Contemporaneous 3D characterization of acute and chronic myocardial I/R injury and response

Nat Commun. 2019 May 24;10(1):2312. doi: 10.1038/s41467-019-10338-2.

Abstract

Cardioprotection by salvage of the infarct-affected myocardium is an unmet yet highly desired therapeutic goal. To develop new dedicated therapies, experimental myocardial ischemia/reperfusion (I/R) injury would require methods to simultaneously characterize extent and localization of the damage and the ensuing inflammatory responses in whole hearts over time. Here we present a three-dimensional (3D), simultaneous quantitative investigation of key I/R injury-components by combining bleaching-augmented solvent-based non-toxic clearing (BALANCE) using ethyl cinnamate (ECi) with light sheet fluorescence microscopy. This allows structural analyses of fluorescence-labeled I/R hearts with exceptional detail. We discover and 3D-quantify distinguishable acute and late vascular I/R damage zones. These contain highly localized and spatially structured neutrophil infiltrates that are modulated upon cardiac healing. Our model demonstrates that these characteristic I/R injury patterns can detect the extent of damage even days after the ischemic index event hence allowing the investigation of long-term recovery and remodeling processes.

Publication types

  • Video-Audio Media

MeSH terms

  • Animals
  • Biopsy
  • Cinnamates / chemistry
  • Coronary Artery Bypass
  • Disease Models, Animal
  • Heart / diagnostic imaging*
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Luminescent Agents / chemistry
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Fluorescence / methods
  • Myocardial Reperfusion Injury / diagnostic imaging*
  • Myocardial Reperfusion Injury / etiology
  • Myocardial Reperfusion Injury / pathology
  • Myocardial Reperfusion Injury / surgery
  • Myocardium / cytology
  • Myocardium / immunology
  • Myocardium / pathology*
  • Neutrophils / immunology

Substances

  • Cinnamates
  • Luminescent Agents
  • Luminescent Proteins
  • red fluorescent protein
  • ethyl cinnamate