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Modification of hyaluronan by cholic acid and the using of these derivatives for carrier applications
Kvaková, Klaudia ; Márová, Ivana (referee) ; Velebný, Vladimír (advisor)
Amphiphilic biopolymers were synthesised using esterification of hyaluronic acid (HA) and cholic acid as hydrophobic segment. First, step of synthesis involves the activation of cholic acid carboxylic moiety by reaction with benzoylchloride and triethylamine (TEA) using as solvent tetrahydrofuran (THF). The formed mixed aliphatic/aromatic anhydride (intermediate) reacted at room temperature with hyaluronic acid catalysed by 4-(dimethylamino)pyridine (DMAP) and triethylamine and using as solvent water/THF. Hydrophobized product was identified as sodium cholyl hyaluronate (HA-CA) was obtained as product of the reaction. Structure of derivative was characterised by Nuclear magnetic resonance (NMR) and Fourier Transform - Infrared Spectroscopy (FT-IR). Size exclusion chromatography (SEC-MALLS) was used to determine molecular weight of derivative to evaluate any possible degradation of hyaluronan during modification. Rheological properties of these substances were also examined and confirmed non-degradation. The hydrophobized hyaluronic acid properties were evaluated for application as drug delivery system. Therefore, two non-polar compounds were encapsulated (coenzyme Q10 and curcumin). The size of formed polymeric micelles was characterised by dynamic light scattering (DLS).
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Preparation and characterization of nanodiamonds modified with biocompatible polymers
Kvaková, Klaudia ; Skoumalová, Petra (referee) ; Cígler, Petr (advisor)
Nanodiamanty (250 nm) boli podrobené oxidácií, a tiež separované podľa veľkosti. Následne bola na povrch nanodiamantov adsorbovaná rozvetvená vrstva polyglycerolu. Táto vrstva vzniká kyslo katalyzovanou ,,ring opening“ polymeráciou glycidolu s hydroxylovými a karboxylovými skupinami povrchu nanodiamantov. Vzniknutá polyglycerolová vrstva zvyšuje stabilitu a biokompatibilitu nanodiamantov. Po vytvorení polyglycerolovej vrstvy bol do reakcie pridaný glycidyl propargyl éter, čím došlo k terminácií polymerácie, v dôsledku zavedenia propargylu do reakcie. Prítomná trojná väzba umožňuje nové modifikácie povrchu, napr. prostredníctvom CuAAC reakcie. S využitím CuAAC reakcie je možné na povrch fluorescenčných nanodiamantov pripojiť manózu (so spojkou obsahujúcou azidoskupinu). Pripravené nanočastice boli charakterizované použitím fyzikálno chemických metód. Distribúcia veľkosti častíc bola meraná pomocou dynamického rozptylu svetla (DLS), a taktiež bola potvrdená obrazovou analýzou mikrografov z elektrónového transmisného mikroskopu (TEM). Vyhodnotené bolo aj správanie sa nanodiamantov v biologických podmienkach (stabilitné testy v 1xPBS, 10xPBS a 100 % FBS) a bol meraný aj zeta potenciál. Nakoniec boli pripravené fluorescenčné diamanty (250 nm) s naviazanou manózou. Vzhľadom k výsledkom termogravimetrie, meraniam DLS a pozorovania pomocou TEM môžeme povedať, pripravené nanodiamanty sú pokryté tenkou vrstvou polyglycerolu. Prítomnosť manózy bola zistená na základe FTIR spektier. Nami pripravené fluorescenčné nanodiamanty obalené vrstvou polyglycerolu sú vhodné pre ďalšie testovanie in vitro a in vivo vďaka ich stabilite v médiách s vysokým obsahom solí ako napr. fosfátový pufor (PBS) a nízkemu viazaniu proteínov vo fetalnom bovinnom sere (FBS).
