Priming integrin alpha 5 promotes the osteogenic differentiation of human periodontal ligament stem cells due to cytoskeleton and cell cycle changes
Priming integrin alpha 5 promotes the osteogenic differentiation of human periodontal ligament stem cells due to cytoskeleton and cell cycle changes
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由于细胞骨架和细胞周期的变化，引发整合素α5促进人牙周膜干细胞的成骨分化
DOI：
10.1016/j.jprot.2018.03.008
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Buling Wu
中科院分区：
文献类型：
--
作者：
He Wang;Jianjia Li;Xiaoyi Zhang;Tingting Ning;D;an Ma;Yihong Ge;Shuaimei Xu;Yilin Hao;Buling Wu
为寻找牙周组织再生的潜在靶点，本研究旨在探讨整合素α5(ITGA5)在人牙周膜干细胞(PDLSCs)中的作用。用Transwell法、细胞计数试剂盒8(CCK8)法、细胞周期分析、碱性磷酸酶(ALP)活性、茜素红染色和免疫印迹等方法研究ITGA5对PDLSC迁移、增殖和成骨分化的影响。通过裸鼠皮下移植细胞和羟基磷灰石/β-磷酸三钙复合体(HA/β-TcP)观察其体内作用。利用iTRAQ蛋白质组学技术对其作用机制进行了初步探讨，并通过免疫印迹和免疫荧光方法进行了验证。我们发现，ITGA5的强制表达促进了PDLSCs的增殖、迁移和成骨能力，而抑制ITGA5的表达则具有相反的作用。粘着斑激酶(FAK)、磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)、丝裂原活化蛋白激酶/细胞外信号调节蛋白激酶1和2(MEK1/2/ERK1/2)的磷酸化在此过程中起着至关重要的作用。ITGA5在PDLSCs中的强制表达增加了体内骨样组织和PDL样组织的形成。蛋白质组学和生物信息学分析显示细胞骨架和细胞周期改变。角蛋白、II型细胞骨架6B(KRT6B)和结蛋白(DES)可区分这一过程，并可作为PDLSC分化的新标记物。牙周炎治疗最有前景的方法之一是利用牙周膜干细胞再生牙周膜。虽然仍有许多障碍需要解决，但PDLSC成骨分化的调控是主要关注的问题。本研究证实了ITGA5启动肽的潜在临床价值，它可能用于PDL组织的修复和再生。这项研究阐明的机制将有助于推动其应用。
To seek a potential target for periodontal tissue regeneration, this study aimed to explore the role of Integrin alpha 5 (ITGA5) in human periodontal ligament stem cells (PDLSCs). Transwell assay, Cell Counting Kit 8 (CCK8) assay, cell cycle assay, alkaline phosphatase (ALP) activity, alizarin red staining, and western blot were used to investigate the effects of ITGA5 on PDLSC migration, proliferation and osteogenic differentiation. The in vivo effect was investigated by nude mice subcutaneous transplantation with cell and hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) complex. The involved mechanism was explored by the iTRAQ proteomic technique and validated by western blot and immunofluorescence. We found that ITGA5forced expression enhanced the proliferation, migration, and osteogenic capacity of PDLSCs, while inhibited ITGA5 expression had the opposite effects. The phosphorylation of focal adhesion kinase (FAK), phosphatidylinositide 3-kinases/protein kinase B (PI3K/AKT), and mitogen-activated protein kinase kinase/extracellular signal-regulated protein kinases 1 and 2 (MEK1/2/ERK1/2) were crucial in this process. Forced expression of ITGA5 in PDLSCs increased osteoid and PDL-like tissue formation in vivo. Proteomic and bioinformatic analysis revealed that cytoskeleton and cell cycle changes were involved. Keratin, type II cytoskeletal 6B (KRT6B) and desmin (DES) may distinguish this process and serve as new markers of PDLSC differentiation.SignificancePeriodontitis is highly prevalent and can impair PDL and teeth functioning. One of the most promising therapies to periodontitis therapies is PDL regeneration by utilizing PDLSCs. While many obstacles remain to be resolved, the regulation of PDLSC osteogenic differentiation is a main concern. The present study demonstrated the potential clinical value of an ITGA5 priming peptide, which may be utilized in PDL tissue repair and regeneration. The mechanism elucidated in this study would help to fuel its application.