Ramirez-Calderon Gustavo, Saleh Abdulelah, Hidalgo Castillo Tania Cecilia, Druet Victor, Almarhoon Bayan, Almulla Latifah, Adamo Antonio, Inal Sahika
Laboratory of Stem Cells and Diseases, Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
Organic Bioelectronics Laboratory, Biological and Environmental Science and Engineering Division, KAUST, Thuwal 23955-6900, Saudi Arabia.
ACS Appl Mater Interfaces. 2024 Dec 11;16(49):66900-66910. doi: 10.1021/acsami.3c18919. Epub 2024 Apr 15.
Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are a promising cell source for cardiac regenerative medicine and modeling. However, hPSC-CMs exhibit immature structural and functional properties compared with adult cardiomyocytes. Various electrical, mechanical, and biochemical cues have been applied to enhance hPSC-CM maturation but with limited success. In this work, we investigated the potential application of the semiconducting polymer poly{[N,N'-bis(2-octyldodecyl)-naphthalene-1,4,5,8-bis(dicarboximide)-2,6-diyl]-alt-5,5'-(2,2'-bithiophene)} (P(NDI2OD-T2)) as a light-sensitive material to stimulate hPSC-CMs optically. Our results indicated that P(NDI2OD-T2)-mediated photostimulation caused cell damage at irradiances applied long-term above 36 μW/mm and did not regulate cardiac monolayer beating (after maturation) at higher intensities applied in a transient fashion. However, we discovered that the cells grown on P(NDI2OD-T2)-coated substrates showed significantly enhanced expression of cardiomyocyte maturation markers in the absence of a light exposure stimulus. A combination of techniques, such as atomic force microscopy, scanning electron microscopy, and quartz crystal microbalance with dissipation monitoring, which we applied to investigate the interface of the cell with the n-type coating, revealed that P(NDI2OD-T2) impacted the nanostructure, adsorption, and viscoelasticity of the Matrigel coating used as a cell adhesion promoter matrix. This modified cellular microenvironment promoted the expression of cardiomyocyte maturation markers related to contraction, calcium handling, metabolism, and conduction. Overall, our findings demonstrate that conjugated polymers such as P(NDI2OD-T2) can be used as passive coatings to direct stem cell fate through interfacial engineering of cell growth substrates.
人多能干细胞衍生的心肌细胞（hPSC-CMs）是心脏再生医学和建模中一种很有前景的细胞来源。然而，与成年心肌细胞相比，hPSC-CMs表现出不成熟的结构和功能特性。人们已经应用了各种电、机械和生化线索来促进hPSC-CM的成熟，但成效有限。在这项工作中，我们研究了半导体聚合物聚{[N,N'-双(2-辛基十二烷基)-萘-1,4,5,8-双(二甲酰亚胺)-2,6-二基]-alt-5,5'-(2,2'-联噻吩)}（P(NDI2OD-T2)）作为一种光敏材料对hPSC-CMs进行光学刺激的潜在应用。我们的结果表明，P(NDI2OD-T2)介导的光刺激在长期辐照度高于36 μW/mm时会导致细胞损伤，并且在以瞬态方式施加的较高强度下不会调节心脏单层跳动（成熟后）。然而，我们发现生长在P(NDI2OD-T2)包被底物上的细胞在没有光暴露刺激的情况下，心肌细胞成熟标志物的表达显著增强。我们应用了诸如原子力显微镜、扫描电子显微镜和带有耗散监测的石英晶体微天平等多种技术来研究细胞与n型涂层的界面，结果表明P(NDI2OD-T2)影响了用作细胞粘附促进剂基质的基质胶涂层的纳米结构、吸附和粘弹性。这种经过修饰的细胞微环境促进了与收缩、钙处理、代谢和传导相关的心肌细胞成熟标志物的表达。总体而言，我们的研究结果表明，诸如P(NDI2OD-T2)之类的共轭聚合物可以用作被动涂层，通过对细胞生长底物进行界面工程来引导干细胞命运。
