Effects Of Low Level Laser On Proliferation,migration And Ros Production In Mouse Fibroblasts

Objective: Low level laser therapy(LLLT)has been widely used as different branches of regenerative medicine due to Non-invasive,drug-free,and safe form of light therapy.However,the complexity of parameters of low level laser hinders its wider clinical application.In this study,the low level mode of the 810 nm Diode laser was used to preliminarily explore the effects of changing power and pulse type on mouse fibroblasts,in order to provide experimental basis and theoretical basis for the application of LLL in skin mucosal wound healing and clinical transformation.Methods: In this study,mouse fibroblasts(L929)were cultured in vitro using 810 nm diode laser with low level mode(E1C 0.1 W,continuous;E1R 0.1 W,intermittent;E2C0.5 W,continuous;E2R 0.5 W,intermittent).The cell migration ability was detected by in vitro scratch test.The cell proliferation ability was detected by CCK-8 assay.ROS fluorescence intensity was detected by inverted fluorescence microscope.The production of ROS was detected by flow cytometry.The statistical analysis was performed via T test and one-way analysis of variance(ANOVA),P < 0.05 was considered to be significant..Results: Scratch results show that E1 C and E2 R significantly increased the cell migration rate compared with the E0 group(P < 0.001).E1 R also enhanced cell migration(P < 0.01).However,there was no significant differences between E2 C and the E0(P > 0.05).CCK-8 results show that the increase in the optical density of E1 R,E1C,E2 R were higher than that of the E0(P < 0.001),while there was no statistical difference between the E2 C and the E0(P > 0.05).The E2 C group did not facilitate cell proliferation.The results of ROS fluorescence analysis and flow cytometry analysis showed that the ROS produced by L929 cells irradiated with different modes of LLL were different.The ROS production of E2 C cells was the highest,followed by E2 R,E1C,E1 R,and the E0 group was the lowest,with a significant difference(P < 0.01).Conclusion: 810 nm Diode Laser 0.1 W,continuous pulse mode and 2 cm away from the wound can maintain the appropriate level of ROS,and can effectively promote cell proliferation and migration.