Cao Yiting, Wang Yiwei, Li Chenlin, Jiang Qian, Zhu Laikuan
Department of Pediatric Dentistry, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China.
Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, Shanghai 200011, P.R. China.
Exp Ther Med. 2021 May;21(5):434. doi: 10.3892/etm.2021.9851. Epub 2021 Feb 26.
The aim of the present study was to investigate the effect of tumor necrosis factor-α (TNF-α) on the proliferation and osteogenesis of human periodontal mesenchymal stem cells (hPDLSCs). Antigen expression in hPDLSCs was detected by flow cytometry. hPDLSCs were divided into four groups: A control group with no TNF-α treatment, and three experimental groups treated with 0.1, 1 and 10 ng/ml TNF-α, respectively. The effect of TNF-α on proliferation of hPDLSCs was detected using a Cell Counting Kit-8 assay. Differentiation into an osteogenic lineage was detected by alkaline phosphatase sand alizarin red staining, and the mRNA and protein expression levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and type I collagen (Col-I) were detected using reverse transcription-quantitative PCR and western blot respectively. Following treatment with 10 ng/ml TNF-α, proliferation was significantly increased compared with an untreated control group (P<0.01). Additionally, there was a significant inhibition of alkaline phosphatase enzyme activity, alizarin red mineralization node size, and in the gene and protein expression levels of osteogenic differentiation markers, including Runx2, OCN and COL-I (all, P<0.05). Taken together, the results indicated that treatment with 10 ng/ml TNF-α promoted the proliferation of hPDLSCs and inhibited osteogenic differentiation of hPDLSCs, providing an experimental basis for regulation of hPDLSC-mediated periodontal tissue regeneration.
本研究的目的是探讨肿瘤坏死因子-α（TNF-α）对人牙周膜间充质干细胞（hPDLSCs）增殖和成骨的影响。通过流式细胞术检测hPDLSCs中的抗原表达。hPDLSCs分为四组：未用TNF-α处理的对照组，以及分别用0.1、1和10 ng/ml TNF-α处理的三个实验组。使用细胞计数试剂盒-8检测TNF-α对hPDLSCs增殖的影响。通过碱性磷酸酶和茜素红染色检测向成骨谱系的分化，分别使用逆转录定量PCR和蛋白质印迹法检测矮小相关转录因子2（Runx2）、骨钙素（OCN）和I型胶原（Col-I）的mRNA和蛋白质表达水平。用10 ng/ml TNF-α处理后，与未处理的对照组相比，增殖显著增加（P<0.01）。此外，碱性磷酸酶活性、茜素红矿化结节大小以及包括Runx2、OCN和COL-I在内的成骨分化标志物的基因和蛋白质表达水平均受到显著抑制（均为P<0.05）。综上所述，结果表明10 ng/ml TNF-α处理促进了hPDLSCs的增殖并抑制了hPDLSCs的成骨分化，为调控hPDLSC介导的牙周组织再生提供了实验依据。
