摘要
背景:研究表明,骨髓间充质干细胞经骨形态发生蛋白诱导可分化为成骨细胞。目的:应用骨髓间充质干细胞、牛骨形态发生蛋白与纤维蛋白胶混合物作为骨移植材料,通过与单纯使用纤维蛋白胶、牛骨形态发生蛋白和骨髓间充质干细胞比较来评价这种移植材料的成骨效果。设计:对比观察实验。单位:广东省第二人民医院,中山大学附属第二医院。材料:实验于2004-01/2006-06在中山大学附属第二医院完成。健康新西兰大白兔由中山大学医学院动物实验中心提供。方法:自兔髂骨抽取骨髓,采用密度梯度离心法在体外培养扩增自体骨髓间充质干细胞。在纤维蛋白胶内加入骨形态发生蛋白制成复合物,再加入骨髓间充质干细胞制成另一种复合物。对12只新西兰大白兔经腹膜外行腰椎间盘切除脊柱融合术。每只动物取L3-63个椎间隙随机接受4种移植方法中的3种:骨髓间充质干细胞+骨形态发生蛋白+纤维蛋白胶、骨髓间充质干细胞+纤维蛋白胶、骨形态发生蛋白+纤维蛋白胶、纤维蛋白胶。术后3月利用放射学和组织学方法观察分析脊柱融合情况。主要观察指标:兔脊柱椎间隙骨痂形成情况。结果:①术后12周时,骨髓间充质干细胞+骨形态发生蛋白+纤维蛋白胶组椎间隙骨痂形成明显较其它组多,组织学上有连续的骨痂形成。骨髓间充质干细胞+纤维蛋白胶、骨形态发生蛋白+纤维蛋白胶组可见前纵韧带后方有编织骨形成,骨量少,不连续。纤维蛋白胶组见纤维组织增生及少量成骨细胞,未见新骨形成。各组均未见纤维蛋白胶残留,椎间盘中有少量椎间盘髓核残余、中间有少量软骨组织形成。②12周时x射线检查显示,各组兔均未造成椎体后缘增生,无椎管狭窄。骨髓间充质干细胞+纤维蛋白胶组节段活动度与骨形态发生蛋白+纤维蛋白胶组、纤维蛋白胶组差异显著(P<0.05);各组间椎体后缘高度丢失差异不显著。结论:自体骨髓间充质干细胞、纤维蛋白胶和骨形态发性蛋白复合体可在椎间关节形成骨性骨痂,融合效果好,组织学方面评价也较其他方法好。
BACKGROUND： Several studies have demonstrated that bone marrow mesenchymal stem cells （MSCs） can differentiate into osteoblasts after induction of bone morphogenetic protein （BMP）. OBJECTIVE： This study was designed to compare the osteogenetic effects between MSCs, bBMP, and fibrin gel alone and their composite. DESIGN： A controlled observation experiment. SETTING： Second People＇s Hospital of Guangdong Province and Second Hospital Affiliated to Sun Yat-sen University. MATERIALS： This study was performed at the Second Hospital Affiliated to Sun Yat-sen University between January 2004 and June 2006. Healthy New Zealand rabbits were provided by the Laboratory Animal Center, Sun Yat-sen University. METHODS： Bone marrow was taken from the iliac bone. By density gradient centrifugation, autologous MSCs were in vitro cultured and amplified. A composite was made of fibrin gel and bovine bone morphogenetic protein （bBMP）. After adding MSCs, another composite was made. Twelve rabbits underwent lumbar L3/4, L4/5, and L5/6 discectomy and interbody fusion via an anterior retroperitoneal approach. Three fusion sites in each animal were randomly assigned into three of four treatments： composite of MSCs and bBMP and fibrin gel （group A）, composite of MSCs and fibrin gel （group B）, composite of bBMP and fibrin gel （group C） and fibrin gel alone （group D）. The interbodies were observed radiologically and histologically 3 months after surgery. MAIN OUTCOME MEASURES： Bone formation in the spinal intervertebral space of rabbits. RESULTS： Twelve weeks after surgery, bone formation was appasently greater in the Group A than in the other groups. Histological findings revealed continuous bone formation. The groups B and C showed equivalent radiological and histological changes. Bone formation was intermittent. Group D had fibrous union, but no bone formation. No residual fibrin gel was found in all groups, but a small amount of residual nucleus gelatinosus with a little cartilaginous tissue was visible in the intervertebral disc. X-ray examination results did not reveal vertebral posterior hyperplasia or spinal stenosis. There was significant difference in range of lumbar motion between the group B and the groups C and D （P 〈 0.05）. No pronounced difference in vertebral posterior height loss existed among the groups. CONCLUSION： The composite of MSCs, fibrin gel and bBMP can promote bone regeneration and achieve osseous spinal fusion.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第34期6763-6766,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
the Natural Science Foundation of Guangdong Province,No.5001754~~
