Abstract
To test a novel early polytherapy treatment strategy targeting mitochondrial bioenergetics, glutamate excitotoxicity, and sterile inflammatory response molecular pathways associated with retinal ganglion cell survival following optic nerve trauma.
Twenty C57BL/6J mice were subjected to sonication-induced traumatic optic neuropathy injury. The control group (n = 10) received intravitreal, retrobulbar, and subcutaneous phosphate buffered saline injections on days 0 and 3 (no repeat retrobulbar vehicle). On day 0, the treatment group (n = 10) received injections of intravitreal interleukin-1 receptor antagonist with ketamine, retrobulbar ropivacaine, and subcutaneous etanercept. Treatment group animals had 1% (wt/vol) N-acetylcysteine ad libitum supplemented in drinking water from day 1. On day 3, intravitreal pan-ephrin receptor antagonist peptide and subcutaneous elamipretide and etanercept injections were given. Pattern electroretinogram assessments continued at weeks 0, 1, 2, 4, 6, 8, 10, and 12. Optical coherence tomography retinal layer thickness was measured on naive, control, and treatment groups at week 12. The whole mount retinas were harvested for retinal ganglion cell quantitation.
At 12 weeks, the averaged retinal ganglion cell density count in the control group was lower (413.37 ± 41.77 cells/mm2) compared with treatment (553.97 ± 18.00 cells/mm2; p < 0.001) and naive (595.94 ± 30.67cells/mm2; p < 0.001) groups. Ganglion cell complex layer thicknesses showed control group (49.29 ± 5.48 μm) thinner than the treated (61.00 ± 2.57 μm; p = 0.004) and naive (67.00 ± 6.12 μm; p = 0.004) groups. No significant difference was seen at 12 weeks between the treated and naive groups. Pattern electroretinogram recordings in the control group revealed a statistically significant decrease in amplitudes for all time points. Apart from week 8, the amplitudes in the treatment group did not significantly differ from the baseline at any time point.
Early combinatorial therapeutic intervention to address disparate molecular pathways following optic nerve trauma effectively halts retinal neurons' progressive structural and functional degeneration.