RRC ID 59890
Author Zorzi GK, Schuh RS, Maschio VJ, Brazil NT, Rott MB, Teixeira HF.
Title Box Behnken design of siRNA-loaded liposomes for the treatment of a murine model of ocular keratitis caused by Acanthamoeba.
Journal Colloids Surf B Biointerfaces
Abstract Acanthamoeba keratitis is an ophthalmic disease with no specific treatment that specially affects contact lens users. The silencing of serine phosphatase (SP) and glycogen phosphorylase (GP) proteins produced by Acanthamoeba has been shown to significantly reduce the cytopathic effect, although no vehicle was proposed yet to deliver the siRNA sequences to the trophozoites. In this study, PEGylated cationic liposomes were proposed and optimized using Box-Behnken design. The influence of DOTAP:DOPE ratio, DSPE-PEG concentration, and siRNA/DOTAP charge ratio were evaluated over both biological response and physicochemical properties of liposomes. The ratio of DOTAP:DOPE had an effect in the trophozoite activity whereas the charge ratio influenced both size and protease activity. The predicted values were very close to the observed values, yielding a formulation with good activity and toxicity profile, which was used in the following experiments. A murine model of ocular keratitis was treated with siGP + siSP-loaded liposomes, as well as their respective controls, and combined treatment of liposomes and chlorhexidine. After 15 days of eight daily administrations, the liposomal complex combined with chlorhexidine was the only treatment able to reverse the more severe lesions associated with keratitis. There was 60% complete regression in corneal damage, with histological sections demonstrating the presence of an integral epithelium, without lymphocytic infiltrate. The set of results demonstrate the efficacy of a combined therapy based on siRNA with classical drugs for a better prognosis of keratitis caused by Acanthamoeba.
Volume 173
Pages 725-732
Published 2019-1-1
DOI 10.1016/j.colsurfb.2018.10.044
PII S0927-7765(18)30740-9
PMID 30384269
MeSH Acanthamoeba / drug effects* Acanthamoeba / enzymology Acanthamoeba / pathogenicity Acanthamoeba Keratitis / parasitology Acanthamoeba Keratitis / pathology Acanthamoeba Keratitis / therapy* Animals Chlorhexidine / pharmacology* Cornea / drug effects Cornea / parasitology Cornea / pathology Disease Models, Animal Drug Administration Schedule Drug Compounding / methods Drug Delivery Systems / methods* Drug Therapy, Combination Factor Analysis, Statistical Fatty Acids, Monounsaturated / chemistry Gene Expression Regulation Glycogen Phosphorylase / antagonists & inhibitors Glycogen Phosphorylase / genetics Glycogen Phosphorylase / metabolism Humans Liposomes / chemistry* Liposomes / metabolism Phosphatidylethanolamines / chemistry Phosphoric Monoester Hydrolases / antagonists & inhibitors Phosphoric Monoester Hydrolases / genetics Phosphoric Monoester Hydrolases / metabolism Polyethylene Glycols / chemistry Protozoan Proteins / antagonists & inhibitors* Protozoan Proteins / genetics Protozoan Proteins / metabolism Quaternary Ammonium Compounds / chemistry RNA, Small Interfering / genetics RNA, Small Interfering / metabolism Rats Rats, Wistar Trophozoites / drug effects* Trophozoites / enzymology Trophozoites / pathogenicity
IF 3.973
Times Cited 3
Human and Animal Cells HCE-T(RCB2280)