From surplus to scientific value: Repurposing vascular tissues for organ bath standardization to de-risk vasoactive liabilities.

INTRODUCTION: The vascular organ bath assay is a well-established classical pharmacology model to characterize vasoactive properties of compounds, offering a platform to elucidate mechanisms of action, optimize dosing, and identify cardiovascular hazards early in the drug selection process and derisking. To address both the regulatory emphasis on detecting drug-induced blood pressure changes and the increasing importance of the 3Rs, repurposed vascular tissue was used to identify key procedural determinants of tissue responsiveness and to support standardization in organ bath studies METHODS: Using a linear mixed effects model, we evaluated the impact of procedural variables on quality control test outcomes and established an optimized and standardized protocol prioritizing efficiency, robustness, and maximal tissue use. Comprehensive tissue characterization was achieved by generating dose-response curves for relevant reference compounds including phenylephrine, acetylcholine, serotonin and gallopamil. Finally, the vasoactive properties of pimobendan were investigated and mechanistic experiments were performed to explore its mechanism of action RESULTS: Overall, sedation method was the most limiting factor identified, as isoflurane markedly attenuated acetylcholine induced relaxation after extended storage of the aortic rings. While the set-up did not capture centrally mediated, humoral or chronic blood pressure effects, we were able to effectively reveal direct vasoactive properties of reference compounds with physiological relevance. Successful mechanistic studies were performed, suggesting a potential role for the endothelium and PVAT in pimobendan induced relaxation.

CONCLUSION: These findings validate the use of repurposed tissues for evaluating pressor effects, support assay harmonization, and demonstrate how thoughtful refinement can reconcile regulatory safety needs with the 3Rs principles in mind.