复合人工骨材料对骨形成影响的实验研究

Experimental study on the effects of artificial bone composite materials on bone formation

目的探讨胶原/羟基磷灰石(hydroxyapatile-collagen,CHA)-骨形态发生蛋白(bone morphogenetic protein,BMP)复合材料的体外成骨效应,以及体内修复兔桡骨缺损的效果。方法采用体外细胞培养技术,将人胚成骨细胞分别与纯羟基磷灰石(hydroxyapatile,HA)、CHA、HA-BMP和CHABMP材料于体外联合培养,观察细胞的生长情况。测定细胞冻融中的细胞增殖率,及碱性磷酸酶(alkline phosphatase,ALP)、骨钙素(osteocalcin,OC)的分泌水平。小鼠股部肌袋内植入,评价其异位骨诱导能力。然后在兔桡骨缺损部位植入,术后2周、4周、8周、16周、24周观察新骨形成规律。结果胶原/羟基磷灰石-骨形态发生蛋白复合材料的细胞增殖率、ALP、OC水平明显高于相应单一材料,异位诱导可见软骨及骨组织,ALP呈阳性。桡骨植入8周、16周时X射线显示骨断端形成紧密的纤维连接,有少量骨痂形成,镜下可见大量成熟板层骨及骨髓组织填充,免疫组织化学(BMP、Ⅰ型胶原、骨胶原)均呈阳性反应,非BMP复合材料未见骨或软骨形成。结论 CHA-BMP复合材料有利于成骨细胞早期黏附、生长,小鼠异位骨诱导,加速兔缺损桡骨的新骨形成,从而促进骨愈合,是一种较理想的骨修复材料。

Objective To explore the osteogenic effect of collagen/hydroxyapatite(CHA)composite of bone morphogenetic protein (BMP)material in vitro and in repairing rabbit radius defect effect.Methods With <br> the technique of cell culture in vitro,the human embryo osteoblast was cultured in vitro with HA,CHA,HA-BMP and CHA-BMP materials respectively and was observed the cell growth in the materials.The cell proliferation rate and secretion of ALP and OC were detected in cell frozen solution.The ability to induce ectopic bone was evaluated in the implant of mouse femoral muscle pouches.The laws of new bone formation were observed in the implantation of <br> rabbit radius defect site after 2,4,8,1 6,24 weeks.Results Cell proliferation rate and the ALP and OC levels of CHA-BMP were significantly higher than the corresponding single material.Ectopic cartilage and bone tissue were induced visible and ALP was positive.X light displayed that bone stump formed dense fiber connection with a small amount of callus formation after 8 and 1 6 weeks.A lot of mature lamellar bone and marrow tissue were filled in the microscope.The immunohistochemistry of BMP,type I collagen and collagen were positive, and there were not bone or cartilage formation in non-BMP composites. Conclusions CHA is beneficial to early adhesion of bone cells,bone morphogenetic protein composite growth,mouse heterotopic bone induction and the acceleration of new bone formation in rabbit radius defect,which is an ideal material for bone repair.