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首页> 外文期刊>American Journal of Physiology >Altered phenotypic gene expression of 10T1/2 mesenchymal cells in nonuniformly stretched PEGDA hydrogels
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Altered phenotypic gene expression of 10T1/2 mesenchymal cells in nonuniformly stretched PEGDA hydrogels

机译：非均匀拉伸PEGDA水凝胶中10T1 / 2间充质细胞表型基因表达的改变

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Disease-related phenotype modulation of many cell types has been shown to be closely related to mechanical loading conditions; for example, vascular smooth muscle cell (SMC) phenotype shift from a mature, contractile state to a proliferative, synthetic state contributes to the formation of neointimal tissue during atherosclerosis and restenosis development and is related to SMC mechanical loading in vivo. The majority of past in vitro cell-stretching experiments have employed simplistic (uniform, uniaxial or biaxial) stretching environments to elucidate mechanobiological pathways involved in phenotypic shifts. However, the in vivo mechanics of the vascular wall consists of highly nonuniform stretch. Here we subjected 10T1/2 murine mesenchymal cells (an SMC precursor) to two- and three-dimensional nonuniform stretch environments. After 24 h of stretch, cells on an elastomeric membrane demonstrated varied proliferation [assessed by 5-bromo-2′-deoxyuridine (BrdU) incorporation] depending on location upon the membrane, with maximal proliferation occurring in a region of high, uniaxial stretch. Cells subjected to a nonuniform stretching regimen within three-dimensional polyethylene glycol diacrylate (PEGDA) hydrogel constructs demonstrated marked changes in mRNA expression of several phenotype-related proteins, indicating a sort of "hybrid" phenotype with contractile and synthetic markers being both upregulated and downregulated. Furthermore, expression levels of mRNAs were significantly different between various locations within the stretched gel. With the proliferation results, these data exhibit the capability of nonuniform stretching devices to induce heterogeneous cell responses, potentially indicative of spatial distributions of disease- related behaviors in vivo.
机译：许多细胞类型的疾病相关表型调节已被证明与机械负荷条件密切相关。例如，血管平滑肌细胞（SMC）表型从成熟的收缩状态转变为增殖的合成状态有助于动脉粥样硬化和再狭窄发展过程中新内膜组织的形成，并且与体内SMC机械负荷有关。过去的大多数体外细胞拉伸实验都采用了简单的（均匀，单轴或双轴）拉伸环境来阐明涉及表型转变的机械生物学途径。然而，血管壁的体内力学由高度不均匀的拉伸组成。在这里，我们使10T1 / 2小鼠间充质细胞（SMC前体）经历了二维和三维非均匀拉伸环境。拉伸24小时后，弹性膜上的细胞表现出不同的增殖[通过5-溴2'-脱氧尿苷（BrdU）掺入评估]，具体取决于膜上的位置，最大的增殖发生在高单轴拉伸区域。在三维聚乙二醇二丙烯酸酯（PEGDA）水凝胶构建物中进行非均匀拉伸方案的细胞显示出几种表型相关蛋白的mRNA表达有明显变化，表明一种“混合”表型，其收缩和合成标记均被上调和下调。此外，在拉伸凝胶内的各个位置之间，mRNA的表达水平显着不同。有了增殖结果，这些数据显示了非均匀拉伸装置诱导异质细胞反应的能力，这可能表明体内疾病相关行为的空间分布。

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