Mineralization of three-dimensional osteoblast cultures is enhanced by the interaction of 1α,25-dihydroxyvitamin D3 and BMP2 via two specific vitamin D receptors
Mineralization of three-dimensional osteoblast cultures is enhanced by the interaction of 1α,25-dihydroxyvitamin D3 and BMP2 via two specific vitamin D receptors
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DOI：
10.1002/term.1770
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发表时间：
2016-01-01
影响因子：
3.3
通讯作者：
Schwartz, Zvi
中科院分区：
文献类型：
--
作者：
Chen, Jiaxuan;Dosier, Christopher R.;Schwartz, Zvi
1 α，25-二羟基维生素D3 [1，25（OH）2D 3]和骨形态发生蛋白-2（BMP 2）都用于刺激成骨细胞分化。1，25（OH）2D 3通过经典的类固醇激素受体机制和通过由两种受体介导的快速反应来调节成骨细胞，所述两种受体是传统的维生素D受体（VDR）和蛋白质二硫化物异构酶家族A成员3（Pdia 3）。1，25（OH）2D 3和BMP 2之间的相互作用，特别是在三维（3D）培养中，以及两种维生素D受体在这种相互作用中的作用还不清楚。我们用1，25（OH）2D 3或BMP 2或1，25（OH）2D 3和BMP 2一起处理野生型（WT）、Pdia 3沉默（Sh-Pdia 3）和VDR沉默（Sh-VDR）前成骨细胞MC 3 T3-E1细胞，并测量2D培养中的成骨细胞标志物表达和3D聚（ε-己内酯）-胶原支架模型中的矿化。定量PCR显示沉默Pdia 3或VDR对成骨细胞标志物的基线表达具有不同的影响。1，25（OH）2D 3 + BMP 2引起WT细胞中成骨细胞标志物表达的协同增加，而沉默Pdia 3或VDR减弱了这种作用。1，25（OH）2D 3 + BMP 2也在两种沉默的细胞系中引起Dlx 5的协同增加。显微计算机断层扫描（CT）显示，未处理的Sh-Pdia 3和Sh-VDR 3D培养物的矿化体积大于WT。1，25（OH）2D 3减少WT和Sh-VDR培养物中的矿物质; BMP 2增加矿化; 1，25（OH）2D 3 + BMP 2引起协同增加，但仅在WT培养物中。SEM显示，与WT细胞相比，沉默细胞在3D培养物中的矿化基质形态不同。这些数据表明1，25（OH）2D 3和BMP 2之间对骨生成和矿物质沉积的协同串扰，涉及VDR和Pdia 3。版权所有（c）2013约翰威利父子有限公司
1 alpha,25-Dihydroxyvitamin D3 [1,25(OH)2D3] and bone morphogenetic protein-2 (BMP2) are both used to stimulate osteoblastic differentiation. 1,25(OH)2D3 regulates osteoblasts through classical steroid hormone receptor mechanisms and through rapid responses that are mediated by two receptors, the traditional vitamin D receptor (VDR) and protein disulphide isomerase family A member 3 (Pdia3). The interaction between 1,25(OH)2D3 and BMP2, especially in three-dimensional (3D) culture, and the roles of the two vitamin D receptors in this interaction are not well understood. We treated wild-type (WT), Pdia3-silenced (Sh-Pdia3) and VDR-silenced (Sh-VDR) pre-osteoblastic MC3T3-E1 cells with either 1,25(OH)2D3, or BMP2, or with 1,25(OH)2D3 and BMP2 together, and measured osteoblast marker expression in 2D culture and mineralization in a 3D poly(epsilon-caprolactone)-collagen scaffold model. Quantitative PCR showed that silencing Pdia3 or VDR had a differential effect on baseline expression of osteoblast markers. 1,25(OH)2D3+BMP2 caused a synergistic increase in osteoblast marker expression in WT cells, while silencing either Pdia3 or VDR attenuated this effect. 1,25(OH)2D3+BMP2 also caused a synergistic increase in Dlx5 in both silenced cell lines. Micro-computed tomography (CT) showed that the mineralized volume of untreated Sh-Pdia3 and Sh-VDR 3D cultures was greater than that of WT. 1,25(OH)2D3 reduced mineral in WT and Sh-VDR cultures; BMP2 increased mineralization; and 1,25(OH)2D3+BMP2 caused a synergistic increase, but only in WT cultures. SEM showed that mineralized matrix morphology in 3D cultures differed for silenced cells compared to WT cells. These data indicate a synergistic crosstalk between 1,25(OH)2D3 and BMP2 toward osteogenesis and mineral deposition, involving both VDR and Pdia3. Copyright (c) 2013 John Wiley & Sons, Ltd.