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6-溴靛玉红-3'-肟调节人牙髓干细胞集落形成、细胞凋亡和向牙源性/成骨分化。

6-Bromoindirubin-3'-Oxime Regulates Colony Formation, Apoptosis, and Odonto/Osteogenic Differentiation in Human Dental Pulp Stem Cells.

机构信息

Dental Stem Cell Biology Research Unit, Faculty of Dentistry, Chulalongkorn University, Bangkok 10330, Thailand.

Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Chulalongkorn University, Bangkok 10330, Thailand.

出版信息

Int J Mol Sci. 2022 Aug 4;23(15):8676. doi: 10.3390/ijms23158676.

DOI:10.3390/ijms23158676
PMID:35955809
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9368902/
Abstract

6-bromoindirubin-3'-oxime (BIO) is a candidate small molecule that effectively modulates Wnt signalling owing to its stable property. The present study investigated the influence of BIO on the odonto/osteogenic differentiation of human dental pulp stem cells (hDPSCs). hDPSCs were treated with 200, 400, or 800 nM BIO, and the effects on hDPSC responses and osteogenic differentiation were assessed. BIO-mediated Wnt activation was confirmed by β-catenin nuclear translocation detected by immunofluorescence staining. BIO attenuated colony formation and cell migration determined by in vitro wound-healing assay. BIO increased early apoptotic cell population evaluated using flow cytometry. For osteogenic induction, BIO promoted alkaline phosphatase (ALP) activity and mineralisation in a dose-dependent manner. , , , , , , and mRNA expression were significantly upregulated. The OPG/RANKL expression ratio was also increased. Further, BIO attenuated adipogenic differentiation as demonstrated by decreased lipid accumulation and adipogenic-related gene expression. Bioinformatic analysis of RNA sequencing data from the BIO-treated hDPSCs revealed that BIO modulated pathways related to autophagy and actin cytoskeleton regulation. These findings demonstrated that BIO treatment promoted hDPSC osteogenic differentiation. Therefore, this small molecule is a strong candidate as a bioactive molecule to enhance dentin repair.

摘要

6-溴靛红-3'-肟（BIO）是一种候选小分子，由于其稳定性，能有效调节 Wnt 信号。本研究探讨了 BIO 对人牙髓干细胞（hDPSCs）牙向/成骨分化的影响。用 200、400 或 800 nM 的 BIO 处理 hDPSCs，评估其对 hDPSC 反应和成骨分化的影响。通过免疫荧光染色检测β-连环蛋白核易位证实了 BIO 介导的 Wnt 激活。BIO 通过体外划痕愈合试验抑制集落形成和细胞迁移。BIO 通过流式细胞术评估增加早期凋亡细胞群体。用于成骨诱导，BIO 以剂量依赖的方式促进碱性磷酸酶（ALP）活性和矿化。 、 、 、 、 、 和 mRNA 表达明显上调。OPG/RANKL 表达比也增加。此外，BIO 抑制脂肪生成分化，表现为脂质积累减少和脂肪生成相关基因表达降低。用 BIO 处理的 hDPSCs 的 RNA 测序数据的生物信息学分析表明，BIO 调节了与自噬和肌动蛋白细胞骨架调节相关的途径。这些发现表明，BIO 处理促进 hDPSC 成骨分化。因此，这种小分子是增强牙本质修复的生物活性分子的有力候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bbf/9368902/ee191ea65e2c/ijms-23-08676-g004.jpg
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