Objective To investigate the expression and significance of S100A9 and TLR4 in vitreous fluid of patients with diabetic retinopathy.Methods A total of 40 patients who underwent vitrectomy at the Second People's Hospital of Hefei from August 2022 to August 2023 were selected.Among them,20 patients with diabetic retinopathy were included as the experimental group,and 20 patients with macular hole or epiretinal membrane were included as the control group.Vitreous fluid samples were collected,and the protein concentrations of S100A9 and TLR4 were measured using the ELISA method.Results The concentration of S100A9 in the experimental group was(5.95±1.55)μg/L,which was significantly higher than that in the control group(2.19±0.76)μg/L,and the difference was statistically significant.The TLR4 concentration in the experimental group was(1.54±0.79)μg/L significantly higher than that in the control group of(0.34±0.18)μg/L,and the difference was statistically significant(P<0.05).There was a posi-tive correlation between S100A9 and TLR4 in the experimental group(r=0.72,P<0.05).Conclusions The in-creased levels of S100A9 and TLR4 in vitreous fluids of diabetic retinopathy suggest that inflammation may be involved in the process of its elevelopment.