Wang Xi, Li Guanghui, Liu Yiming, Yu Weiwei, Sun Qiang
Department of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Biomed Rep. 2017 May;6(5):519-524. doi: 10.3892/br.2017.881. Epub 2017 Mar 28.
To investigate the biocompatibility of the biomaterial, polylactic acid (PLA) with stem cells from human exfoliated deciduous teeth (SHED) and its induction of mineralization as a type of scaffold material. To determine the impacts of biomaterial PLA on proliferation and mineralization of SHED, the expression of surface molecules of SHED isolated and cultured was detected by flow cytometry. In addition, cell proliferation was measured using MTT and Edu assays, and the evaluation of mineralized differentiation was performed using Alizarin Red S staining. In addition, the expression levels of osteogenic marker genes were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. SHED were successfully isolated and identified. The MTT and Edu results indicated that the proliferation of SHED cultured in PLA and normal medium was not significantly different. The Alizarin Red S staining demonstrated that the mineralization capability was significantly higher in the SHED that were cultured in PLA medium. Furthermore, RT-qPCR and western blot analyses indicated that the expression levels of osteogenic marker genes were higher in the SHED cultured in PLA medium. These results suggested that PLA possesses good biocompatibility with SHED and may effectively induce the mineralization of SHED and serve as a scaffold material.
为研究生物材料聚乳酸(PLA)与人乳牙脱落干细胞(SHED)的生物相容性及其作为一种支架材料对矿化的诱导作用。为确定生物材料PLA对SHED增殖和矿化的影响,通过流式细胞术检测分离培养的SHED表面分子的表达。此外,使用MTT和Edu检测法测量细胞增殖,并使用茜素红S染色进行矿化分化评估。另外,通过逆转录定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹分析测量成骨标记基因的表达水平。SHED成功分离并鉴定。MTT和Edu结果表明,在PLA和正常培养基中培养的SHED增殖无显著差异。茜素红S染色表明,在PLA培养基中培养的SHED矿化能力显著更高。此外,RT-qPCR和蛋白质免疫印迹分析表明,在PLA培养基中培养的SHED中成骨标记基因的表达水平更高。这些结果表明,PLA与SHED具有良好的生物相容性,可能有效诱导SHED矿化并作为支架材料。