Enhancing anti-calcification efficacy in veterinary cardiovascular surgeries: evaluating short-term ethanol’s role in glutaraldehyde fixed pericardial tissues in rats

Abstract

Autologous pericardial tissues are utilized in veterinary cardiovascular surgeries due to their accessibility and effectiveness. To enhance handling and biomechanical properties, glutaraldehyde (GA) fixation is applied. However, GA fixation can induce calcification, leading to tissue failure. This study aimed to establish an optimal rapid anti-calcification protocol by integrating ethanol treatment with the proven effective GA concentration and fixation time, facilitating application from collection to utilization. Pericardia were fixed with 0.625% GA for 20 min and subjected to ethanol treatment for 0 (group A, control), 20 (group B), and 30 minutes (group C). The treated tissues underwent mechanical test and were implanted subcutaneously in 3-week-old male rats for 7 weeks before extraction, followed by calcium analysis and histological examination via hematoxylin and eosin staining. No significant differences in mechanical properties were observed among the groups. The ethanol-treated groups (groups B and C; p < 0.05) exhibited significantly lower calcium levels than control (group A). Microscopy confirmed collagen and elastic fibers preservation, without significant immune cell variance. However, higher fibrocyte presence was noted in the ethanol-treated groups. This study presents a rapid anti-calcification protocol combining ethanol treatment with optimal GA fixation, suitable for direct surgical use of autologous tissues. Further research is necessary for long-term efficacy evaluation.