Hyaluronan, a linear glycosaminoglycan comprising D-N-acetylglucosamine and D-glucuronic acid, is the main component of the extracellular matrix. Its influence on cell proliferation, migration, inflammation, signalling, and other functions, depends heavily on its molecular weight and chemical modification. Unsaturated HA oligosaccharides are available in defined length and purity. Their potential therapeutic utility can be further improved by chemical modification, e. g., reduction. No synthesis of such modified oligosaccharides, either stepwise or by hyaluronan cleavage, has been reported yet. Here we show a three-step synthesis (esterification, depolymerization and reduction) of unsaturated even numbered hyaluronan oligosaccharides with carboxylates and the reducing terminus reduced to an alcohol. Particular oligosaccharides were synthesised. The modified oligosaccharides are not cleaved by mammalian or bacterial hyaluronidase and do not affect the growth of mouse and human fibroblasts. Further, MTT and NRU viability tests showed that they inhibit the growth of human colon carcinoma cells HT-29 by 20-50 % in concentrations 500-1000 μg/mL. Interestingly, this effect takes place regardless of CD44 receptor expression and was not observed with unmodified HA oligosaccharides. These compounds could serve as enzymatically stable building blocks for biologically active substances.
- MeSH
- antigeny CD44 metabolismus MeSH
- buňky HT-29 MeSH
- cytostatické látky * farmakologie chemie chemická syntéza MeSH
- fibroblasty účinky léků MeSH
- hyaluronoglukosaminidasa * metabolismus antagonisté a inhibitory MeSH
- kyselina hyaluronová * chemie farmakologie MeSH
- lidé MeSH
- myši MeSH
- oligosacharidy * chemie farmakologie MeSH
- proliferace buněk * účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Nanofibrous materials are used in drug delivery as carriers of active ingredients. These can be incorporated into the materials with various electrospinning methods that differ mainly in the way spinning solutions are prepared. Each method affects primarily the encapsulation efficiency and distribution of active ingredients in the materials. This study focuses on the incorporation of octenidine dihydrochloride (OCT) and triclosan (TRI) into nanofibrous materials electrospun from native hyaluronic acid emulsions, dispersions, and blends. OCT had no substantial effect on fiber morphology, which is affected by the solvent system. All OCT encapsulation efficiencies were comparable (approximately 90%). TRI encapsulation efficiencies varied greatly depending on the method used. Merely 3% of TRI was encapsulated when it was spun from a dispersion. Encapsulation efficiency was higher, and TRI was incorporated in clusters when an emulsion was used. The best result was achieved with a blend, in which case 96% of TRI was encapsulated.
- MeSH
- antiinfekční látky lokální chemie MeSH
- emulze chemie MeSH
- kyselina hyaluronová chemie MeSH
- nanovlákna chemie MeSH
- Publikační typ
- časopisecké články MeSH
Nanofibrous materials produced from natural polymers have wide range of potential uses in regenerative medicine. This paper focuses on preparation of nanofibrous layers produced from intentionally hydrophobized derivatives of hyaluronan, which is known for its ability to promote wound healing. This structural modification of hyaluronan expands the range of potential uses of this promising material, which is otherwise limited due to the hydrophilic nature of hyaluronic acid. The aim of this research was preparation of nanofibrous material that would retain its fibrous structure and dimensional stability even after getting into contact with an aqueous medium, which is impossible to achieve with layers composed solely of native hyaluronan. As a result, such material would be able to retain its breathability and good mechanical properties when both dry and wet. Furthermore, all prepared materials were proved non-toxic for cells. This self-supporting nanofibrous matrix can be used as a scaffold, or porous wound dressing.
- Publikační typ
- časopisecké články MeSH
Nonwoven textiles are used extensively in the field of medicine, including wound healing, but these textiles are mostly from conventional nondegradable materials, e.g., cotton or synthetic polymers such as polypropylene. Therefore, we aimed to develop nonwoven textiles from hyaluronan (HA), a biocompatible, biodegradable and nontoxic polysaccharide naturally present in the human body. For this purpose, we used a process based on wet spinning HA into a nonstationary coagulation bath combined with the wet-laid textile technology. The obtained HA nonwoven textiles are soft, flexible and paper like. Their mechanical properties, handling and hydration depend on the microscale fibre structure, which is tuneable by selected process parameters. Cell viability testing on two relevant cell lines (3T3, HaCaT) demonstrated that the textiles are not cytotoxic, while the monocyte activation test ruled out pyrogenicity. Biocompatibility, biodegradability and their high capacity for moisture absorption make HA nonwoven textiles a promising material for applications in the field of wound healing, both for topical and internal use. The beneficial effect of HA in the process of wound healing is well known and the form of a nonwoven textile should enable convenient handling and application to various types of wounds.
