Hyaluronic Acid Based Nanofibrous Materials Stable in Aquaeous Environment for Incorporation of Active Substances

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ICE Krakow

ICE Krakow

ul. Marii Konopnickiej 17 30-302 Kraków


Bardoňová, Lenka ( Contipro a.s.; (2) Energy and Environmental Technology Centre, VŠB–Technical University of Ostrava)


Nanofibrous materials have the potential to be used in medicine, the most mentioned areas being topical applications, e.g., skin wound healing. Promising materials are biopolymers such as hyaluronic acid (HA), which is a part of the extracellular matrix and plays an important role in the inflammatory and granulating phase of wound healing. Nevertheless, HA is highly hydrophilic and nanofibrous materials of HA dissolve in water immediately, which excludes the applications requiring longer degradation periods. This problem can be overcome by the covalent crosslinking of HA or hydrophobization by modifying the polymer chain.

Nanofibrous materials based on a hydrophobized HA derivative – lauroyl hyaluronan (L-HA), a photo-cross-linkable derivative – furanyl hyaluronan (F-HA), or composites with different ratios of F-HA:L-HA were prepared by electrospinning. Materials containing F-HA were further crosslinked in a UV crosslinker for 60 minutes. The morphology of nanofibrous mats was studied by SEM in the dry state and after the immersion into phosphate buffer solution (PBS). The non-enzymatic and enzymatic degradation study was performed, respectively, in sodium acetate buffer (pH 5.3) containing bovine serum albumin (BSA) and in sodium acetate buffer with BSA and bovine testicular hyaluronidase. The effect of the nanofibrous mats' extracts on the viability of 3T3 fibroblasts and migration of HaCaT keratinocytes (scratch test) was assessed. NHDF cell adhesion on nanofibrous materials incubated in partially heparinized blood was also monitored. Active substances (octenidine dihydrochloride – OCT, triclosan - TRI) were incorporated into the composite materials. The release profiles and solubilization of the nanofibrous matrix were studied in PBS and PBS+BSA.

The diameter of the prepared fibers ranged from 130 nm (F-HA) to 650 nm (L-HA). The nanofibrous structure was preserved even after 72 hours in PBS for materials with the equal ratio of F-HA:L-HA (C1), or with a majority of F-HA (C2), thus maintaining the porosity required for gas exchange, fluid drainage, etc. The nanofibers consisting mainly of L-HA eventually fused into a film. The materials from F-HA exhibited the fastest degradation, and the degradation rate decreased with decreasing ratio of F-HA in the sample. None of the extracts from the nanofibrous mats was cytotoxic, neither affected the keratinocyte migration. The NHDF cells adhered and proliferated on samples C1 and C2. TRI was released into PBS gradually, but burst release occurred in PBS+BSA. On the other hand, OCT was not released into PBS and released gradually into PBS+BSA, simultaneously with the solubilization of the polymers.

Nanofibrous mats based on HA derivatives that are mechanically stable in aqueous environments were prepared by electrospinning. Materials gradually degraded in the media enriched with proteins and enzymes were not cytotoxic and did not influence cell migration. Cellular adhesion and proliferation were observed on mats preserving their fibrous structure in the wet state. Nanofibrous materials made of the combination of F-HA and L-HA were further modified by the incorporation of OCT or TRI. The release rate of the incorporated active substances depended on their physicochemical characteristics and dissolution medium.


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