Division of Biomedical Engineering, Department of Life Science, Faculty of New Sciences and Technologies, University of Tehran, Tehran 439957131, Iran.
Division of Biomedical Engineering, Department of Life Science, Faculty of New Sciences and Technologies, University of Tehran, Tehran 439957131, Iran.
Tissue Cell. 2023 Apr;81:101996. doi: 10.1016/j.tice.2022.101996. Epub 2022 Dec 13.
In the development of vascular tissue engineering, particularly in the case of small diameter vessels, one of the key obstacles is the blockage of these veins once they enter the in vivo environment. One of the contributing factors to this problem is the aberrant proliferation and migration of vascular smooth muscle cells (VSMCs) from the media layer of the artery to the interior of the channel. Two distinct phenotypes have been identified for smooth muscle cells, namely synthetic and contractile. Since the synthetic phenotype plays an essential role in the unusual growth and migration, the aim of this study was to convert the synthetic phenotype into the contractile one, which is a solution to prevent the abnormal growth of VSMCs. To achieve this goal, these cells were subjected to electrical signals, using a 1000 μA sinusoidal stimulation at 10 Hz for four days, with 20 min duration per 24 h. The morphological transformations and changes in the expression of vimentin, nestin, and β-actin proteins were then studied using ICC and flow cytometry assays. Also, the expression of VSMC specific markers such as smooth muscle myosin heavy chain (SMMHC) and smooth muscle alpha-actin (α-SMA) were evaluated using RT-PCR test. In the final phase of this study, the sheep decellularized vessel was employed as a scaffold for seeding these cells. Based on the results, electrical stimulation resulted in some morphological alterations in VSMCs. Furthermore, the observed reductions in the expression levels of vimentin, nestin and β-actin proteins and increase in the expression of SMMHC and α-SMA markers showed that it is possible to convert the synthetic phenotype to the contractile one using the studied regime of electrical stimulation. Finally, it can be concluded that electrical stimulation can significantly affect the phenotype of VSMCs, as demonstrated in this study.
在血管组织工程的发展中，特别是在小直径血管的情况下，其中一个关键障碍是这些静脉进入体内环境后会被阻塞。造成这个问题的一个因素是血管平滑肌细胞（VSMCs）从动脉的中层异常增殖和迁移到通道内部。平滑肌细胞已经确定了两种不同的表型，即合成型和收缩型。由于合成型表型在异常生长和迁移中起着至关重要的作用，本研究的目的是将合成型表型转化为收缩型表型，这是防止 VSMCs 异常生长的一种解决方案。为了实现这一目标，这些细胞受到电信号的刺激，使用 1000 μA 的正弦波刺激，频率为 10 Hz，持续 20 分钟，每天 24 小时。然后使用 ICC 和流式细胞术检测 vimentin、nestin 和 β-actin 蛋白的表达变化。此外，还通过 RT-PCR 试验评估了 VSMC 特异性标志物，如平滑肌肌球蛋白重链（SMMHC）和平滑肌α-肌动蛋白（α-SMA）的表达。在本研究的最后阶段，羊去细胞血管被用作接种这些细胞的支架。基于这些结果，电刺激导致 VSMCs 发生一些形态变化。此外，观察到 vimentin、nestin 和 β-actin 蛋白的表达水平降低，SMMHC 和 α-SMA 标志物的表达增加，表明使用研究的电刺激方案可以将合成型表型转化为收缩型表型。最后，可以得出结论，电刺激可以显著影响 VSMCs 的表型，正如本研究所示。
