Experimental Study On Lumber Intertransverse Process Spinal Fusion With Tissue Engineered Bone (TEB) Constructed By Xenogeneic Deproteinized Cancellous Bone Scaffold

Objective (1) To study the physical–chemical and mechanical property of xenogeneic deproteinized cancellous bone. (2) Cellular compatibility and biological action of MSCs on xenogeneic deproteinized cancellous bone. (3) To explore the intensity of immune reaction sparked by xenogeneic deproteinized cancellous bone implanted at intertransverse. (4) lumber intertransverse process spinal fusion in goals with TEB constructed by xenogeneic deproteinized cancellous bone.Methods (1) we made deproteinized bone derived from pig femoral cancellous bone by physical and chemical treatments. Their morphological features, constitute components and mechanical properties were examined by scanning electron microscopy,x-rays diffraction analysis,mechanical experimental instrument and so on. (2) 5 ml bone marrow harvested aseptically from ilium of goal was applied to density gradient centrifugation to isolate MSCs. Then the cells were cultured-expanded in Dulbecco modified Eagle medium plus 10% fetal bovine serum. When the cells were 90% confluency, they were passaged. At the second passage, the MSCs were cultivated for 14 days in the condition of 1×10-7mol/L dexamethasone, 1×10-2mol/Lβ-glycerophosphate, and 50mg/L ascorbic acid, to be induced into osteoblastic cells, which were then labeled for 1 hour with 5-bromo-2 deoxyuridine (Brdu) by delivering them into serum-free DMEM containing 0.25% Brdu. Eventually, these labeled cells were transferred at a sum of 2×106 cells onto xenogeneic deproteinized cancellous bone. They were incubated for 7 days at 37 degrees Celsiusin in a humidified 5 percent carbon-dioxide environment. The cells-material complex was observed under Inverted phase contrast microscope in order to evaluate the adhesion and growth of osteoblasts. cell viability, ALP activity were measured, cell cycle was analysed by flow cytometer. (3) 8 male goals were divided into experimental and controlled group according to transplant materials. materials include xenogeneic deproteinized cancellous bone and fresh xenogeneic cancellous bone. T lymphocyte subsets CD3+,CD4+,CD8+ and CD28+ in...