北京生物医学工程

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

作者：李练兵张其清侯志伟朱一剑张丹妍吕静赵乐天杨智曦朱明华马明福

单位：                      重庆市人口和计划生育技术研究院国家卫生计生委出生缺陷与生殖健康重点实验室(重庆 400020)        

关键词：胶原/羟基磷灰石；骨形态发生蛋白；骨细胞培养；骨缺损

分类号：

出版年·卷·期（页码）：2014·33·3（240-246）

摘要：

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CHA-BMP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 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 rabbit radius defect site after 2, 4, 8, 16, 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 16 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.

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