Preparation and characterization of hyaluronic acid-cyclosporine A nano micelles as novel treatment for Dry Eye
Fereshte Khalili1 *, Mozhgan Rezaei Kanavi2 , Hamid akbari javar3 , Rassoul Dinarvand4
- Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
- Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
- Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran b Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran,Leicester School of Pharmacy, De Montfort University, Leicester, UK
Abstract: Investigating the efficacy of hyaluronic acid (HA) - cyclosporine A (CyA) nano-micelles, a novel drug delivery system, in rabbit models of dry eye syndrome (DES).
Methods: After characterization of HA-CyA nano-micelles by evaluating particle size, zeta potential, critical micelle concentration (CMC), morphology, and enzymatic-triggered CyA release in the presence of esterase, in vitro toxicity and irritancy were confirmed by hen’s egg chorioallantoic membrane and MTT using human corneal epithelial cells, respectively. Ocular toxicity of nano-micelle was investigated by clinical examinations, histology, and TUNEL assay.
Experimental model of dry eye was induced by topical1.0% atropine sulfate followed after 15 min by treatment with HA, Restasis (CyA) and nano-micelle in individual groups, three times a day for 7 days. In addition to performing Schirmer’s test and tear break-up time test (TBUT), occurrence of superficial punctate keratopathy (SPK) was also investigated in the study groups. Using ELISA, the amounts of tear TNF-α and interleukin1-β were measured. Histopathological examinations of corneoscleral tissues along with conjunctiva were also performed.
Results: A nano-micelle was designed with particle size of 67 nm, polydispersity index of 0.221, zeta potential of -13.98 mV, and 88% release rate in-vitro. It was not toxic in in-vitro and in-vivo investigations. TBUT values and tear volume were decreased in untreated group while these indices improved significantly in groups treated with nano-micelle in a promising manner. Nano-micelle increased mean tear volume from 4.83 to 11.50 mm (P=.0001) and mean TBUT values from 16 s to 32.5 s (P=.0001) compared with untreated group. In contrast to presence of severe SPK in untreated eyes, no significant dotted staining was observed in nano-micelle and Restasis groups. The most reduction of tear TNF-α and interleukin1-β was observed in nano-micelle treated eyes. Histopathological examinations disclosed a saw-tooth appearance of irregular corneal epithelium in HA and untreated eyes, a finding that was not observed in nano-micelle and Restasis groups. Immunohistochemical investigations disclosed lowest expression of TNF-α and interleukin1-β in micelle group.
Conclusion: These outcomes propose that HA-CyA nano-micelle has a therapeutic effect in rabbit model of dry eye and might be a potential treatment for DES.
