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Periostin promotes migration, proliferation, and differentiation of human periodontal ligament mesenchymal stem cells

机译:骨蛋白促进人牙周韧带间充质干细胞的迁移,增殖和分化

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Overview: Periostin (POSTN) is critical to bone and dental tissue morphogenesis, postnatal development, and maintenance; however, its roles in tissue repair and regeneration mediated by human periodontal ligamentmesenchymal stemcells (PDLSCs) remain unclear. The present study was designed to evaluate the effects of POSTN on hPDLSCs in vitro. Materials and Methods: hPDLSCs were isolated and characterized by their expression of the cell surface markers CD44, CD90, CD105, CD34, and CD45. Next, 100 ng/mL recombinant human POSTN protein (rhPOSTN) was used to stimulate the hPDLSCs. Lentiviral POSTN shRNA was used to knockdown POSTN. The cell counting kit-8 (CCK8) and scratch assay were used to analyze cell proliferation and migration, respectively. Osteogenic differentiation was investigated using an alkaline phosphatase (ALP) activity assay, alizarin staining, and quantitative calcium analysis and related genes/protein expression assays. Results: Isolated hPDLSCs were positive for CD44, CD90, and CD105 and negative for CD34 and CD45. In addition, 100 ng/mL rhPOSTN significantly accelerated scratch closure, and POSTN-knockdown cells presented slower closure at 24 h and 48 h. Furthermore, the integrin inhibitor Cilengitide depressed the scratch closure that was enhanced by POSTN at 24 h. The CCK8 assay showed that 100 ng/mL rhPOSTN promoted hPDLSC proliferation. Moreover, 100 ng/mL rhPOSTN increased the expression of RUNX2, OSX, OPN, OCN, and VEGF and enhanced ALP activity and mineralization. POSTN silencing decreased the expression of RUNX2, OSX, OPN, OCN, and VEGF and inhibited ALP activity and mineralization. Conclusions: POSTN accelerated the migration, proliferation, and osteogenic differentiation of hPDLSCs.
机译:概述:Periostin(Postn)对骨骼和牙科组织形态发生,产后开发和维护至关重要;然而,它在由人牙周韧带梗死梗塞梗塞细胞(PDLSCs)介导的组织修复和再生中的作用仍然尚不清楚。本研究旨在评估Postn对体外HPDLSCS的影响。材料和方法:分离HPDLSCs,其特征在于它们的细胞表面标志物CD44,CD90,CD105,CD34和CD45的表达。接下来,使用100ng / ml重组人员Postn蛋白(RhPostn)刺激HPDLSC。慢病毒Postn shrna被用来击倒postn。用于分别分析细胞增殖和迁移的细胞计数试剂盒-8(CCK8)和划痕测定。使用碱性磷酸酶(ALP)活性测定,茜素染色和定量钙分析以及相关基因/蛋白表达测定来研究骨质发生分化。结果:分离的HPDLSCs对于CD44,CD90和CD105是阳性的,CD34和CD45负阴性。此外,100ng / ml rhPostn显着加速划痕闭合,并且在24小时和48小时时呈现出较慢的封闭件。此外,整联蛋白抑制剂Cilengitide抑制了由Postn在24小时增强的划痕闭合。 CCK8测定表明,100ng / ml rhPostn促进了HPDLSC增殖。此外,100ng / ml rhPostn增加了Runx2,OSX,OPN,OCN和VEGF的表达以及增强的ALP活性和矿化。 Postn沉默降低了runx2,osx,opn,OCN和VEGF的表达,并抑制ALP活性和矿化。结论:Postn加速了HPDLSCs的迁移,增殖和成骨分化。

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