An Optimized Protocol for Guidewire-Induced Aortic Valve Stenosis Model in Mice.

Aortic valve stenosis (AVS) is a major cardiovascular disease with increasing prevalence in aging populations, yet the scarcity of diseased valve tissue severely limits mechanistic studies and therapeutic development. This protocol establishes and optimizes a reproducible mouse model of AVS by improving upon the Honda guidewire-induced valve injury method through controlled hemorrhage management. Male and female C57BL/6 mice undergo guidewire-induced aortic valve injury; the procedure is refined by a custom 3D-printed vascular support platform that achieves precise hemorrhage control during guidewire insertion and withdrawal. Outcome measures include 24 h survival, echocardiographic assessment at 4 weeks, and histopathological validation. The optimized protocol achieves a 95% 24 h survival rate (19 of 20 animals); among survivors, 94.7% (18 of 19) meet the predefined hemodynamic success criterion at 4 weeks, yielding an overall model success rate of 90% (18 of 20 animals), confirmed by echocardiography and histological analysis. This standardized model provides a reliable, reproducible platform for investigating mechanisms of valve degeneration and evaluating candidate therapeutic interventions.