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Guzel Bikbova, Toshiyuki Oshitari, Shuichi Yamamoto; In vitro study of potential neuroprotective and regenerative agents for diabetic retinopathy. Invest. Ophthalmol. Vis. Sci. 2016;57(12):No Pagination Specified.
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© ARVO (1962-2015); The Authors (2016-present)
To determine what combination of therapeutic agents has neuroprotective abilities for diabetic retinopathy.
All of the procedures were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. Retinal explants of 7 adult SD rats were three-dimensionally cultured in collagen gel, and incubated in serum free media, AGEs, AGEs+100 µM citicoline, AGEs+10 ng/ml NT-4, AGEs+100 µM TUDCA, AGEs+100 µM citicoline + TUDCA (doublet), AGEs+100 µM citicoline+TUDCA+10 ng/ml NT-4 (triplet). The number of regenerating neurites was counted under a phase-contrast microscope after 7 days of culture. After counting, the retinal explants were fixed, cryosectioned, and stained with TUNEL and DAPI. The ratio of TUNEL-positive cells to the number of DAPI-stained nuclei in the ganglion cell layer was calculated. Statistical analyses were performed by one-way ANOVA.
The number of neurites was significantly higher in retinas incubated with triplets than in any of the other groups (P<0.0001), and the number of TUNEL-positive cells was fewer than in any of the other groups (P<0.0001). In retinas culture in the doublets, the number of neurits was higher than that of single agent groups (P<0.0001), and the number of TUNEL-positive cells were fewer than in the single agent group (P<0.0001).
Media with two agents (doublet) without NT-4 and that with three agents with NT-4 significantly increased the number of regenerated neurites and decreased the number of TUNEL-positive cells. Thus, these combinations of agents should be considered for neuroprotective and regenerative therapy for diabetic retinopathy.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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