The unique potential of fullerene C60 for various biological applications has ignited significant interest. However, its inherent non-polarity poses a critical challenge for its effective integration within biological systems. This study delves into the intricate physicochemical characteristics of the innovative [C60 + NO] complex using density functional theory and time-dependent density functional theory. The computational analyses encompass molecular charge, surface electrostatic potential, and dipole moment evaluations. Impressively, the dipole moment of the [C60 + NO] complex significantly increases to 12.92 D. Meticulous surface analysis reveals a subtle interplay between molecular structures, indicating weak interactions. The analysis of the absorption spectrum unveils a noteworthy red-shift of 200 nm subsequent to complex formation. To elucidate the electron transfer mechanisms, we explore photo-induced electron transfer through CAM-B3LYP. This exploration elucidates intricate pathways governing electron transfer, with complementary insights gleaned from Marcus theory's outputs, especially the Gibbs free energy of electron transfer. Changes in the physicochemical properties of approaching C60 and NO molecules reveal interesting results compared to separate molecules. These findings resonate profoundly in the context of potential biological and pharmaceutical utilization. With implications for the biomedical area, the outcomes linked to the [C60 + NO] complex kindle optimism for pioneering biomedical applications.
Cellular localization of carbon nanomaterials in cancer cells is essential information for better understanding their interaction with biological targets and a crucial factor for further evaluating their biological properties as nanovehicles or nanotherapeutics. Recently, increasing efforts to develop promising fullerene nanotherapeutics for cancer nanotechnology have been made. However, the main challenge regarding studying their cellular effects is the lack of effective methods for their visualization and determining their cellular fate due to the limited fluorescence of buckyball scaffolds. Herein, we developed a method for cellular localization of nonfluorescent and water-soluble fullerene nanomaterials using the in vitro click chemistry approach. First, we synthesized a triple-bonded fullerene probe (TBC60ser), which was further used as a starting material for 1,3-dipolar cycloaddition using 3-azido-7-hydroxycoumarin and sulfo-cyanine5 azide fluorophores to create fluorescent fullerene triazoles. In this work, we characterized the structurally triple-bonded [60]fullerene derivative and confirmed its high symmetry (Th) and the successful formation of fullerene triazoles by spectroscopic techniques (i.e., ultraviolet-visible, fluorescence, and Fourier transform infrared spectroscopies) and mass spectrometry. The created fluorescent fullerene triazoles were successfully localized in the MCF-7 breast cancer cell line using fluorescent microscopy. Overall, our findings demonstrate that TBC60ser localizes in the lysosomes of MCF-7 cells, with only a small affinity to mitochondria.
- MeSH
- fullereny * chemie farmakologie MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- nádory prsu * farmakoterapie MeSH
- nanostruktury * chemie MeSH
- triazoly MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Fullerene derivatives with hydrophilic substituents have been shown to exhibit a range of biological activities, including antiviral ones. For a long time, the anti-HIV activity of fullerene derivatives was believed to be due to their binding into the hydrophobic pocket of HIV-1 protease, thereby blocking its activity. Recent work, however, brought new evidence of a novel, protease-independent mechanism of fullerene derivatives' action. We studied in more detail the mechanism of the anti-HIV-1 activity of N,N-dimethyl[70]fulleropyrrolidinium iodide fullerene derivatives. By using a combination of in vitro and cell-based approaches, we showed that these C70 derivatives inhibited neither HIV-1 protease nor HIV-1 maturation. Instead, our data indicate effects of fullerene C70 derivatives on viral genomic RNA packaging and HIV-1 cDNA synthesis during reverse transcription-without impairing reverse transcriptase activity though. Molecularly, this could be explained by a strong binding affinity of these fullerene derivatives to HIV-1 nucleocapsid domain, preventing its proper interaction with viral genomic RNA, thereby blocking reverse transcription and HIV-1 infectivity. Moreover, the fullerene derivatives' oxidative activity and fluorescence quenching, which could be one of the reasons for the inconsistency among reported anti-HIV-1 mechanisms, are discussed herein.
