应用骨髓基质细胞及组织工程方法修复兔桡骨实验性缺损(英文)

Bone marrow stromal cell and tissue engineering in repairing experimental bone defects of rabbit radius

背景:应用干细胞移植及组织工程的方法修复骨缺损是近年来研究的热点。目的:观察应用自体骨髓基质细胞的组织工程骨修复骨缺损的效果。设计:左右侧对照实验。单位:哈尔滨医科大学第一临床医学院实验中心。材料:10～个月新西兰大白兔12只,雌雄不限,体质量2～2.5kg。方法:实验于2002-06/2003-06在哈尔滨医科大学第一临床医学院实验中心完成。将兔自体骨髓基质细胞经淋巴细胞分离液分离培养、传代扩增。取兔12只行桡骨中段截取1.5cm骨质,造成完全骨缺损。于左侧缺损处植入载有骨髓基质细胞的胶原海绵,右侧植入单纯胶原海绵对照。12周后麻醉状态处死动物,比较基质细胞和胶原海绵复合物移植与单纯胶原移植治疗骨缺损的疗效。X射线片评价标准按骨缺损修复分期方法(分为0～5级,5级为骨缺损全部被新骨替代,0级为无新骨修复)。主要观察指标:兔桡骨缺损处大体观察,X射线观察,组织化学观察及电镜观察结果。结果:纳入12只兔均进入结果分析。①两侧兔桡骨缺损处大体观察:第12周实验组骨缺损处骨痂坚实突出于骨缺损处,与断端连续性好。对照侧无连续骨痂通过骨缺损处,断端间有纤维组织连接。②两侧兔桡骨缺损处X射线观察:在第12周实验侧有连续骨痂通过骨缺损处,髓腔通畅,塑形不完全。对照侧无连续骨痂通过断端。③两侧兔桡骨缺损处组织学观察:第12周实验侧骨缺损处可见大量成骨细胞及新生基质。对照侧仅在断端间见少量骨细胞,断端处为纤维组织充填。④兔桡骨缺损电镜观察:实验侧成骨细胞接近正常骨细胞可见大量丰富扩张内质网,蛋白合成旺盛,细胞器含量丰富。结论:骨髓基质细胞作为一种骨源干细胞,具有良好的成骨作用,经过扩增可以用作种子细胞,通过组织工程的方法可以修复骨缺损性疾病。

BACKGROUND：Stem cell transplantation and tissue engineering in re- pairing bone defects is a hotspots of recent study. OBJECTIVE：To observe the therapeutic effect of engineering repair on bone defect by auto-transplantation of bone marrow stromal cells（BMSCs） DESIGN：Left-right comparative study SETTING：Experimental center of the First Affiliated Hospital of Harbin Medical University MATERIALS ：Twelve New Zealand rabbits with birth age of 10 days to 2 months were selected,male or female with body mass of 2 to 2.5 kg. METHODS ：The experiment was conducted in the Experimental Center of First Clinieal Medical College, Harbin Medical University from June 2002 to June 2003. Self-BMSCs were separated for subculture. 1.5 cm bone was intercepted at middle of radius in 12 rabbits so as to simulate complete bone defect. Then, the left radius defect was filled with collagen sponge carrying BMSCs （ experimental side）,which was replaced by simple collagen sponge in the right side（control side）. Twelve weeks later, rabbits were put to death and the outcomes of both sides were compared.X-ray assessment was accorded to the standardized stage of bone defect repair （bone repair was graded into 0 to 5 grades,grade 5 implies that bone defect has been completely replaced by new bone,grade 0 implies that no new bone repair）. MAIN OUTCOME MEASURES ：The general observations of rabbit radius defeets,X-ray scanning, histological and electro-microscopic observations. RESULTS： ①The general observations of bilateral rabbit radius defects： At week 12, callus became strong and protruded to bone defects in experimental side,well connecting with broken ends. While broken ends in control group were only connected by fibrous tissue and no continuous callus could be observed passing through. ②X-ray scanning：At week 12, callus was found continuously crossing through the bone defect of experimental side, marrow cavity was smooth ,but molding was incomplete. While in control side, no continuous callus could be observed passing through the broken ends. ③Histological observations： At week 12, a great deal of osteoblasts and new stroma could be observed in bone defect of experimental side, but only a few of osteocytes appeared in the broken ends of control sides which were connected by fibrous tissue.④Electro-microscopic observations： The osteoblast observed in the experimental side seems normal and was rich in enlarged endoplasmic reticula energetic in ,protein synthesis and abundant in organelle. CONCLUSION：As osteogenetic cells, BMSCs possess better osteogenesis property. They ean be used as seed cells in bone defect repair by using bone-engineering techniques.