Ispitivanje kriozaštitnog učinka komercijalno dostupnih medija za pohranu rožnica

master's thesis

• Title (english): An examination of the cryoprotective effect of commercially available preservation media
• Author: Josipa Stjepanović
• Mentor: Ivan Pepić (mentor)
• Committee member: Ivan Pepić (predsjednik povjerenstva)
• Committee member: Dubravka Vitali Čepo (član povjerenstva)
• Committee member: Višnja Drinovac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical technology); Zagreb
• Defense date and country: 2018-09-21, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Sve raširenijom potrošnjom oftalmičkih lijekova dolazi do povećane potrebe za pretkliničkim istraživanjima lijekova. Nedostaci ex vivo modela rožnice u svrhu ispitivanja permeabilnosti djelatne tvari preko barijere rožnice su osiguravanje primjerenog broja animalnih rožnica te brzo provođenje pokusa. Budući da je epitel glavna barijera prijenosu djelatnih tvari, a standardnom pohranom humanih rožnica stvaraju se velika epitelna oštećenja, takva pohrana nije primjenjiva za ispitivanja permeabilnosti na animalnim rožnicama. Cilj ovog diplomskog rada je ispitati primarnu i sekundarnu zaštitu rožnica kriopohranjenih u složenim medijima kao kriozaštitnim sredstvima korištenjem različitih protokola zamrzavanja i odmrzavanja. Primjereno kriozaštićene rožnice mogu poslužiti kao vrijedan alat za pretklinička istraživanja. Očuvanost i integritet epitelne barijere rožnice procijenjeni su pomoću vrijednosti izmjerenog transepitelnog električnog otpora. Rezultati ukazuju na oštećenje i narušavanje epitelne barijere tijekom 48 sati kriopohrane u komercijalno dostupnim medijima za pohranu rožnice, CorneaMax® i CorneaJet®. Najslabija primarna zaštita opažena je provođenjem protokola zamrzavanja u tekućem dušiku pri -196°C, dok su više vrijednosti opažene postupnim zamrzavanjem rožnica. Medij CorneaJet® kao kriozaštitno sredstvo pokazao je bolju ukupnu sekundarnu zaštitu rožnica tijekom kriopohrane u odnosu na medij CorneaMax®.

Abstract (english)

The increased use of ophthalmic medications leads to an increased demand for preclinical drug research. The disadvantages of the use of ex vivo corneal model for examining the permeability of active substance through corneal barrier are ensuring an adequate number of animal corneas and the immediate conduct of the experiment. Since epithelium is the main barrier to the transfer of active substances, and the standard storage of human corneas causes serious epithelial damage, that type of storage is not suitable for permeability tests on animal corneas. The aim of this thesis is to examine the primary and secondary protection of cornea cryopreserved in complex media as cryoprotective agents, by using different freezing and thawing protocols. Properly cryoprotected corneas can be used as a valuable tool for preclinical research in ophthalmology. The preservation and integrity of epithelial barrier were estimated by measuring the transepithelial electrical resistance. The results indicate damage and deterioration of the epithelial barrier during 48 hours of cryopreservation in two commercially available corneal storage media, CorneaMax® i CorneaJet®. The lowest primary protection has been proved by the application of the liquid nitrogen freezing protocol at -196°C, while higher values have been obtained by gradual freezing. CorneaJet® medium has shown better overall secondary protection as a cryoprotective agent compared to CorneaMax® medium.

• Keywords: kriopohrana; epitel; transepitelni električni otpor
• Keywords (english): cryopreservation; epithelium; transepithelial electrical resistance
• Language: croatian
• URN:NBN: urn:nbn:hr:163:824421
• 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: 2019-09-06 15:03:32