Experimental Studies On The Construction And Neovascularation Of The Nano Cellular Tissue Engineering Bone

With the appearance and development of bone tissue engineering, it maybe become the new and ideal therapy of the bone defect. With the more and more achievement has been harvested about the seeding cells and the scaffolds, the cellular tissue engineering bone has been constructed successfully in vitro. Moreover the experiments have been attempt to repair the bone defects by the cellular tissue engineering bone. But it is important for the tissue engineering bone to be vascularized , just the same as the blood circulation is necessary for the normal skeleton. Therefore, it is the key point for the tissue engineering bone to have the vascularization, when it is to be transplanted in vivo. Now our research explored the construction and neovascularization of the tissue engineering bone about five works.PART I: The experimental study on the inducing rabbits bone marrow stem cells into osteoblast in vitro.Objective To study the bone tissue osteogenesis ability of bone marrow stem cells (BMSCs) in vitro in order to offer the ideal seed cells for tissue engineering.Methods The BMSCs were separated by gradient centrifugation on percoll from rabbit bone marrow, then observing and testing the biologic characters of BMSCs induced by culturing with a conditional medium for thirty days.Results Osteoblast can be induced from BMSCs by conditional medium, moreover can secret ALP and format the calcific nodules.Conclusion BMSCs can be induced successfully to osteoblast in vitro and can be used as ideal seed cells for bone tissue engineering.PART II : The Study on Cultuting and Facial Type of Renal Microvascularl Endothelial Cell in VitroObjective To explore the methods of culturing and identify the renal microvascularExperimental Studies on the construction and neovascularation of the nano cellular tissue engineering bone Abstractendothelial cell in vitro and study it's facial type in orther to offer the ideal seeding cell for bone tissue engineer.Methods first,to acquire the highly pured glomeruli by three steps grads filtrated net; next,to culture the microvascular endothelial cell in vitro with the transplant methods of isolated glomerali. moreover,the renal microvascular endothelial cell were identified by irnmunohistochemical SP methods and observed by transmission electron microscope and studied the CD31 - CD3^ CD44 antigen by indirect immunofluorescence methods.Results The highly pured renal microvascular endothelial cell were cultured in vitro and maintained eleven passages .and factor VIII related antigen were positive by immunohistochemical SP methods, and Weibel-Palade bodies were observed by transmission electron microscope and it has been shown that CD31 , CD34 were positive and CD44 was negative with indirect immunofluorescence methods.Conclusion Renal microvascular endothelial cell can be cultured successfully in vitro by this methods, and CD31, CD34 antigen have been identified on the renal microvascular endothelial cell.PART III : The study on the influence of biological characteristics of osteoblasts cocultured with endothelial cells in vitro.Objects To explore the influence of biological characteristics of osteoblasts cocultured with endothelial cells in vitro.Methods Osteoblasts induced from BMSCs were cocultured with renal microvascular endothelial cells directively or indirectively, osteogenesis were detecte by inverted microscope, cell count method, alkaline phosphatase activity assay respectively.Results The proliferation of the two kinds cells are all increased significantly than the single cell was cultured whenever cocultured directively or indirectively; The ALP activity of osteoblast was also increased significantly than without culturing with endothelial cells.Conclusions There were good cellular compatibility between the osteoblasts and endothelial cells when the two kinds cells were cocultrued in vitro, moreover, enhance the proliferation and the functions of the two kinds cells.Experimental Studies on the construction and neovascularation of the nano cellular...