Influence of Substrate Stiffness on the Phenotype of Heart Cells
Influence of Substrate Stiffness on the Phenotype of Heart Cells
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DOI：
10.1002/bit.22647
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发表时间：
2010-04-15
影响因子：
3.8
通讯作者：
Radisic, Milica
中科院分区：
文献类型：
--
作者：
Bhana, Bashir;Iyer, Rohin K.;Radisic, Milica
成年心肌细胞（CM）几乎没有再生能力，这激发了努力设计可以模仿天然心肌功能和机械性能的心脏组织。尽管已经探索了矩阵刚度对单个CM的影响，但对单层和组织水平的研究的关注较少。这项研究的目的是表征底物机械刚度对心脏细胞表型和功能特性的影响。将新生大鼠心脏细胞接种到胶原蛋白涂层的聚丙烯酰胺（PA）底物上，杨氏模量为3、22、50和144 kPa。没有PA的胶原蛋白涂层玻璃盖玻片具有有效的“无限”刚度的表面。天然新生大鼠心脏组织的局部弹性模量范围为4.0至11.4 kPa（平均值为6.8 kPa），对于天然成年大鼠心脏组织的范围为11.9至46.2 kPa（平均值25.6 kPa），以激励我们选择的选择上述PA凝胶刚度。总体而言，最低的刚度PA底物（3 kPa）表现出最低的激发阈值（ET； 3.5 +/- 0.3 V/cm），肌钙蛋白I I染色（52％呈阳性染色的面积），但细胞密度降低，收缩力（0.18 +/- 0.1 mn/mm 2）和细胞伸长（长宽比= 1.3-1.4）。较高的刚度（144 kPa）PA底物表现出降低的肌钙蛋白I染色（30％阳性染色面积），成纤维细胞密度增加（70％阳性染色面积）和较差的电兴奋性。发现与本地成年大鼠心肌（22-50 kPa）相当的刚度中等刚度PA PA底物对于心脏细胞的形态和功能是最佳的，具有上伸长率（纵横比> 4.3），合理的ET（范围为3.95 +//-范围0.8至4.4 +/- 0.7 v/cm），高收缩力开发（范围从0.52 +/- 0.2到1.60 +/- 0.6 Mn/mm（2））和发达的条纹，都与分化的表型一致。生物技术。 Bioeng。 2010; 105：1148-1160。 （c）2009年Wiley期刊公司
Adult cardiomyocytes (CM) retain little capacity to regenerate, which motivates efforts to engineer heart tissues that can emulate the functional and mechanical properties of native myocardium. Although the effects of matrix stiffness on individual CM have been explored, less attention was devoted to studies at the monolayer and the tissue level. The purpose of this study was to characterize the influence of substrate mechanical stiffness on the heart cell phenotype and functional properties. Neonatal rat heart cells were seeded onto collagen-coated polyacrylamide (PA) substrates with Young's moduli of 3, 22, 50, and 144 kPa. Collagen-coated glass coverslips without PA represented surfaces with effectively "infinite" stiffness. The local elastic modulus of native neonatal rat heart tissue was measured to range from 4.0 to 11.4 kPa (mean value of 6.8 kPa) and for native adult rat heart tissue from 11.9 to 46.2 kPa (mean value of 25.6 kPa), motivating our choice of the above PA gel stiffness. Overall, by 120 h of cultivation, the lowest stiffness PA substrates (3 kPa) exhibited the lowest excitation threshold (ET; 3.5 +/- 0.3 V/cm), increased troponin I staining (52% positively stained area) but reduced cell density, force of contraction (0.18 +/- 0.1 mN/mm 2), and cell elongation (aspect ratio = 1.3-1.4). Higher stiffness (144 kPa) PA substrates exhibited reduced troponin I staining (30% positively stained area), increased fibroblast density (70% positively stained area), and poor electrical excitability. Intermediate stiffness PA substrates of stiffness comparable to the native adult rat myocardium (22-50 kPa) were found to be optimal for heart cell morphology and function, with superior elongation (aspect ratio > 4.3), reasonable ET (ranging from 3.95 +/- 0.8 to 4.4 +/- 0.7 V/cm), high contractile force development (ranging from 0.52 +/- 0.2 to 1.60 +/- 0.6 mN/mm(2)), and well-developed striations, all consistent with a differentiated phenotype. Biotechnol. Bioeng. 2010; 105: 1148-1160. (C) 2009 Wiley Periodicals, Inc.