兰州大学机构库 >学院待认领
骨髓间充质干细胞复合纤维蛋白凝胶修复兔关节软骨缺损
Alternative TitleRepair of Articular Cartilage Defects by Using Composite of Bone Marrow Mesenchymal Stem Cells and Fibrin Glue in Rabbit
刘文忠
Thesis Advisor孙正义 ; 王栓科
2005-05-22
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword骨髓间充质干细胞 纤维蛋白凝胶 修复 软骨缺损
Abstract目的:探讨纤维蛋白凝胶与骨髓间充质干细胞、转化生长因子-β1的复合物对兔膝关节软骨全层缺损的修复机理。 方法:35只(70个关节)3~4月龄青紫蓝兔,平均体重为2.7㎏(2.4~3.0kg),随机分为4组,实验组A组(n=20):纤维蛋白凝胶+骨髓间充质干细胞+转化生长因子-β1;对照组B组(n=20):纤维蛋白凝胶+骨髓间充质干细胞;对照组C组(n=20):纤维蛋白凝胶+转化生长因子-β1;空白对照组D组(n=10):单纯钻孔。自大转子处抽取骨髓4~5ml,密度梯度离心法分离间充质干细胞(MSCs)并行体外培养扩增,用骨髓间充质干细胞特异性标记CD44抗体和造血系细胞特异性表面标记CD34抗体对分离扩增的细胞行免疫细胞化学方法鉴定。依分组将复合物分别填充到软骨缺损处,12周后取标本,进行大体及HE染色组织学观察,行甲苯胺蓝染色,Ⅱ型胶原免疫组化染色鉴定修复组织的性质,并行改良pineda法半定量评分后统计学处理。 结果:对扩增的骨髓间充质干细胞行其特异性标记CD44染色为阳性,而造血系细胞特异性表面标记CD34则为阴性。12周后见:实验组缺损由类透明软骨或透明软骨修复,Ⅱ型胶原免疫组化染色阳性,甲苯胺蓝异染性明显,修复面较平整光滑;而对照组则以纤维软骨及纤维组织修复,Ⅱ型胶原免疫组化染色阴性。改良pineda法半定量组织学评分实验组总分平均为6.50,较对照组有显著性差异。 结论:应用密度梯度离心法能够分离、培养、扩增大量的骨髓间充质干细胞,将骨髓间充质干细胞、纤维蛋白凝胶及转化生长因子-β1的复合物移植到兔关节软骨缺损处,能够生成透明软骨而修复软骨缺损。
Other AbstractObjective To explore the repair of articular cartilage defects by using bone-derived mesenchymal stem cells(MSCs), fibrin glue(FG) and transforming growth factor-β1(TGF-β1) in rabbit. Methods 70 knee joints of 35 qingzilan rabbits of three to four months old and with an average weight of 2.7kg were used in this experiment. All rabbits were divided into four groups at random. Experimental group A(n=20): the defects were filled with MSCs, FG and TGF-β1; Control group B(n=20): the defects were filled with MSCs and FG; Control group C (n=20): the defects were filled with FG and TGF-β1; Control group D (n=10): the defects were left only. The autologous mesenchymal stem cells derived from bone marrow were isolated with the lymphocyte separate liquid by density gradient centrifuge and expanded in vitro. The cultured MSCs were investigated with antibody of anti-CD44 and anti-CD34 by using immunohistochemical staining. The graft were implanted according to various groups respectively. The neo-cartilage were investigated macroscopically and analysed with a semi-quantitative score system by staining with haematoxylin and eosin , toluidine blue and collagen type Ⅱ at the end of 12 weeks after implantation. Results The cultured MSCs showed immunohistochemical positive staining with antibody of anti-CD44 to specific antigen MSCs and negative staining of anti-CD34 to specific antigen of the hematopoietic lineages. The defects area of articular cartilage was filled with hyaline neo-cartilage and hyaline-like cartilage in the experiment group in 12 weeks and possess smooth surface .While the defects area of articular cartilage was replaced with fibrous tissue and fibrous cartilage in the control group at the same time.Histologically, the neo-cartilage which showed positive staining of toluidine blue and collagen type Ⅱ was similar to the surrounding cartilage in the experiment group in 12 weeks.But in the control group,neo-tissue showed negative staining of collagen type Ⅱ and weakly positive staining of toluidine blue at the same time. The average gross score that with improved pineda’ s semi-quantitative score system was 6.50 in the experiment group in 12 weeks.It had the significant difference statistically between the experiment group and the control group. Conclusion A large number of bone-derived mesenchymal stem cells could be acquired by density gradient centrifuge with the lymphocyte separate liquid and expanded in vitro. The articular cartilage ...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223380
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
刘文忠. 骨髓间充质干细胞复合纤维蛋白凝胶修复兔关节软骨缺损[D]. 兰州. 兰州大学,2005.
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