Příprava a charakterizace nanodiamantů modifikovaných biokompatibilními polymery
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Vysoké učení technické v Brně. Fakulta chemická
Abstract
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).
Nanodiamonds (NDs, 250 nm) were oxidized, separated by size and the polyglycerol layer was polymerized on the ND's surface. The polymerization was achieved by acid catalyzed ring opening multi-branching polymerization of glycidol with the hydroxyl and carboxylic groups formed, during oxidation on the NDs surface. The polyglycerol layer increases the stability and biocompatibility of the NDs. After the coating, a terminal propargyl group was introduced by glycidyl propargyl ether addition. The presence of the alkyne group enables further modification using copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC-click). Employing CuAAC click reaction, a mannose (with azide linker) was attached to the NDs surface. Thus prepared NPs were characterised from the physicochemical point of view. The size distribution was measured by dynamic light scattering (DLS), and it was further confirmed by transmission electron microscopy (TEM). Inertness towards biological conditions was evaluated (stability tests in 1xPBS, 10xPBS and 100 % FBS) and zeta potential was measured. Next, fluorescent nanodiamonds (FNDs, 250 nm) with attached D-mannose were prepared. According to the thermogravimetry results, DLS measurement and TEM images, it was determined, that NDs are covered by thin polyglycerol layer. Presence of glycidol coating was further confirmed by stability tests. Presence of attached D-mannose was supported by Fourier-transform infrared spectroscopy (FTIR). Prepared polyglycerol coated NDs are well suited for in vitro and in vivo testing. That was proven by their stability in high salt concentration media such as phosphate buffer saline (PBS) and low protein binding properties in fetal bovine serum (FBS).
Nanodiamonds (NDs, 250 nm) were oxidized, separated by size and the polyglycerol layer was polymerized on the ND's surface. The polymerization was achieved by acid catalyzed ring opening multi-branching polymerization of glycidol with the hydroxyl and carboxylic groups formed, during oxidation on the NDs surface. The polyglycerol layer increases the stability and biocompatibility of the NDs. After the coating, a terminal propargyl group was introduced by glycidyl propargyl ether addition. The presence of the alkyne group enables further modification using copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC-click). Employing CuAAC click reaction, a mannose (with azide linker) was attached to the NDs surface. Thus prepared NPs were characterised from the physicochemical point of view. The size distribution was measured by dynamic light scattering (DLS), and it was further confirmed by transmission electron microscopy (TEM). Inertness towards biological conditions was evaluated (stability tests in 1xPBS, 10xPBS and 100 % FBS) and zeta potential was measured. Next, fluorescent nanodiamonds (FNDs, 250 nm) with attached D-mannose were prepared. According to the thermogravimetry results, DLS measurement and TEM images, it was determined, that NDs are covered by thin polyglycerol layer. Presence of glycidol coating was further confirmed by stability tests. Presence of attached D-mannose was supported by Fourier-transform infrared spectroscopy (FTIR). Prepared polyglycerol coated NDs are well suited for in vitro and in vivo testing. That was proven by their stability in high salt concentration media such as phosphate buffer saline (PBS) and low protein binding properties in fetal bovine serum (FBS).
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Citation
KVAKOVÁ, K. Příprava a charakterizace nanodiamantů modifikovaných biokompatibilními polymery [online]. Brno: Vysoké učení technické v Brně. Fakulta chemická. 2019.
Document type
Document version
Date of access to the full text
Language of document
en
Study field
Chemie pro medicínské aplikace
Comittee
prof. Ing. Miloslav Pekař, CSc. (předseda)
prof. RNDr. Ivana Márová, CSc. (místopředseda)
doc. Ing. Pavel Diviš, Ph.D. (člen)
doc. Ing. Petr Dzik, Ph.D. (člen)
doc. Ing. Stanislav Obruča, Ph.D. (člen)
prof. RNDr. Zbyněk Zdráhal, Dr. (člen)
Date of acceptance
2019-05-28
Defence
Student prezentoval řešenou práci, její teoretická východiska a vlastní výsledky, zodpověděl dotazy položené oponentem a následně byly poleženy dotazy členů komise. Diviš: Jaká je výhoda použití nanodiamantů proti jiným nanočásticím?Pekař: Jaký je význam zkratky 10xPBS?Márová: Jaká je kinetika distribuce a vylučování nanodiamantů v těle?Obruča: Jaký je váš vztah k řešitelskému pracovišti? Už tam pracujete? Student prokázal výbornou orientaci v dané problematice a schopnost samostatné prezentace získaných výsledků.
Result of defence
práce byla úspěšně obhájena
Document licence
Standardní licenční smlouva - přístup k plnému textu bez omezení