Calcifying nanoparticles promote mineralization in vascular smooth muscle cells: implications for atherosclerosis.
Calcifying nanoparticles promote mineralization in vascular smooth muscle cells: implications for atherosclerosis.
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DOI：
10.2147/ijn.s63189
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发表时间：
2014
影响因子：
8
通讯作者：
Miller VM
中科院分区：
文献类型：
--
作者：
Hunter LW;Charlesworth JE;Yu S;Lieske JC;Miller VM
磷酸钙矿物质和蛋白质的纳米复合物（钙化纳米颗粒[CNP]）可作为矿物质伴侣。因此，CNP可能是软组织钙化过程的结果和原因。本研究确定了CNP是否可以在体外增加动脉血管平滑肌细胞的钙化。直径为210 nm的CNP从人血清中体外增殖。猪主动脉平滑肌细胞在不存在（对照）或存在2 mM磷酸盐（[P]阳性钙化对照）的培养基中培养长达28天，或单次暴露于CNP 3天后培养。透射电子显微镜用于表征CNP并检查其细胞摄取。通过光学显微镜和von Kossa染色观察钙沉积，并通过比色法定量。通过共聚焦显微镜对活/死染色细胞进行定量，同时通过荧光标记暴露的磷脂酰丝氨酸检查细胞凋亡。CNP，以及较小的钙晶体，通过透射电子显微镜观察到的第3天，在CNP处理的，但不是P处理的细胞。到第28天，在CNP和P处理的细胞的多细胞结节内可见相似量的钙沉积。在所有处理下，细胞凋亡随细胞密度增加而增加。CNP处理增加了与矿化多细胞结节相关的凋亡小体和细胞碎片的密度。外源性CNP在体外被主动脉平滑肌细胞摄取，并增强平滑肌来源的凋亡小体在矿化位点的积累。因此，CNP可加速血管钙化。
Nano-sized complexes of calcium phosphate mineral and proteins (calcifying nanoparticles [CNPs]) serve as mineral chaperones. Thus, CNPs may be both a result and cause of soft tissue calcification processes. This study determined if CNPs could augment calcification of arterial vascular smooth muscle cells in vitro. CNPs 210 nm in diameter were propagated in vitro from human serum. Porcine aortic smooth muscle cells were cultured for up to 28 days in medium in the absence (control) or presence of 2 mM phosphate ([P] positive calcification control) or after a single 3-day exposure to CNPs. Transmission electron-microscopy was used to characterize CNPs and to examine their cellular uptake. Calcium deposits were visualized by light microscopy and von Kossa staining and were quantified by colorimetry. Cell viability was quantified by confocal microscopy of live-/dead-stained cells and apoptosis was examined concurrently by fluorescent labeling of exposed phosphatidylserine. CNPs, as well as smaller calcium crystals, were observed by transmission electron-microscopy on day 3 in CNP-treated but not P-treated cells. By day 28, calcium deposits were visible in similar amounts within multicellular nodules of both CNP- and P-treated cells. Apoptosis increased with cell density under all treatments. CNP treatment augmented the density of apoptotic bodies and cellular debris in association with mineralized multicellular nodules. Exogenous CNPs are taken up by aortic smooth muscle cells in vitro and potentiate accumulation of smooth-muscle-derived apoptotic bodies at sites of mineralization. Thus, CNPs may accelerate vascular calcification.