Engineering an in vitro organotypic model for studying cardiac hypertrophy

Colloids Surf B Biointerfaces. 2018 May 1:165:355-362. doi: 10.1016/j.colsurfb.2018.02.036. Epub 2018 Feb 27.

Abstract

Neonatal cardiomyocytes cultured on flat surfaces are commonly used as a model to study cardiac failure of diverse origin. A major drawback of such a system is that the cardiomyocytes do not exhibit alignment, organization and calcium transients, similar to the native heart. Therefore, there is a need to develop in vitro platforms that recapitulate the cellular microenvironment of the murine heart as organotypic models to study cardiovascular diseases. In this study, we report an engineered platform that mimics cardiac cell organization and function of the heart. For this purpose, microscale ridges were fabricated on silicon using ultraviolet lithography and reactive ion etching techniques. Physical characterization of the microstructures was done using scanning electron microscopy and atomic force microscopy. Cardiomyocytes grown on these micro-ridges showed global parallel alignment and elliptical nuclear morphology as observed in the heart. Interestingly, calcium currents traversed the engineered cardiomyocytes in a coordinated and directional manner. Moreover, the cardiomyocytes on the engineered substrates were found to be responsive to hypertrophic stimuli, as observed by the expression of a fetal gene, atrial natriuretic peptide and increase in calcium transients upon agonist treatment. Taken together, our work demonstrates that micro-ridges can be used to obtain cardiomyocyte response in vitro, which closely resembles mammalian heart.

Keywords: Calcium transients; Cardiac hypertrophy; Cardiomyocytes; Micro-ridges.

MeSH terms

  • Animals
  • Animals, Newborn
  • Atrial Natriuretic Factor / genetics
  • Atrial Natriuretic Factor / metabolism
  • Calcium / metabolism*
  • Cardiomegaly / chemically induced
  • Cardiomegaly / metabolism
  • Cardiomegaly / pathology*
  • Connectin / genetics
  • Connectin / metabolism
  • Gene Expression
  • Heart / drug effects
  • Heart / physiopathology*
  • Models, Biological
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology*
  • Phenylephrine / pharmacology
  • Primary Cell Culture
  • Rats
  • Rats, Sprague-Dawley
  • Rats, Wistar
  • Silicon / chemistry
  • Silicon / radiation effects
  • Surface Properties
  • Tissue Engineering / methods*
  • Tissue Scaffolds*
  • Ultraviolet Rays

Substances

  • Connectin
  • Phenylephrine
  • Atrial Natriuretic Factor
  • Calcium
  • Silicon