Objectives
Although heart valve transplantation delivers living, growth-capable valves, its clinical implementation is restricted by transplant logistics and donor availability. We developed a preservation solution for the ex vivo storage of living valves that maintains tissue viability for at least 7 weeks.
Methods
Porcine pulmonary roots (N = 25) were dissected, treated with a clinically used antibiotic cocktail, and stored in a custom-made preservation solution (Valve Preservation Solution, VPS) or Hanks’ Balanced Salt Solution with serum for up to 7 weeks. Tissues were preserved in normothermia (37 °C, 5% CO2) and hypothermia (4 °C, control group). Tissue preservation was monitored weekly by evaluating viability (AlamarBlue), metabolic activity (media glucose), histologic tissue preservation (hematoxylin and eosin, Movat Pentachrome stain), and valve cell phenotype (immunostaining).
Results
Cold-stored valves (VPS and Hanks’ Balanced Salt Solution) demonstrated significantly reduced viability within 1 to 2 weeks ex vivo. In contrast, normothermic storage in VPS preserved leaflet viability for 7 weeks, with consistent glucose metabolism. Immunostaining of normothermic VPS-stored valve leaflets showed a quiescent cell phenotype with little expression of alpha-smooth muscle actin or proliferation markers (relative to baseline), and baseline-level leaflet Vimentin expression. Despite preserved cellular viability, leaflet microarchitectural integrity was only maintained for 3 weeks of ex vivo storage.
Conclusions
Normothermic preservation of living valves can support tissue viability for at least 7 weeks ex vivo. With additional preclinical validation, stored living valves could act as a tissue source for heart valve transplantation, with key advantages in enhanced availability and resource-efficiency.
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