- MeSH
- buňky 3T3 MeSH
- hojení ran * MeSH
- kyselina hyaluronová chemie MeSH
- lidé MeSH
- myši MeSH
- obvazy * MeSH
- testování materiálů * MeSH
- textilie * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hyaluronan (HA) is widely used for eye drops as lubricant to counteract dry eye disease. High and low molecular weight HA are currently used in ophthalmology. However, a large portion of the current literature on friction and lubrication addresses articular (joint) cartilage. Therefore, eye drops compositions based on HA and its derivatized forms are extensively characterized providing data on the tribological and mucoadhesive properties. The physiochemical properties are investigated in buffers used commonly in eye drops formulations. The tribological investigation reveals that amphiphilic HA-C12 decreases the friction coefficient. At the same time, the combination of trehalose/HA or HAC12 enhances up to eighty-fold the mucoadhesiveness. Thus, it is predicted a prolonged residence time on the surface of the eye. The incorporation of trehalose enhances the protection of human keratinocytes (HaCaT) cells, as demonstrated in an in-vitro cell-desiccation model. The presence of trehalose increases the friction coefficient. Medium molecular weight HA shows significantly lower friction coefficient than high molecular weight HA. This research represents a first, wide array of features of diverse HA forms for eye drops contributing to increase the knowledge of these preparations. The results here presented also provide valuable information for the design of highly performing HA-formulations addressing specific needs before preclinic.
- MeSH
- adhezivita MeSH
- buněčné linie keratinocytů HaCaT MeSH
- filtrace MeSH
- hlen účinky léků MeSH
- kyselina hyaluronová chemická syntéza chemie farmakologie MeSH
- lékové transportní systémy * MeSH
- lidé MeSH
- lubrikace * MeSH
- nefelometrie a turbidimetrie MeSH
- oči účinky léků MeSH
- oční roztoky farmakologie MeSH
- protonová magnetická rezonanční spektroskopie MeSH
- reologie MeSH
- sterilizace MeSH
- tření MeSH
- viskozita MeSH
- vysoušení MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Due to their large active surface, high loading efficiency, and tunable dissolution profiles, nanofibrous mats are often cited as promising drug carriers or antimicrobial membranes. Hyaluronic acid has outstanding biocompatibility, but it is hydrophilic. Nanofibrous structures made from hyaluronan dissolve immediately, making them unsuitable for controlled drug release and longer applications. We aimed to prepare a hyaluronan-based antimicrobial nanofibrous material, which would retain its integrity in aqueous environments. Self-supporting nanofibrous mats containing octenidine dihydrochloride or triclosan were produced by electrospinning from hydrophobized hyaluronan modified with a symmetric lauric acid anhydride. The nanofibrous mats required no cross-linking to be stable in PBS for 7 days. The encapsulation efficiency of antiseptics was nearly 100%. Minimal release of octenidine was observed, while up to 30% of triclosan was gradually released in 72 h. The nanofibrous materials exhibited antimicrobial activity, the fibroblast viability was directly dependent on the antiseptic content and its release.
- MeSH
- antibakteriální látky chemie farmakologie toxicita MeSH
- buňky 3T3 MeSH
- hydrofobní a hydrofilní interakce MeSH
- iminy chemie farmakologie toxicita MeSH
- kyselina hyaluronová chemie farmakologie toxicita MeSH
- léky s prodlouženým účinkem chemie farmakologie toxicita MeSH
- mikrobiální testy citlivosti MeSH
- myši MeSH
- nanovlákna chemie toxicita MeSH
- nosiče léků chemie farmakologie toxicita MeSH
- Pseudomonas aeruginosa účinky léků MeSH
- pyridiny chemie farmakologie toxicita MeSH
- Staphylococcus aureus účinky léků MeSH
- triclosan chemie farmakologie toxicita MeSH
- uvolňování léčiv MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Wound dressings with silver have been shown to be cytotoxic in vitro. However, the extrapolation of this cytotoxicity to clinical settings is unclear. We applied dressings with various forms of silver on porcine skin ex vivo and investigated silver penetration and DNA damage. We assessed antimicrobial efficacy, cytotoxicity to skin cells, and immune response induced by the dressings. All dressings elevated the DNA damage marker γ-H2AX and the expression of stress-related genes in explanted skin relative to control. This corresponded with the amount of silver in the skin. The dressings reduced viability, induced oxidative stress and DNA damage in skin cells, and induced the production of pro-inflammatory IL-6 by monocytes. The oxidative burst and viability of activated neutrophils decreased. The amount of silver released into the culture medium varied among the dressings and correlated with in vitro toxicity. However, antimicrobial efficiencies did not correlate strongly with the amount of silver released from the dressings. Antimicrobial efficiency and toxicity are driven by the form of silver and the construction of dressings and not only by the silver concentration. The damaging effects of silver dressings in ex vivo skin highlight the importance of thorough in vivo investigation of silver dressing toxicity.
