate of the secondary inner layer is 0.7m L/H, the flow rate of the secondary outer layer is 0.9m L/H, the flow rate of the outermost layer is 1.1m L/H, the voltage is 22kV, the receiving distance is 24cm, and the spinning time is 25H, so that an electrospun fiber membrane is obtained;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 3 days, and then packaged and sterilized.
Example 16
(1) Adding 1.0g of gelatin into 10ml of trifluoroethanol solvent, and magnetically stirring at room temperature for 12 hours to obtain a solution A with the gelatin mass concentration of 0.10 g/ml;
(2) adding 0.25g of celecoxib into the solution A, and magnetically stirring for 12 hours at room temperature to obtain a solution B, wherein the mass ratio of the celecoxib to the gelatin in the solution B is 25/100;
(3) adding 0.25g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution C with the mass concentration of the polylactic acid of 0.25 g/ml;
(4) adding 0.75g of gelatin into the solution C, and magnetically stirring for 6 hours at room temperature to obtain a solution D, wherein the mass ratio of the polylactic acid to the gelatin in the solution D is 25/75;
(5) adding 0.01g of PD-ECGF into the solution D, and magnetically stirring for 6 hours at room temperature to obtain a solution E, wherein the mass ratio of the PD-ECGF in the solution E to the total mass of the polylactic acid and the gelatin is 1/100;
(6) adding 0.5g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring for 9 hours at room temperature to obtain a solution F with the mass concentration of the polylactic acid of 0.05 g/ml;
(7) adding 0.5G of gelatin into the solution F, and magnetically stirring for 6 hours at room temperature to obtain a solution G, wherein the mass ratio of the polylactic acid to the gelatin in the solution G is 50/50;
(8) adding 0.35G of carbon nano tubes into the solution G, and uniformly stirring to obtain a solution H, wherein the ratio of the mass of the carbon nano tubes in the solution H to the total mass of the polylactic acid and the gelatin is 35/100;
(9) adding 0.8g of polylactic acid into 10ml of trifluoroethanol solvent, and magnetically stirring at room temperature for 12h to obtain a solution I with the mass concentration of the polylactic acid of 0.08g/ml
(10) Adding 0.2g of gelatin into the solution I, and magnetically stirring for 10 hours at room temperature to obtain a solution J, wherein the mass ratio of the polycaprolactone to the gelatin in the solution J is 80/20;
(11) adding 0.01g of IGF into the solution J, and magnetically stirring for 10min at room temperature to obtain a solution K with the total mass concentration of the high molecules being 0.10g/ml, wherein the ratio of the mass of the IGF in the solution K to the total mass of the polylactic acid and the gelatin is 1/100;
(12) respectively adding the solution B, the solution E, the solution H and the solution K into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 100rpm, the flow rate of the innermost layer of the spinning solution is 0.1m L/H, the flow rate of the secondary inner layer is 0.1m L/H, the flow rate of the secondary outer layer is 0.1m L/H, the flow rate of the outermost layer is 0.5m L/H, the voltage is 30kV, the receiving distance is 20cm, and the spinning time is 30H, so that an electrospun fiber membrane is obtained;
(13) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 7 days, and then packaged and sterilized.
The following is a drug release chart of the examples
Application example
The application part is as follows: mild to moderate bone defects in the skull, ulna, radius, femur, etc
The application method comprises the following steps: the bone defect part and the surrounding tissues are separated cleanly, tiny bone fragments at the defect part are removed, the defect part is fixed and then is wrapped by a film material with proper tightness, so that the purposes of preventing the tissues from growing into the affected part and providing nutrient substances and growth factors required by defect repair for the defect part are achieved.
The application effect is as follows: on days 1-3 after the defect occurred: the antibacterial and anti-inflammatory drugs are released firstly, so that the possible inflammation or infection at the defect part can be effectively prevented; then releasing a blood vessel promoting drug, and basically completing the reconstruction of the blood vessel at the defect position from day 3 to day 20, so that the blood vessel can play a role of transporting required nutrients for the defect position and discharging metabolic waste; after day 20, the bone drug was allowed to be released slowly, cells were induced to differentiate osteogenically, and new bone was essentially formed after day 40.