- MeSH
- fullereny metabolismus farmakologie MeSH
- genom virový účinky léků MeSH
- genové produkty gag - virus lidské imunodeficience metabolismus MeSH
- HEK293 buňky MeSH
- HIV-1 účinky léků genetika metabolismus fyziologie MeSH
- látky proti HIV metabolismus farmakologie MeSH
- lidé MeSH
- nukleokapsida - proteiny metabolismus MeSH
- reverzní transkripce MeSH
- RNA virová metabolismus MeSH
- svlékání virového obalu účinky léků MeSH
- vazba proteinů MeSH
- virion metabolismus MeSH
- zabalení virového genomu účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Doxorubicin (DOX) is one of the most frequently used anticancer drugs in breast cancer treatment. However, clinical applications of DOX are restricted, largely due to the fact that its action disturbs the pro/antioxidant balance in both cancerous and non-cancerous cells. The aim of this study was to investigate the influence of fullerene (C60) in cell treatment by DOX on the proliferation of human breast cancer cells (MCF-7), concentration of metallothionein (MT) and superoxide dismutase (SOD), and SOD activity in these cells. The use of C60 in complexes with DOX causes a change in the level of cell proliferation of about 5% more than when caused by DOX alone (from 60⁻65% to 70%). The use of C60 as a DOX nanotransporter reduced the MT level increase induced by DOX. C60 alone caused an increase of SOD1 concentration. On the other hand, it led to a decrease of SOD activity. C60 in complex with DOX caused a decrease of the DOX-induced SOD activity level. Exposure of MCF-7 cells to DOX-C60 complexes results in a decrease in viable cells and may become a new therapeutic approach to breast cancer. The effects of C60 in complexes with DOX on MCF-7 cells included a decreased enzymatic (SOD activity) and nonenzymatic (MT) antioxidant status, thus indicating their prooxidant role in MCF-7 cells.
- MeSH
- antitumorózní látky farmakologie MeSH
- doxorubicin farmakologie MeSH
- fullereny chemie MeSH
- lidé MeSH
- metalothionein metabolismus MeSH
- MFC-7 buňky MeSH
- nanočástice chemie MeSH
- proliferace buněk účinky léků MeSH
- superoxid dismutáza 1 metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The clinical use of doxorubicin (DOX) is limited by dose-related cardiomyopathy, which becomes more prevalent with increasing cumulative doses of the drug. Complexes of fullerene with DOX were designed and studied using biophysical methods. The ability of DOX to release from fullerene at different pHs was analyzed. It has been shown that the size of the fullerene-DOX complexes was ∼280 nm. The zeta potential for fullerene was -30 mV, for DOX -8 mV, and for fullerene-DOX conjugates -24 mV. Drug release was studied by CE with LIF detection. When fullerene-DOX conjugates were separated in a pH 7.5 buffer, 43% of all DOX signals were derived from free DOX, with the signal increasing as pH decreased. At pH 5.25, all DOX had been released and 100% of the signal was derived from free DOX. The release of DOX from complexes with fullerene at lower pH can be used in targeted antineoplastic therapy, resulting in lower toxicity for less acidic non-target tissue.
- MeSH
- antibiotika antitumorózní terapeutické užití MeSH
- doxorubicin terapeutické užití MeSH
- elektroforéza kapilární MeSH
- fluorescenční barviva chemie MeSH
- fullereny chemie MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- nádory prsu farmakoterapie MeSH
- nanočástice chemie MeSH
- nosiče léků chemie MeSH
- optické zobrazování MeSH
- povrchové vlastnosti MeSH
- uvolňování léčiv MeSH
