siRNA-loaded liposomes: Inhibition of encystment of Acanthamoeba and toxicity on the eye surface
| Autor: | Helder Ferreira Teixeira, Nathalya Tesch Brazil, Kathrin Faber, Marilise Brittes Rott, Giovanni K. Zorzi |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male Cell Survival 030106 microbiology Protozoan Proteins Acanthamoeba Eye Biochemistry Cell Line Cornea 03 medical and health sciences Glycogen phosphorylase Mice In vivo Drug Discovery medicine Animals Humans Viability assay Particle Size RNA Small Interfering Incubation Pharmacology Liposome biology Chemistry Organic Chemistry Glycogen Phosphorylase biology.organism_classification medicine.disease Molecular biology 030104 developmental biology Acanthamoeba keratitis Toxicity Liposomes Molecular Medicine RNA Interference |
| Zdroj: | Chemical biologydrug design. 90(3) |
| ISSN: | 1747-0285 |
| Popis: | Current treatments for Acanthamoeba keratitis are unspecific. Because of the presence of the resilient cyst form of the parasite, the infection is persistent. Silencing the key protein of cyst formation, glycogen phosphorylase, has shown potential for reducing encystment processes of the Acanthamoeba trophozoite. However, a suitable carrier to protect and deliver siRNA sequences is still needed. Dotap DOPE:DSPE-PEG liposomes were prepared by three different techniques and used to associate a therapeutic siRNA sequence. Liposomes prepared by film hydration followed by membrane extrusion were considered the most adequate ones with average size of 250 nm and zeta potential of +45 mV, being able to associate siRNA for at least 24 hr in culture medium. siRNA-liposomes could inhibit up to 66% of the encystment process. Cell viability studies demonstrated MTT reduction capacity higher than 80% after 3 hr incubation with this formulation. After 24 hr of incubation, LDH activity ranged for both the formulations from around 4% to 40%. In vivo tolerance studies in mice showed no macroscopic alteration in the eye structures up to 24 hr after eight administrations during 1 day. Histological studies showed regular tissue architecture without any morphological alteration. Overall, these results suggest that the formulations developed are a promising new strategy for the treatment of ocular keratitis caused by Acanthamoeba spp. |
| Databáze: | OpenAIRE |
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