Claims (4)
1. The multilayer coaxial fiber bone repair membrane material is characterized in that:
(1) the outermost fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural macromolecules as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural macromolecules is 0/100-20/80, and the mass ratio of the antibacterial and anti-inflammatory drugs to the total mass of the mixture of the degradable aliphatic polyester and the degradable natural macromolecules which can be prepared by the outermost matrix material is 1/100-30/100;
(2) the secondary outer layer fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural high polymer as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural high polymer is 20/80-50/50, so that the mass ratio of the bone drug to the total mass of the mixture of degradable aliphatic polyester and degradable natural high polymer which can be obtained by the secondary outer layer matrix material is 1/100-40/100;
(3) the secondary inner layer fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural high polymer as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural high polymer is 50/50-80/20, so that the mass ratio of the bone drug to the total mass of the mixture of degradable aliphatic polyester and degradable natural high polymer which can be obtained by the secondary inner layer matrix material is 1/100-40/100;
(4) the innermost layer fiber matrix takes a mixture of degradable aliphatic polyester and degradable natural high polymer as a matrix material, wherein the mass ratio of the degradable aliphatic polyester to the degradable natural high polymer is 80/20-100/0, so that the mass ratio of the bone drug to the total mass of the mixture of degradable aliphatic polyester and degradable natural high polymer which can be degraded by the innermost layer matrix material is 1/100-40/100.
2. The multilayer coaxial fiber bone repair membrane material of claim 1, wherein the degradable aliphatic polyester comprises: one or a mixture of more than two of polylactic acid, polycaprolactone, polylactic acid-glycolic acid copolymer, polylactic acid-caprolactone copolymer and polylactic acid-glycolic acid-caprolactone copolymer; the degradable natural polymer comprises: one or more of type I collagen, gelatin, chitosan, and fibroin.
3. The multilayer coaxial fiber bone repair membrane material of claim 1, wherein the drug loaded into the outermost matrix comprises penicillin, cephamycins, tetracyclines, chloramphenicol, macrolides, lincomycin, fluoroquinolones, nitroimidazoles, polypeptides and quaternary ammonium salt antibacterial drugs, aspirin, indomethacin, naproxen, diclofenac, ibuprofen, nimesulide, celecoxib anti-inflammatory drugs, the drug loaded into the second outer matrix comprises vascular endothelial growth factor VEGF, platelet derived endothelial growth factor PD-ECH, heparanase, angiogenin angs, cyclooxygenase COX-2, hypoxia inducible factor-1, DFO, erythropoietin Epo, β -elemene angiogenins, the drug loaded into the second and innermost matrices comprises hydroxyapatite, graphene oxide, carbon nanotubes, tricalcium phosphate, ovine glycosides, bioglasses and/or growth factors, the bioglasses comprises 45S5, phospho-active glass, bioactive glass factors such as bioactive glass cutting factor, osteogenic growth factor, insulin growth factor, IGF-like, or one or more transforming growth factors.
