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  • Engineering of fluorescent vaginal Lactobacillus species for their monitoring in nanofibers and cell models = Razvoj fluorescentnih vaginalnih vrst iz rodu Lactobacillus za njihovo spremljanje v nanovlaknih in celičnih modelih : doctoral dissertation
    Stojanov, Spase
    Vaginal infections are one of the main reasons why women seek medical help. They are associated with decreased number of lactobacilli and overgrowth of opportunistic pathogens. Restoring the normal ... vaginal microbiota with lactobacilli as probiotics can help cure the infections. However, the lack of techniques for their functional studies and the need for an appropriate delivery system hinders their use as probiotics. In the dissertation, we propose two solutions for overcoming the deficiencies. First, genetic engineering of three vaginal lactobacilli (Lactobacillus crispatus, Lactobacillus gasseri and Lactobacillus jensenii), and control Lactiplantibacillus plantarum, enabled expression of fluorescent proteins, i.e. infrared fluorescent protein (IRFP), green fluorescent protein (GFP), red fluorescent protein (mCherry) and blue fluorescent protein (mTagBFP2), thereby facilitating distinction and quantification of lactobacilli. Second, vaginal lactobacilli have been successfully incorporated into polyethylene oxide (PEO) nanofibers with addition of different excipients, thereby developing a promising delivery system. Alginate and sucrose showed the highest stimulation of growth of lactobacilli in vitro and were included in the formulations. Interaction between excipients and bacteria was assessed thermally and spectroscopically. All the species survived electrospinning, with L. gasseri demonstrating the highest survival, followed by L. jensenii and L. crispatus. The species were viable in all formulations after 56 days of storage. The lactobacilli retained their fluorescence after incorporation and after their release from nanofibers. We therefore used engineered lactobacilli expressing mCherry to study the release kinetics, where PEO nanofibers loaded with lactobacilli were completely dissolved after 30?45 min. Lactobacilli have also adhered to Caco-2 epithelial cells as observed with a confocal microscope. To conclude, this dissertation describes innovative technology for tracking vaginal lactobacilli and their successful incorporation into electrospun nanofibers. The results hold a great promise for the development of a novel medicine against vaginal infections.
    Type of material - dissertation ; adult, serious
    Publication and manufacture - Ljubljana : [S. Stojanov], 2022
    Language - english
    COBISS.SI-ID - 124677891

    Link(s):

    Repository of the University of Ljubljana – RUL
    Digitalna knjižnica Slovenije - dLib.si

    Dostop z namenskih računalnikov v prostorih NUK



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