The modification of biomaterial surfaces has become increasingly relevant in the context of ongoing advancements in tissue engineering applications and the development of tissue-mimicking polymer materials. In this study, we investigated the layer-by-layer (LbL) deposition of polyelectrolyte multilayer protein reservoirs consisting of poly-l-lysine (PLL) and hyaluronic acid (HA) on the hydrophobic surface of poly(glycerol sebacate) (PGS) elastomer. Using the methods of isothermal titration calorimetry and surface plasmon resonance, we systematically investigated the interactions between the polyelectrolytes and evaluated the deposition process in real time, providing insight into the phenomena associated with film assembly. PLL/HA LbL films deposited on PGS showed an exceptional ability to incorporate bone morphogenetic protein-2 (BMP-2) compared to other growth factors tested, thus highlighting the potential of PLL/HA LbL films for osteoregenerative applications. The concentration of HA solution used for film assembly did not affect the thickness and topography of the (PLL/HA)10 films, but had a notable impact on the hydrophilicity of the PGS surface and the BMP-2 release kinetics. The release kinetics were successfully described using the Weibull model and hyperbolic tangent function, underscoring the potential of these less frequently used models to compare the protein release from LbL protein reservoirs.
This study illustrates the synthesis of functionalized carbon quantum dots (CQDs) by the one-pot pyrolysis method. The functionalization agent used in CQD synthesis was poly l- lysine (PLL). Various physicochemical techniques were employed to confirm the successful formation of PLLCQD including High resolution transmission electron microscopy (HR-TEM), UV-Vis spectroscopy, fluorescence spectroscopy; Atomic force microscopy (AFM), X-ray Photoelectron Spectroscopy (XPS) and X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy. The size of PLLCQD was confirmed by HRTEM and AFM. The synthesized PLLCQD shows bright blue fluorescence and has a quantum yield of 19.35%. The highest emission band was observed at 471nm when excited to 370nm. The prepared PLLCQD exhibited excellent antibacterial activity against Escherichia coli and Staphylococcus aureus with inhibition zone 7-20 mm. The concentrations of 0.9 to 0.1gmL-1 were studied to determine minimum inhibitory concentration (MIC) by the agar well diffusion assay method. MIC of 0.2gml -1 concentration of PLLCQD is achieved. The anti-angiogenic activity of PLLCQD was determined using (Chick Chorioallantoic Membrane) CAM assay. CAM assay is a reliable in -vivo model to study angiogenesis also; many stimulators and inhibitors have been examined by this method. This study proves higher antibacterial efficiency of PLLCQD over non functionalized CQD. PLLCQD was successfully employed in bio-imaging of the bacterial cell through fluorescence microscopy. Further, PLLCQD displayed cytotoxic effect on endothelial cells and inhibited blood vessel formation in the CAM model.
Filmy jako krycí prostředky představují jednu z možností v terapii ran. K jejich výrobě lze použít různé polymery. V současné době se výzkum zaměřuje na materiály přírodního původu, pro lidský organismus přívětivější, které jsou v řadě případů schopné zapojit se aktivně do procesu hojení rány. K takovým patří polyaminokyseliny bakteriálního původu, látky biodegradabilní, netoxické, skýtající velký potenciál pro uplatnění nejen ve zdravotnictví. Z hlediska formulace filmového krytí na rány jsou v této skupině zajímavé kyselina poly-γ-glutamová (PGA), jako látka filmotvorná, a poly-ε-lysin (PL) charakteristický antimikrobiálním účinkem. Proto cílem našeho experimentu bylo připravit filmy tvořené PGA nebo kombinací PGA a PL s přísadou různých změkčovadel. Filmy se připravovaly metodou odpaření rozpouštědla a následně se hodnotily jejich organoleptické (vzhled, barva, průhlednost, snadnost manipulace), fyzikálně-chemické (tloušťka, hustota, opacita, pH výluhu i povrchové pH) i mechanické vlastnosti (pevnost v tahu a odolnost proti protržení). Výsledkem byly filmy vykazující vzájemnou kompatibilitu obou polymerů s vyhovujícími vlastnostmi z hlediska aplikace na ránu.
