RESUMEN
A fully automated sequential injection system was tested in terms of its application in liberation testing, and capabilities and limitations were discussed for clotrimazole liberation from three semisolid formulations. An evaluation based on kinetic profiles obtained in short and longer sampling intervals and steady-state flux values were applied as traditional methods. The obtained clotrimazole liberation profile was faster in the case of Delcore and slower for Clotrimazol AL and Canesten cream commercial formulations. The steady-state flux values for the tested formulations were 52 µg cm-2 h-1 for Canesten, 35 µg cm-2 h-1 for Clotrimazol AL, and 7.2 µg cm-2 h-1 for Delcore measured in 4 min sampling intervals. A simplified approach for the evaluation of the initial rate based on the gradient between the second and third sampling points was used for the first time and was found to correspond well with the results of the conventional methods. A comparison based on the ratio of the steady-state flux and the initial rate values for Canesten and Clotrimazol AL proved the similarity of the obtained results. The proposed alternative was successfully implemented for the comparison of short-term kinetic profiles. Consequently, a faster and simpler approach for dissolution/liberation testing can be used.
Asunto(s)
Antifúngicos/análisis , Automatización de Laboratorios/métodos , Clotrimazol/análisis , Análisis de Inyección de Flujo/métodos , Composición de Medicamentos , Liberación de Fármacos , Cinética , Crema para la PielRESUMEN
Due to its native origin, excellent biocompatibility and biodegradability, hyaluronan (HA) represents an attractive polymer for superparamagnetic iron oxide nanoparticles (SPION) coating. Herein, we report HA polymeric micelles encapsulating oleic acid coated SPIONs, having a hydrodynamic size of about 100 nm and SPION loading capacity of 1-2 wt %. The HA-SPION polymeric micelles were found to be selectively cytotoxic toward a number of human cancer cell lines, mainly those of colon adenocarcinoma (HT-29). The selective inhibition of cell growth was even observed when the SPION loaded HA polymeric micelles were incubated with a mixture of control and cancer cells. The selective in vitro inhibition could not be connected with an enhanced CD44 uptake or radical oxygen species formation and was rather connected with a different way of SPION intracellular release. While aggregated iron particles were visualized in control cells, nonaggregated solubilized iron oxide particles were detected in cancer cells. In vivo SPION accumulation in intramuscular tumor following an intravenous micelle administration was confirmed by magnetic resonance (MR) imaging and histological analysis. Having a suitable hydrodynamic size, high magnetic relaxivity, and being cancer specific and able to accumulate in vivo in tumors, SPION-loaded HA micelles represent a promising platform for theranostic applications.
Asunto(s)
Antineoplásicos/administración & dosificación , Compuestos Férricos/administración & dosificación , Ácido Hialurónico/administración & dosificación , Nanopartículas del Metal/administración & dosificación , Micelas , Polímeros/administración & dosificación , Animales , Antineoplásicos/química , Células CACO-2 , Portadores de Fármacos/administración & dosificación , Portadores de Fármacos/química , Compuestos Férricos/química , Células HCT116 , Humanos , Ácido Hialurónico/química , Células MCF-7 , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Nanopartículas del Metal/química , Ratones , Polímeros/química , Ratas Endogámicas BN , Ratas Endogámicas Lew , Células 3T3 Swiss , Resultado del Tratamiento , Ensayos Antitumor por Modelo de Xenoinjerto/métodosRESUMEN
In our previous research, we concluded that polymeric micelles based on hyaluronic acid are able to penetrate into the deeper layers of skin tissue. The aim of this work was to characterize the mechanisms involved in the uptake by skin cells, which is important for understanding the influence of the carrier composition on the drug penetration. To reach this goal, we used micelles encapsulating curcumin made of oleyl-hyaluronan (HAC18:1) and hexyl-hyaluronan (HAC6) covalently linked with fluorescent Nile Blue. This labeling enabled us to track the micelle-forming derivative and also micelle payload into the keratinocytes and fibroblasts by fluorescent microscopy and flow cytometry. The regulation of both the passive and active cellular uptake was used to determine the mechanism of micelle internalization. Furthermore, the changes of membrane fluidity were measured for these derivatives by FRAP. Using these methods we concluded that carriers entered the cells using both active and passive transport. Passive transport was facilitated by the affinity of the carrier to the cell membrane, especially in the case of HAC18:1 carrier, which changed significantly the membrane fluidity. The active transport was dependent on cell type, but mainly driven by the clathrin-mediated endocytosis and macropinocytosis. Surprisingly, the main HA receptor, CD44, was not involved in the uptake. We can conclude that these carrier systems could be used for the local transport of active substances or hydrophobic drugs into the skin cells using the advantage of passive transport of oleyl-HA derivative.
