【作者】 钱奇春；

【导师】 刘彦普；

【作者基本信息】 第四军医大学 ， 口腔临床医学， 2002， 硕士

【摘要】 骨缺损修复是多年来医学界尚待解决的难题之一，人们一直在寻求一种理想的修复方法，骨组织工程学的兴起，为临床骨缺损修复提供了新的思路。 本研究以人骨髓基质细胞(human bone marrow stromal cell,HBMSC)作为种子细胞，研究其体外培养条件下的生物学特性；探讨了重组人骨形成蛋白-2(recombinant human bone morphogenetic protein-2,rhBMP-2)和碱性成纤维细胞生长因子(basic fibroblast growth factor,b-FGF)对HBMSC的生物学作用；同时以珊瑚(coral)为支架材料，形成HBMSC/coral复合人工骨，观察HBMSC/coral在无免疫动物皮下异位移植后的成骨作用。 方法：穿刺抽吸人HBMSC，进行体外培养，采用细胞形态学观察、生长曲线测定、扫描电镜(SEM)、碱性磷酸酶(ALP)染色、钙染色等方法观察HBMSC的生物学特性；采用MTT法检测、碱性磷酸酶(ALP)测定法探讨rhBMP-2和b-FGF对HBMSC的生物学作用；采用大体观察、SEM、组织学染色等方法评价新骨形成情况。 结果：HBMSC可以在体外适宜条件下培养，经诱导可向成骨细胞转化；rhBMP-2和b-FGF在一定条件下，可促进HBMSC增殖并向成骨细胞 第。军区人甘硕士砂位估土 转化；HBMSClcoral在无免疫动物皮下异位成骨作用明显，随时间延长， 成骨量增加且更加成熟。 结论：HBMSC取材方便、损伤小、倍增迅速、可定向分化，可以作 为骨组织工程的种子细胞；hBMP－二和bFGF对HBMSC具有定向诱导分 化作用；HBMSC儿m 在无免疫动物体内成骨活跃，显示出良好的应用前 景。更多还原

【Abstract】 Reconstructive, orthopedic, and plastic surgeons are continually faced with challenge of replacing bone lost by trauma, tumor or congenital abnormalities. Bone tissue engineering of autologous bone seems to be a promising method for repairing bone defects in clinical practice.Materials: In this study, human bone marrow stromal cells(HBMSC) were used as seed cells of bone tissue engineering. The biological characteristic of HBMSC cultured in vitro was studied. The biological effects of recombinant human bone morphogenetic protein-2 (rhBMP-2 ) and basic fibroblast growth factor( b-FGF)on HBMSC were also investigated. To test the feasibility of the compound artificial bone in vivo, using coral as scaffold , the HBMSC/coral composites were implanted subcutantously into athymic nude mice.Method: HBMSC obtained from human iliac crest was cultured in different medium. During culture, the growth and the biological characteristic of HBMSC were observed by cell morphological survey, cell growth curve, scanning electron microscope(SEM), alkaline phosphatase(ALP) stains and Von Kossa stains. The biological effects of rhBMP-2 and b-FGF on HBMSC were measured with methylthiazoltetrazolium (MTT) method and ALP measurement kit. The effectiveness of new bone formation was evaluated by means of gross, histological observation and SEM examination.Result: HBMSC can be cultured in vitro and induced into osteoblast. rhBMP-2 and b-FGF can promote the proliferation and differentiation into osteoblast of HBMSC. In immunodeficiency animal subcutantous tissue, the ectopic osteogenesis of HBMSC/coral composite was obvious, and the new bone formation increased and became more mature in pace with time.Conclusion: HBMSC can be used as seeded cells in bone tissue engineering, made with only a small amount of aspired marrow from patient with little host morbidity. rhBMP-2 and b-FGF can promote the proliferation and differentiation of BMSC. The HBMSC/coral composite can form bone tissue in vivo, showing a promising prospect for clinical practice.更多还原