Film wound dressings represent one of the options in wound therapy. Various polymers can be used for their production. Currently, research focuses on materials of natural origin, more friendly to the human body, which are in many cases able to participate actively in the wound healing process. These include polyamino acids of bacterial origin, substances that are biodegradable, non-toxic, and have a great potential for an application not only in the medical field. From the point of view of film wound dressing formulation, poly-γ-glutamic acid (PGA), as a film-forming agent, and poly-ε-lysine (PL), characterized by antimicrobial activity, are of interest from this group. Therefore, the aim of our experiment was to prepare films consisting of PGA or a combination of PGA and PL with the addition of different plasticizers. The films were prepared by solvent evaporation method and then evaluated for their organoleptic (appearance, colour, transparency, ease of handling), physicochemical (thickness, density, opacity, surface pH), and mechanical properties (tensile strength and tear resistance). As a result, films showing mutual compatibility between the two polymers were obtained, with satisfactory properties for wound application.
- Klíčová slova
- filmové krytí,
- MeSH
- hojení ran * MeSH
- kyselina polyglutamová farmakologie terapeutické užití MeSH
- lidé MeSH
- objevování léků MeSH
- polylysin farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
Background: Poly-l-lysine (PLL) enhances nanoparticle (NP) uptake, but the molecular mechanism remains unresolved. We asked whether PLL may interact with negatively charged glycoconjugates on the cell surface and facilitate uptake of magnetic NPs (MNPs) by tumor cells. Methods: PLL-coated MNPs (PLL-MNPs) with positive and negative ζ-potential were prepared and characterized. Confocal and transmission electron microscopy was used to analyze cellular internalization of MNPs. A colorimetric iron assay was used to quantitate cell-associated MNPs (MNPcell). Results: Coadministration of PLL and dextran-coated MNPs in culture enhanced cellular internalization of MNPs, with increased vesicle size and numbers/cell. MNPcell was increased by eight- to 12-fold in response to PLL in a concentration-dependent manner in human glioma and HeLa cells. However, the application of a magnetic field attenuated PLL-induced increase in MNPcell. PLL-coating increased MNPcell regardless of ζ-potential of PLL-MNPs, whereas magnetic force did not enhance MNPcell. In contrast, epigallocatechin gallate and magnetic force synergistically enhanced PLL-MNP uptake. In addition, heparin, but not sialic acid, greatly reduced the enhancement effects of PLL; however, removal of heparan sulfate from heparan sulfate proteoglycans of the cell surface by heparinase III significantly reduced MNPcell. Conclusion: Our results suggest that PLL-heparan sulfate proteoglycan interaction may be the first step mediating PLL-MNP internalization by tumor cells. Given these results, PLL may facilitate NP interaction with tumor cells via a molecular mechanism shared by infection machinery of certain viruses.
- MeSH
- buněčná membrána metabolismus MeSH
- dextrany chemie metabolismus MeSH
- endoteliální buňky pupečníkové žíly (lidské) MeSH
- gliom farmakoterapie patologie MeSH
- HeLa buňky MeSH
- heparansulfát proteoglykany chemie metabolismus MeSH
- lidé MeSH
- magnetické nanočástice aplikace a dávkování chemie MeSH
- magnetické pole MeSH
- nádorové buněčné linie MeSH
- polylysin chemie metabolismus farmakokinetika MeSH
- polysacharid-lyasy metabolismus MeSH
- transmisní elektronová mikroskopie MeSH
- železo metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Cationic colloidal gold nanorods (GNRs) have a great potential as a theranostic tool for diverse medical applications. GNRs' properties such as cellular internalization and stability are determined by physicochemical characteristics of their surface coating. GNRs modified by (16-mercaptohexadecyl)trimethylammonium bromide (MTAB), MTABGNRs, show excellent cellular uptake. Despite their promise for biomedicine, however, relatively little is known about the cellular pathways that facilitate the uptake of GNRs, their subcellular fate and intracellular persistence. Here we studied the mechanism of cellular internalization and long-term fate of GNRs coated with MTAB, for which the synthesis was optimized to give higher yield, in various human cell types including normal diploid versus cancerous, and dividing versus nondividing (senescent) cells. The process of MTABGNRs internalization into their final destination in lysosomes proceeds in two steps: (1) fast passive adhesion to cell membrane mediated by sulfated proteoglycans occurring within minutes and (2) slower active transmembrane and intracellular transport of individual nanorods via clathrin-mediated endocytosis and of aggregated nanorods via macropinocytosis. The expression of sulfated proteoglycans was the major factor determining the extent of uptake by the respective cell types. Upon uptake into proliferating cells, MTABGNRs were diluted equally and relatively rapidly into daughter cells; however, in nondividing/senescent cells the loss of MTABGNRs was gradual and very modest, attributable mainly to exocytosis. Exocytosed MTABGNRs can again be internalized. These findings broaden our knowledge about cellular uptake of gold nanorods, a crucial prerequisite for future successful engineering of nanoparticles for biomedical applications such as photothermal cancer therapy or elimination of senescent cells as part of the emerging rejuvenation approach.