- MeSH
- buněčné linie MeSH
- infekce v ráně MeSH
- kůže chemie cytologie účinky léků MeSH
- lidé MeSH
- obvazy škodlivé účinky MeSH
- poškození DNA * MeSH
- prasata MeSH
- stříbro toxicita MeSH
- techniky tkáňových kultur MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This work concerns the chemical modification of medium molecular weight hyaluronan for ophthalmic applications. The synthesis of amphiphilic HA with dodecanoyl moities was carried out under mild aqueous conditions. Perfect control of the degree of substitution was obtained by varying the molar ratio of activated fatty acid used in the reaction feed. Moreover, the preparation of the derivatives was optimized to achieve the desired degree of substitution (DS = 9.0 ± 0.2 %). The prepared hyaluronan derivatives were water-soluble and exhibited self-associating properties (amphiphilicity). The structure of the prepared derivatives was elucidated by NMR spectroscopy, rheology, turbidity, SEC-MALLS, and gas chromatography (GC). The hydrophobic moieties increase the solution viscosity by physical crosslinking. Low concentration of HAC12 is needed to prepare highly viscous solutions with potential use for ophthalmic applications. Amphiphilic HA kept the biocompatibility of hyaluronan. The degree of substitution and Mw of the amphiphilic HA controls the sterilization by filtration. The protection against desiccation was tested using human keratinocytes (HaCaT) cells lines.
- MeSH
- buněčné linie keratinocytů HaCaT MeSH
- buňky NIH 3T3 MeSH
- hydrofobní a hydrofilní interakce MeSH
- kyselina hyaluronová chemie MeSH
- kyseliny laurové chemie MeSH
- lidé MeSH
- molekulová hmotnost MeSH
- muciny chemie MeSH
- myši MeSH
- povrchové napětí účinky léků MeSH
- příprava léků metody MeSH
- reologie metody MeSH
- syndromy suchého oka farmakoterapie MeSH
- viabilita buněk účinky léků MeSH
- viskozita účinky léků MeSH
- zvířata MeSH
- zvlhčující oční kapky chemie farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In this work, a hybrid copolymer consisting of poly(3-hydroxybutyrate) grafted to hyaluronic acid (HA) was synthesised and characterised. Once formed, the P(3HB)-g-HA copolymer was soluble in water allowing a green electrospinning process. The diameters of nanofibres can be tailored by simply varying the Mw of polymer. The optimization of the process allowed to produce fibres of average diameter in the range of 100-150 nm and low polydispersity. The hydrophobic modification has not only increased the fibre diameter, but also the obtained layers were homogenous. At the nanoscale, the hybrid copolymer exhibited an unusual hairy topography. Moreover, the hardness and tensile properties of the hybrid were found to be superior compared to fibres made of unmodified HA. Particularly, this reinforcement was achieved at the longitudinal direction. Additionally, this work reports the use in the composition of a water-soluble copolymer containing photo cross-linkable moieties to produce insoluble materials post-electrospinning. The derivatives as well as their nanofibrous mats retain the biocompatibility of the natural polymers used for the fabrication.
- MeSH
- biokompatibilní materiály * chemická syntéza chemie MeSH
- biomechanika MeSH
- hydrofobní a hydrofilní interakce MeSH
- hydroxybutyráty chemická syntéza chemie MeSH
- kyselina hyaluronová chemie MeSH
- nanovlákna chemie MeSH
- polyestery chemická syntéza chemie MeSH
- polymery chemická syntéza chemie MeSH
- poskytování zdravotní péče MeSH
- tkáňové podpůrné struktury chemie MeSH
- vstřebatelné implantáty * MeSH
- zdravotnické prostředky MeSH
- Publikační typ
- časopisecké články MeSH
