An Investigation into the Effects of Carbon Black Nanoparticles on Erythrocytes and Leukocytes In Vitro.
University of Wales Institute Cardiff
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The purpose of this laboratory investigation was to discover if the nanoparticle Carbon Black had any significant effect on the membranes of horse erythrocytes and leukocytes, in order to hypothesise the effect of Carbon Black on the human equivalent. Human leukocytes were later utilised to determine if they were activated by Carbon Black exposure at different dilutions. Erythrocytes were incubated with two different dilutions of Carbon Black in phosphate buffered saline (PBS), spun, and the supernatant was saved and measured spectrophotometrically for haemolysis. The absorbance values were converted by an equation to give a percentage haemolysis value for each dilution. The activation of horse leukocytes was examined microscopically after incubation with different dilutions of Carbon Black in PBS. The number of activated white cells out of 100 counted was converted into a percentage, which demonstrated the percentage activation. Immunofluorescent microscopy was later performed on human leukocytes to see if they were activated by exposure to Carbon Black. These were incubated with Carbon Black dilutions and observed on slides after cytospinning by immunofluorescent microscopy. By separating the white blood cells into two distinct groups, mononuclear leukocytes and polymorphonuclear leukocytes, it was possible to see if either or both groups were activated, and hypothesise the reasons for this. It was found that leukocytes were activated by Carbon Black when examined microscopically, and this could indicate that Carbon Black could initiate an immune response after recognition as a foreign body. This could create cytokine and free radical release, which in large amounts could be detrimental to health. It was also found that there was haemolysis present after erythrocyte incubation with Carbon Black; however this was shown by statistical analysis not to be significantly linked to amount of Carbon Black or length of exposure. Therefore, more testing would be essential in order to show if there is a significant link between Carbon Black exposure and damage to erythrocytes, as the amount of results collected by this investigation is insufficient to confirm if the haemolysis present was due to the Carbon Black or other factors. Leukocyte activation was shown to have occurred; however further testing of leukocyte activation, and quantification of activation, would be useful to determine if this changes with different Carbon Black levels in solution.
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