Biomimetic Microstructure Morphology in Electrospun Fiber Mats is Critical for Maintaining Healthy Cardiomyocyte Phenotype
- 310 Downloads
Despite recent advances in biomimetic substrates, there is still only limited understanding of how the extracellular matrix (ECM) functions in the maintenance of cardiomyocyte (CM) phenotype. In this study, we designed electrospun substrates inspired by morphologic features of non-failing and failing human heart ECM, and examined how these substrates regulate phenotypes of adult and neonatal rat ventricular CMs (ARVM and NRVM, respectively). We found that poly(ε-caprolactone) fiber substrates designed to mimic the organized ECM of a non-failing human heart maintained healthy CM phenotype (evidenced by cell morphology, organized actin/myomesin bands and expression of β-MYH7 and SCN5A.1 and SCN5A.2) compared to both failing heart ECM-mimetic substrates and tissue culture plates. Moreover, culture of ARVMs and NRVMs on aligned substrates showed differences in m- and z-line alignment; with ARVMs aligning parallel to the ECM fibers and the NRVMs aligning perpendicular to the fibers. The results provide new insight into cardiac tissue engineering by illustrating the importance models that mimic the cardiac ECM microenvironment in vitro.
KeywordsElectrospinning Poly(ε-caprolactone) Cell–matrix interaction Cardiomyocyte Cell phenotype
This study was supported by NIH HL091465, NSF DMR 1006558, U0100398, and AHA 13GRNT16690019. The authors would also like to acknowledge the use of resources at the Vanderbilt Institute of Nanoscale Science and Engineering (VINSE), a facility renovated under NSF ARI-R2 DMR-0963361.
Conflict of interest
Rutwik Rath, Jung Bok Lee, Truc-Linh Tran, Sean F. Lenihan, Cristi L. Galindo, Yan Ru Su, Tarek Absi, Leon M. Bellan, Douglas B. Sawyer, and Hak-Joon Sung declare that they have no conflicts of interest.
Human and Animal Rights and Informed Consent
All human subject research was carried out in accordance with the guidelines of Vanderbilt IRB and human research protection program and approved by approved by the Vanderbilt Institutional Review Board. All animal studies were carried out in accordance with the guidelines of the Vanderbilt University Animal Care and Use Committee and approved by the Vanderbilt Institutional Review Board.
- 3.Bhana, B., R. K. Iyer, W. L. Chen, R. Zhao, K. L. Sider, M. Likhitpanichkul, C. A. Simmons, and M. Radisic. Influence of substrate stiffness on the phenotype of heart cells. Biotechnol. Bioeng. 105:1148–1160, 2010.Google Scholar
- 6.Bursac, N., M. Papadaki, R. J. Cohen, F. J. Schoen, S. R. Eisenberg, R. Carrier, G. Vunjak-Novakovic, and L. E. Freed. Cardiac muscle tissue engineering: toward an in vitro model for electrophysiological studies. Am. J. Physiol. 277:H433–H444, 1999.Google Scholar
- 10.Ellingsen, O., A. J. Davidoff, S. K. Prasad, H. J. Berger, J. P. Springhorn, J. D. Marsh, R. A. Kelly, and T. W. Smith. Adult rat ventricular myocytes cultured in defined medium: phenotype and electromechanical function. Am. J. Physiol. 265:H747–H754, 1993.Google Scholar
- 14.Galindo, C. L., E. Kasasbeh, A. Murphy, S. Ryzhov, S. Lenihan, F. A. Ahmad, P. Williams, A. Nunnally, J. Adcock, Y. Song, F. E. Harrell, T. L. Tran, T. J. Parry, J. Iaci, A. Ganguly, I. Feoktistov, M. K. Stephenson, A. O. Caggiano, D. B. Sawyer, and J. H. Cleator. Anti-remodeling and anti-fibrotic effects of the neuregulin-1beta glial growth factor 2 in a large animal model of heart failure. J. Am. Heart Assoc. 3:e000773, 2014.CrossRefGoogle Scholar
- 16.Gupta, M. K., J. M. Walthall, R. Venkataraman, S. W. Crowder, D. K. Jung, S. S. Yu, T. K. Feaster, X. Wang, T. D. Giorgio, C. C. Hong, F. J. Baudenbacher, A. K. Hatzopoulos, and H.-J. Sung. Combinatorial polymer electrospun matrices promote physiologically-relevant cardiomyogenic stem cell differentiation. PLoS One 6:e28935, 2011.CrossRefGoogle Scholar
- 18.Herron, T. J., F. S. Korte, and K. S. McDonald. Loaded shortening and power output in cardiac myocytes are dependent on myosin heavy chain isoform expression. Am. J. Physiol. Heart Circ. Physiol. 281(3):H1217–H1222, 2001.Google Scholar
- 21.Inserte, J., V. Hernando, M. Ruiz-Meana, M. Poncelas-Nozal, C. Fernandez, L. Agullo, C. Sartorio, U. Vilardosa, and D. Garcia-Dorado. Delayed phospholamban phosphorylation in post-conditioned heart favours ca2+ normalization and contributes to protection. Cardiovasc. Res. 103:542–553, 2014.CrossRefGoogle Scholar
- 33.Modis, L. Organization of the extracellular matrix. Taylor & Francis: CRC Press, 1990.Google Scholar
- 42.Stout, D. A., J. Yoo, A. N. Santiago-Miranda, and T. J. Webster. Mechanisms of greater cardiomyocyte functions on conductive nanoengineered composites for cardiovascular application. Int. J. Nanomed. 7:5653–5669, 2012.Google Scholar