3D Printing Tissue Engineered Artificial Bone And Biological Performance Research

Objective: Preparation of nano-hydroxyapatite(n-HA)/polycaprolactone(PCL)tissue engineered artificial bone with three-dimensional porous structure based on 3D printing technology.The characterization of tissue engineered artificial bone,its biological performance in vivo and in vitro,and its ability to repair bone defects in vivo are preliminarily evaluated.Methods:1.Preparation and characterization of 3D printing tissue engineered artificial bone.Using different ratios of nano-hydroxyapatite(nHA)/polycaprolactone(PCL)as a matrix,prepare nano-hydroxyapatite(nHA)/polycaprolactone(PCL)with a three-dimensional porous structure based on 3D printing technology.Characterize the physical and chemical properties,mechanical strength,composition and structural characteristics of artificial bone.2.Research on the biological properties of rabbit bone marrow mesenchymal stem cells combined with tissue engineered artificial bone in vitro.Cultivate rabbit bone marrow mesenchymal stem cells and inoculate them on tissue engineered artificial bone for in vitro culture,observe cell morphology and adhesion.Culture rabbit bone marrow mesenchymal stem cells in the extract of tissue engineering artificial bone medium,and pass the cell proliferation experiment,cell mineralization experiment to evaluate the biological performance of tissue artificial bone in vitro.3.Research on the biological properties of rabbit bone marrow mesenchymal stem cells combined with tissue engineered artificial bone in vivo.The physiological saline extract of tissue engineered artificial bone was injected into rabbits,and the biological safety of tissue engineered artificial bone in vivo was analyzed through pyrogen test and acute toxicity test.Through the establishment of a rabbit femoral bone defect model,the rabbit bone marrow mesenchymal stem cell composite tissue engineering artificial bone was implanted in the bone defect.At the 8th week after operation,the biological performance and bone defect repair ability of rabbit bone marrow mesenchymal stem cell composite tissue engineered artificial bone were preliminarily evaluated through specimen observation,X-ray detection,and histological examination.Result: 1.Preparation and characterization of 3D printing tissue engineered artificial bone.Preparation of nano-hydroxyapatite(n-HA)/polycaprolactone(PCL)tissue engineered artificial bone with three-dimensional porous structure based on 3D printing technology,through characterization and analysis,SEM shows that nanohydroxyapatite On the surface of the tissue engineered artificial bone,a loose and dispersed microporous structure with different pore sizes is formed.EDS spectrometer,FTIR,XRD,TGA indicate that nano-hydroxyapatite and polycaprolactone are physically combined in the composite artificial bone.At the same time,the chemical structure of the material will not change during the preparation process.The hydrophilicity and porosity of tissue engineered artificial bone increase with the increase of nano-hydroxyapatite content(P<0.05),and the mechanical properties of compressive strength increase with the increase of polycaprolactone content in the composition(P <0.05).2.Research on the biological properties of rabbit bone marrow mesenchymal stem cells combined with tissue engineered artificial bone in vitro.Through cell morphology observation,cell proliferation experiments and cell mineralization experiments,it was found that after 7 days of cultured rabbit bone marrow mesenchymal stem cells on nanohydroxyapatite/polycaprolactone tissue engineering artificial bone,the extracellular matrix covered the tissue engineering artificial bone.Bone,cell proliferation is remarkable.The results of cell proliferation experiments showed that with the extension of culture time,rabbit bone marrow mesenchymal stem cells showed higher proliferation ability in the composite artificial bone extract containing hydroxyapatite(P<0.05).A large number of calcium nodules were formed in the cell mineralization experiment,indicating that the nano-hydroxyapatite/polycaprolactone tissue engineered artificial bone has good cytocompatibility and the ability to promote bone differentiation for rabbit bone marrow mesenchymal stem cells.3.Research on the biological properties of rabbit bone marrow mesenchymal stem cells combined with tissue engineered artificial bone in vivo.Pyrogen test and acute toxicity test found no significant increase in body temperature performance,the acute toxicity test of the heart,liver,kidney HE staining was found in bone tissue engineering acute toxic change in the body,indicating biocompatible artificial bone tissue engineering It has good performance and meets the product standards of safe biological materials.In the rabbit femoral large segment bone defect model,the results of specimen observation,X-ray detection,and histological examination at the 8th week after surgery showed that the rabbit bone marrow mesenchymal stem cell composite tissue engineered artificial bone was found in the implanted bone defect.New bone was formed and bone defects were repaired well,indicating that the nano-hydroxyapatite/polycaprolactone tissue engineered artificial bone has good bone defect repair capabilities.Conclusion: 1.In this research,a nano-hydroxyapatite(n-HA)/polycaprolactone(PCL)issue engineered artificial bone with suitable microporous structure was prepared by3 D printing technology.Through the characterization and analysis of tissue engineering artificial bone,the results show that it has good physical and chemical properties,certain mechanical strength,and has composition and structural characteristics close to natural bone.2.Research on the biological properties of rabbit bone marrow mesenchymal stem cells combined with tissue engineered artificial bone in vitro.The results show that it has the ability to support the adhesion and proliferation of bone marrow mesenchymal stem cells,and has the ability to promote the osteogenic differentiation of bone marrow mesenchymal stem cells.Possess good cytocompatibility and osteoinductivity.3.Research on the biological properties of rabbit bone marrow mesenchymal stem cells combined with tissue engineered artificial bone in vivo.That is its biological safety and bone defect repair ability in animal models.The results show that it has relatively good biocompatibility in vivo and in vitro,is safe and has no significant toxicity,and has good bone defect repair capabilities,which lays a good experimental foundation for further experimental research.