College of Stomatology, Chongqing Medical University, 426# Songshibei Road, Yubei District, Chongqing, 401147, People's Republic of China.
Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, People's Republic of China.
BMC Oral Health. 2023 Apr 6;23(1):203. doi: 10.1186/s12903-023-02905-3.
Human dental pulp stem cells (hDPSCs) may be the best choice for self-repair and regeneration of teeth and maxillofacial bone tissue due to their homogeneous tissue origin, high proliferation and differentiation rates, and no obvious ethical restrictions. Recently, several studies have shown that extracellular matrix (ECM) proteins can effectively regulate the proliferation and differentiation fate of mesenchymal stem cells (MSCs). However, the role of elastin microfibril interface-located protein-1 (EMILIN-1), a new ECM glycoprotein, in osteo/odontogenic differentiation of hDPSCs has not been reported. The aim of this study was to explore the effect of EMILIN-1 during osteo/odontogenic differentiation of hDPSCs.
hDPSCs were cultured in osteo/odontogenic induction medium. qPCR and Western blot analysis were performed to detect osteo/odonto-specific genes/proteins expression as well as the expression of EMILIN-1. After knockdown of Emilin-1 in hDPSCs with small interfering RNA and exogenous addition of recombinant human EMILIN-1 protein (rhEMILIN-1), Cell Counting Kit-8 assay, alkaline phosphatase staining, alizarin red S staining, qPCR and Western blot were performed to examine the effect of EMILIN-1 on proliferation and osteo/odontogenic differentiation of hDPSCs.
During the osteo/odontogenic induction of hDPSCs, the expression of osteo/odonto-specific genes/proteins increased, as did EMILIN-1 protein levels. More notably, knockdown of Emilin-1 decreased hDPSCs proliferation and osteo/odontogenic differentiation, whereas exogenous addition of rhEMILIN-1 increased them.
These findings suggested that EMILIN-1 is essential for the osteo/odontogenic differentiation of hDPSCs, which may provide new insights for teeth and bone tissue regeneration.
人牙髓干细胞（hDPSCs）由于组织来源均一、增殖分化率高、无明显伦理限制等特点，可能是牙齿和颌面骨组织自我修复和再生的最佳选择。最近的几项研究表明，细胞外基质（ECM）蛋白可以有效地调节间充质干细胞（MSCs）的增殖和分化命运。然而，弹性蛋白微纤维界面定位蛋白 1（EMILIN-1）作为一种新的 ECM 糖蛋白，在 hDPSCs 成骨/成牙分化中的作用尚未报道。本研究旨在探讨 EMILIN-1 在 hDPSCs 成骨/成牙分化中的作用。
hDPSCs 在成骨/成牙诱导培养基中培养。采用 qPCR 和 Western blot 分析检测成骨/成牙特异性基因/蛋白的表达以及 EMILIN-1 的表达。用小干扰 RNA 敲低 hDPSCs 中的 Emilin-1 后，用重组人 EMILIN-1 蛋白（rhEMILIN-1）外源性处理，用细胞计数试剂盒-8 法、碱性磷酸酶染色、茜素红 S 染色、qPCR 和 Western blot 分析检测 EMILIN-1 对 hDPSCs 增殖和成骨/成牙分化的影响。
在 hDPSCs 的成骨/成牙诱导过程中，成骨/成牙特异性基因/蛋白的表达增加，同时 EMILIN-1 蛋白水平也增加。更值得注意的是，敲低 Emilin-1 降低了 hDPSCs 的增殖和成骨/成牙分化能力，而外源性添加 rhEMILIN-1 则增加了这些能力。
这些发现表明 EMILIN-1 对于 hDPSCs 的成骨/成牙分化是必需的，这可能为牙齿和骨组织再生提供新的思路。
