Evaluation of apoptotic caspase levels in estimation of the wound age

Abstract

AbstractObjectives:We aimed to investigate the potential use of the expression of apoptotic signaling pathway genes of rat in skin wound age estimation.Material and methods:For this purpose, we formed cutting tool injuries using a scalpel in an experimental model. Then, we assessed Caspase 3, 8 and 9 mRNA levels by using quantitative real-time PCR and protein levels by using immunohistochemistry in rat skin wounds. In addition, we used TUNEL assay to detect apoptotic cells.Results:We observed that Caspase 3 mRNA level significantly increased (2.1±0.4 folds) on day 3 (p<0.05) and Caspase 8 mRNA level significantly increased (1.8±0.2 folds) on day 5 (p<0.05). Caspase 9 mRNA level increased (1.9±0.1 folds) on day 3 and (2.5±0.4 folds) on day 5 (p<0.05). The percentage values of polymorphonuclear leukocytes (PMNLs) and inflammatory mononuclear cells (IMCs) were observed after immunohistochemical staining by Caspase 3, 8, 9 antibodies. Our immunohistochemistry results were found to be consistent with the mRNA results observed. We reported a statistically significant increase in Caspase 3, 8 and 9-positive cells on days 3 and 5 after immunohistochemical staining as well.Conclusion:Our results suggest that time-dependent features of apoptotic factors might offer a potential tool in estimating wound age.