| Objective:The in vitro photothermal heating performance of fulvic acids(FA)with different concentrations under 808nm laser irradiation,and select the parameters suitable for the subsequent photothermal treatment experiment.To investigate the in vitro inhibitory effect of FA on KB cells of oral epidermoid carcinoma,and to study the photothermal therapeutic effect of FA combined with 808 nm laser on KB cells.It provides the data basis for the follow-up related basic experiments,and provides the theoretical basis for the clinical application of fulvic acid in oral epidermoid carcinoma.Methods:The 808nm laser was used to irradiate FA,the laser power density was adjusted to 0.5,1,2,3W/cm2,and the FA concentration was 0.25,0.5,1,2,3mg/ml,and the temperature change was recorded by infrared thermal imager.The FA cells were co-cultured with KB cells of oral epidermoid carcinoma,and the cultured medium containing FA was added at 0.25,0.5,1,2 and 3 mg/ml,and the control group without FA was used to detect the cytotoxicity by CCK8 assay.The KB cells treated jointly by FA and laser irradiation at 808-nm wavelength were further divided into three groups:0.5 W/cm~2laser+1 mg/ml FA group,1 W/cm~2laser+0.5 mg/ml FA group,and 2W/cm~2laser+0.25 mg/ml FA group.The culture media containing KB cells in the three groups were treated with FA at different concentrations that were irradiated with lasers at different power densities for 10 min.While cells with irradiation at different power densities for 10 min alone without FA treatment were established as corresponding control groups,respectively.Its photothermal therapeutic effect on cells was examined through CCK-8 and live-dead cell staining experiments.Results:Under 808 nm laser irradiation,the temperature of FA with different concentrations increased,and gradually reached the peak and remained basically unchanged.Laser power density is 0.5W/cm~2+FA concentration is 1mg/ml;The laser power density is 1W/cm~2+FA concentration is 0.5mg/ml;The laser power density is2W/cm~2+FA concentration is 0.25mg/ml,and the final temperature of these three parameters is in the ideal tumor temperature sensitive range(43～50℃),which can be used as a choice for subsequent photothermal therapy experiments.After co culturing KB cells with FA at concentrations of 0.25,0.5,1,2 and 3 mg/m L for 24h,the results of the cytotoxicity test showed a significant decrease in cell viability in a dose-dependent manner(P<0.05).Furthermore,following the co-culture of KB cells with FA at 0.25,0.50,and 1.00 mg/ml for 24 h,cells were then divided into three groups of A,B,and C to receive another treatment of laser irradiation for 10 min,the results of photothermal treatment showed that the cell survival rates were(3.48±0.53)%,(7.42±0.79)%and(8.11±0.98)%,respectively,which were statistically significant(P<0.001)compared with the control pore cell survival rates(100±1.71)%.After FA combined with laser irradiation of KB cells,the staining of live and dead cells showed a large number of dead cells emitting red fluorescence in the visual field,and only a small number of live cells labeled green.Conclusion:FA has excellent photo thermal conversion performance;FA can inhibit the proliferation of KB cells in tongue cancer;FA combined with low energy laser can generate heat and effectively kill KB cells of tongue cancer. |
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