- MeSH
- buněčná membrána účinky léků metabolismus MeSH
- endocytóza účinky léků fyziologie MeSH
- exocytóza * účinky léků fyziologie MeSH
- konfokální mikroskopie MeSH
- kultivační média MeSH
- kvartérní amoniové sloučeniny chemická syntéza chemie MeSH
- lidé MeSH
- lyzozomy účinky léků MeSH
- mikroskopie elektronová rastrovací MeSH
- nádorové buněčné linie MeSH
- nanotrubičky analýza chemie MeSH
- polylysin chemie farmakokinetika MeSH
- proliferace buněk účinky léků MeSH
- proteoglykany chemie metabolismus MeSH
- průtoková cytometrie MeSH
- stabilita léku MeSH
- sulfhydrylové sloučeniny chemie MeSH
- techniky syntetické chemie MeSH
- zlato chemie farmakokinetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The parasitic mite Varroa destructor is a major pest of the western honeybee, Apis mellifera. The development of acaricide resistance in Varroa populations is a global issue. Discriminating concentrations of acaricides are widely used to detect pest resistance. Two methods, using either glass vials or paraffin capsules, are used to screen for Varroa resistance to various acaricides. We found the glass vial method to be useless for testing Varroa resistance to acaridices, so we developed a polypropylene vial bioassay. This method was tested on tau-fluvalinate-, acrinathrin-, and amitraz-resistant mite populations from three apiaries in Czechia. Acetone was used as a control and technical grade acaricide compounds diluted in acetone were applied to the polypropylene vials. The solutions were spread on the vial surface by rolling the vial, and were then evaporated. Freshly collected Varroa females were placed in the vials and the mortality of the exposed mites was measured after 24 h. The Varroa populations differed in mortality between the apiaries and the tested compounds. Mites from the Kyvalka site were resistant to acrinathrin, tau-fluvalinate, and amitraz, while mites from the Postrizin site were susceptible to all three acaricides. In Prelovice apiary, the mites were susceptible to acrinathrin and amitraz, but not to tau-fluvalinate. The calculated discriminating concentrations for tau-fluvalinate, acrinathrin, and amitraz were 0.66, 0.26 and 0.19 µg/mL, respectively. These results indicate that polyproplyne vial tests can be used to determine discriminating concentrations for the early detection of acaricide resistant Varroa. Finally, multiple-resistance in Kyvalka may indicate metabolic resistance.
- MeSH
- akaricidy * MeSH
- fixní kombinace léků MeSH
- kontrola klíšťat * MeSH
- nitrily * MeSH
- polylysin analogy a deriváty MeSH
- pyrethriny * MeSH
- toluidiny * MeSH
- Varroidae * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Coprecipitation of FeCl2 and FeCl3 with aqueous ammonia was used to prepare iron oxide nanoparticles dispersible in aqueous medium. Oxidation of the particles with sodium hypochlorite then yielded maghemite (γ-Fe2 O3 ) nanoparticles which were coated with two types of coating -d-mannose or poly(l-lysine) (PLL) as confirmed by FTIR analysis. The particles were <10 nm according to transmission electron microscopy. Their hydrodynamic particle size was ∼180 nm (by dynamic light scattering). The d-mannose-, PLL-coated, and neat γ-Fe2 O3 particles as well as commercial Resovist® were used to label rat macrophages. The viability and contrast properties of labeled macrophages were compared. PLL-coated γ-Fe2 O3 nanoparticles were found optimal. The labeled macrophages were injected to rats monitored in vivo by magnetic resonance imaging up to 48 h. Transport of macrophages labeled with PLL-γ-Fe2 O3 nanoparticles in rats was confirmed. Tracking of macrophages using the developed particles can be used for monitoring of inflammations and cell migration in cell therapy.