- velikost částic MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cíl: Cílem této práce bylo vyvinout nanoléčivo založené na specifickém transportu pomocí oligonukleotidové próby (komplementární k BRCA1 genové mutaci) s předpokládaným účinkem cytostatika bez výrazných toxických efektů k nenádorovým buňkám. Použitým cytostatikem bylo antracyklinové antibiotikum doxorubicin. Přes svou terapeutickou účinnost však vykazuje vysokou kardiotoxicitu. Možnost, jak zvýšit terapeutické okno, představují nanotransportéry. Metodika: Doxorubicin a oligonukleotidová próba byly navázány k fullerenům. Doba modifikace nanotransportérů byla 24 h. Nanotransportér byl následně charakterizován pomocí biofyzikálních technik (SEM, dynamického rozptylu světla, spektrální a elektrochemické metody). Pomocí elektrochemické a fluorescenční analýzy byl vzniklý komplex nanotransportéru potvrzen. Výsledky: Doxorubicin se záporně nabitým povrchem interaguje s kladně nabitými fullereny elektrostatickou interakcí a tato interakce byla potvrzena na základě elektronové mikroskopie (SEM), elektrochemicky (změna potenciálu byla 35 mV) a měřením dynamického rozptylu světla (změna ζ-potenciálu byla 22 mV). Ke komplexu byla amidovou vazbou připojena oligonukleotidová proba, která je komplementární k mutované sekvenci BRCA genu. Pokles fluorescenčního signálu nanokonstruktu o 80 % indikoval navázání oligonukleotidu. Pro prokázání funkčnosti byla navržena magnetická zlatá nanočástice modifikovaná komplementární sekvencí k testovanému nanotransportéru. Závěr: Předpokládáme, že navržený nanokonstrukt bude možné využít pro DNA cílené směřování protinádorového léčiva k buňkám s mutací v genu BRCA. Nanotransportér má tyto základní vlastnosti: a) fulleren vykazuje vysokou afinitu k buňce a proniká cytoplazmatickou membránou; b) doxorubicin se uvolní do cytoplazmy v nádorových buňkách díky nízkému pH; c) celý konstrukt je směřován cíleně na mutovanou sekvenci genu BRCA; d) zlatá nanočástice zesiluje cytotoxický efekt
Objective: Main goal of this project was to develop a nanotransporter based on a targeted delivery using oligonucleotide probe, which is complementary to a mutated BRCA1 gene sequence with the expected effect of a cytostatic without significant toxic effects. The mentioned cytostatic is often used doxorubicin. Despite its therapeutic efficacy, however, it exhibits high cardiotoxicity. Nanotransporters represent the possibility of increasing the therapeutic window. Methods: Doxorubicin and oligonucleotide probe were bound to fullerenes. The modification time was 24 hours. The nanotransporter was subsequently characterized by biophysical techniques (SEM, dynamic light scattering, spectral and electrochemical methods.) Using the electrochemical and fluorescence analysis the formation of nanotransporter was confirmed. Results: Doxorubicin with a negatively charged surface interacts with positively charged fullerenes by electrostatic interaction and this interaction was confirmed by on the basis of electron microscopy (SEM), electrochemically (change of potential was 35 mV) and by measurement of dynamic light scattering (change of ζ-potential was 22 mV). Oligonucleotide probe, which is complementary to the BRCA mutated gene sequence, was bound to the complex by an amide bond. The drop of the fluorescence signal by 80% indicated binding of the oligonucleotide. To demonstrate the functionality, a magnetic gold nanoparticle, modified by the complementary sequence to the tested nanotransporter, was developed. Conclusion: We assume, that the proposed nanotransporter will be used for DNA targeted delivery of the antitumor drug to the cells with BRCA mutated genes. The nanotransporter has these basic characteristics: a) fullerene has a high affinity to the cell and penetrates the cell membrane; b) doxorubicin is released into cytoplasm in tumour cells due to low pH; c) the entire construct is targeted to the BRCA mutated gene sequence; d) the gold nanoparticle enhances the cytotoxic effect.
