Carbon Nanotubes Instruct Physiological Growth and Functionally Mature Syncytia: Nongenetic Engineering of Cardiac Myocytes
Carbon Nanotubes Instruct Physiological Growth and Functionally Mature Syncytia: Nongenetic Engineering of Cardiac Myocytes
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DOI：
10.1021/nn4002193
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发表时间：
2013-07-01
期刊：
影响因子：
17.1
通讯作者：
Mestroni, Luisa
中科院分区：
文献类型：
--
作者：
Martinelli, Valentina;Cellot, Giada;Mestroni, Luisa
心肌组织工程是目前最现实的心脏修复策略之一。我们最近发现碳纳米管支架能够促进心肌细胞的细胞分裂和成熟。在这里，我们测试的假设，即碳纳米管支架促进心肌细胞的生长和成熟，通过改变基因表达程序，实现细胞电生理特性和改善网络和成熟的功能合胞体。在我们的研究中，我们结合联合收割机显微镜，生物和电生理方法，钙成像，以验证是否新生大鼠心室肌细胞培养的基板上的多壁碳纳米管获得生理上更成熟的表型相比，控制（明胶）。我们发现，碳纳米管基质刺激诱导基因表达谱分化和生理生长，α-肌球蛋白重链增加2倍（P < 0.001），肌浆网Ca 2 + ATP酶2a上调。相反，病理性肥大的标志物保持不变（β-肌球蛋白重链、骨骼α-肌动蛋白、心房利钠肽）。这些修饰被连接蛋白-43基因表达、间隙连接和功能性合胞体的增加所证实。此外，碳纳米管似乎对苯肾上腺素的病理刺激发挥保护作用。最后，碳纳米管上的心肌细胞表现出更成熟的合胞体和细胞内钙信号传导的电生理表型。因此，碳纳米管与心肌细胞相互作用具有促进生理生长和功能成熟的能力。这些特性在目前令人烦恼的组织工程领域是独一无二的，并为心脏修复的创新疗法的发展提供了前所未有的前景。
Myocardial tissue engineering currently represents one of the most realistic strategies for cardiac repair. We have recently discovered the ability of carbon nanotube scaffolds to promote cell division and maturation in cardiomyocytes. Here, we test the hypothesis that carbon nanotube scaffolds promote cardiomyocyte growth and maturation by altering the gene expression program, implementing the cell electrophysiological properties and improving networking and maturation of functional syncytia. In our study, we combine microscopy, biological and electrophysiological methodologies, and calcium imaging, to verify whether neonatal rat ventricular myocytes cultured on substrates of multiwall carbon nanotubes acquire a physiologically more mature phenotype compared to control (gelatin). We show that the carbon nanotube substrate stimulates the induction of a gene expression profile differentiation and physiological growth, with a 2-fold increase of a-myosin heavy chain (P < 0.001) and upregulation of sarcoplasmic reticulum Ca2+ ATPase 2a. In contrast, markers of pathological hypertrophy remain unchanged (beta-myosin heavy chain, skeletal a-actin, atrial natriuretic peptide). These modifications are paralleled by an increase of connexin-43 gene expression, gap junctions and functional syncytia. Moreover, carbon nanotubes appear to exert a protective effect against the pathologic stimulus of phenylephrine. Finally, cardiomyocytes on carbon nanotubes demonstrate a more mature electrophysiological phenotype of syncytia and intracellular calcium signaling. Thus, carbon nanotubes interacting with cardiomyocytes have the ability to promote physiological growth and functional maturation. These properties are unique in the current vexing field of tissue engineering, and offer unprecedented perspectives in the development of innovative therapies for cardiac repair.