Asunto(s)
Sistemas de Liberación de Medicamentos , Ácido Hialurónico/administración & dosificación , Polímeros/administración & dosificación , Absorción Cutánea , Administración Tópica , Células Cultivadas , Curcumina/administración & dosificación , Endocitosis , Fibroblastos/metabolismo , Humanos , Queratinocitos/metabolismo , Lisosomas/metabolismo , Piel/metabolismoRESUMEN
A better understanding of in vivo behavior of nanocarriers is necessary for further improvement in their development. Here we present a novel approach, where both the matrix and the drug can be analyzed by LCMS/MS after one sample handling. The developed method was applied for the comparison of pharmacokinetic profile of free and encapsulated doxorubicin (DOX) in oleyl hyaluronan (HA-C18:1) polymeric micelles. The results indicated that nanocarriers were rapidly dissociated upon in vivo administration. Despite this fact, the administration of encapsulated DOX led to its longer circulation time and enhanced tumor targeting. This effect was not observed injecting blank HA-C18:1 micelles followed by unencapsulated DOX. Biodistribution studies and molecular weight estimation of the carrier matrix indicated relatively high stability of HA-C18:1 ester bond in bloodstream and complete elimination of the derivative within 72 h. The proposed methodology provides a novel strategy to elucidate the pharmacokinetic behavior of polysaccharide-based drug delivery systems.
Asunto(s)
Doxorrubicina/química , Portadores de Fármacos/química , Ácido Hialurónico/química , Micelas , Animales , Cromatografía Liquida , Doxorrubicina/farmacocinética , Liberación de Fármacos , Femenino , Ratones , Peso Molecular , Espectrometría de Masas en Tándem , Distribución TisularRESUMEN
Development of delivery systems which allow real-time visual inspection of tumors is critical for effective therapy. Near-infrared (NIR) fluorophores have a great potential for such an application. To overcome NIR dyes short blood circulation time and increase tumor accumulation, a NIR dye, cypate, was associated with oleyl hyaluronan, which can self-assemble into polymeric aggregates. The cypate association with oleyl hyaluronan was performed either by a covalent linkage, or physical entrapment. The two systems were compared for tumor targeting and contrast enhancement using BALB/c mice bearing 4T1 breast cancer tumors. Independently on the way of cypate association, it took more than 24â¯h from intravenous administration to detect NIR signal in tumors and the tumors were clearly visualized for 2 following weeks without substrate reinjection. Covalently linked cypate generated 2-3 fold stronger fluorescence signal than physically loaded cypate. This study demonstrates the potential of HA matrix to be used as carrier of contrast agents for non-invasive long-term tumor visualization.
RESUMEN
In this work, amphiphilic hyaluronic acid (HA) was synthesized by the chemical bonding of steroids. Particularly, succinyl cholesterol (SCH), cholic acid (CA), deoxycholic acid (DOCA), and 18ß-glycyrrhetinic acid (GA) were activated by benzoyl chloride towards the esterification reaction of HA in water. The degree of substitution can be controlled by varying the feed ratio of mixed anhydride to HA and up to 25% (mol/mol) can be obtained. Due to mild reaction conditions, no degradation of the polysaccharide was observed. The prepared amphiphilic polymers were characterized by NMR, infrared spectroscopy (FT-IR) and SEC/MALLS, as well as turbidity and size of the aggregates. The developed system is proposed for the delivery of hydrophobic drugs; for this purpose, curcumin, vitamin E and coenzyme Q10 were used as hydrophobic models; these molecules were loaded into the conjugates with high efficiency. The loading capacity was a function of degree of substitution. Furthermore, the biocompatibility of the derivatives and the cellular uptake of the delivery system enabled us to demonstrate the potential of the prepared delivery systems.
