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Določanje velikosti lipidnih veziklov pripravljenih z ekstruzijo
ID Zagorc, Urška (Author), ID Kogej, Ksenija (Mentor) More about this mentor... This link opens in a new window

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Abstract
Študije veziklov so pomembne za boljše razumevanje delovanja organizmov, saj imajo vezikli številne pomembne naloge. Med drugim skrbijo za medcelično komunikacijo in transport snovi. Vezikli so sestavljeni iz lipidnega dvosloja, v katerem je navadno ujeta tekočina skupaj z makromolekulami kot so proteini in RNA. V zadnjih letih se znanstveniki veliko ukvarjajo s pripravo umetnih veziklov ali liposomov, ki jih lahko izkoriščamo za različne namene. Ena od tehnik priprave velikih umetnih veziklov, zgrajenih iz enega lipidnega dvosloja, je ekstruzija. V diplomskem delu smo se posvetili veziklom iz treh različnih lipidov: iz jajčnega lecitina, lipida 1,2-dioleoil-sn-glicero-3-fosfatidiletanolamin ter mešanice lipidov 1,2-dioleoil-sn-glicero-3-fosfatidiletanolamin in 1,2-di-palmitoil-sn-glicero-3-fosfatidilholin. S pomočjo ekstruzije smo pripravili vezikle različnih velikosti lipida 1,2-dioleoil-sn-glicero-3-fosfatidiletanolamin. Velikost veziklov smo določali z metodo dinamičnega sipanja svetlobe, kjer po Stokes-Einsteinovi enačbi preko izmerjenega translacijskega difuzijskega koeficienta izračunamo hidrodinamski radij delcev. Po prvi ekstruziji so bile vse izmerjene velikosti radijev delcev občutno prevelike, kar smo pripisali nečistočam v ekstrudorju. Ostanek jajčnega lecitina smo zato ponovno ekstrudirali z drugim ekstrudorjem in dobili vezikle željene velikosti, kljub temu pa se pri ponovnih ekstruzijah lipida 1,2-dioleoil-sn-glicero-3-fosfatidiletanolamin velikosti delcev niso skladale z velikostjo por uporabljenega filtra za ekstruzijo. Po ponovni ekstruziji že ekstrudiranih vzorcev se je namreč vzpostavila približno enaka velikost hidrodinamskega radija ne glede na uporabljen filter.

Language:Slovenian
Keywords:vezikli, ekstruzija, dinamično sipanje svetlobe
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2020
PID:20.500.12556/RUL-119534 This link opens in a new window
COBISS.SI-ID:28344067 This link opens in a new window
Publication date in RUL:09.09.2020
Views:1775
Downloads:194
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Secondary language

Language:English
Title:Measuring the size of the lipid vesicles made with extrusion
Abstract:
Studies of vesicles are important to better understand the functioning of living organisms, as they have a wide range of important roles like cell-cell communication and transport of substances. They are composed of a phospholipid bilayer consisted of water and macromolecules like proteins and RNA. Lately scientists have been working on preparation of artificial vesicles or liposomes which can be used for many different purposes. One of the techniques to prepare large vesicles formed from one lipid bilayer is the extrusion technique. We used extrusion for liposomes preparation with the aim of synthesizing vesicles of different sizes, composed of three different phospholipid bilayers. For the structure of phospholipid bilayers, we used natural or synthetic surfactants which reduce the surface tension. We made vesicles from egg lecithin, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine lipid, and a mixture of the 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine lipids. We measured the size of vesicles via dynamic light scattering and thus we were able to calculate the hydrodynamic radius of the particles with the help of measured translation diffusion coefficient (using the Stokes-Einstein equation). According to the first extrusion, all the measured sizes of the particles were too large, presumably due to the impurities in the extruder. We have repeated extrusion with the egg lecithin residues using another cleaner extruder. The vesicles were of the right size. However, with repeated extrusions of 2-dioleoyl-sn-glycero-3-phosphoethanolamine lipid, sizes of vesicles did not agree with the pore size of filters that were used for the extrusion. After another extrusion of samples that had already been extruded, the hydrodynamic radius of vesicles wase almost the same, irrespective of the filter used.

Keywords:vesicles, extrusion, dynamic light scattering

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