20.500.12556/RUL-181368 Vpliv vgradnje laktobacilov v liposome z antioksidanti na njihovo preživetje v polimernih nanovlaknih Effect of liposomal encapsulation of Lactobacilli with antioxidants on their viability in polymer nanofibers Vaginalna mikrobiota ima ključno vlogo pri ohranjanju zdravja žensk, pri čemer prevladujoče bakterije rodu Lactobacillus zagotavljajo kisel pH in zavirajo rast patogenih mikroorganizmov. Zmanjšana prisotnost laktobacilov je povezana s povečano pojavnostjo vaginalnih okužb, ki so pogosto ponavljajoče in težko obvladljive. Namen magistrske naloge je bil razviti in ovrednotiti nanovlakna za vgradnjo modelnega potencialno probiotičnega seva Lactobacillus jensenii LJE6 z namenom izboljšanja njegove stabilnosti in preživetja med pripravo in shranjevanjem. V začetni fazi smo izvedli teste adhezije L. jensenii LJE6 na mucin ter rastne eksperimente v različnih formulacijah medijev, ki so služili kot osnova za nadaljnji razvoj nanovlaken. Laktobacile smo vgrajevali v različna nanovlakna na osnovi polietilenoksida (PEO) in natrijevega alginata (ALG). Nanovlakna smo pripravili z elektrostatskim sukanjem in pri tem prilagajali parametre postopka, kot sta pretok in napetost. Preučevali smo vpliv dodatkov antioksidantov (glutationa in kurkumina) ter liposomov z ali brez kurkumina na potek elektrostatskega sukanja, morfologijo nanovlaken in živost vgrajenih bakterij. S testiranjem živosti smo ovrednotili vpliv formulacije in pogojev shranjevanja na stabilnost vgrajenih laktobacilov. Gojenje seva L. jensenii LJE6 v z mucinom obogatenem De Man-Rogosa-Sharpe gojišču (MRS) je statistično značilno povečalo njegovo adhezijo na mucin v primerjavi z običajnim MRS gojiščem (p < 0,001). Kot najprimernejše polimerno ogrodje se je izkazala kombinacija PEO in ALG, ki je ugodno vplivala na rast bakterij. Vgradnja bakterij v liposome ni vplivala na njihovo živost (p > 0,05), je pa prispevala k zaščiti bakterij v nadaljnjih fazah izdelave nanovlaken. Največji padec živosti bakterij je bil zaznan med elektrostatskim sukanjem nanovlaken, pri čemer je bil ta padec najmanjši pri formulacijah z vgrajenimi liposomi. Podaljšanje časa elektrostatskega sukanja je povzročilo večji upad živosti bakterij tako pri osnovni formulaciji (p < 0,05) kot pri formulaciji z liposomi (p < 0,001). Slednje so izkazale boljše ohranjanje živosti bakterij tudi med dolgoročnim shranjevanjem, zlasti v anaerobnih pogojih in pri nižjih temperaturah, medtem ko antioksidanti niso prispevali k povečanju preživetja laktobacilov (p > 0,05). Na podlagi rezultatov lahko zaključimo, da uporaba liposomov izboljša zaščito seva L. jensenii LJE6 med procesom izdelave nanovlaken in med njihovim shranjevanjem. The vaginal microbiota plays a key role in maintaining women's health, with dominant bacteria of the genus Lactobacillus maintaining an acidic pH and inhibiting the growth of pathogenic microorganisms. A reduced presence of lactobacilli is associated with an increased incidence of vaginal infections, which are often recurrent and difficult to manage. The aim of this master’s thesis was to develop and evaluate nanofibers for the incorporation of a model potentially probiotic strain Lactobacillus jensenii LJE6, in order to improve its stability and survival during preparation and storage. In the initial phase, adhesion tests to mucin and growth experiments in different media formulations were performed, providing the basis for the subsequent development of nanofibers. Lactobacilli were incorporated into various nanofibers based on polyethylene oxide (PEO) and sodium alginate (ALG). Nanofibers were prepared by electrospinning, with process parameters such as flow rate and applied voltage being adjusted. The effects of antioxidant additives (glutathione and curcumin) as well as liposomes with or without curcumin on the electrospinning process, nanofiber morphology, and the viability of incorporated bacteria were investigated. Viability testing was used to evaluate the effects of formulation and storage conditions on the stability of the incorporated lactobacilli. Cultivation of L. jensenii LJE6 in mucin-enriched De Man-Rogosa-Sharpe medium (MRS) significantly increased its adhesion to mucin compared with standard MRS medium (p < 0,001). A combination of PEO and ALG proved to be the most suitable polymeric matrix, as it had a favorable effect on bacterial growth. Incorporation of bacteria into liposomes did not affect their viability (p > 0,05), but it contributed to bacterial protection in subsequent stages of nanofiber production. The greatest decrease in bacterial viability was observed during the electrospinning process, with the smallest reduction observed in formulations containing liposomes. Prolongation of the electrospinning time resulted in a greater decline in bacterial viability in both the basic formulation (p < 0,05) and the liposome-containing formulation (p < 0,001). The latter also demonstrated improved preservation of bacterial viability during long-term storage, particularly under anaerobic conditions and at lower temperatures, whereas antioxidants did not contribute to enhanced survival of lactobacilli (p > 0,05). Based on these results, it can be concluded that the use of liposomes improves the protection of L. jensenii LJE6 during nanofiber fabrication and subsequent storage. nanovlakna elektrostatsko sukanje vaginalna mikrobiota Lactobacillus jensenii probiotiki nanofibers electrospinning vaginal microbiota Lactobacillus jensenii probiotics true false false [K. Vrbinc] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-04-03 08:45:08 2026-04-03 08:45:20 2026-05-05 11:05:35 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 2027-04-02 615.45.014:579(043.2) 124949 274930435 31914.pdf 31914.pdf 1 E84F489A6CC8ECAC99396A279A0A30C1 a15506dd4280c2ca066e908031ef0b2359394605ff5e4f10a11e691f9c2fc851 840b8be3-2f28-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=231763 Fakulteta za farmacijo 0 0 0