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ACCEPTED MANUSCRIPT Statistical analysis was performed using Prism 6 (GraphPad Software, San Diego, CA). Data were analysed using Tukey's multiple comparisons test, with P<0.05 considered to represent a statistically significant difference. All data is represented as the mean ± SEM. 3. Results 3.1 670nm light suppressed cytokine expression following PD in vivo We examined the expression of Il-1β and Ccl2 in rat retinas following photo-oxidative damage (PD) and assessed the effect of 670nm light treatment. The expression of Il-1β and Ccl2 increased significantly in PD retinas compared to dim-reared controls (P<0.05, Figure 1A, B). 670nm light-treated animals had significantly reduced levels of Il-1β and Ccl2 expression compared to untreated PD retinas (P<0.05). Il-1β and Ccl2 expression was comparable between 670nm-treated dim-reared animals and dim-reared controls (without 670nm treatment or PD, Figure 1A, B). For localisation of Ccl2 in the PD retinas, we examined Ccl2 expression using in situ hybridisation (Figure 1C-F). In PD retinas (Figure 1E) a large number of Ccl2-positive cells were apparent in the inner nuclear layer, where the nuclei of Müller cells reside. Conversely, PD retinas treated with 670 nm light had fewer Ccl2-positive cells in the INL compared to untreated PD retinas (Figure 1F). Quantification of the number of Ccl2-positive cells in the INL demonstrated that 670nm light significantly suppressed cells in the INL to express Ccl2 post-PD compared to untreated PD retinas (P<0.05, Figure 1G). 3.2 670nm light reduced stress in primary Müller cells co-cultured with damaged 661W cells We assessed Müller cell changes under photoreceptor damage-initiated stress, and investigated whether treatment with 670nm light had an effect on these changes. Co-culturing 12 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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