Elevated extracellular calcium levels induce smooth muscle cell matrix mineralization in vitro.
BACKGROUND
Hyperphosphatemia, elevated calcium x phosphorus product (Ca x P), and calcium burden, major causes of vascular calcification, are correlated with increased cardiovascular morbidity and mortality in dialysis patients.


METHODS
To address the underlying mechanisms responsible for these findings, we have utilized an in vitro human smooth muscle cell (HSMC) model of vascular calcification. Previous studies using this system demonstrated enhanced calcification of HSMC cultures treated with phosphorus levels in the hyperphosphatemic range, and implicated a sodium-dependent phosphate cotransport-dependent mechanism in this effect. In the present study, we examine the effect of increasing calcium concentrations on HSMC calcification in vitro.


RESULTS
Increasing calcium to levels observed in hypercalcemic individuals increased mineralization of HSMC cultures under normal phosphorus conditions. Importantly, at these total calcium concentrations, ionized calcium levels increased from 1.2 mmol/L to 1.7 mmol/L, consistent with levels observed physiologically in normocalcemic and hypercalcemic individuals, respectively. Furthermore, increasing both calcium and phosphorus levels led to accelerated and increased mineralization in the cultures. Calcium-induced mineralization was dependent on the function of a sodium-dependent phosphate cotransporter, since it was inhibited by phosphonoformic acid (PFA). While elevated calcium did not affect short-term phosphorus transport kinetics, long-term elevated calcium treatment of HSMCs induced expression of the sodium-dependent phosphate cotransporter, Pit-1.


CONCLUSIONS
These studies suggest that elevated calcium may stimulate HSMC mineralization by elevating Ca x P product and enhancing the sodium-dependent phosphate cotransporter-dependent mineralization pathway previously observed in HSMCs.
Yang
Hsueh
H
Department of Bioengineering, University of Washington, Seattle, Washington 98195, USA.
Curinga
Gabrielle
G
Giachelli
Cecilia M
CM
D001011:Aorta / Q000166:cytology; D002113:Calcification, Physiologic / Q000502:physiology; D002114:Calcinosis / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D006801:Humans; D006934:Hypercalcemia / Q000378:metabolism; D066298:In Vitro Techniques; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010758:Phosphorus / Q000378:metabolism / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology
D010758:Phosphorus; D002118:Calcium