| 米修奎 李彦 余善超 胡文军.miR-26a修饰的人牙周膜干细胞成骨分化能力的研究[J].,2016,16(3):445-448 |
| miR-26a修饰的人牙周膜干细胞成骨分化能力的研究 |
| The Study of Human Periodontal Ligament StemCells OsteogenicDifferentiation Capacity after Modification with miR-26a |
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| DOI： |
| 中文关键词: 人牙周膜干细胞 成骨分化 miR-26a 细胞培养 |
| 英文关键词: Human periodontal ligament stemcells Osteogenic differentiation miR-26a Cell culture |
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| 中文摘要: |
| 目的：研究人牙周膜干细胞（hPDLSCs）在转染miR-26a 后成骨分化的促进效果。方法：从因正畸拔除的无龋坏、无牙周疾病
离体牙的牙根中部牙周膜组织中分离、胶原酶消化，进行人牙周膜干细胞的体外培养。使用脂质体2000进行miRNA转染，采用
激光共聚焦观察转染效果；MTT 方法检测转染后的细胞活力；成骨诱导后使用实时定量PCR技术检测miRNA 修饰的hPDLSCs
成骨分化相关基因的表达；采用BCIP/NBT、天狼星红、茜素红S分别对碱性磷酸酶、胶原以及钙化进行染色观察。结果：采用脂质
体2000 能够成功转染hPDLSCs，且转染效率较高；转染后的细胞活力有所下降，但仍在80%以上；转染后的细胞在经成骨诱导分
化过程中骨钙素（OCN）和骨桥蛋白（OPN）基因表达显著上调，同时碱性磷酸酶活性、胶原分泌以及钙化能力均得到显著提升。结
论：miR-26a 可以用于修饰hPDLSCs以提高其成骨分化能力。 |
| 英文摘要: |
| Objective:To investigate the osteogenic differentiation capacity of human periodontal ligament stem cells (hPDLSCs)
after modification with miR-26a.Methods:The periodontal ligament tissues from no-caries-no-periodontal-lesions teeth obtained from
orthodontic treatment were isolated and digested with collagenase to perform the in vitro culture. The miRNA transfection was performed
by lipofectamine 2000 according to the manufacturer's instructions. Confocal laser scanning microscope was used to observe the transfection
efficiency and the cell viability was monitored by MTT assay. The real time quantitative PCR was used to check the osteogenic differentiation
alteration after miRNA modification. Meanwhile, the alkaline phosphatase activity, collagen secretion and calciumdeposition
were visualized by BCIP/NBT, Sirius Red, Alizarin Red S staining，respectively.Results:The lipofectamine 2000 was able to transfect
hPDLSCs and the transfection efficiency was remarkable. Cell viability was suppressed after transfection, but it was still more than 80
percent. During the osteogenic induction, cells modified with miR-26a could enormously improve the osteocalcin (OCN) and osteopontin
(OPN) mRNA level. In the meanwhile, the alkaline phosphatase activity, collagen secretion and calcium deposition were all augmented
significantly.Conclusion:The miR-26a can be used to modify hPDLSCs in order to improve their osteogenic differentiation ability. |
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