Gong Ting, Heng Boon Chin, Xu Jianguang, Zhu Shaoyue, Yuan Changyong, Lo Edward Chin Man, Zhang Chengfei
Department of Endodontology, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong, China.
Department of Biological Sciences, Sunway University, Bandar Sunway, Selangor Darul Ehsan, Malaysia.
J Biomed Mater Res A. 2017 Apr;105(4):1083-1093. doi: 10.1002/jbm.a.36003. Epub 2017 Feb 2.
Dental stem cells can serve as a potential source of functional endothelial cells for tissue engineering applications, but the endothelial-lineage differentiation efficiency is rather low even with growth factors and mechanical stimuli, which greatly limits their clinical applications. This is partly due to the deficiency of standard two-dimensional (2-D) culture systems, which is unable to recapitulate the three-dimensional (3-D) in vivo milieu that is rich in extracellular matrix. Hence, we extracted decellularized extracellular matrix from human umbilical vein endothelial cells (HUVECs-DECM) to provide a bioactive substratum conducive to the endothelial differentiation of dental stem cells. Compared to cells plated on tissue culture polystyrene (TCP), stem cells from exfoliated deciduous teeth (SHED) cultured on the HUVECs-DECM demonstrated more regular arrangement and elongated morphology. HUVECs-DECM significantly enhanced the rapid adhesion and proliferation rates of SHED, as demonstrated by WST-8 assay and immunocytochemistry indicating higher expression levels of vinculin by newly adherent SHED on HUVECs-DECM versus TCP. In addition, there was twofold to fivefold higher mRNA expression levels of endothelial-specific markers CD31 and VEGFR-2 in SHED after seven days of culture on DECM versus TCP. Functional testing with in vitro matrigel angiogenesis assay identified more capillary-like structure formation with significantly higher tubule length in SHED induced by DECM versus TCP. Hence, the results of this study provide a better understanding of the unique characteristics of cell-specific ECM and demonstrated the potential use of HUVECs-DECM as a culture substratum conducive for stimulating the endothelial differentiation of SHED for therapeutic angiogenic applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1083-1093, 2017.
牙干细胞可作为组织工程应用中功能性内皮细胞的潜在来源，但即使有生长因子和机械刺激，其向内皮谱系的分化效率仍相当低，这极大地限制了它们的临床应用。部分原因是标准的二维（2-D）培养系统存在缺陷，该系统无法重现富含细胞外基质的三维（3-D）体内环境。因此，我们从人脐静脉内皮细胞中提取了脱细胞细胞外基质（HUVECs-DECM），以提供有利于牙干细胞内皮分化的生物活性基质。与接种在组织培养聚苯乙烯（TCP）上的细胞相比，在HUVECs-DECM上培养的乳牙脱落干细胞（SHED）排列更规则，形态更细长。WST-8检测和免疫细胞化学表明，与TCP相比，新黏附在HUVECs-DECM上的SHED中纽蛋白表达水平更高，这表明HUVECs-DECM显著提高了SHED的快速黏附率和增殖率。此外，与TCP相比，SHED在DECM上培养7天后，内皮特异性标志物CD31和VEGFR-2的mRNA表达水平高出两到五倍。体外基质胶血管生成试验的功能测试表明，与TCP相比，DECM诱导的SHED形成了更多的毛细血管样结构，且小管长度显著更长。因此，本研究结果有助于更好地理解细胞特异性细胞外基质的独特特性，并证明HUVECs-DECM作为一种培养基质，有潜力用于刺激SHED的内皮分化，以用于治疗性血管生成应用。© 2017威利期刊公司。《生物医学材料研究杂志》A部分：105A：1083 - 1093，2017年。
