雍志军1 贾帅军2 韩林章3 周翔1 户刚1 郝赋1 刘建1△.富血小板纤维蛋白对兔骨髓间充质干细胞成软骨分化的影响*[J].,2014,14(8):1407-1410 |
富血小板纤维蛋白对兔骨髓间充质干细胞成软骨分化的影响* |
Effect of Platelet-rich Fibrin on Chondrogenic Differentiation of RabbitBone Marrow Mesenchymal StemCells* |
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DOI: |
中文关键词: 富血小板纤维蛋白 骨髓间充质干细胞 成软骨分化 组织工程 |
英文关键词: Platelet-rich fibrin Bone marrow mesenchymal stemcells Chondrogenic differentiation Tissue engineering |
基金项目:国家高技术研究发展计划“863 计划”(434112082);国家自然科学基金项目(50675108) |
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中文摘要: |
摘要目的:观察自体富血小板纤维蛋白(platelet-rich fibrin, PRF)对体外培养的兔骨髓间充质干细胞(Bone marrow mesenchymal
stemcells, BMSCs)成软骨分化的影响。方法:兔心脏采血制备PRF,电镜观察其超微结构;分离培养兔BMSCs,取第3代细胞用于
实验,分为PRF组、阳性对照组、空白对照组。诱导培养21d 后,对三组细胞分别进行形态学观察,成软骨鉴定染色(甲苯胺蓝、II
型胶原免疫组化染色),软骨相关基因表达检测(Ⅱ型胶原、Aggrecan、SOX9)。结果:PRF 组和阳性对照组中BMSCs经诱导后,细
胞由长梭形变为三角形、多角形、圆形;甲苯胺蓝、II 型胶原免疫组化染色均为阳性;Ⅱ型胶原、Aggrecan、SOX9基因表达水平均
较高,两组比较无统计学差异,空白对照组未见相关分化现象。结论:PRF在体外可促进兔BMSCs 成软骨分化,可作为自体生物
材料,在构建组织工程软骨中发挥更好的作用。 |
英文摘要: |
ABSTRACT Objective:To investigate the effect of autologous platelet rich fibrin (PRF) on chondrogenic differentiation of rabbit
bone marrow mesenchymal stemcells (BMSCs)in vitro .Methods: Blood extracted from rabbit's heart was used to prepare PRF, and the
ultrastructure of PRF was observed by electron microscope. BMSCs were isolated and cultured ex vivo. BMSCs at passage 3 were
divided into three groups: PRF group, positive control group and blank control group. After 2 weeks of culture, the morphology of cells in
each group were observed by phase-contrast microscope, the phenotype was identified with toluidine blue staining and collagen Ⅱ
immunocytochemistry, and the gene expression levels of collagen type II, Aggrecan, SOX 9 were detected by Real-time PCR.
Results:Phase-contrast microscope showed that inducing BMSCs of PRF group and positive control group grew form long spindle to triangle,
polygon, and circle. The toluidine blue staining and collagen Ⅱ immunocytochemistry of two groups were positive, gene expression
levels of collagen type II, Aggrecan, and SOX 9 were high, there was no statistical difference between two groups. However, cells in the
control group were not detected these changes. Conclusion:PRF could promote chondrogenic differentiation of BMSCs in vitro. It can be
used as autologous biological materials, and it will play an important role in constructing tissue engineering cartilage. |
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