Asunto(s)
Antioxidantes/química , Sistemas de Liberación de Medicamentos , Diseño de Fármacos , Ácido Hialurónico/química , Esteroides/química , Animales , Antioxidantes/farmacología , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ácido Hialurónico/farmacología , Interacciones Hidrofóbicas e Hidrofílicas , Ratones , Conformación Molecular , Células 3T3 NIH , Esteroides/farmacología , Relación Estructura-ActividadRESUMEN
Acyl derivatives of hyaluronan (acyl-HA) are promising materials for biomedical applications. Depending on the acyl length and the degree of substitution, these derivatives range from self-assembling water-soluble polymers to materials insoluble in aqueous environments. The behaviour of acyl-HA was studied in solution, but little attention was paid to the solid state, despite its importance for applications such as medical device fabrication. We thus used X-ray scattering and electron microscopy to explore the solid-state nano-structure of acyl-HA. The set of samples included various substituents, substitution degrees and molecular weights. The obtained data showed that all studied acyl-HA materials contained structures with dimensions on the order of nanometres that were not present in unmodified HA. The size of the nanostructures increased with the acyl length, while the degree of substitution and molecular weight had negligible effects. We suggest that the observed nanostructure corresponds to a distribution of hydrophobic domains in a hydrophilic matrix of unmodified HA segments.
RESUMEN
A lignite humic acid (HA) was fractionated by preparative high performance size-exclusion chromatography (HPSEC) in seven different size-fractions. The size-fractions were characterized by cross polarization (CP) magic angle spinning (MAS) (13)C NMR spectroscopy and a further analytical HPSEC elution under UV and fluorescence detection. The alkyl hydrophobic components mainly distributed in the largest molecular-size-fraction, whereas the amount of oxidized carbons increased with decreasing size of fractions. Cross polarization time (T(CH)) and proton spin-lattice relaxation time in the rotating frame (T(1rho)(H)) were measured from variable contact time (VCT) experiments. The bulk HA was characterized by the shortest T(CH) values and the longest T(1rho)(H) values which suggested, respectively, one. an aggregation of components in a large conformation that favored a fast H-C cross polarization, and, two. consequent steric hindrances that prevented fast local molecular motions and decreased proton relaxation rates. Conversely, the separated size-fractions showed longer T(CH) values and shorter T(1rho)(H) values than the bulk HA, thereby indicating that they were constituted by a larger number of mobile molecular conformations. The UV and fluorescence absorptions were both low in the large size-fractions that mainly contained alkyl carbons, whereas they increased in the olephinic- and aromatic-rich fractions with intermediate molecular-size, and decreased again in the smaller fractions which were predominantly composed by oxidized carbons. These results support the supramolecular structure of humic substances and indicate that the observed variation in conformational distribution in humic association may be used to explain environmental processes with additional precision.
Asunto(s)
Sustancias Húmicas/análisis , Espectroscopía de Resonancia Magnética , Monitoreo del Ambiente , Contaminantes Ambientales , Lignina/química , Tamaño de la Partícula , Análisis EspectralRESUMEN
This work reports the synthesis and characterisation of new amphiphilic hyaluronan (HA) grafted with poly(3-hydroxyalkanoates) (PHAs) conjugates. Hydrolytic depolymerisation of PHAs was used for the synthesis of defined oligo(3-hydroxyalkanoates)-containing carboxylic terminal moieties. A kinetic study of the depolymerisation was followed to prepare oligomers of required molecular weight. PHAs were coupled with hydroxyl groups of HA mediated by N, N'-carbonyldiimidazole (CDI) or HSTU Tetramethyl-O-(N-succinimidyl) uronium hexafluorophosphate. For the first time, the covalent bonding of oligo derivatives of P(3-hydroxybutyrate), P(3-hydroxyoctanoate), P(3-hydroxyoctanoate-co-3-hydroxydecanoate) and P(3-hydroxyoctanoate-co-3-hydroxydecanoate-co-3-hydroxydodecanoate) and HA was achieved by "grafting to" strategy. Achieved grafting degree was a function of hydrophobicity of PHA, Mw and polarity of the solvent. The most suitable reaction conditions were observed for oligo (3-hydroxybutyrate) grafted to HA (grafting degree of 14%). Graft copolymers were characterized by FT-IR, NMR, DSC and SEC-MALLS. Graft copolymers can be physically loaded with hydrophobic drugs and may serve as drug delivery system.
