Experimental Study On Applicating Porites In Construction Of Tissue Engineered Bone

Tissue engineering is an interdisciplinary science of co-development and combination of modem cell biology, biomaterial science and engineering. The establishment of tissue engineering is a breakthrough over conventional organ transplantation to restore the function and structure of tissues lost to the injury and disease. Tissue engineering, embryonic stem cell and genetic engineering together are regarded as the three marvels in life science development history.Bone tissue has been successfully reconstructed in lab. In the light of present knowledge, the formation and repair of bone with tissue engineering require study on: (1)a source of pluripotentional mesenchymal stem cells with strongly ability of proliferation and differentiation; (2)immunology of tissue engineered bone and establishment of osteoblast bank; (3)introduce the metal rack to bone tissue engineering. The present study is based on these requirements and contains three main parts.Parti Biok>g^cal characteristic research of marnnvmesencfavnial stem ceOs l.Cuttrvation and identification of human mesenchymal stem cells Objective To create the model of human mesenchymal stem cells (HMSCs) separation and culture in vitro. Methods Adult bone marrow about 20ml were drawed by puncture. After centrifuging, HMSCs were harvested, cultivated aoi transferred The growm and the characteristics of HMSCs were observed through cell morphobgical surey, cell growth and proKferationand ALP activity. The cells of fourth passage were cultured continuously in mineral condition for 30 days, the mineral nodules were observed by Von-Kossa staining. Results HMSCs were cultured in vitro and could be transferred for 5-7 passages steadily. The cells of fourth passage were ALP positive. When cultured up to 30 days, \bn-Kossa staining showed there was calcium deposition in the nodules. Conclusion The HMSCs were cultured successfully in vitro and can be induced to osteoblast cells. It wfll be a kind of excellent seeding-cells for bone tissue engineering.ZJScanning electron microscope study of osteoblast cultivated with coral in vitro Objective To investigate osteoblasts growth and adhesion to natural coral Methods Tencuboid porous Xisha Islands coral ( 5mm X 5mm X 5mm) were processed Then they were cultivated in vitro simultaneously with osteoblast derived from mesenchymal stem cells of rabbit for 2 days, 4 days, 7 days, 14 days and 28days.The morphogenetic pattern and number of anchored cells, capability of cleavage and proliferation, and osteoblasts function were observed under scanning electron microscope. Results From 2 days, cells grew into the porous coral The number of cells enhanced greatly with time lasting. To 14-28 days, cells overgrew and many secretory collagen fiber bundles and mineralized nods were observed Coadusioo Xisha Islands natural coral have good histocompatibility with the osterblasts as carrier of tissue engineering bone in vitro.3.The biological effects of combination of BMP and bPGF to bone mesenchymal stems cellsObjective To investigate the effects of BMP and bFGF on bone mesenchymal stems cells transformation and protiferatioa Methods Subcuturing the 4th passage of BMSCs into 96 pore plate and exposed to rhBMP2 and (or) bFGF of different concentratioa Cell proliferation, alkaline phosphate activity (ALP) activity and total cellular protein were measured Results The results clearly demonstrated that bFGF could promote the proliferation of BMSCs, but the content of ALP and total celhuar protein were suppressed RhBMP2 could enhance total cellular protein of BMSCs significantly at bw dose in a dose responsible manner, and increase cell proliferation. Synergistic action to the proliferation and differentiation of BMSCs was found Conclusion BFGF and BMP can enhance the BMSCs differentiated into osteoblast It indicates that BMSCs accompanied with cooperation of them might be an applicable method for bone defect repair. Part II Construction of tissue engineered bone and immunological study l.Tissue engineering bone with higher porosity po...