The distribution of metals across the environment is increasingly becoming a major concern as they not only pollute the environment but also pose a danger to humans and animals. Human exposure to heavy metals often occurs as a combination of metals the synergistic effects of which can be more toxic than a single metal. The aim of this study was to investigate the effects that the metals mercury (Hg), nickel (Ni) and manganese (Mn) alone and in combination have on erythrocyte morphology and other components of the coagulation system using the haemolysis assay, scanning electron microscopy (SEM), and confocal laser scanning microscopy. Human blood was exposed to the heavy metals ex vivo, and percentage haemolysis was determined. Ultrastructural analysis of erythrocytes, platelets and fibrin networks was performed using SEM. Analysis of phosphatidylserine (PS) flip-flop was determined using confocal laser scanning microscopy. At the highest concentration of 10,000× the World Health Organization safety limit, all the metals caused haemolysis. The results showed that the exposure of erythrocytes to Hg alone and in combination with other metals displayed more haemolysis compared to Ni and Mn alone and in combination. Components of the coagulation system showed ultrastructural changes, including the formation of echinocytes and the activation of platelets with all single metals as well as the combinations. Confocal laser scanning microscopy analysis showed the presence of PS on the outer surface of the echinocytes that were exposed to metals alone and in combination. It can, therefore, be concluded that these heavy metals have a negative impact on erythrocytes and the coagulation system.
049 Simultaneous assessment of histopathology and direct immunofluorescence in pemphigus vulgaris using ex vivo confocal laser scanning microscopy
