All-trans-retinoic acid (atRA) is a potent ligand that regulates gene expression and is used to treat several skin disorders. Hyaluronic acid (HA) was previously conjugated with atRA (HA-atRA) to obtain a novel amphiphilic compound. HA-atRA forms micelles that incorporate hydrophobic molecules and facilitate their transport through the skin. The aim of this study was to determine the influence of HA-atRA on gene expression in skin cells and to compare it with that of unbound atRA. Gene expression was investigated using microarrays and a luciferase system with a canonical atRA promoter. HA-atRA upregulated gene expression similarly to atRA. However, HA-atRA activated the expression of cholesterol metabolism genes, unlike atRA. Further investigation using HPLC and filipin III staining suggested that the treated cells induced cholesterol synthesis to replenish the cholesterol removed from the cells by HA-atRA. HA modified with oleate (HA-C18:1) removed cholesterol from the cells similarly to HA-atRA, suggesting that the cholesterol removal stemmed from the amphiphilic nature of the two derivatives. HA-atRA induces retinoid signaling. Thus, HA-atRA could be used to treat skin diseases, such as acne and psoriasis, where the combined action of atRA signaling and anti-inflammatory cholesterol removal may be potentially beneficial.
Although silver is an efficient antimicrobial and is a widely used antiseptic in wound healing, previous studies have reported the cytotoxic in vitro effects of silver dressings. Moreover, few studies have addressed the distribution of silver in chronic wounds. The study compares the healing of chronic wounds treated with a standard-of-care silver dressing (Ag-CMC) and a dressing containing antiseptic octenidine (OCT-HA). Biopsies were taken from two wound areas before the commencement of treatment (baseline), after 2 weeks and after 6 weeks (the end of the study). We analyzed the histopathologic wound-healing score, silver distribution, and expression of selected genes. The wound-healing score improved significantly in the wounded area treated with OCT-HA after 2 weeks compared to the baseline and the Ag-CMC. The Ag-CMC wound areas improved after 6 weeks compared to the baseline. Moreover, collagen maturation and decreases in the granulocyte and macrophage counts were faster in the OCT-HA parts. Treatment with OCT-HA resulted in less wound slough. The silver, visualized via autometallography, penetrated approximately 2 mm into the wound tissue and associated around capillaries and ECM fibers, and was detected in phagocytes. The metallothionein gene expression was elevated in the Ag-CMC wound parts. This exploratory study determined the penetration of silver into human chronic wounds and changes in the distribution thereof during treatment. We observed that silver directly affects the cells in the wound and elevates the metallothionein gene expression. Octenidine and hyaluronan dressings provide a suitable alternative to silver and carboxymethyl cellulose dressings without supplying silver to the wound.
- MeSH
- antiinfekční látky farmakologie MeSH
- hojení ran účinky léků MeSH
- lidé středního věku MeSH
- lidé MeSH
- obvazy statistika a číselné údaje MeSH
- popálení farmakoterapie MeSH
- pyridiny farmakologie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- stříbro farmakologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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.
- MeSH
- aplikace lokální MeSH
- endocytóza MeSH
- fibroblasty metabolismus MeSH
- keratinocyty metabolismus MeSH
- kožní absorpce * MeSH
- kultivované buňky MeSH
- kurkumin aplikace a dávkování MeSH
- kůže metabolismus MeSH
- kyselina hyaluronová aplikace a dávkování MeSH
- lidé MeSH
- lyzozomy metabolismus MeSH
- polymery aplikace a dávkování MeSH
- systémy cílené aplikace léků * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Polysaccharides meet several criteria for a suitable biomaterial for tissue engineering, which include biocompatibility and ability to support the delivery and growth of cells. Nevertheless, most of these polysaccharides, for example dextran, alginate, and glycosaminoglycans, are highly soluble in aqueous solutions. Hyaluronic acid hydrophobized by palmitic acid and processed to the form of wet-spun fibers and the warp-knitted textile scaffold is water non-soluble, but biodegradable material, which could be used for the tissue engineering purpose. However, its surface quality does not allow cell attachment. To enhance the biocompatibility the surface of palmitoyl-hyaluronan was roughened by freeze drying and treated by different cell adhesive proteins (fibronectin, fibrinogen, laminin, methacrylated gelatin and collagen IV). Except for collagen IV, these proteins covered the fibers uniformly for an extended period of time and supported the adhesion and cultivation of dermal fibroblasts and mesenchymal stem cells. Interestingly, adipose stem cells cultivated on the fibronectin-modified scaffold secreted increasing amount of HGF, SDF-1, and VEGF, three key growth factors involved in cardiac regeneration. These results suggested that palmitoyl-hyaluronan scaffold may be a promising material for various applications in tissue regeneration, including cardiac tissue repair. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1488-1499, 2018.
- MeSH
- biokompatibilní materiály chemie MeSH
- buněčná adheze MeSH
- buněčné linie MeSH
- fibronektiny chemie MeSH
- kmenové buňky cytologie MeSH
- kultivované buňky MeSH
- kyselina hyaluronová chemie MeSH
- kyselina palmitová chemie MeSH
- lidé MeSH
- povrchové vlastnosti MeSH
- proliferace buněk MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this work, we report on the preparation of a novel biodegradable textile scaffold made of palmitoyl-hyaluronan (palHA). Monofilament fibres of palHA with a diameter of 120μm were prepared by wet spinning. The wet-spun fibres were subsequently processed into a warp-knitted textile. To find a compromise between swelling in water and degradability of the final textile scaffold, a series of palHA derivatives with different degrees of substitution of the palmitoyl chain was synthesized. Freeze-drying not only provided shape fixation, but also speeded up scaffold degradation in vitro. Fibronectin, fibrinogen, laminin and collagen IV were physically adsorbed on the textile surface to enhance cell adhesion on the material. The highest amount of adsorbed cell-adhesive proteins was achieved with fibronectin (89%), followed by fibrinogen (81%). Finally, textiles modified with fibronectin or fibrinogen both supported the adhesion and proliferation of normal human fibroblasts in vitro, proving to be a useful cellular scaffold for tissue engineering.
- MeSH
- biokompatibilní materiály chemie metabolismus farmakologie MeSH
- buněčná adheze účinky léků MeSH
- fibroblasty cytologie účinky léků MeSH
- hydrofobní a hydrofilní interakce * MeSH
- kyselina hyaluronová chemie metabolismus farmakologie MeSH
- lidé MeSH
- povrchové vlastnosti MeSH
- textilie * MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