RESUMEN
The physicochemical properties and biological functions of hyaluronan (HA) are closely related to its molecular weight (MW) and molecular weight distribution (MWD). Therefore, it is crucially important to provide a reliable characterization of these parameters for proper use of HA and its degradation products in both chemical and clinical fields. In this study, we present a novel method for the preparation of HA fragments of defined size with narrow molecular weight distribution. The HA fractionation was performed using an anion-exchange chromatography and is applicable either after enzymatic or chemical hydrolysis of polymeric HA. Isolated fractions with a molecular weight ranging from 3000-420,000gmol-1 were analyzed by size exclusion chromatography with multi-angle laser light scattering (SEC-MALLS). Hundred-milligram scale HA fragments were obtained from 5g hyaluronan starting material. Independently on weight-average molecular weight (Mw), the polydispersity index (PDI) of the HA fractions was less than 1.23. The fractionation methodology can be easily up-scaled and is applicable on any negatively charged polymers. We have also found that PDI is insufficient to characterize almost monodisperse fractions and for proper material characterization we proposed a new characteristic termed "distribution angle", calculated from the slope of the cumulative molecular weight distribution curve. Compared to PDI, the distribution angle reflects more efficiently changes in size distribution and thus is highly recommended to be used along with Mw determination of any polymer.
Asunto(s)
Ácido Hialurónico/química , Cromatografía en Gel , Hidrólisis , Peso Molecular , PolímerosRESUMEN
Nanosized materials offer promising strategy for topical drug delivery due to their enhancing effect on drug percutaneous transport across the stratum corneum barrier. In this work, polymeric micelles made from hydrophobized hyaluronic acid (HA) were probed for skin delivery. Compared to non-polymeric micelle solutions containing similar drug amount, in vitro skin penetration analysis indicated 3 times larger deposition of drug in the epidermis and 6 times larger drug deposition in the dermis after 5h of topical treatment in Franz diffusion cells. The drug deposition was further increased with prolonged time of topical treatment. Laser confocal microscopy revealed the accumulation of both, the HA forming the vehicle and the payload, in the epidermis and dermis. Although fluorescent labeling of the HA would suggest co-transport of the HA and the drug, loading FRET pair dyes in the micellar core clearly demonstrated gradual micelle disruption with increasing skin depth. Transcellular penetration was the predominant pathway for the loaded drug. The HA polymeric micelles also demonstrated increased bioactivity of loaded compound in vitro and in vivo. In addition, the loaded micelles were found to be stable in cream formulations and thus they have great potential for topical applications for cosmetic and pharmaceutical purposes.
Asunto(s)
Portadores de Fármacos/química , Ácido Hialurónico/química , Micelas , Absorción Cutánea , Adulto , Animales , Línea Celular , Liberación de Fármacos , Humanos , Técnicas In Vitro , Persona de Mediana Edad , Polímeros , Crema para la Piel , PorcinosRESUMEN
In this work, hyaluronan (HA) was grafted by a novel and an efficient mixed anhydrides methodology with (hetero)-aryl and aliphatic acrylic moieties suitable for cross-linking. A precise control of stoichiometry was achieved. Derivatives with degree of substitution (DS) below 20% did not show self-crosslinking. Due to mild reaction conditions, a negligible degradation of the polysaccharide was obtained. The influence of the feed components on the reaction efficiency and DS were studied up to 200 g/batch. The structure of the modified HA was characterized by Infrared Spectroscopy, Nuclear Magnetic Resonance, SEC-MALS and chromatographic analyses. Enzymatic degradation of derivatives was performed and isolated dimers demonstrated to be non-cytotoxic. The feasibility of the grafted HA for electrospinning with subsequent photo-crosslinking to avoid nanofibers water dissolution was demonstrated. The biocompatibility of the material, its degradation products, and the formation of honeycomb porous structures also proved the potential of the material for future in vivo applications.
