Proliferation and odontogenic differentiation of BMP2 gene‑transfected stem cells from human tooth apical papilla: an in vitro study.
Proliferation and odontogenic differentiation of BMP2 gene‑transfected stem cells from human tooth apical papilla: an in vitro study.
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DOI：
10.3892/ijmm.2014.1862
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发表时间：
2014-10
影响因子：
5.4
通讯作者：
Zheng J
中科院分区：
文献类型：
--
作者：
Zhang W;Zhang X;Ling J;Liu W;Zhang X;Ma J;Zheng J
根尖乳头干细胞（Stem cells from the apical papilla，SCAP）具有成牙潜能，在恒牙牙根牙本质发育过程中起着关键作用。人骨形态发生蛋白2（BMP 2）是一个参与调控牙源性干细胞向牙本质分化的基因。然而，关于BMP 2基因对SCAP增殖和牙源性分化的影响知之甚少。本研究旨在探讨慢病毒介导的BMP 2基因转染人SCAP（SCAP/BMP 2）的体外牙向分化潜能。采用酶解法分离人牙根尖乳头，分离SCAP。SCAP的成骨和成脂分化特性证实了其具有多向分化潜能。流式细胞术（FCM）检测SCAP细胞表型。通过细胞计数试剂盒-8（CCK-8）分析空白载体转染的SCAP（SCAP/Vector）和SCAP/BMP 2的增殖状态。采用定量聚合酶链反应（qPCR）检测两组细胞碱性磷酸酶（ALP）、骨钙素（OCN）、牙本质涎磷蛋白（DSPP）和牙本质基质蛋白1（DMP 1）等牙源性基因的表达。转染后第16天进行ALP染色和茜素红（alizarinred，AR）染色。CCK-8、qPCR、ALP和AR染色的体外结果表明：i）SCAP/BMP 2具有与SCAP/Vector相当的增殖率; ii）与SCAP/Vector相比，SCAP/BMP 2通过上调ALP、OCN、DSPP和DMP 1基因而呈现出显著更好的分化成牙本质细胞的潜力; iii）与SCAP/Vector相比，SCAP/BMP 2形成更多的ALP颗粒和矿化沉积物。结果表明，慢病毒介导的BMP 2基因转染可增强人SCAP的体外牙向分化能力。
Stem cells from the apical papilla (SCAP) have odontogenic potential, which plays a pivotal role in the root dentin development of permanent teeth. Human bone morphogenetic protein 2 (BMP2) is a well-known gene that participates in regulating the odontogenic differentiation of dental tissue-derived stem cells. However, little is known regarding the effects of the BMP2 gene on the proliferation and odontogenic differentiation of SCAP. This study aimed to evaluate the odontogenic differentiation potential of lentiviral-mediated BMP2 gene-transfected human SCAP (SCAP/BMP2) in vitro. SCAP were isolated by enzymatic dissociation of human teeth apical papillae. The multipotential of SCAP was verified by their osteogenic and adipogenic differentiation characteristics. The phenotype of SCAP was evaluated by flow cytometry (FCM). The proliferation status of the blank vector-transfected SCAP (SCAP/Vector) and SCAP/BMP2 was analyzed by a cell counting kit-8 (CCK-8). Odontogenic genes, including alkaline phosphatase (ALP), osteocalcin (OCN), dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) of the two groups of cells were evaluated by quantitative polymerase chain reaction (qPCR). ALP staining and alizarin red (AR) staining of the cells was performed on the 16th day after transfection. In vitro results of CCK-8, qPCR, ALP and AR staining demonstrated that: i) SCAP/BMP2 had a comparable proliferation rate to SCAP/Vector; ii) SCAP/BMP2 presented significantly better potential to differentiate into odontoblasts compared to SCAP/Vector by upregulating ALP, OCN, DSPP and DMP1 genes; iii) more ALP granules and mineralized deposits were formed by SCAP/BMP2 as compared to SCAP/Vector. The results suggested that lentiviral-mediated BMP2 gene transfection enhances the odontogenic differentiation capacity of human SCAP in vitro.
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