- MeSH
- buněčný tracking metody MeSH
- kontrastní látky * chemie farmakologie MeSH
- krysa rodu rattus MeSH
- magnetická rezonanční tomografie metody MeSH
- makrofágy radiografie MeSH
- nanočástice chemie MeSH
- polylysin * chemie farmakologie MeSH
- velikost částic MeSH
- železité sloučeniny * chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Dendrimers are well-defined macromolecules whose highly branched structure is reminiscent of many natural structures, such as trees, dendritic cells, neurons or the networks of kidneys and lungs. Nature has privileged such branched structures for increasing the efficiency of exchanges with the external medium; thus, the whole structure is of pivotal importance for these natural networks. On the contrary, it is generally believed that the properties of dendrimers are essentially related to their terminal groups, and that the internal structure plays the minor role of an 'innocent' scaffold. Here we show that such an assertion is misleading, using convergent information from biological data (human monocytes activation) and all-atom molecular dynamics simulations on seven families of dendrimers (13 compounds) that we have synthesized, possessing identical terminal groups, but different internal structures. This work demonstrates that the scaffold of nanodrugs strongly influences their properties, somewhat reminiscent of the backbone of proteins.
- MeSH
- aza sloučeniny chemie farmakologie MeSH
- biokompatibilní materiály chemie farmakologie MeSH
- bisfosfonáty chemie farmakologie MeSH
- dendrimery chemie farmakologie MeSH
- inhibitory kostní resorpce chemie farmakologie MeSH
- lidé MeSH
- molekulární struktura MeSH
- monocyty účinky léků MeSH
- nanočástice chemie MeSH
- polylysin chemie farmakologie MeSH
- polypropyleny chemie farmakologie MeSH
- průtoková cytometrie MeSH
- silany chemie farmakologie MeSH
- simulace molekulární dynamiky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Dendritic poly(L-lysines) (DGL) constitute promising nanomaterials applicable as a nonviral gene-delivery vector. In this study, we evaluate the transfection abilities of four DGL generations with special emphasis on the systematic description of the relationship of how generation (i.e., molecule size) affects the transfection efficacy. Using Hep2 cells, we demonstrated that the capability of unmodified DGL to deliver plasmid is of a magnitude lower than that of jetPEI. On the other hand, employing the Hep2 cell line stably transduced with eGFP, we observed that DGL G5 delivers the siRNA oligonucleotide with the same efficiency as Lipofectamine 2000. In further experiments, it was shown that DGL affords excellent ability to bind DNA, protect it against DNase I attack, and internalize it into cells.
- MeSH
- buněčné linie MeSH
- deoxyribonukleasa I metabolismus MeSH
- lidé MeSH
- lipidy MeSH
- malá interferující RNA MeSH
- molekulární sekvence - údaje MeSH
- oligonukleotidy metabolismus farmakokinetika MeSH
- plazmidy farmakokinetika MeSH
- polylysin chemie farmakokinetika MeSH
- sekvence nukleotidů MeSH
- transfekce metody MeSH
- viabilita buněk účinky léků MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zelené fluorescenční proteiny genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The present paper deals with the preparation and characterization of a conjugate of isoniazid (INH) with the block copolymer methoxypoly(ethylene glycol)-b-poly(L-lysine) (mPEG-b-PLL). The structure of the conjugate (mPEG-b-PLL-INH) was verified by means of (1)H NMR, GPC, infrared spectroscopy, elemental analysis and powder X-ray diffraction. The conjugate contains six l-lysine units with five INH molecules, which are attached by means of pH-sensitive amidine bond. Under in vitro conditions, the conjugate is hydrolyzed and isoniazid is released (pH 4; 37 °C; t(1/2) ≈ 10 h).
