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Abstract: PO0320

SNF472 Inhibits Heart Valve Calcification in a Novel In Vitro Method Using Porcine Whole Leaflets

Session Information

Category: Bone and Mineral Metabolism

  • 401 Bone and Mineral Metabolism: Basic

Authors

  • Salcedo, Carolina, Sanifit Therapeutics, Palma, Balearic Islands, Spain
  • Zabirnyk, Arsenii, University of Oslo, Oslo, Norway
  • Perez, Maria del mar, Sanifit Therapeutics, Palma, Balearic Islands, Spain
  • Blasco Ferrer, Marc, Sanifit Therapeutics, Palma, Balearic Islands, Spain
  • Stensløkken, Kåre-Olav, Oslo University Hospital, Oslo, Norway
  • Ferrer, Miquel D., Sanifit Therapeutics, Palma, Balearic Islands, Spain
  • Vaage, Ingvar Jarle, University of Oslo, Oslo, Norway
Background

Chronic kidney disease and end-stage kidney disease (ESKD) patients are prone to develop calcific aortic or mitral valve disease. Currently, there exists no pharmacological treatment to prevent or stop the calcification process of aortic valves that causes aortic stenosis, and the models to study this process in vitro and in vivo are scarce. SNF472 is an inhibitor of calcium phosphate crystallization in development for the treatment of calciphylaxis and cardiovascular calcification in ESKD patients. The aims of this study were to develop a robust in vitro model of induced calcification in whole aortic valve leaflets suitable for testing potentially inhibitory drugs, and to test the effects of SNF472 in this model.

Methods

Aortic valve leaflets from commercial pig hearts were dissected free and randomized between experimental groups. Whole leaflets were cultured in individual wells. Two growth media were used for cultivation: standard growth medium and an antimyofibroblastic growth medium. The latter was employed to inhibit contraction of the leaflet into a ball-like structure. Calcification was induced in the growth media by supplementation with an osteogenic medium. Leaflets were cultivated for four weeks and medium was changed every third day. To block calcification, SNF472 was used at concentrations between 1 and 100 µM. Calcium amount in leaflets after four weeks was measured by inductively coupled plasma optical emission spectroscopy.

Results

Osteodifferentiation with calcium accumulation was in principle absent when standard medium was used. However, when the antimyofibroblastic medium was used, a strong calcium accumulation was induced (p=0.006 compared to controls), and this was blocked in a dose-dependent manner by the calcification inhibitor SNF472 (p=0.008), with an EC50 of 3.3 µM.

Conclusion

Cultured whole leaflets of porcine aortic valves are a new in vitro model to study calcification of heart valves. This model will be useful for studying the basic mechanisms of valve calcification and to test pharmacological approaches to inhibit calcification. The latter was shown by SNF472, which strongly inhibited calcification in this model of aortic valve disease.

Funding

  • Commercial Support