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| CEMP1基因修饰骨质疏松大鼠BMSCs与β-TCP体外复合培养实验 |
| Co-culture of CEMP1 gene modified osteoporotic rat BMSCs with β-TCP in vitro |
| 投稿时间：2018-11-01 修订日期：2019-03-06 |
| DOI： |
| 中文关键词: 牙骨质蛋白1 骨髓基质细胞 骨质疏松 慢病毒 β-TCP |
| 英文关键词:CEMP1 BMSCs Osteoporosis Lentiviral vector β-TCP |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 中文摘要: |
| 目的 探讨牙骨质蛋白1（cementum protein 1，CEMP1）基因修饰骨质疏松大鼠骨髓基质细胞(bone marrow stromal cells, BMSCs)与β-磷酸三钙（β-tricalcium phosphate, β-TCP）体外复合培养的可行性。方法 体外培养骨质疏松大鼠BMSCs，将携带有CEMP1基因的慢病毒LV-CEMP1-EGFP转染BMSCs后倒置荧光显微镜观察，流式细胞仪检测转染率，实时荧光定量PCR检测转染后BMSCs中CEMP1基因的表达，Western Blot及免疫组织化学染色检测BMSCs中CEMP1蛋白的表达情况。将CEMP1基因修饰的BMSCs与β-TCP体外复合培养，倒置荧光显微镜和扫描电镜观察细胞的生长状况。结果 LV-CEMP1-EGFP具有较高的转染率，CEMP1基因修饰的BMSCs高表达CEMP1。倒置荧光显微镜和扫描电镜结果显示CEMP1基因修饰BMSCs在β-TCP上生长良好，分泌胞外基质。结论 CEMP1基因修饰骨质疏松大鼠BMSCs与β-TCP复合培养，有望应用于骨质疏松状态下的牙周组织再生研究。 |
| 英文摘要: |
| Objective To evaluate the feasibility of co-culture of cementum protein 1(CEMP1) gene modified osteoporotic rat bone marrow stromal cells (BMSCs) with β-TCP in vitro. Methods BMSCs of osteoporotic rats were isolated and cultured in vitro and transfected with lentiviral vector carrying CEMP1 gene (LV-CEMP1-EGFP) and detected under inverted fluorescence microscope. Transfection efficiency was detected by flow cytometry 72 hours after transfection. Expression of CEMP1 in BMSCs were detected by real time PCR, Western blot and immunohistochemistry. LV-CEMP1-EGFP transfected cells were co-cultured with β-TCP and was observed under inverted fluorescence microscope and scanning electron microscope (SEM).
Results: LV-CEMP1-EGFP has high transfection rate and CEMP1 was overexpressed in BMSCs. Fluorescent microscope and SEM demonstrated that CEMP1 gene modified BMSCs could migrate into β-TCP and secrete extracellular matrix. Conclusion Recombinant lentiviral vector containing human CEMP1 gene can be transfected into osteoporotic rat BMSCs efficiently. CEMP1 gene modified BMSCs can be combined with β-TCP and grow well. The composite might be benefit for periodontal tissue regeneration with osteoporosis. |
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