Effect of TNF-α on the proliferation and osteogenesis of human periodontal mesenchymal stem cells.
Effect of TNF-α on the proliferation and osteogenesis of human periodontal mesenchymal stem cells.
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DOI：
10.3892/etm.2021.9851
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发表时间：
2021-05
影响因子：
2.7
通讯作者：
Zhu L
中科院分区：
文献类型：
--
作者：
Cao Y;Wang Y;Li C;Jiang Q;Zhu L
本研究旨在探讨肿瘤坏死因子-α（TNF-α）对人牙周膜间充质干细胞（hPDLSCs）增殖和成骨的影响。流式细胞术检测hPDLSCs中抗原表达。将hPDLSCs分为4组：对照组，不加TNF-α处理; 3个实验组，分别加0.1、1和10 ng/ml TNF-α处理。采用Cell Counting Kit-8法检测TNF-α对hPDLSCs体外增殖的影响。碱性磷酸酶和茜素红染色检测成骨细胞的分化情况，逆转录-定量PCR检测Runx 2、骨钙素（OCN）和I型胶原（Col-I）mRNA和蛋白表达水平。10 ng/ml TNF-α处理后，细胞增殖明显高于未处理对照组（P<0.01）。此外，碱性磷酸酶活性、茜素红矿化结节大小以及成骨分化标志物Runx 2、OCN和COL-I的基因和蛋白表达水平均受到显著抑制（均P<0.05）。以上结果表明，10 ng/ml TNF-α促进hPDLSC体外增殖，抑制hPDLSC向成骨细胞分化，为hPDLSC介导的牙周组织再生调控提供了实验依据。
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 in vitro 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 in vitro and inhibited osteogenic differentiation of hPDLSCs, providing an experimental basis for regulation of hPDLSC-mediated periodontal tissue regeneration.
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