One Hour Reperfusion is Enough to Assess Function and Infarct Size With TTC Staining in Langendorff Rat Model

Abstract

There is not general agreement concerning the optimal time of reperfusion necessary to assess myocardial function and necrosis on isolated perfused heart model. Nevertheless, the study of cardioprotection (especially, pre- and postconditioning) requires a reliable and standardized assessment of myocardial necrosis. The objective of this study was thus to evaluate whether 1 h of reperfusion was sufficient to assess rat heart viability on Langendorff preparation. Isolated rat hearts (n = 30) underwent 40 min of global normothermic ischemia followed by 60 or 120 min Langendorff reperfusion. In each group, hearts were also randomly assigned into the 2 following sub-groups: postconditioning (PostC, consisting in 2 episodes of 30 s ischemia and 30 s reperfusion at the onset of reperfusion), and control (no intervention). Coronary flow, heart rate, dP/dt and rate-pressure-product were measured. Myocardial necrosis was assessed by TTC staining and LDH, CK release analysis. Our results indicated that heart function tended to slightly decrease between 60 min and 120 min reperfusion. Infarct size was identical at 60 min and 120 min reperfusion, averaging 33-34% of total LV area in controls versus 17% in PostC (p < 0.001 between control and PostC groups). Similarly, the maximum of enzymatic releases (CK and LDH) measured in coronary effluents was at 60 min of reperfusion, followed by a progressive decrease at 90 min and 120 min. As expected, postconditioning limited enzymatic releases whatever the studied time of reperfusion. In conclusion, we showed that prolonged reperfusion beyond 60 min was not useful for function assessment and did not change infarct size measurement, on Langendorff rat model of ischemia-reperfusion.