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处理牙本质浸提液对牙髓细胞分化方面作用的研究

Research on the Effect of Extract of Dentin Matrix on the Differentiation of Dental Pulp Cells

  • 摘要: 目的 观察人处理牙本质基质(human treated dentin matrix,hTDM)浸提液对人牙髓细胞(human dental pulp cells,hDPCs)成牙和成神经方向分化的影响,为有效利用细胞外基质分子参与牙髓牙本质复合体的修复提供新线索。 方法 体外分离培养hDPCs,免疫荧光及流式细胞术鉴定细胞组织来源;成脂、成骨及成神经诱导培养基诱导培养hDPCs,鉴定细胞多向分化能力。制备hTDM浸提液,诱导培养hDPCs 7 d后,实时荧光定量PCR(qRT-PCR)分析细胞成牙和成神经方向分化的基因的改变。 结果 qRT-PCR结果显示被诱导后hDPCs不同程度的增强表达ALP、OPN、OCN、BSP、DMP-1、DSP、β-Ⅲ tubulin。 结论 浸提的方法能够有效收集牙本质基质中的细胞外基质分子;浸提液能提供良好的诱导微环境诱导hDPCs成牙和成神经方向分化,这有利于牙髓牙本质复合体的修复。

     

    Abstract: Objective To investigate the effects of human treated dentin matrix (hTDM) extracellular matrix molecules on odontogenetic and neural differentiation of human dental pulp cells (hDPCs) with an aim to find an effective method to collect extracellular matrix molecules to contribute to reparation dental-pulp complex with dentin defects. Methods hDPCs were obtained and biological characteristics such as source of cells and multi-differentiation potentials were assessed using immunofluorescence and flow cytometry. Fabrication of hTDM extracts and hDPCs was induced with it for 1 week. The odontogenetic differentiation associated genes were tested by qRT-PCR. Results qRT-PCR results showed that cells were higher expression of odontogenetic differentiation associated genes ALP, OPN, OCN, BSP, DMP-1, DSP, β-Ⅲ tubulin. Conclusion The method of extracting extracellular matrix molecules from dentin matrix was effective. The extract liquid provides a suitable microenvironment for odontogenetic and neural differentiation of hDPCs and contributes to reparation dental-pulp complex.

     

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