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PURPOSE To determine the possible impact of wearing N95 respirator or surgical masks on retinal vessel diameters and choroidal thickness in healthy healthcare workers. METHODS Diameters of peripapillary retinal arteries and veins and choroidal thickness values at the foveal center and at 1000 μm distances from the foveal center in both nasal and temporal directions were measured before mask wearing using a spectral-domain optical coherence tomography. After four hours (h) of N95 or surgical mask wearing vessel diameter and choroidal thickness measurements were repeated. RESULTS A total of 52 eyes from 52 participants (28 F [53.8%]; 24 M [46.2%]) were enrolled in this study. The mean age of patients was 34.58 ± 5.24 years (25-44 years). The diameters of all measured arteries [inferior temporal artery (p = 0.003), superior temporal artery (p < 0.001), inferior nasal artery (p = 0.003), and superior nasal artery (p = 0.004)] and veins,with the exception of superior nasal vein, (inferior temporal vein (p = 0.031), superior temporal vein (p = 0.027), inferior nasal vein (p < 0.001), and superior nasal vein (p = 0.063)] increased significantly after four hour use of N95 respirators and surgical maskswhen compared to baseline. There was also a significant diameter increase of the superior temporal (p < 0.001), inferior nasal veins (p < 0.001), and superior temporal artery (p = 0.037) for N95 respirators and surgical masks use, respectively. The differences in central subfoveal, temporal, and nasal choroidal thickness between baseline and after 4 h use of N95 respirators were statistically significant (From 366.73 ± 70.81 μm to 381.23 ± 69.29μm,p < 0.001 for the subfoveal; from 324.00 ± 64.13μm to 335.40 ± 61.35 μm, p = 0.007 for the temporal; from 297.40 ± 68.18 μm to 308.23 ± 74.51μm, p = 0.002 for the nasal thicknesses). Choroidal thickness values were also increased with surgical mask use. But only the increase in central subfoveal thickness was statistically significant (From 366.78 ± 71.00 μm to 372.58 ± 76.56 μm, p = 0.031 for the central subfoveal; from 297.42 ± 68.35 μmto 302.79 ± 73.05 μm, p = 0.068 for the nasal; from 324.01 ± 64.21μm to 330.33 ± 65.84, p = 0.117 for the temporal thicknesses). CONCLUSION With four hours use of N95 respirators or surgical face masks, retinal vessel diameters and choroidal thicknesses showed an increase in comparison to baseline measurements. Hemodynamic changes seen secondary to hypercapnia due to prolonged use of N95 respirators or surgical masks may also be observed in the retinal and choroidal circulation. Furthermore, the use of face masks should be taken into account while assessing the retinal microvasculature.