Relationship between irradiation time of riboflavin-ultraviolet A scleral collagen cross-linking and retinal injury

Objective

To investigate the effects of riboflavin-ultraviolet A scleral collagen cross-linking on the retina under different irradiation time, and to determine the safe irradiation time.

Methods

Sixty healthy New Zealand white rabbits were randomly divided into control group (0 minute group), 10 minutes group, 20 minutes group, 30 minutes group and 40 minutes irradiation group according to the irradiation time, with 12 rabbits in each group.The left eye was irradiated with riboflavin-ultraviolet A scleral collagen (370 nm, 10 mW/cm²). The histopathological change of retina was observed by light microscope and transmission electron microscope and compared among different groups.The concentration of MDA and the activities of SOD, CAT and GSH-Px in retinal tissue were detected by corresponding kits.The expression levels of SOD and CAT proteins in retinal tissue were detected by Western blot method.The study protocol was approved by the Binzhou Medical University Laboratory Animal Ethical Committee (No.2017-80). The use and care of animals complied with the statement of ARVO and the Regulation on the Management of Laboratory Animal Quality of China.

Results

Under the light microscope, the structure of the retinas in the control group was orderly arranged.Under the transmission electron microscope, the lamellar structure in the inner segment and the mitochondrial structure in the outer segment of the photoreceptor cells were intact, and the mitochondrial ridge was continuous in the control group.There was no obvious difference in retinal morphology between the 10 minutes irradiation group and the control group under both the light microscope and the transmission electron microscope, and the retinal damage became more severe with the prolongation of irradiation time.The concentration of MDA in the retina of each group was elevated gradually with the increase of irradiation time, and the difference was statistically significant (F=65.25, P<0.05). The concentration of MDA was (11.31±1.84), (14.94±1.04)and (18.25±1.42)nmol/mgprot in the 20 minutes, 30 minutes and 40 minutes irradiation groups respectively, which were significantly higher than (1.13±0.02)nmol/mgprot in the control group (all at P<0.05). The MDA concentration in 20 minutes, 30 minutes and 40 minutes irradiation groups was increased successively, showing statistically significant differences (all at P<0.05). With the prolongation of irradiation time, the activities of SOD, CAT and GSH-Px as well as the expression levels of SOD and CAT proteins were significantly decreased gradually (F=44.09, 34.18, 35.60, 115.75, 78.86; all at P<0.05). The differences between the control group and 20 minutes, 30 minutes, 40 minutes irradiation groups, and the differences among 20 minutes, 30 minutes, 40 minutes irradiation groups were statistically significant (all at P<0.05).

Conclusions

Riboflavin-ultraviolet A 10 mW/cm² scleral collagen cross-linking irradiation for 10 minutes is safe.Excessive irradiation time can cause damage to the retina of rabits.

Collagen cross-linking; Sclera/drug effects; Riboflavin/pharmacology; Ultraviolet ray/adverse effects; Photoreceptor cells/drug effects; Photosensitizing agents/toxicity; Radiation injuries, experimental/pathology, ultrastructure; Lipid peroxidation