Photodynamic damage study of HeLa cell line using ALA |
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Authors: | AlSalhi M S Atif M AlObiadi A A Aldwayyan A S |
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Institution: | (1) Departamento de Química, FFCLRP, Universidade de São Paulo, Av. dos Bandeirantes 3900, 14040-901 Ribeirão Preto, São Paulo, Brazil;(2) LPBC, UMR CNRS 7033 and Université Pierre et Marie Curie, Paris, France; |
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Abstract: | The present study evaluates the photodynamic damage with 5-aminolevulinic acid (5-ALA) using HeLa as experimental model. HeLa
cell line was irradiated with red light (He-Ne laser, λ = 632.8 CW nm). The influence of different incubation times and concentrations
of 5-ALA, different irradiation doses and various combinations of photosensitizer and light doses on the cellular viability
of HeLa cells were studied. The optimal uptake of photosensitizer ALA in HeLa cells was investigated by means of PpIX fluorescence
intensity by exciting the HeLa cell suspension at 450 nm and a detection wavelength set at 690 nm. Cells viability was determined
by means of trypan blue solution. The spectrometric measurements showed that the maximal cellular uptake of 5-ALA occurred
after 4 h in vitro incubation. We found that the combination with 5-ALA and laser irradiation leads to time/concentration-dependent
increase of cells death and also energy doses-dependent enlarge the cells death. The fluorescence intensity after PDD of carcinoma
cells reduce when compared with the control group. The fluorescence emission spectral profiles after PDD of carcinoma cells
showed a dip around 425–525 nm when compared with the control group. This may be due to the damage of mitochondria component
of cells. The percentage of HeLa cells after PDD shows that the percentage of cells survival rate as function of laser dose
(power). Hence it is clear that at 200 μg/ml ALA and 20 mW laser irradiation, more than 70% of HeLa cells were dead after
15 min. |
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