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Atsuhiko Fukuto, Soohyun Kim, Brooke L. Gates, Laura Van Winkle, Kent E. Pinkerton, Sara M Thomasy; The impact of zinc oxide and vanadium pentoxide nanoparticles on corneal epithelial wound healing in vitro and in vivo. Invest. Ophthalmol. Vis. Sci. 2019;60(9):914. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Engineered nanomaterials (ENMs) are promising carriers for ophthalmic drug delivery. However, the impact of ENMs on the ocular surface, particularly corneal wound healing, is understudied. Thus, the purpose of this study is to evaluate the impact of two ENMs – zinc oxide (ZnO) and vanadium pentoxide (V2O5) nanoparticles (NPs) on corneal epithelial cell viability and migration in vitro, as well as epithelial wound repair in vivo utilizing a rabbit model.
ZnO-NPs (50 nm) and V2O5-NPs (100 nm) were studied at concentrations ranging from 0.05-250 µg/mL. Immortalized human corneal epithelial (hTCEpi) cells were co-cultured for 24 h with ZnO-NPs or V2O5-NPs and cell viability was assessed using two assays - MTT and calcein-AM. Gold nanoparticle (15 nm; 5 µg/mL), 0.1% saponin and distilled water were used as negative, positive and vehicle controls, respectively. A round wound healing assay with a monolayer of hTCEpi cells was performed using ZnO-NPs (0.05-10 µg/mL) and V2O5-NPs (0.05-7.5 µg/mL). For in vivo studies, an 8 mm epithelial debridement was performed in the right eye only of 18 rabbits who were then treated with ZnO-NPs (50 µg/mL, n=6), V2O5-NPs (50 µg/mL, n=6) or balanced salt solution (BSS, n=6) in both eyes four times/day. Fluorescein stain and digital photography were performed twice daily to monitor the wound area until complete healing had occurred. The semiquantitive preclinical ocular toxicology scoring (SPOTS) system was used to access the anterior segment with a hand-held slit lamp biomicroscope.
Cell viability using either MTT or calcein-AM assay was significantly (P < 0.001) decreased with ZnO-NPs at ≥5 µg/mL, V2O5-NPs at ≥ 5µg/mL. Migration of the hTCEpi cells was significantly inhibited (P < 0.05) following incubation with ZnO-NPs at ≥ 5µg/mL and V2O5-NPs at ≥ 5 µg/mL. Corneal epithelial wound closure rate was significantly slower (P < 0.001) in rabbits treated with ZnO-NPs, but not with V2O5-NPs. Corneal opacity was significantly denser and larger (P < 0.05) in rabbits treated with ZnO-NPs.
Both ZnO-NPs and V2O5-NPs decrease corneal epithelial cell viability and migration in a concentration-dependent manner in vitro but only ZnO-NPs impaired corneal epithelial wound closure in vivo. Given the common use of ZnO-NPs in cosmetics and sunscreen, further study of the impact of these ENMs on corneal wound healing is warranted.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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