4. A method of preparing a multilayer coaxial fibrous bone repair membrane material according to any one of claims 1 to 3, characterized by the steps of:
(1) dissolving degradable aliphatic polyester in an organic solvent, magnetically stirring at room temperature for 6-12h to obtain a solution A with the mass concentration of the degradable aliphatic polyester being 0.00-0.02g/m L, and obtaining a pure solvent which is the solution A when the content of the degradable aliphatic polyester is 0;
(2) adding degradable natural polymers into the solution A, and magnetically stirring for 6-12h at room temperature to obtain a solution B with the mass concentration of the degradable natural polymers being 0.08-0.10g/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution B is 0/100-20/80;
(3) adding the antibacterial drug 1 into the solution B, and magnetically stirring for 6-12h at room temperature to obtain a solution C with the matrix material concentration of 0.1g/m L, wherein the ratio of the mass of the antibacterial drug 1 in the solution C to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-30/100;
(4) dissolving the degradable aliphatic polyester in an organic solvent, and magnetically stirring for 6-12h at room temperature to obtain a solution D with the mass concentration of the degradable aliphatic polyester being 0.02-0.05g/m L;
(5) adding degradable natural polymers into the solution D, and magnetically stirring for 6-12h at room temperature to obtain a solution E with the mass concentration of the degradable natural polymers being 0.05-0.08g/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution E is 20/80-50/50;
(6) adding a blood vessel promoting substance 2 into the solution E, and magnetically stirring for 6-12h at room temperature to obtain a solution F with the matrix material mass concentration of 0.10g/m L, wherein the ratio of the mass of the blood vessel promoting substance 2 in the solution F to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-40/100;
(7) dissolving the degradable aliphatic polyester in an organic solvent, and magnetically stirring for 6-12h at room temperature to obtain a solution G with the mass concentration of the degradable aliphatic polyester being 0.02-0.05G/m L;
(8) adding degradable natural polymers into the solution G, and magnetically stirring for 6-12H at room temperature to obtain a solution H with the mass concentration of the degradable natural polymers being 0.05-0.08G/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution H is 50/50-80/20;
(9) adding bone substance 3 into the solution H, and magnetically stirring at room temperature for 6-12H to obtain solution I with a matrix material concentration of 0.1g/m L, wherein the ratio of the mass of the bone substance 3 to the total mass of the degradable aliphatic polyester and the degradable natural polymer in the solution I is 1/100-40/100;
(10) dissolving degradable aliphatic polyester in an organic solvent, and magnetically stirring for 6-12h at room temperature to obtain a solution J with the mass concentration of the degradable aliphatic polyester being 0.08-0.10g/m L;
(11) adding degradable natural polymers into the solution J, magnetically stirring for 6-12h at room temperature to obtain a solution K with the mass concentration of the degradable natural polymers being 0.00-0.02g/m L, wherein the mass ratio of the degradable aliphatic polyesters to the degradable natural polymers in the solution K is 80/20-100/0, and when the mass of the degradable natural polymers is 0, the step (11) is omitted, and the solution K is the solution J substantially;
(12) adding bone substance 4 into the solution K, magnetically stirring at room temperature for 6-12h to obtain solution L with matrix material concentration of 0.1g/m L, wherein the ratio of the mass of the bone substance 4 in the solution L to the total mass of the degradable aliphatic polyester and the degradable natural polymer is 1/100-40/100;
(13) respectively adding the solution C, the solution F, the solution I and the solution L into propellers corresponding to the outermost layer, the secondary outer layer, the secondary inner layer and the innermost layer of the fiber for electrostatic spinning, taking a stainless steel roller as a receiving device, wherein the rotation speed of the roller is 100-600rpm, the flow rate of the innermost layer of the spinning solution is 0.1-1m L/h, the flow rate of the secondary inner layer is 0.5-1.5m L/h, the flow rate of the secondary outer layer is 0.5-1.5m L/h, the flow rate of the outermost layer is 1-3m L/h, the voltage is 15-30kV, the receiving distance is 15-30cm, and the spinning time is 5-30h to obtain an electrospun fiber membrane;
(14) after the electrostatic spinning is finished, the spinning film is placed in a fume hood at room temperature for 2-7 days, and then packaged and sterilized.
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| CN108144116A (en) * | 2018-02-26 | 2018-06-12 | 丁文铃 | A kind of novel antibacterial artificial ligament |
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| CN108744061B (en) * | 2018-06-28 | 2021-05-11 | 广州贝奥吉因生物科技有限公司 | BMP-2-loaded strontium-doped hydroxyapatite/silk fibroin/heparin composite scaffold and preparation method and application thereof |
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