Heng Boon Chin, Zhu Shaoyue, Xu Jianguang, Yuan Changyong, Gong Ting, Zhang Chengfei
Comprehensive Dental Care, Endodontics, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong.
Comprehensive Dental Care, Endodontics, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong.
Tissue Cell. 2016 Apr;48(2):133-43. doi: 10.1016/j.tice.2015.12.004. Epub 2015 Dec 25.
A major bottleneck to the therapeutic applications of dental pulp stem cells (DPSC) are their limited proliferative capacity ex vivo and tendency to undergo senescence. This may be partly due to the sub-optimal in vitro culture milieu, which could be improved by an appropriate extracellular matrix substratum. This study therefore examined decellularized matrix (DECM) from stem cells derived from human exfoliated deciduous teeth (SHED) and periodontal ligament stem cells (PDLSC), as potential substrata for DPSC culture. Both SHED-DECM and PDLSC-DECM promoted rapid adhesion and spreading of newly-seeded DPSC compared to bare polystyrene (TCPS), with vinculin immunocytochemistry showing expression of more focal adhesions by newly-adherent DPSC cultured on DECM versus TCPS. Culture of DPSC on SHED-DECM and PDLSC-DECM yielded higher proliferation of cell numbers compared to TCPS. The qRT-PCR data showed significantly higher expression of nestin by DPSC cultured on DECM versus the TCPS control. Osteogenic differentiation of DPSC was enhanced by culturing on PDLSC-DECM and SHED-DECM versus TCPS, as demonstrated by alizarin red S staining for mineralized calcium deposition, alkaline phosphatase assay and qRT-PCR analysis of key osteogenic marker expression. Hence, both SHED-DECM and PDLSC-DECM could enhance the ex vivo culture of DPSC under both non-inducing and osteogenic-inducing conditions.
牙髓干细胞（DPSC）治疗应用的一个主要瓶颈是其体外增殖能力有限且有衰老倾向。这可能部分归因于体外培养环境欠佳，而合适的细胞外基质底物可能会改善这一情况。因此，本研究检测了来源于人脱落乳牙的干细胞（SHED）和牙周膜干细胞（PDLSC）的脱细胞基质（DECM），作为DPSC培养的潜在底物。与裸聚苯乙烯（TCPS）相比，SHED-DECM和PDLSC-DECM均能促进新接种的DPSC快速黏附与铺展，纽蛋白免疫细胞化学显示，与在TCPS上培养相比，在DECM上培养的新黏附DPSC表达更多的黏着斑。与TCPS相比，在SHED-DECM和PDLSC-DECM上培养DPSC，细胞数量增殖更高。qRT-PCR数据显示，与TCPS对照相比，在DECM上培养的DPSC巢蛋白表达显著更高。与TCPS相比，在PDLSC-DECM和SHED-DECM上培养可增强DPSC的成骨分化，这通过茜素红S染色检测矿化钙沉积、碱性磷酸酶测定以及关键成骨标志物表达的qRT-PCR分析得以证实。因此，在非诱导和成骨诱导条件下，SHED-DECM和PDLSC-DECM均可增强DPSC的体外培养。
