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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
引用本文:雍志军,贾帅军,韩林章,周翔,户刚,郝赋,刘建.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].生物磁学,2014(8):1407-1410.
作者姓名:雍志军  贾帅军  韩林章  周翔  户刚  郝赋  刘建
作者单位:[1]第四军医大学西京医院骨科,陕西西安710032 [2]武警陕西省总队医院骨科,陕西西安710054 [3]解放军总后勤部机关第一门诊部,北京100842
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)
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(Bonemarrowmesenchymalstemcells,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PIuF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。

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

Effect of Platelet-rich Fibrin on Chondrogenic Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells
Institution:YONG Zhi-jun, JIA Shuai-jun, HAN Lin-zhang, ZHOU Xian, HU Gang, HAO Fu, L1U Jian (1 Department of orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China; 2 Department of orthopedics, Shaanxi Provincial Corps Hospital, Chinese People's Armed Police Forces, Xi'an, Shaanxi, 710054, China; 3 First Clinic Department ofGLD ofPLA, Beijing, 100842, China)
Abstract:Objective:To investigate the effect of autologous platelet rich fibrin (PRF) on chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells (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 rive. 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 ha vitro. It can be used as autologous biological materials, and it will play an important role in constructing tissue engineering cartilage.
Keywords:Platelet-rich fibrin  Bone marrow mesenchymal stem cells  Chondrogenic differentiation  Tissue engineering
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