Abstract

Permeation and subcellular localization of photosensitizer in photodynamic therapy leads to precisely explaining the biological effects in the clinic. We aim to study cell permeation and subcellular localization of azulene between inflamed and normal human peripheral blood mononuclear cells (hPBMCs). 1, 10, 100 and, 1,000 mM (in 1%v/v methanol+deionized water) of azulene were incubated into hPBMCs and observed under a confocal fluorescence microscope. After the optimum concentration was determined, the permeation of this concentration of azulene was observed at 5, 10, 15, 20, 25 and, 30 minutes in either inflamed or normal hPBMCs. The subcellular localization was conducted by staining with 500 nM Mitotracker Red CMXRos and Lysotracker Green DND-26 followed the manufacturer's instruction for localization determination in mitochondria and lysosome, respectively, compared with the position of azulene in hPBMCs. The result shows that the longer the incubation time, the higher the permeation. In inflamed cells, the maximum permeation was at 25 minutes which gave the emission intensity close to 30 minutes. In normal PBMCs, the maximum permeation of azulene was in 20 minutes and azulene signal could not be detected at 30 minutes. The permeation of azulene in inflamed hPBMCs is higher than in normal hPBMCs. Regarding subcellular localization, indifference between mitochondria and lysosome localization of azulene in both inflamed and normal hPBMCs was demonstrated. In conclusion, the permeation of azulene was higher in inflamed hPBMCs compared with normal hPBMCs. With the limitation of this study, azulene appears to localize at both mitochondria and lysosome.

Keywords: Azulene; Permeation; Subcellular Localization; Peripheral Blood Mononuclear Cells; Inflammation

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