Intrafibrillar-silicified collagen scaffolds enhance the osteogenic capacity of human dental pulp stem cells
Intrafibrillar-silicified collagen scaffolds enhance the osteogenic capacity of human dental pulp stem cells
复制标题
纤维内硅化胶原支架增强人牙髓干细胞的成骨能力
DOI：
10.1016/j.jdent.2014.03.016
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发表时间：
2014-07-01
影响因子：
4.4
通讯作者：
Chen, Ji-hua
中科院分区：
文献类型：
--
作者：
Niu, Li-na;Sun, Jia-qi;Chen, Ji-hua
目的：本研究旨在探讨纤维内硅化胶原支架（intrapibrillar-silicified collagen scaffolds，ISCS）对人牙髓干细胞（human dental pulp stem cells，hDPSCs）成骨分化的影响。流式细胞仪检测细胞周期和细胞凋亡，检测hDPSCs的存活率。采用逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法检测hDPSCs成骨标志基因和蛋白的表达水平。碱性磷酸酶（ALP）染色和茜素红S法检测hDPSCs的ALP活性和钙沉积能力。此外，将hDPSC和支架皮下植入裸鼠体内8周。结果：ISCS对hDPSCs无不良影响，能促进hDPSCs增殖，并能显著上调hDPSCs成骨相关基因和蛋白的表达。与ISCS共培养的hDPSC在ODM中表现出最高的ALP活性和钙沉积。结论：与NCS相比，硅化胶原支架可显著促进hDPSCs的增殖、成骨分化和矿化。该研究表明，结合使用hDPSC和ISCS来促进骨样组织形成是临床骨修复和再生的有希望的方法。(C)2014爱思唯尔有限公司版权所有。
Objectives: The present study investigated the effects of intrafibrillar-silicified collagen scaffolds (ISCS) on the osteogenic differentiation of human dental pulp stem cells (hDPSCs) in vitro and in vivo.Methods: The hDPSCs were co-cultured with ISCS or nonsilicified collagen scaffolds (NCS) in control medium (CM) or osteogenic differentiation medium (ODM). Cell cycle and cell apoptosis were analyzed with flow cytometry to measure the viability of hDPSCs. Reverse transcription-polymerase chain reaction (RT-PCR) and western blotting were used to evaluate the expression levels of osteogenic marker genes and proteins of hDPSCs. Alkaline phosphatase (ALP) staining and alizarin red S assay were used to evaluate the ALP activity of hDPSCs and their calcium deposition potential. In addition, hDPSCs and scaffolds were implanted subcutaneously in nude mice for 8 weeks. Harvested tissues were immunohistochemically stained for osteocalcin (OCN) expression from hDPSCs, and stained with alizarin red S for examination of their calcium deposition in vivo.Results: The ISCS had no adverse effect on hDPSCs, promoted their proliferation, and significantly up-regulated the expression of osteogenesis-related genes and proteins. The hDPSCs co-cultured with ISCS in ODM exhibited the highest ALP activity and calcium deposition in vitro. The ISCS promoted the OCN expression and calcium deposition of hDPSCs after ectopic transplantation in vivo.Conclusions: Intrafibrillar-silicified collagen scaffolds significantly promoted the proliferation, osteogenic differentiation and mineralization of hDPSCs, when compared with NCS. This study demonstrates combining the use of hDPSCs and ISCS to promote bone-like tissue formation is a promising approach for clinical bone repair and regeneration. (C) 2014 Elsevier Ltd. All rights reserved.