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The Effect of Combinaterial Application of BMP-2 and BMP-7 on Osteoblasts Biological Activies in Rat

Author: YiDongSheng
Tutor: LiJie
School: Dalian Medical University
Course: Surgery
Keywords: Osteoblast Bone morphogenetic protein Alkalinity phos phatase Cell increment Bone calciumelement
CLC: R68
Type: Master's thesis
Year: 2009
Downloads: 103
Quote: 0
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Abstract


Objective:①The aim of this study was to develop a simple and rapid way to culture a large number of pure osteoblasts with high survival rate,which would be a foundation of bone tissue engineering.②The effects of osteoblast`s biology activity were observed with using incorporation of BMP-2 and BMP-7 .Methods:①5 days old SD rats received an aseptic surgery to get its calvaria bone.An improved second enzymatic digestion method were used to separate osteoblasts. To observe the osteoblast activity distribution,the inverted phase contrast microscope observation, alkaline phosphatase staining, mineralized nodule identification and micro-titration (MTT) method were used after a culture cycle.②Cells were cultured with BMP-2 and BMP-7 respectively and bothly.After 48h,MTT was used to assay cell proliferation,and the ALP activity and OCN content were determined from cell lysate.Results:①The inverted phase contrast microscope morphological observation showed that non-adherent cells were round, and the growth of adherent cells were irregular, fusiform, triangular or polygonal.Some of them were monocytes,with 1 ~ 3 nucleoli;The stain of alkaline phosphate acid enzymes and calcified nodule stained was positive (positive rate of≥98.06%);MTT method and morphological observation showed that 10 d was the strongest activity time after osteoblasts were cultured.②The neonatal rat osteoblast proliferation was stimulated with using BMP-2 and BMP-7,so was the incorporation of BMP-2 and BMP-7.osteoblast ALP activity and OCN contents were significantly increased after using BMP-2 and BMP-7 respectively and bothly. Treatment with BMP-2 and BMP-7HA resulted in signifcant improvement when compared to BMP-2 and BMP-7 respectively treatmented group (P<0.05)Conclusions:①These results suggest that the modified secondary enzymatic digestion can be able to access a large number of osteoblasts in a short time,and the cultured cells were with typical osteoblast morphology and function.②The biological activity of osteoblasts was significantly enhanced with incorporation of BMP-2 and BMP-7,which means more replantable cells could be provide for bone defects,nonunion repair,promote fracture healing,and spinal fusion, that methods could also provide a foundation for bone tissue engineering.

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CLC: > Medicine, health > Surgery > Orthopaedic Surgery ( movement system diseases,orthopedic surgery )
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