Nanoanalytical analysis of bisphosphonate-driven alterations of microcalcifications using a 3D hydrogel system and in vivo mouse model. Proc Natl Acad Sci U S A 2021 Apr 06;118(14)
Date
04/03/2021Pubmed ID
33795519Pubmed Central ID
PMC8040669DOI
10.1073/pnas.1811725118Scopus ID
2-s2.0-85103807262 (requires institutional sign-in at Scopus site) 14 CitationsAbstract
Vascular calcification predicts atherosclerotic plaque rupture and cardiovascular events. Retrospective studies of women taking bisphosphonates (BiPs), a proposed therapy for vascular calcification, showed that BiPs paradoxically increased morbidity in patients with prior acute cardiovascular events but decreased mortality in event-free patients. Calcifying extracellular vesicles (EVs), released by cells within atherosclerotic plaques, aggregate and nucleate calcification. We hypothesized that BiPs block EV aggregation and modify existing mineral growth, potentially altering microcalcification morphology and the risk of plaque rupture. Three-dimensional (3D) collagen hydrogels incubated with calcifying EVs were used to mimic fibrous cap calcification in vitro, while an ApoE-/- 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.
Author List
Ruiz JL, Hutcheson JD, Cardoso L, Bakhshian Nik A, Condado de Abreu A, Pham T, Buffolo F, Busatto S, Federici S, Ridolfi A, Aikawa M, Bertazzo S, Bergese P, Weinbaum S, Aikawa EAuthor
Amirala Bakhshiannik PhD, MS Postdoctoral Researcher 4 in the Physiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsCalcinosis
Cells, Cultured
Diphosphonates
Extracellular Vesicles
Finite Element Analysis
Humans
Hydrogels
In Vitro Techniques
Mice
Mice, Knockout, ApoE
Plaque, Atherosclerotic
Vascular Calcification