- MeSH
- antibiotika antitumorózní MeSH
- doxorubicin aplikace a dávkování terapeutické užití toxicita MeSH
- fullereny terapeutické užití MeSH
- geny BRCA1 MeSH
- geny BRCA2 MeSH
- kovové nanočástice využití MeSH
- lidé MeSH
- mutace genetika MeSH
- nádory prsu * terapie MeSH
- nanomedicína * MeSH
- oligonukleotidy terapeutické užití MeSH
- pilotní projekty MeSH
- systémy cílené aplikace léků MeSH
- zlato terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Photodynamic therapy (PDT) has garnered immense attention as a minimally invasive clinical treatment modality for malignant cancers. However, its low penetration depth and photodamage of living tissues by UV and visible light, which activate a photosensitizer, limit the application of PDT. In this study, monodisperse NaYF4 :Yb3+ /Er3+ nanospheres 20 nm in diameter, that serve as near-infrared (NIR)-to-visible light converters and activators of a photosensitizer, were synthesized by high-temperature co-precipitation of lanthanide chlorides in a high-boiling organic solvent (octadec-1-ene). The nanoparticles were coated with a thin shell (≈3 nm) of homogenous silica via the hydrolysis and condensation of tetramethyl orthosilicate. The NaYF4 :Yb3+ /Er3+ @SiO2 particles were further functionalized by methacrylate-terminated groups via 3-(trimethoxysilyl)propyl methacrylate. To introduce a large number of reactive amino groups on the particle surface, methacrylate-terminated NaYF4 :Yb3+ /Er3+ @SiO2 nanospheres were modified with a branched polyethyleneimine (PEI) via Michael addition. Aluminum carboxyphthalocyanine (Al Pc-COOH) was then conjugated to NaYF4 :Yb3+ /Er3+ @SiO2 -PEI nanospheres via carbodiimide chemistry. The resulting NaYF4 :Yb3+ /Er3+ @SiO2 -PEI-Pc particles were finally modified with succinimidyl ester of poly(ethylene glycol) (PEG) in order to alleviate their future uptake by the reticuloendothelial system. Upon 980 nm irradiation, the intensive red emission of NaYF4 :Yb3+ /Er3+ @SiO2 -PEI-Pc-PEG nanoparticles completely vanished, indicating efficient energy transfer from the nanoparticles to Al Pc-COOH, which generates singlet oxygen (1 O2 ). Last but not least, NaYF4 :Yb3+ /Er3+ @SiO2 -PEI-Pc-PEG nanospheres were intratumorally administered into mammary carcinoma MDA-MB-231 growing subcutaneously in athymic nude mice. Extensive necrosis developed at the tumor site of all mice 24-48 h after irradiation by laser at 980 nm wavelength. The results demonstrate that the NaYF4 :Yb3+ /Er3+ @SiO2 -PEI-Pc-PEG nanospheres have great potential as a novel NIR-triggered PDT nanoplatform for deep-tissue cancer therapy.
- MeSH
- antitumorózní látky chemická syntéza chemie farmakologie MeSH
- erbium chemie farmakologie MeSH
- experimentální nádory farmakoterapie patologie MeSH
- fluoridy chemie farmakologie MeSH
- fotochemoterapie * MeSH
- fotosenzibilizující látky chemická syntéza chemie farmakologie MeSH
- indoly chemie farmakologie MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- molekulární struktura MeSH
- myši nahé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nanokuličky chemie MeSH
- oxid křemičitý chemie farmakologie MeSH
- proliferace buněk účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- yterbium chemie farmakologie MeSH
- ytrium chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We studied the surface properties and cytocompatibil-ity of a grafted biopolymer, poly(3-hydroxybutyrate) (P3HB). P3HB was exposed to an inert argon plasma dis-charge, then grafted by 1,1 '-biphenyl-4,4 '-dithiol (BFD) and finally grafted by golden nanoparticles (nanospheres and nanorods). The surface properties were studied using multiple methods – goniometry, atomic force microscopy and X-ray photoelectron spectroscopy. Cytocompatibility was determined in vitro by studying adhesion, prolifera-tion and viability of vascular smooth muscle cells (VSMCs) from the aorta of Rattus norvegicus. The cyto-compatibility was compared for pristine, modified P3HB and standard tissue culture polystyrene (TCPS). Our re-sults show that surface morphology and wettability are affected by both plasma discharge and nanoparticles graft-ing. These changes suggest that the adhesion and prolifera-tion of VSMCs is enhanced more on the plasma modified and grafted substrate.
Relative concentrations of six isomeric Eu@C 72 -one based on the IPR C 72 cage (i.e., obeying the isolated-pentagon rule, IPR), two cages with a pentagon-pentagon junction (symmetries C 2 and C 2 v ), a cage with one heptagon, a cage with two heptagons, and a cage with two pentagon-pentagon fusions-are DFT computed using the Gibbs energy in a broad temperature interval. It is shown that the two non-IPR isomers with one pentagon-pentagon junction prevail at any relevant temperature and exhibit comparable populations. The IPR-satisfying structure is disfavored by both energy and entropy.
- Klíčová slova
- magnetické zlaté nanočástice,
- MeSH
- doxorubicin * MeSH
- fullereny * MeSH
- geny BRCA2 MeSH
- lidé MeSH
- magnetické nanočástice MeSH
- mutace MeSH
- nádory prsu farmakoterapie genetika MeSH
- nosiče léků * chemická syntéza MeSH
- oligodeoxyribonukleotidy MeSH
- protein BRCA2 genetika MeSH
- zlato MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
