Faculty of Dentistry, Department of Periodontology, Selcuk University, Konya, Turkey.
Research Center of Dental Faculty, Selcuk University, Konya, Turkey.
J Periodontal Res. 2021 Aug;56(4):774-781. doi: 10.1111/jre.12876. Epub 2021 Mar 17.
BACKGROUND/OBJECTIVES: In this in vitro study, the effects of Stromal cell-derived factor-1 (SDF-1) was evaluated on the periodontal ligament-Mesenchymal Stem Cells (pdl-MSCs) functions.
Real-time cell analyzer-single plate (RTCA-SP) was employed for proliferation, and RTCA-dual purpose (DP) was utilized for pdl-MSCs migration potential treated with different SDF-1 concentrations (0, 0.1, 1, 10, 100, 200, and 400 ng/ml). Based on the dose-response findings, 10 ng/ml SDF-1 was used for further mRNA experiments. RNAs isolated at 6 and 24 h were checked using quantitative RT-PCR for mineralized tissue-associated genes including type I collagen (COL I), osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor 2 (Runx2). cRNA was synthesized for 6 h, and whole-genome array analysis was performed for over 47.000 probes. Data were subjected to quantile normalization before analysis.
Increased proliferation and migration were observed in pdl-MSCs treated with 0.1, 1, and 10 ng/ml SDF-1. Increased COL I was observed at both time points: 6 and 24 h. While there was no significant change for OCN, OPN, and Runx2 at 6 h, SDF-1 up-regulated OCN and OPN, but down-regulated Runx2 mRNA expressions at 24 h. IL-8 and ESM1 genes were differentially expressed over twofold when the pdl-MSCs were exposed to SDF-1 at whole-genome array analysis. IL-8 induction was confirmed with RT-PCR.
Findings of this study displayed that SDF-1 modulated pdl-MSCs which were important for periodontal regeneration, inducing migration and proliferation, and regulating extracellular matrix synthesis in favor of the formation of new attachment.
背景/目的:在这项体外研究中,评估基质细胞衍生因子-1(SDF-1)对牙周韧带间充质干细胞(pdl-MSCs)功能的影响。
采用实时细胞分析-单孔(RTCA-SP)检测不同 SDF-1 浓度(0、0.1、1、10、100、200 和 400ng/ml)处理后 pdl-MSCs 增殖和迁移能力,采用 RTCA-双用途(DP)检测 pdl-MSCs 迁移能力。根据剂量反应结果,选用 10ng/ml SDF-1 进行进一步的 mRNA 实验。6h 和 24h 时提取 RNA,用定量 RT-PCR 检测矿化组织相关基因,包括Ⅰ型胶原(COL I)、骨钙素(OCN)、骨桥蛋白(OPN)和 runt 相关转录因子 2(Runx2)。6h 时合成 cRNA,进行全基因组芯片分析,检测超过 47000 个探针。分析前进行分位数归一化。
pdl-MSCs 经 0.1、1 和 10ng/ml SDF-1 处理后增殖和迁移增加。6h 和 24h 时 COL I 均增加。虽然 OCN、OPN 和 Runx2 在 6h 时没有明显变化,但 SDF-1 在 24h 时上调 OCN 和 OPN,下调 Runx2 mRNA 表达。全基因组芯片分析显示,当 pdl-MSCs 暴露于 SDF-1 时,IL-8 和 ESM1 基因的表达差异超过两倍。通过 RT-PCR 证实了 IL-8 的诱导。
本研究结果显示,SDF-1 调节了 pdl-MSCs,这对牙周再生很重要,可诱导迁移和增殖,并调节细胞外基质合成,有利于新附着的形成。