| 慢病毒介导的Sox9基因在兔骨髓间充质干细胞的过表达促进软骨损伤修复 |
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Received:February 20, 2014
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| 作者 | Author | 单位 | Unit | E-Mail |
| 王震 |
WANG Zhen |
山西医科大学第二医院骨科, 山西 太原 030001 |
Department of Orthopaedics, the Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China |
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| 梁大川 |
LIANG Da-chuan |
山西医科大学第二医院骨科, 山西 太原 030001 |
Department of Orthopaedics, the Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China |
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| 白洁玉 |
BAI Jie-yu |
山西医科大学第二医院骨科, 山西 太原 030001 |
Department of Orthopaedics, the Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China |
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| 康宁 |
KANG Ning |
山西医科大学第二医院骨科, 山西 太原 030001 |
Department of Orthopaedics, the Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China |
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| 冯军宇 |
FENG Jun-yu |
山西医科大学第二医院骨科, 山西 太原 030001 |
Department of Orthopaedics, the Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China |
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| 杨自权 |
YANG Zi-quan |
山西医科大学第二医院骨科, 山西 太原 030001 |
Department of Orthopaedics, the Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China |
yzqonline@126.com |
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| 期刊信息：《中国骨伤》2015年28卷，第5期，第433-440页 |
| DOI：10.3969/j.issn.1003-0034.2015.05.011 |
| 基金项目:国家自然科学基金资助项目(编号:30973048);国际科技合作项目(编号:2010DFA32450);国家人事部及山西省人事厅留学回国人员科技活动择优资助项目;山西省留学基金项目(编号:107);山西省自然科学基金资助项目(编号:2010011050-6) |
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目的:明确在体内Sox9基因过表达的兔骨髓间充质干细胞对于关节软骨损伤修复的作用。
方法:以慢病毒介导的Sox9基因转染兔骨髓间充质干细胞(BMSCs),体外检测软骨特异性分子,将新西兰大白兔24只48个膝关节随机分为3组,动物麻醉后,双侧股骨滑车处的关节面上用直径4 mm的钻头钻孔,深度3 mm,穿透软骨下骨,造成全层关节软骨损伤,将转染后的细胞植入体内用以修复全层关节软骨损伤,实验组植入BMSCs-(Lenti-Sox9-EGFP)-藻酸钙复合物,实验对照组植入BMSCs-藻酸钙复合物,空白对照组只钻孔。术后6、12周分别进行光镜、电镜观察,以及HE、免疫组织化学染色检测软骨的修复程度。
结果:经Sox9基因转染后的细胞在3 d时,Sox9基因表达最高,随后下降。转染后3 d,Ⅱ型胶原开始表达,到14 d时达到最高。表明Sox9过表达启动了兔骨髓间充质干细胞的软骨分化。组织学观察显示,实验组术后6周缺损处有透明软骨样组织填充,术后12周缺损处软骨和软骨下骨修复良好。两对照组,缺损处由纤维组织填充。免疫组织化学显示,修复组织内Ⅱ型胶原,免疫组化染色结果阳性强于两对照组。组织学评分结果显示实验组软骨损伤修复各时间点效果明显优于两对照组,差异有统计学意义。
结论:Sox9基因过表达的兔骨髓间充质干细胞(BMSCs)促进软骨损伤的修复。 |
| [关键词]:Sox9基因 骨髓间充质干细胞 转染 软骨 兔 |
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| Overexpression of Sox9 gene by the lentiviral vector in rabbit bone marrow mesenchymal stem cells for promoting the repair of cartilage defect |
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Abstract:
Objective:To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.
Methods:Rabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile,the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope,electron microscope,HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.
Results:At 3 days after the transfection,Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection,the expression of collagen type Ⅱ began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation,the defects in the experimental group was filled with hyaline like cartilage tissue,12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively,the defects were filled with fibrous tissues in control groups. Meanwhile,immunohistochemical staining of sections with type Ⅱ collagen antibodies showed the proteins in the regenerated tissue stained positive for type Ⅱ collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.
Conclusion:Overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect. |
| KEYWORDS:Sox9 gene Bone marrow mesenchymal stem cells (BMSCs) Transfection Cartilage Rabbit |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 王震,梁大川,白洁玉,康宁,冯军宇,杨自权.慢病毒介导的Sox9基因在兔骨髓间充质干细胞的过表达促进软骨损伤修复[J].中国骨伤,2015,28(5):433~440 |
| 英文格式: | WANG Zhen,LIANG Da-chuan,BAI Jie-yu,KANG Ning,FENG Jun-yu,YANG Zi-quan.Overexpression of Sox9 gene by the lentiviral vector in rabbit bone marrow mesenchymal stem cells for promoting the repair of cartilage defect[J].zhongguo gu shang / China J Orthop Trauma ,2015,28(5):433~440 |
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