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Fluorescent Nanoprobes Emitting in Near-Infrared Region for Biomedical Applications
Kvaková, Klaudia ; Cígler, Petr (advisor) ; Bogdanowicz, Robert (referee) ; Poláková, Kateřina (referee)
The primary focus of this thesis is to conduct an in-depth investigation into the capabilities of near-infrared photoluminescent (PL) nanoprobes and their application in biomedicine. Specifically, this research explores the potential of fluorescent nanodiamonds (FNDs) and gold nanoclusters (AuNCs). First, the thesis examined the potential of FNDs for in vivo visualization of mice sentinel lymph nodes (SLNs). The study observed that FNDs modified with mannose exhibited greater uptake by macrophages and improved retention within SLNs compared to non- mannosylated FNDs. In addition, FNDs possess a remarkable capacity to accurately measure cellular temperature as was demonstrated by local temperature variation measurements associated to hippocampal neurons firing. Furthermore, the study explored the potential of AuNCs as viable tools for cellular imaging. A simple synthetic approach was developed to create AuNCs with enhanced stability, ligand-based PL enhancement, and prolonged fluorescence lifetime. The thesis also describes the reversible photo- and thermal-induced effects on the PL properties of the synthesized AuNCs. Additionally, the enhancement of AuNCs' PL through their combination with plasmonic nanostructures, specifically gold nanorods, was described. Lastly, a successful visualization of the...
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Preparation and characterization of nanodiamonds modified with biocompatible polymers
Kvaková, Klaudia ; Skoumalová, Petra (referee) ; Cígler, Petr (advisor)
Nanodiamanty (250 nm) boli podrobené oxidácií, a tiež separované podľa veľkosti. Následne bola na povrch nanodiamantov adsorbovaná rozvetvená vrstva polyglycerolu. Táto vrstva vzniká kyslo katalyzovanou ,,ring opening“ polymeráciou glycidolu s hydroxylovými a karboxylovými skupinami povrchu nanodiamantov. Vzniknutá polyglycerolová vrstva zvyšuje stabilitu a biokompatibilitu nanodiamantov. Po vytvorení polyglycerolovej vrstvy bol do reakcie pridaný glycidyl propargyl éter, čím došlo k terminácií polymerácie, v dôsledku zavedenia propargylu do reakcie. Prítomná trojná väzba umožňuje nové modifikácie povrchu, napr. prostredníctvom CuAAC reakcie. S využitím CuAAC reakcie je možné na povrch fluorescenčných nanodiamantov pripojiť manózu (so spojkou obsahujúcou azidoskupinu). Pripravené nanočastice boli charakterizované použitím fyzikálno chemických metód. Distribúcia veľkosti častíc bola meraná pomocou dynamického rozptylu svetla (DLS), a taktiež bola potvrdená obrazovou analýzou mikrografov z elektrónového transmisného mikroskopu (TEM). Vyhodnotené bolo aj správanie sa nanodiamantov v biologických podmienkach (stabilitné testy v 1xPBS, 10xPBS a 100 % FBS) a bol meraný aj zeta potenciál. Nakoniec boli pripravené fluorescenčné diamanty (250 nm) s naviazanou manózou. Vzhľadom k výsledkom termogravimetrie, meraniam DLS a pozorovania pomocou TEM môžeme povedať, pripravené nanodiamanty sú pokryté tenkou vrstvou polyglycerolu. Prítomnosť manózy bola zistená na základe FTIR spektier. Nami pripravené fluorescenčné nanodiamanty obalené vrstvou polyglycerolu sú vhodné pre ďalšie testovanie in vitro a in vivo vďaka ich stabilite v médiách s vysokým obsahom solí ako napr. fosfátový pufor (PBS) a nízkemu viazaniu proteínov vo fetalnom bovinnom sere (FBS).
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Modification of hyaluronan by cholic acid and the using of these derivatives for carrier applications
Kvaková, Klaudia ; Márová, Ivana (referee) ; Velebný, Vladimír (advisor)
Amphiphilic biopolymers were synthesised using esterification of hyaluronic acid (HA) and cholic acid as hydrophobic segment. First, step of synthesis involves the activation of cholic acid carboxylic moiety by reaction with benzoylchloride and triethylamine (TEA) using as solvent tetrahydrofuran (THF). The formed mixed aliphatic/aromatic anhydride (intermediate) reacted at room temperature with hyaluronic acid catalysed by 4-(dimethylamino)pyridine (DMAP) and triethylamine and using as solvent water/THF. Hydrophobized product was identified as sodium cholyl hyaluronate (HA-CA) was obtained as product of the reaction. Structure of derivative was characterised by Nuclear magnetic resonance (NMR) and Fourier Transform - Infrared Spectroscopy (FT-IR). Size exclusion chromatography (SEC-MALLS) was used to determine molecular weight of derivative to evaluate any possible degradation of hyaluronan during modification. Rheological properties of these substances were also examined and confirmed non-degradation. The hydrophobized hyaluronic acid properties were evaluated for application as drug delivery system. Therefore, two non-polar compounds were encapsulated (coenzyme Q10 and curcumin). The size of formed polymeric micelles was characterised by dynamic light scattering (DLS).
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