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. 2021 Sep 21;13(9):1527.
doi: 10.3390/pharmaceutics13091527.

Multifunctional Nanofibrous Dressing with Antimicrobial and Anti-Inflammatory Properties Prepared by Needle-Free Electrospinning

Laura Victoria Schulte-Werning  1 , Anjanah Murugaiah  1 , Bhupender Singh  2 , Mona Johannessen  2 , Rolf Einar Engstad  3 , Nataša Škalko-Basnet  1 , Ann Mari Holsæter  1
Affiliations

Multifunctional Nanofibrous Dressing with Antimicrobial and Anti-Inflammatory Properties Prepared by Needle-Free Electrospinning

Laura Victoria Schulte-Werning et al. Pharmaceutics. .

Abstract

An active wound dressing should address the main goals in wound treatment, which are improved wound healing and reduced infection rates. We developed novel multifunctional nanofibrous wound dressings with three active ingredients: chloramphenicol (CAM), beta-glucan (βG) and chitosan (CHI), of which βG and CHI are active nanofiber-forming biopolymers isolated from the cell walls of Saccharomyces cerevisiae and from shrimp shells, respectively. To evaluate the effect of each active ingredient on the nanofibers' morphological features and bioactivity, nanofibers with both βG and CHI, only βG, only CHI and only copolymers, polyethylene oxide (PEO) and hydroxypropylmethylcellulose (HPMC) were fabricated. All four nanofiber formulations were also prepared with 1% CAM. The needle-free NanospiderTM technique allowed for the successful production of defect-free nanofibers containing all three active ingredients. The CAM-containing nanofibers had a burst CAM-release and a high absorption capacity. Nanofibers with all active ingredients (βG, CHI and CAM) showed a concentration-dependent anti-inflammatory activity, while maintaining the antimicrobial activity of CAM. The promising anti-inflammatory properties, together with the high absorption capacity and antimicrobial effect, make these multifunctional nanofibers promising as dressings in local treatment of infected and exuding wounds, such as burn wounds.

Keywords: NanospiderTM; anti-inflammatory activity; antimicrobial activity; chloramphenicol; electrospinning; nanofiber.

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Conflict of interest statement

Rolf Einar Engstad is employed at Biotec BetaGlucans. The company Biotec BetaGlucans had role in the providing Soluble β-1.3/1.6-glucan (SBG®) to this current work. Rolf Einar Engstad has no economical or commercial interest to disclaim. The other authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Representative SEM images for each nanofiber formulation. Fiber diameter distribution was determined by measurement of 300 single-fiber diameters for each batch. The overall mean diameter can be found above the diameter distribution (n = 3). (A): βG-CHI-nf, (B): βG-CHI-CAM-nf, (C): βG-nf, (D): βG-CAM-nf, (E): CHI-nf, (F): CHI-CAM-nf, (G): Copol-nf, (H): Copol-CAM-nf. Abbreviations: βG (β-glucan), CHI (chitosan), Copol (co-polymers: polyethylene oxide and hydroxypropylmethylcellulose), CAM (chloramphenicol), nf (nanofiber).
Figure 2
Figure 2
Cumulative release (%) of chloramphenicol from nanofibers during a 6 h test-period in a Franz diffusion setup. Abbreviations: βG (β-glucan), CHI (chitosan), Copol (co-polymers: polyethylene oxide and hydroxypropylmethylcellulose), CAM (chloramphenicol), nf (nanofibers). Results are presented as mean ± SD (n = 3).
Figure 3
Figure 3
Relative cell viability (%) of (A) HaCaT cells and (B) macrophages (RAW 264.7) after 24 h incubation at 37 °C and exposure to nanofibers (dissolved in concentrations of 125, 250 and 1000 µg/mL) and chloramphenicol (CAM) (in concentrations of 1.25, 2.5 and 10 µg/mL). Results are presented as mean ± SD (n = 3). Abbreviations: βG (β-glucan), CHI (chitosan), Copol (co-polymers: polyethylene oxide and hydroxypropylmethylcellulose), CAM (chloramphenicol), nf (nanofiber).
Figure 4
Figure 4
Antibacterial activity of chloramphenicol-containing nanofibers (containing 30 µg of chloramphenicol per fiber) compared to a standard 30 µg chloramphenicol disc as positive control and no-CAM containing nanofibers as negative control (*). Results are expressed as the mean inhibition zone (mm) ± SD (n = 3). Abbreviations: βG (β-glucan), CHI (chitosan), Copol (co-polymers: polyethylene oxide and hydroxypropylmethylcellulose), CAM (chloramphenicol), nf (nanofiber).
Figure 5
Figure 5
NO production (%) of LPS-induced macrophages (RAW 264.7 cells) after 24 h exposure to nanofibers in three different concentrations (12.5, 25 and 100 µg/mL) compared to untreated cells. Abbreviations: βG (β-glucan), CHI (chitosan), Copol (co-polymers: polyethylene oxide and hydroxypropylmethylcellulose), CAM (chloramphenicol), nf (nanofiber). Results are presented as mean ± SD (n = 3). Formulations marked with * are statistically significant (p < 0.05) compared to untreated LPS-stimulated macrophages.
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