Evaluation of Different Decellularization Protocols for Obtaining and Characterizing Canine Cardiac Extracellular Matrix

• Published in: Biomedicines Vol. 12; no. 6; p. 1190
• Main Authors: da Silva, Izabela Gabriela Rodrigues, Miglino, Maria Angelica, de Souza, Samara Silva, Buchaim, Daniela Vieira, Buchaim, Rogerio Leone
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 27.05.2024; MDPI
• Subjects: Analysis; canine heart; Cardiovascular diseases; Collagen; Coronary artery disease; decellularization; Electron microscopes; Extracellular matrix; Freezing; Heart attacks; Heart diseases; Heart failure; Mortality; Myocardium; Pandemics; Preservation; Protocol; Regenerative medicine; scaffolds; Scanning electron microscopy; Sodium; Sodium lauryl sulfate; Sulfates; Surface active agents; Tissue engineering; Transplants & implants; Trypsin; Veterinary medicine; Brazil; United Kingdom > UK; United States > US; Germany; tissue engineering; canine heart; decellularization; scaffolds; regenerative medicine; myocardium; extracellular matrix; translational science; transplants
• ISSN: 2227-9059; 2227-9059
• DOI: 10.3390/biomedicines12061190

Cardiovascular diseases are considered the leading cause of mortality globally; even with low mortality in dogs, such diseases are described in the same way in companion animals and humans. This study aimed to devise an effective decellularization protocol for the canine myocardium through the association of physical, chemical, and enzymatic methods, assessing resultant alterations in the myocardial extracellular matrix to obtain a suitable scaffold. Two canine hearts were collected; the samples were sectioned into ±1 cm fragments, washed in distilled water and 1× PBS solution, and followed by treatment under four distinct decellularization protocols. Sodium Dodecyl Sulfate (SDS) 1% 7 days + Triton X-100 1% for 48 h (Protocol I); Sodium Dodecyl Sulfate (SDS) 1% 5 days + Triton X-100 1% for 48 h (Protocol II); Trypsin 0.05% for 1 h at 36 °C + freezing -80 °C overnight + Sodium Dodecyl Sulfate (SDS) 1% for 3 days, Triton-X-100 for 48 h hours (Protocol III); 0.05% trypsin for 1 h at 36 °C + freezing at -80 °C overnight + 1% Sodium Dodecyl Sulfate (SDS) for 2 days + 1% Triton-X-100 for 24 h (Protocol IV). After analysis, Protocols I and II showed the removal of cellular content and preservation of extracellular matrix (ECM) contents, unlike Protocols III and IV, which retracted the ECM and removed essential elements of the matrix. In theory, although Protocols I and II have similar results, Protocol II stands out for the preservation of the architecture and components of the extracellular matrix, along with reduced exposure time to reagents, making it the recommended protocol for the development of a canine myocardial scaffold.