Zhu Shao Yue, Yuan Chang Yong, Lin Yi Fan, Liu Hao, Yang Yan Qi, Wong Hai Ming, Zhang Cheng Fei, Wang Peng Lai, Gu Min
Discipline of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, The University of Hong Kong, Hong Kong, China.
Orthodontic Department, Affiliated Stomatological Hospital of Xuzhou Medical University, Xuzhou, China.
Stem Cells Int. 2021 Jul 24;2021:8859902. doi: 10.1155/2021/8859902. eCollection 2021.
Pericytes play an important role in forming functional blood vessels and establishing stable and effective microcirculation, which is crucial for vascular tissue engineering. The slow ex vivo expansion rate, limited proliferative capacity, and variability of tissue-specific phenotypes would hinder experimental studies and clinical translation of primary pericytes. In this study, the angiogenic and pericyte functions of stem cells from human exfoliated deciduous teeth (SHEDs) and postnatal human dental pulp stem cells (DPSCs) were investigated.
Osteogenic and adipogenic induction assays were performed to evaluate the mesenchymal potential of SHEDs, DPSCs, and pericytes. An in vitro Matrigel angiogenesis assay was conducted to reveal the ability of SHEDs, DPSCs, and pericytes to stabilize vascular-like structures. Quantitative real-time polymerase chain reaction (RT-qPCR) was performed to evaluate mRNA expression. Flow cytometry, western blotting, and immunostaining were used to assess the protein expression. Wound healing and transwell assays were performed to evaluate the migration ability of SHEDs, DPSCs, and pericytes.
The osteogenic and adipogenic induction assays showed that SHEDs, DPSCs, and pericytes exhibited similar stem cell characteristics. The mRNA expression levels of PDGFR-, -SMA, NG2, and DEMSIN in SHEDs and DPSCs cultured in EC medium were significantly higher than those in the control groups on day 7 ( < 0.05), but significantly higher than those in the pericytes group on day 14 ( < 0.05). Flow cytometry showed that high proportions of SHEDs and DPSCs were positive for various pericyte markers on day 7. The DPSCs, SHEDs, and pericytes displayed strong migration ability; however, there was no significant difference among the groups ( > 0.05).
The SHEDs and DPSCs display a profile similar to that of pericytes. Our study lays a solid theoretical foundation for the clinical use of dental pulp stem cells as a potential candidate to replace pericytes.
周细胞在形成功能性血管和建立稳定有效的微循环中发挥着重要作用，这对血管组织工程至关重要。原代周细胞体外扩增速率缓慢、增殖能力有限以及组织特异性表型的变异性会阻碍其实验研究和临床转化。在本研究中，对人脱落乳牙干细胞（SHEDs）和人出生后牙髓干细胞（DPSCs）的血管生成及周细胞功能进行了研究。
进行成骨和成脂诱导试验以评估SHEDs、DPSCs和周细胞的间充质潜能。开展体外基质胶血管生成试验以揭示SHEDs、DPSCs和周细胞稳定类血管结构的能力。进行定量实时聚合酶链反应（RT-qPCR）以评估mRNA表达。采用流式细胞术、蛋白质印迹法和免疫染色来评估蛋白质表达。进行伤口愈合试验和Transwell试验以评估SHEDs、DPSCs和周细胞的迁移能力。
成骨和成脂诱导试验表明，SHEDs、DPSCs和周细胞表现出相似的干细胞特征。在第7天，培养于内皮细胞培养基中的SHEDs和DPSCs中血小板衍生生长因子受体（PDGFR）、α-平滑肌肌动蛋白（α-SMA）、神经胶质抗原2（NG2）和双皮质素（DEMSIN）的mRNA表达水平显著高于对照组（P<0.05），但在第14天显著高于周细胞组（P<0.05）。流式细胞术显示，在第7天，高比例的SHEDs和DPSCs对各种周细胞标志物呈阳性。DPSCs、SHEDs和周细胞均表现出较强的迁移能力；然而，各组之间无显著差异（P>0.05）。
SHEDs和DPSCs表现出与周细胞相似的特征。我们的研究为牙髓干细胞作为替代周细胞的潜在候选者的临床应用奠定了坚实的理论基础。