RESUMEN
Polymeric micelles are attractive drug delivery systems for intravenously administered nonpolar drugs. Although physical parameters like size, shape and loading capacity are considered as the most important for their efficiency, here we demonstrate that the effects of serum protein interaction and characteristics of loaded compound cannot be neglected during the micelle development and design of experimental set up. Polymeric micelles prepared from amphiphilic hyaluronic acid grafted with short (hexanoic) and long fatty acids (oleic) were tested after loading with two different hydrophobic models, Nile red and curcumin. The composition of micelles affected mainly the loading capacity. Both encapsulated compounds behaved differently in the in vitro cell uptake, which was also influenced by serum concentration, where serum albumin was found to be the primary destabilizing component. This destabilization was found to be influenced by polymeric micelle concentration. Thus, the chemical structure of micelle, the properties of non-covalently loaded substance and serum albumin/polymeric micelle ratio modulate the in vitro intracellular uptake of drugs loaded in nanocarriers.
Asunto(s)
Portadores de Fármacos/metabolismo , Ácido Hialurónico/metabolismo , Líquido Intracelular/metabolismo , Micelas , Polímeros/metabolismo , Albúmina Sérica/metabolismo , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Portadores de Fármacos/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Células HCT116 , Células HT29 , Humanos , Ácido Hialurónico/administración & dosificación , Líquido Intracelular/efectos de los fármacos , Polímeros/administración & dosificación , Unión Proteica/fisiología , Albúmina Sérica/administración & dosificaciónRESUMEN
Novel hydrophobized hyaluronan (HA) derivatives, containing ω-phenylalkanoic acids (ω-PAA, 4-phenylbutyric acid, 6-phenylhexanoic, 8-phenyloctanoic or 11-tolylundecanoic acids) were prepared by esterification. Mixed anhydrides obtained after reaction of the carboxyl acid moiety and benzoyl chloride were found to be active acylating agents, affording hydrophobized HA in good yield and under mild conditions. The reactivity of the aromatic fatty acids towards esterification has decreased with the increasing length of the aliphatic spacer between the aromatic substituent and carboxylic acid moiety. The novel HA derivatives self-assembled from very low concentrations and were found to be non-cytotoxic. The potential use of ω-phenylalkanoic acids grafted-HA towards drug delivery applications was demonstrated by hydrophobic drugs (resveratrol and retinyl palmitate) encapsulation. The drug loading capacity of the novel HA derivatives was significantly improved most likely because of πâ¯π interactions between the micelle core and loaded hydrophobic aromatic compound.
Asunto(s)
Ácidos Grasos/química , Ácidos Grasos/síntesis química , Ácido Hialurónico/química , Interacciones Hidrofóbicas e Hidrofílicas , Micelas , Polímeros/química , Animales , Supervivencia Celular/efectos de los fármacos , Técnicas de Química Sintética , Ácidos Grasos/toxicidad , Ratones , Células 3T3 NIHRESUMEN
In this study, hyaluronan (HA) was grafted with alpha-linolenic acid (αLNA) by benzoyl mixed anhydrides methodology, which allowed the derivatization of HA under mild reaction conditions. The reaction was optimized and transferred from laboratory to semi-scale production. The derivative revealed an unexpected cytotoxicity after oven drying and storage at 40°C. For this reason, the storage conditions of sodium linolenyl hyaluronate (αLNA-HA) were optimized in order to preserve the beneficial effect of the derivative. Oven, spray dried and lyophilized samples were prepared and stored at -20°C, 4°C and 25°C up to 6 months. A comprehensive material characterization including stability study of the derivative, as well as evaluation of possible changes on chemical structure and presence of peroxidation products were studied by Nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR), gas chromatography-mass spectrometry (GC-MS), thermogravimetric analysis (TGA) and complemented with assessment of in vitro viability on mouse fibroblasts NIH-3T3. The most stable αLNA-HA derivative was obtained after spray drying and storage at ambient temperature under inert atmosphere. The choice of inert atmosphere is recommended to suppress oxidation of αLNA supporting the positive influence of the derivative on cell viability. The encapsulation of hydrophobic drugs of αLNA-HA were also demonstrated.
