Marcello Rattazzi1, Elisa Bertacco2, Laura Iop3, Susanna D'Andrea2, Massimo Puato2, Giacomo Buso2, Valerio Causin4, Gino Gerosa3, Elisabetta Faggin2, Paolo Pauletto5. 1. Department of Medicine, University of Padova, Italy; Medicina Interna Iˆ, Ca' Foncello Hospital, Treviso, Italy. Electronic address: marcello.rattazzi@unipd.it. 2. Department of Medicine, University of Padova, Italy. 3. Department of Cardiac, Thoracic and Vascular Sciences, University of Padova, Italy. 4. Department of Chemical Sciences, University of Padova, Italy. 5. Department of Medicine, University of Padova, Italy; Medicina Interna Iˆ, Ca' Foncello Hospital, Treviso, Italy.
Abstract
OBJECTIVES: Pyrophosphate (PPi) is a potent inhibitor of ectopic mineralization but its role during aortic valve calcification is not known. METHODS: Anti-calcific effect of PPi was investigated by using an in vitro model of serum-driven calcification of collagen sponges and decellularized porcine aortic valve leaflets. Bovine interstitial valve cells (VIC), seeded either within the collagen matrices or in transwell chambers, were used to test cellular ability to inhibit serum-induced calcification. PPi metabolism was investigated in clonal VIC harboring different calcifying potential. RESULTS: In a cell-free system, high serum levels induced a dose-dependent calcification of type I collagen matrices which was prevented by PPi and ATP supplementation. Blockade of serum-driven calcification by PPi and ATP was also observed when using decellularized porcine aortic valve leaflets. A similar anti-calcific effect was also seen for bovine VIC, either statically seeded into the collagen matrices or co-cultured by using a transwell system. However, when we performed co-culture experiments by using clonal VIC harboring different calcifying potential, we observed that the subset of cells acquiring a pro-calcific profile lost the ability to protect the collagen from serum-driven calcification. Pro-calcific differentiation of the clonal VIC was accompanied by increase in ALP along with significant reduction in NPP activity and ATP/PPi extracellular accumulation. These changes were not observed in the clonal subtype with lower propensity towards calcification. CONCLUSIONS: We showed that PPi and ATP are potent inhibitors of serum-driven calcification of collagen matrix and that their extracellular accumulation is reduced in calcifying VIC.
OBJECTIVES:Pyrophosphate (PPi) is a potent inhibitor of ectopic mineralization but its role during aortic valve calcification is not known. METHODS: Anti-calcific effect of PPi was investigated by using an in vitro model of serum-driven calcification of collagen sponges and decellularized porcine aortic valve leaflets. Bovine interstitial valve cells (VIC), seeded either within the collagen matrices or in transwell chambers, were used to test cellular ability to inhibit serum-induced calcification. PPi metabolism was investigated in clonal VIC harboring different calcifying potential. RESULTS: In a cell-free system, high serum levels induced a dose-dependent calcification of type I collagen matrices which was prevented by PPi and ATP supplementation. Blockade of serum-driven calcification by PPi and ATP was also observed when using decellularized porcine aortic valve leaflets. A similar anti-calcific effect was also seen for bovine VIC, either statically seeded into the collagen matrices or co-cultured by using a transwell system. However, when we performed co-culture experiments by using clonal VIC harboring different calcifying potential, we observed that the subset of cells acquiring a pro-calcific profile lost the ability to protect the collagen from serum-driven calcification. Pro-calcific differentiation of the clonal VIC was accompanied by increase in ALP along with significant reduction in NPP activity and ATP/PPi extracellular accumulation. These changes were not observed in the clonal subtype with lower propensity towards calcification. CONCLUSIONS: We showed that PPi and ATP are potent inhibitors of serum-driven calcification of collagen matrix and that their extracellular accumulation is reduced in calcifying VIC.
Authors: Brian R Lindman; Marie-Annick Clavel; Patrick Mathieu; Bernard Iung; Patrizio Lancellotti; Catherine M Otto; Philippe Pibarot Journal: Nat Rev Dis Primers Date: 2016-03-03 Impact factor: 52.329
Authors: Shira G Ziegler; Carlos R Ferreira; Elena Gallo MacFarlane; Ryan C Riddle; Ryan E Tomlinson; Emily Y Chew; Ludovic Martin; Chen-Ting Ma; Eduard Sergienko; Anthony B Pinkerton; José Luis Millán; William A Gahl; Harry C Dietz Journal: Sci Transl Med Date: 2017-06-07 Impact factor: 17.956
Authors: Hiu-Gwen Tsang; Lin Cui; Colin Farquharson; Brendan M Corcoran; Kim M Summers; Vicky E Macrae Journal: Mol Med Rep Date: 2017-11-27 Impact factor: 2.952
Authors: Vivien M Hsu; Eszter Kozák; Qiaoli Li; Márta Bocskai; Naomi Schlesinger; Ann Rosenthal; Scott T McClure; László Kovács; László Bálint; Szilvia Szamosi; Gabriella Szücs; Mary Carns; Kathleen Aren; Isaac Goldberg; András Váradi; John Varga Journal: Rheumatology (Oxford) Date: 2022-03-02 Impact factor: 7.046