20.500.12556/RUL-170694 Vpliv lipidov iz arheje Aeropyrum pernix K1 na fizikalno-kemijske lastnosti liposomov, sestavljenih iz sfingomielina in holesterola Effect of lipids from the archaeon Aeropyrum pernix K1 on physico-chemical properties of liposomes composed of sphingomyelin and cholesterol Liposomske sisteme lahko izboljšamo z arhealnimi lipidi, katerih edinstvena struktura omogoča preživetje arhejam v ekstremnih pogojih. Z dodatkom lipidov arheje Aeropyrum pernix K1 smo optimizirali priznano liposomsko formulacijo sfingomielina in holesterola. Analiza dinamičnega sipanja svetlobe je pokazala, da že majhni dodatki (2 mol%) arhealnih lipidov omogočajo oblikovanje monodisperznih liposomov s premerom pod 100 nm s sonikacijo ter znižujejo zeta potencial na vrednosti, ki zagotavljajo koloidno stabilnost. Meritve anizotropije fluorescentnih sond so razkrile, da arhejski lipidi sorazmerno z dodatkom zvišujejo fluidnost liposomov. Arhealni lipidi omogočajo tudi popolno reverzibilnost morfoloških parametrov liposomov po dodatku EDTA ob destabilizaciji s 4 mM Ca²⁺, kar kaže na ohranjanje oblike liposomov ob prisotnosti kalcijevih ionov. Sproščanje kalceina iz liposomov z dodatkom arhejskih lipidov pri povišani temperaturi in dolgotrajnem shranjevanju pri nižjih temperaturah je bilo primerljivo ali boljše kot pri kontrolnih vzorcih brez arhealnih lipidov. Poleg tega je dodatek 2 mol% arhealnih lipidov omogočil izvrstno kapsulacijsko učinkovitost (95 %) aktivnega polnjenja z vinkristinom ter izboljšal biološko stabilnost liposomov v primerjavi z arheosomi, sestavljenimi izključno iz arhejskih lipidov, kar smo potrdili s pretočno citometrijo in fluorescenčno mikroskopijo. Liposomal systems can be improved by archaeal lipids, whose unique structure enables archaeans to survive in extreme conditions. By adding lipids from the archaeon Aeropyrum pernix K1, we optimized the established liposomal formulation of sphingomyelin and cholesterol. Dynamic light scattering analysis revealed that even small additions (2 mol%) of archaeal lipids allow for the formation of monodisperse liposomes with a mean diameter below 100 nm via sonication, while also reducing the zeta potential to values that ensure colloidal stability. Fluorescence probe anisotropy showed that archaeal lipids proportionally increase liposomal fluidity as their concentration increases. Archaeal lipids also enable complete reversibility of the morphological parameters of liposomes after destabilization with 4 mM Ca²⁺ followed by the addition of EDTA, indicating the liposomal integrity is preserved in the presence of calcium ions. The release of calcein from liposomes with added archaeal lipids at elevated temperatures and during long-term storage at lower temperatures was comparable to, or better than in control samples without archaeal lipids. Furthermore, the addition of 2 mol% archaeal lipids facilitated an excellent encapsulation efficiency (95%) for active loading of vincristine and improved the biological stability of liposomes compared to archaeosomes composed solely of archaeal lipids, as confirmed by flow cytometry and fluorescence microscopy. Aeropyrum pernix aktivno polnjenje arheje biološke membrane dostavni sistemi fizikalno-kemijska stabilnost fluidnost membran fosfolipidi holesterol kalcein kapsulacija liposomi sfingomielin stabilnost in vitro vinkristin active loading Aeropyrum pernix archaea biological membranes calcein cholesterol drug delivery encapsulation liposomes membrane fluidity phospholipids physico-chemical stability sphingomyelin stability in vitro vincristine true false false [J. Kejžar] Slovenski jezik Angleški jezik Doktorsko delo/naloga 2025-07-13 07:16:14 2025-07-13 07:16:22 2025-07-15 03:31:57 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 620.3:577.352:582.233 257804 242370563 48387.pdf 48387.pdf 1 85864E07D6FEFA0C29810ACECCD083AF 99ff21c95aeee3ede74ab168d76c612351af55cd53f30dfa4803361146e86d53 97cf3502-5fa6-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=212738 Biotehniška fakulteta 0 0 0