Nanoanalytical analysis of bisphosphonate-driven alterations of microcalcifications using a 3D hydrogel system and in vivo mouse model
Jessica Ruiz, Joshua D. Hutcheson, Luís Cardoso, Amirala Bakhshian Nik, Alexandra Condado de Abreu, Tan Pham, Fabrizio Buffolo, Sara Busatto, Stefania Federici, Andrea Ridolfi, Masanori Aikawa, Sérgio Bertazzo, Paolo Bergese, Sheldon Weinbaum, Elena Aïkawa
Abstract
mouse was used as a model of atherosclerosis in vivo. EV aggregation and formation of stress-inducing microcalcifications was imaged via scanning electron microscopy (SEM) and atomic force microscopy (AFM). In both models, BiP (ibandronate) treatment resulted in time-dependent changes in microcalcification size and mineral morphology, dependent on whether BiP treatment was initiated before or after the expected onset of microcalcification formation. Following BiP treatment at any time, microcalcifications formed in vitro were predicted to have an associated threefold decrease in fibrous cap tensile stress compared to untreated controls, estimated using finite element analysis (FEA). These findings support our hypothesis that BiPs alter EV-driven calcification. The study also confirmed that our 3D hydrogel is a viable platform to study EV-mediated mineral nucleation and evaluate potential therapies for cardiovascular calcification.