Regulating orientation and phenotype of primary vascular smooth muscle cells by biodegradable films patterned with arrays of microchannels and discontinuous microwalls
Regulating orientation and phenotype of primary vascular smooth muscle cells by biodegradable films patterned with arrays of microchannels and discontinuous microwalls
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DOI：
10.1016/j.biomaterials.2010.04.059
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发表时间：
2010-08-01
期刊：
影响因子：
14
通讯作者：
Chan-Park, Mary B.
中科院分区：
文献类型：
--
作者：
Cao, Ye;Poon, Yin Fun;Chan-Park, Mary B.
已知体外培养的血管平滑肌细胞（vSMC）表现出表型增生。这种可塑性在血管组织工程中可能非常有用。合成表型对于组织支架上的细胞增殖是必要的，但细胞最终必须呈现静止、收缩的表型以实现正常的血管功能。 vSMC 表型的体外控制一直具有挑战性。这项研究表明，具有不连续壁的微通道支架可以支持原代vSMC增殖，并且当细胞在通道内达到汇合时，将细胞表型转变为更大的收缩性并促进细胞排列。进行了彻底的时间分辨研究，以表征收缩蛋白 α-肌动蛋白、钙调蛋白、肌球蛋白重链 (MHC) 和平滑蛋白的表达作为时间和微通道支架上初始细胞密度的函数。结果一致表明，在微通道基底上培养的原代vSMC基本上与微壁平行排列，只有当细胞达到汇合时才变得更加伸长并显着增加其收缩蛋白的表达。 MHC 免疫染色在汇合后的微图案细胞中可见，但在平坦基底细胞或非汇合微图案细胞中则不然，这进一步验证了汇合通道受限的 vSMC 的收缩性增加。汇合的扁平培养物中沉积的弹性蛋白和胶原蛋白的总量高于汇合的微图案培养物中的沉积总量，这也证实了通道受限细胞的更高收缩性。这些结果表明，我们的微通道薄膜可以触发原代 vSMC 从增殖状态转变为汇合时更具收缩性的表型。 (C) 2010 Elsevier Ltd. 保留所有权利。
Vascular smooth muscle cells (vSMCs) cultured in vitro are known to exhibit phenotype hyperplasticity. This plasticity is potentially very useful in tissue engineering of blood vessels. The synthetic phenotype is necessary for cell proliferation on the tissue scaffold but the cells must ultimately assume a quiescent, contractile phenotype for normal vascular function. In vitro control of vSMC phenotype has been challenging. This study shows that microchannel scaffolds with discontinuous walls can support primary vSMC proliferation and, when the cells reach confluence inside the channels, transform the cell phenotype towards greater contractility and promote cell alignment. A thorough time-resolved study was undertaken to characterize the expression of the contractile proteins alpha-actin, calponin, myosin heavy chain (MHC) and smoothelin as a function of time and initial cell density on microchannel scaffolds. The results consistently indicate that primary vSMCs cultured on the microchannel substrate substantially align parallel to the microwalls, become more elongated and significantly increase their expression of contractile proteins only when the cells reach confluence. MHC immunostaining was visible in the micropatterned cells after confluence but not in flat substrate cells or non-confluent micropatterned cells, which further verifies the increased contractility of the confluent channel-constrained vSMCs. The higher total amount of deposited elastin and collagen in confluent flat cultures than in confluent micropatterned cultures also provides confirmation of the higher contractility of the channel-constrained cells. These results establish that our microchanneled film can trigger the switch of primary vSMCs from a proliferative state to a more contractile phenotype at confluence. (C) 2010 Elsevier Ltd. All rights reserved.