北京生物醫(yī)學(xué)工程

復(fù)合人工骨材料對骨形成影響的實驗研究

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

作者：李練兵張其清侯志偉朱一劍張丹妍呂靜趙樂天楊智曦朱明華馬明福

單位：                      重慶市人口和計劃生育技術(shù)研究院國家衛(wèi)生計生委出生缺陷與生殖健康重點實驗室(重慶 400020)        

關(guān)鍵詞：膠原/羥基磷灰石；骨形態(tài)發(fā)生蛋白；骨細(xì)胞培養(yǎng)；骨缺損

分類號：

出版年·卷·期（頁碼）：2014·33·3（240-246）

摘要：

目的探討膠原/羥基磷灰石(hydroxyapatile-collagen, CHA)-骨形態(tài)發(fā)生蛋白(bone morphogenetic
protein, BMP)復(fù)合材料的體外成骨效應(yīng)，以及體內(nèi)修復(fù)兔橈骨缺損的效果。方法采用體外細(xì)胞培養(yǎng)技術(shù)，
將人胚成骨細(xì)胞分別與純羥基磷灰石(hydroxyapatile, HA)、CHA、HA-BMP和CHA-BMP材料于體外聯(lián)合培養(yǎng)，
觀察細(xì)胞的生長情況。測定細(xì)胞凍融中的細(xì)胞增殖率，及堿性磷酸酶(alkline phosphatase, ALP)、骨鈣素
(osteocalcin, OC)的分泌水平。小鼠股部肌袋內(nèi)植入，評價其異位骨誘導(dǎo)能力。然后在兔橈骨缺損部位植
入，術(shù)后2周、4周、8周、16周、24周觀察新骨形成規(guī)律。結(jié)果膠原/羥基磷灰石-骨形態(tài)發(fā)生蛋白復(fù)合材料
的細(xì)胞增殖率、ALP、OC水平明顯高于相應(yīng)單一材料，異位誘導(dǎo)可見軟骨及骨組織，ALP呈陽性。橈骨植入8
周、16周時X射線顯示骨斷端形成緊密的纖維連接，有少量骨痂形成，鏡下可見大量成熟板層骨及骨髓組織
填充，免疫組織化學(xué)(BMP、I型膠原、骨膠原)均呈陽性反應(yīng)，非BMP復(fù)合材料未見骨或軟骨形成。結(jié)論 CHA
-BMP復(fù)合材料有利于成骨細(xì)胞早期黏附、生長，小鼠異位骨誘導(dǎo)，加速兔缺損橈骨的新骨形成，從而促進(jìn)骨
愈合，是一種較理想的骨修復(fù)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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