Asunto(s)
Portadores de Fármacos , Ácido Hialurónico , Ácido alfa-Linolénico , Animales , Supervivencia Celular/efectos de los fármacos , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacología , Estabilidad de Medicamentos , Ácido Hialurónico/química , Ácido Hialurónico/farmacocinética , Ácido Hialurónico/farmacología , Ratones , Células 3T3 NIH , Ácido alfa-Linolénico/química , Ácido alfa-Linolénico/farmacocinética , Ácido alfa-Linolénico/farmacologíaRESUMEN
The present work describes a novel and efficient method of synthesis of amphiphilic hyaluronan (HA) by esterification with alkyl fatty acids. These derivatives were synthesized under mild aqueous and well controlled conditions using mixed aliphatic aromatic anhydrides. These anhydrides characterized by the general formula RCOOCOC6H2Cl3 can be easily prepared by the reaction of the corresponding fatty acid (R) with 2,4,6-trichlorobenzoyl chloride (TCBC) in the presence of triethylamine. The aliphatic aromatic anhydrides RCOOCOC6H2Cl3 then react with the polysaccharide and enable the synthesis of aliphatic acid esters of HA in good yields. No hydrolytic degradation of hyaluronic acid could be observed. Parameters controlling the degree of esterification were systematically studied. Fatty acids with different chain lengths can be introduced applying this methodology. The degree of substitution was decreasing with increasing length of hydrophobic chain. The reaction products were fully characterized by Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), SEC-MALLS and chromatographic analyses. Although the esterified HA products exhibited aggregation in solution as demonstrated by NMR, microscopy and rheology, they were still water-soluble.
Asunto(s)
Anhídridos/química , Ácidos Grasos/química , Ácido Hialurónico/síntesis química , Tensoactivos/síntesis química , Esterificación , Ácido Hialurónico/química , Hidrocarburos Aromáticos/química , Espectroscopía Infrarroja por Transformada de Fourier , Tensoactivos/químicaRESUMEN
The positive effect of humic acids on the growth of plant roots is well known, however, the mechanisms and role of their physical structure in these processes have not been fully explained yet. In this work, South-Moravian lignite was oxidized by means of nitric acid and hydrogen peroxide to produce a set of regenerated humic acids. The elemental composition, solid state stability and solution characteristics were determined and correlated in vitro with their biological activity. A modified hydroponic method was applied to determine the effects of their potassium salts on Zea mays seedlings roots with respect to the plant weight, root length, root division, and starch and protein content. The relations between the determined parameters were evaluated through Principal Component Analysis and Pearson's correlation coefficients. The results indicated that the most important factor determining the biological activity of South-Moravian lignite potassium humates is related to the nature of self-assemblies, while the chemical composition had no direct connection with the root growth of Zea mays seedlings. It was demonstrated a controlled processing that provided humic substances with different chemical and physicochemical properties and variable biological activity.
RESUMEN
Physical and chemical structure of paclitaxel (PTX) was studied after its incorporation into polymeric micelles made of hyaluronic acid (HA) (Mw=15 kDa) grafted with C6 or C18:1 acyl chains. PTX was physically incorporated into the micellar core by solvent evaporation technique. Maximum loading capacity for HAC6 and HAC18:1 was determined to be 2 and 14 wt.%, respectively. The loading efficiency was higher for HAC18:1 and reached 70%. Independently of the derivative, loaded HA micelles had spherical size of approximately 60-80 nm and demonstrated slow and sustained release of PTX in vitro. PTX largely changed its form from crystalline to amorphous after its incorporation into the micelle's interior. This transformation increased PTX sensitivity towards stressing conditions, mainly to UV light exposure, during which the structure of amorphous PTX isomerized and formed C3C11 bond within its structure. In vitro cytotoxicity assay revealed that polymeric micelles loaded with PTX isomer had higher cytotoxic effect to normal human dermal fibroblasts (NHDF) and human colon carcinoma cells (HCT-116) than the same micelles loaded with non-isomerized PTX. Further observation indicated that PTX isomer influenced in different ways cell morphology and markers of cell cycle. Taken together, PTX isomer loaded in nanocarrier systems may have improved anticancer activity in vivo than pure PTX.
