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Calcification in vitro: experimental factors that induce nanocrystal formation (1077.9)
Author(s) -
Hortells Luis,
Sosa Cecilia,
Millán Ángel,
Sorribas Victor
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1077.9
Subject(s) - calcification , pathogenesis , incubation , chemistry , in vitro , calcium , endocrinology , medicine , biochemistry
Medial vascular calcification (VC) of arteries is frequently observed in chronic kidney disease and diabetes. Pathogenesis in VC is frequently studied on primary cultures of aortic smooth muscle cells. However, several experimental factors can affect the rate of calcification in vitro, which can be misleading in the research of VC pathogenesis. The aim of this work has been the identification of these factors and to improve the adequacy of the model. Calcification was studied as a function of Pi concentration (1‐3 mM), incubation time and media composition (MEM vs. DMEM). We have determined the rate of calcium deposition, medium pH, nucleation rate, degree of crystallization, cell death and osteogene expression. Calcification was more intense with DMEM than MEM due to increased basicity induced by higher bicarbonate content in DMEM. Cell death (as LDH leakage) correlated and preceded calcification. Particle nucleation depended on pH and Pi concentration, and was already observed after 2h (DMEM) or 5h (MEM) of incubation with the cells. Osteogene expression and crystal structure were also modified by the same experimental conditions.