Distribution of selected elements in calcific human aortic valves studied by microscopy combined with SR-µXRF: influence of lipids on progression of calcification.

Calcified heart valves display a significant imbalance in tissue content of trace and essential elements. The valvular calcification is an age-related process and there are data suggesting involvement of lipids. We studied elemental composition and lipid distribution in three distinct regions of calcified human aortic valves, representing successive stages of the calcific degeneration: normal, thickened (early lesion) and calcified (late lesion), using SR-µXRF (Synchrotron Radiation Micro X-Ray Fluorescence) for elemental composition and Oil Red O (ORO) staining for demonstration of lipids. Two-dimensional SR-µXRF maps and precise point spectra were compared with histological stainings on consecutive valve sections to prove topographical localization and colocalization of the examined elements and lipids. In calcified valve areas, accumulation of calcium and phosphorus was accompanied by enhanced concentrations of strontium and zinc. Calcifications preferentially developed in lipid-rich areas of the valves. Calcium concentration ratio between lipid-rich and lipid-free areas was not age-dependent in early lesions, but showed a significant increase with age in late lesions, indicating age-dependent intensification of lipid involvement in calcification process. The results suggest that mechanisms of calcification change with progression of valve degeneration and with age.

[Effect of the dispersion of calcium deposits on allogenic aortic valves durability. Mineralization phases]. / Wplyw rozproszenia zlogów wapnia na trwalosc allogennych aortalnych zastawek serca. Fazowosc mineralizacji.

This investigation was aimed at comparison of calcium content and calcium dispersion in allogenic aortic valve leaflets removed due to dysfunction, to establish the influence of both parameters on graft durability. Calcification was assessed histochemically (von Kossa) as well as physicochemically using atomic absorption spectroscopy (AAS). The morpho-metric data (leaflet area involved in the calcification process) were obtained by computer-assisted image analysis system. The dry weight content of leaflet calcium and phosphorus were assessed by atomic absorptive spectroscopy (AAS) and Ca/P ratio was calculated. Calcium dispersion coefficient (Dc) was established according to the formula: Dc = 1/Ca(c)/Ap, where Ca(c) = calcium dry weight concentration; Ap = percent of leaflet area involved in calcification. We found biphasic correlation between calcium concentration and area involved in calcification. The first one was characterized by rising dispersion of calcium deposits while for the second one saturation with hydroxyapatite of formerly calcified areas was predominant, negatively influencing graft durability. Allograft durability was correlated with calcium dispersion (Dc) (p<0.001), while no significant correlation was found with calcium concentration. Decreased Dc was characteristic for 93.8% of low durability grafts (<11.6 years). Our results suggest that lowered calcium dispersion decreasing allograft lifetime and is a better predictor of allograft durability than the total calcium content.