Asunto(s)
Antineoplásicos Fitogénicos/administración & dosificación , Portadores de Fármacos/administración & dosificación , Ácido Hialurónico/administración & dosificación , Micelas , Paclitaxel/administración & dosificación , Antineoplásicos Fitogénicos/química , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Portadores de Fármacos/química , Fibroblastos/efectos de los fármacos , Células HCT116 , Humanos , Ácido Hialurónico/química , Interacciones Hidrofóbicas e Hidrofílicas , Isomerismo , Paclitaxel/química , Polímeros/administración & dosificación , Polímeros/químicaRESUMEN
Silver has been used since time immemorial in different chemical form to treat burns, wounds and several different infections caused by pathogenic bacteria, advancement of biological process of nanoparticles synthesis is evolving into a key area of nanotechnology. The current study deals with the green synthesis, characterization, and evaluation of the biological activity and cell viability of hyaluronan fibers with incorporated silver nanoparticles (HA-Ag NPs). Hyaluronan fiber was prepared by the dissolving of sodium hyaluronate (HA) in aqueous alkaline solution to prepare a transparent solution, which was used for the preparation of fibers by a wet-spinning technique. Consequently, hyaluronan fiber was used as capping and stabilizing agent for the preparation of fibers with silver nanoparticles. HA-Ag NPs were confirmed by transmission electron microscopy, dynamic light scattering, UV/VIS spectroscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy, thermal analysis, nuclear magnetic resonance, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. HA-Ag NPs showed high antibacterial activity of against Staphylococcus aureus and Escherichia coli. Cell viability tests indicated that hyaluronan, hyaluronan fibers and hyaluronan fibers with silver nanoparticles were non-toxic on the cell growth. Two different particles size of Ag NPs (10, 40 nm) had not any toxicity till the concentration limit. These tests were performed using mouse fibroblast cell line 3T3.
Asunto(s)
Antibacterianos/química , Antibacterianos/farmacología , Ácido Hialurónico/química , Ácido Hialurónico/farmacología , Nanopartículas del Metal/química , Plata/química , Células 3T3 , Animales , Antibacterianos/toxicidad , Supervivencia Celular/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Ácido Hialurónico/toxicidad , Ratones , Tamaño de la Partícula , Solventes/química , Staphylococcus aureus/efectos de los fármacosRESUMEN
Native hyaluronan (HA) has been oxidized to polyaldehyde polymers with a degree of substitution (DS) of up to 50%. Two different procedures enabling the control of the degree of substitution were followed in this study. Selective oxidation of primary hydroxyl groups of N-acetyl-D-glucosamine of hyaluronan was performed either in an aqueous solution containing AcNH-TEMPO/NaBr/NaOCl or in an aprotic solvent containing Dess-Martin periodinane (DMP). It was found that a change of reaction parameters (reaction time and temperature, type of catalyst, oxidant-to-HA ratio, presence of nitrogen, buffer type, and concentration) had an influence on the degree of substitution and molecular weight. The derivatives were characterized by MS, NMR spectroscopy, and SEC-MALLS. Degradation of hyaluronic acid by the oxidant was observed and confirmed by SEC. The effect of oxidized derivatives of hyaluronan on cells was studied by means of NIH 3T3 fibroblast viability, which indicates that prepared hyaluronan polyaldehydes are biocompatible and suitable for medical applications and tissue engineering. The function of polyaldehyde as precursor for other modification was illustrated in the reaction with lysine.