title: Survival-Related Autophagic Activity Versus Procalcific Death in Cultured Aortic Valve Interstitial Cells Treated With Critical Normophosphatemic-Like Phosphate Concentrations.
abstract: Valve dystrophic calcification is a common disorder affecting normophosphatemic subjects. Here, cultured aortic valve interstitial cells (AVICs) were treated 3 to 28 days with phosphate (Pi) concentrations spanning the normal range in humans (0.8, 1.3, and 2.0 mM) alone or supplemented with proinflammatory stimuli to assess possible priming of dystrophic-like calcification. Compared with controls, spectrophotometric analyses revealed marked increases in calcium amounts and alkaline phosphatase activity for 2.0-mM-Pi-containing cultures, with enhancing by proinflammatory mediators. Ultrastructurally, AVICs treated with low/middle Pi concentrations showed an enormous endoplasmic reticulum (ER) enclosing organelle debris, so apparently executing a survival-related atypical macroautophagocytosis, consistently with ultracytochemical demonstration of ER-associated acid phosphatase activity and decreases in autophagosomes and immunodetectable MAP1LC3. In contrast, AVICs cultured at 2.0-mM Pi underwent mineralization due to intracellular release and peripheral layering of phospholipid-rich material acting as hydroxyapatite nucleator, as revealed by Cuprolinic Blue and von Kossa ultracytochemical reactions. Lack of immunoblotted caspase-3 cleaved form indicated apoptosis absence for all cultures. In conclusion, fates of cultured AVICs were crucially driven by Pi concentration, suggesting that serum Pi levels just below the upper limit of normophosphatemia in humans may represent a critical watershed between macroautophagy-associated cell restoring and procalcific cell death.
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Bonetti
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Della Mora
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Magali
authors.initials: M
authors.lastname: Contin
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Gregoraci
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Franco
authors.initials: F
authors.lastname: Tubaro
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Marchini
authors.affiliation: Departments of Experimental and Clinical Medicine (AB, ADM, MC, MM, FO), Medical and Biological Sciences (GG), and Agricultural, Food, Environmental and Animal Sciences (FT), University of Udine, Udine, Italy.
authors.forename: Fulvia
authors.initials: F
authors.lastname: Ortolani
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001021:Aortic Valve / Q000166:cytology* / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D001343:Autophagy; D002114:Calcinosis / Q000378:metabolism / Q000473:pathology*; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002470:Cell Survival; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D010710:Phosphates / Q000378:metabolism*
chemical_list: D010710:Phosphates; D000469:Alkaline Phosphatase; D002118:Calcium
keywords: aortic valve interstitial cells; autophagy; calcific valve disease; ectopic calcification; electron microscopy; phosphatemia