Prikladnost in vitro modela rožnice za ispitivanje biokompatibilnosti oftalmičkih in situ gelirajućih sustava

master's thesis

• Title (english): The suitability of in vitro corneal epithelial model for studying the biocompatibility of ophthalmic in situ gelling systems
• Author: Mirta Nenadić
• Mentor: Jasmina Lovrić (mentor)
• Committee member: Jasmina Lovrić (predsjednik povjerenstva)
• Committee member: Ivan Pepić (član povjerenstva)
• Committee member: Dubravka Vitali Čepo (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical technology); Zagreb
• Defense date and country: 2017-07-20, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Bioraspoloživost lijeka u oku je vrlo niska pri topikalnoj primjeni konvencionalnih kapi za oko zbog brze eliminacije djelatne tvari s površine oka. Glavna karakteristika in situ gelirajućih sustava je produljenje zadržavanja pripravka na površini oka te posljedično povećanje bioraspoloživosti lijeka. Cilj ovog diplomskog rada bio je ispitati prikladnost staničnih modela epitela rožnice u ispitivanju biokompatibilnosti in situ gelirajućih sustava koji se temelje na poloksamerima i kitozanu. U tu svrhu korištena je imortalizirana stanična linija humanih epitelnih stanica rožnice, HCE-T, te in situ gelirajući sustav koji se sastoji od poloksamera P407 (14,2%, m/m), P188 (17%, m/m) te kitozana (0,25 %, m/m).Citotoksičnost in situ gelirajućeg sustava ispitivana je na monosloju humanih epitelnih stanica rožnice HCE-T te na in vitro modelima rožnice temeljenih na dvosloju i višesloju humanih epitelnih stanica HCE-T. Nakon tretiranja stanica s formulacijom, stanična vijabilnost procjenjivala se MTT testom. Citotoksični učinak nije zabilježen pri izlaganju monosloja HCE-T stanica in situ gelirajućem sustavu tijekom 5, 10 i 15 minuta. In vitro model epitela rožnice omogućio je određivanje utjecaja kitozana prisutnog u in situ gelirajućem sustavu na otvaranje čvrstih veza između epitelnih stanica rožnice mjerenjem transepitelnog električnog otpora. Citotoksični učinak nije zabilježen pri izlaganju in vitro modela epitela rožnice in situ gelirajućem sustavu tijekom 30 minuta. In vitro modeli epitela rožnice pokazali su se manje osjetljivim u usporedbi s monoslojem stanica te su stoga prikladniji modeli u ispitivanju citotoksičnosti in situ gelirajućih formulacija u razvoju.

Abstract (english)

When conventional eye drops are applied topically, the bioavailability of the drug in the eye is very low due to the rapid elimination of the active substance. The main characteristic of in situ gelling systems is the prolongation of the retention time on the surface of the eye and consequently the increase in the bioavailability of the drug. The aim of this diploma thesis was to investigate the suitability of in vitro corneal epithelial models for the evaluation of the biocompatibility of in situ gelling system that are based on poloxamers and chitosan. For this purpose, the immortalized corneal epithelial cell line, HCE-T, and an in situ gel system consisting of P407 (14,2%, m/m), P188 (17%, m/m) and chitosan (0,25%, m/m) were used.
The cytotoxicity of the in situ gelling system was investigated on a monolayer of a human corneal epithelial cell line, HCE-T and on in vitro corneal models based on bilayer and multilayer of HCE-T cells. After treating cells with the formulation, cell viability was evaluated by MTT test. The cytotoxic effect was not observed after a monolayer of HCE-T cells was exposed to the in situ gel system for 5, 10 and 15 minutes. The in vitro corneal epithealial model enabled the determination of the influence of chitosan present in the in situ gel system on the opening of tight junctions between the corneal epithelial cells by measuring the transepithelial electric resistance. The cytotoxic effect was not observed during the 30-minute exposure of the in vitro corneal epithelial models to the in situ gelling system. In vitro corneal epithelial models proved less sensitive in comparison to monolayer cells and are, therefore, more suitable models for studying cytotoxicity of the in situ gelling formulations.

• Keywords: in situ gelirajući sustavi; oftalmička primjena; citotoksičnost; HCE-T; kitozan; Pluronic F127; Pluronic F68
• Keywords (english): in situ gel systems; ophtalmic application; cytotoxicity; HCE-T; chitosan; Pluronic F127; Pluronic F68
• Language: croatian
• URN:NBN: urn:nbn:hr:163:964645
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-01-25 13:15:58