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. 2021 Dec 22:9:tkab041.
doi: 10.1093/burnst/tkab041. eCollection 2021.

Antibacterial polysaccharide-based hydrogel dressing containing plant essential oil for burn wound healing

Huanhuan Wang  1 , Yang Liu  1 , Kun Cai  1 , Bin Zhang  1 , Shijie Tang  2 , Wancong Zhang  2 , Wenhua Liu  1
Affiliations

Antibacterial polysaccharide-based hydrogel dressing containing plant essential oil for burn wound healing

Huanhuan Wang et al. Burns Trauma. .

Abstract

Background: Polysaccharide-based hydrogels have been developed for many years to treat burn wounds. Essential oils extracted from aromatic plants generally exhibit superior biological activity, especially antibacterial properties. Studies have shown that antibacterial hydrogels mixed with essential oils have great potential for burn wound healing. This study aimed to develop an antibacterial polysaccharide-based hydrogel with essential oil for burn skin repair.

Methods: Eucalyptus essential oil (EEO), ginger essential oil (GEO) and cumin essential oil (CEO) were employed for the preparation of effective antibacterial hydrogels physically crosslinked by carboxymethyl chitosan (CMC) and carbomer 940 (CBM). Composite hydrogels were prepared and characterized using antimicrobial activity studies, Fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, gas chromatography-mass spectrometery, rheological analysis, viscosity, swelling, water loss rate and water vapor transmission rate studies. In addition, the biocompatibility of hydrogels was evaluated in vivo by cytotoxicity and cell migration assays and the burn healing ability of hydrogels was tested in vivo using burn-induced wounds in mice.

Results: The different essential oils exhibited different mixing abilities with the hydrogel matrix (CMC and CBM), which caused varying levels of reduction in essential oil hydrogel viscosity, swelling and water vapor transmission. Among the developed hydrogels, the CBM/CMC/EEO hydrogel exhibited optimal antibacterial activities of 46.26 ± 2.22% and 63.05 ± 0.99% against Staphylococcus aureus and Escherichia coli, respectively, along with cell viability (>92.37%) and migration activity. Furthermore, the CBM/CMC/EEO hydrogel accelerated wound healing in mouse burn models by promoting the recovery of dermis and epidermis as observed using a hematoxylin-eosin and Masson's trichrome staining assay. The findings from an enzyme-linked immunosorbent assay demonstrated that the CBM/CMC/EEO hydrogel could repair wounds through interleukin-6 and tumor necrosis factor-α downregulation and transforming growth factor-β, vascular endothelial growth factor (VEGF) and epidermal growth factor upregulation.

Conclusions: This study successfully prepared a porous CBM/CMC/EEO hydrogel with high antibacterial activity, favorable swelling, optimal rheological properties, superior water retention and water vapor transmission performance and a significant effect on skin repair in vitro and in vivo. The results indicate that the CBM/CMC/EEO hydrogel has the potential for use as a promising burn dressing material for skin burn repair.

Keywords: Antibacterial activity; Burn; Carboxymethyl chitosan; Eucalyptus essential oil; Hydrogel; Wound healing.

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Figures

Figure 1.
Figure 1.
Schematic diagram of antibacterial polysaccharide-based hydrogels preparation. CBM carbomer940, CEO cumin essential oil, EO essential oil
Figure 2.
Figure 2.
SEM images of the (a) CBM/CMC, (b) CBM/CMC/EEO, (c) CBM/CMC/GEO and (d) CBM/CMC/CEO hydrogels. CBM Carbomer 940, CMC Carboxymethyl chitosan, EEO Eucalyptus essential oil, GEO ginger essential oil, CEO cumin essential oil, SEM scanning electron microscope
Figure 3.
Figure 3.
The characteristics of morphosis parameter of CBM/CMC (black line), CBM/CMC/GEO (red line), CBM/CMC/CEO (blue line) and CBM/CMC/EEO (pink line) hydrogels. (a) FT-IR spectra of the four kinds of hydrogels. (b) XRD patterns of the four kinds of hydrogels. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, GEO ginger essential oil, CEO cumin essential oil, XRD X-ray diffraction
Figure 4.
Figure 4.
Storage (G′) and loss (G″) moduli of the CBM/CMC, CBM/CMC/GEO, CBM/CMC/CEO and CBM/CMC/EEO hydrogels as a function of angular frequency. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, GEO ginger essential oil, CEO cumin essential oil
Figure 5.
Figure 5.
Various physical parameters of CBM/CMC (black line), CBM/CMC/GEO (red line), CBM/CMC/CEO (blue line) and CBM/CMC/EEO (pink line) hydrogels. (a) Viscosity of the four kinds of hydrogels. (b) Swelling rate of the four kinds of hydrogels. (c) Water loss rate of the four kinds of hydrogels. (d) Water vapor transmission rate (WVTR) rate of the four kinds of hydrogels. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, GEO ginger essential oil, CEO cumin essential oil
Figure 6.
Figure 6.
Cell viability of the L929 cells treated with different concentrations of the hydrogel extracts after 24 h. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, GEO ginger essential oil, CEO cumin essential oil
Figure 7.
Figure 7.
In vitro cell experiments of the various hydrogels. (a) The percentage of the covered area after treatment with different hydrogel extracts. (b) The images of the L929 cell migration after treatment. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, GEO ginger essential oil, CEO cumin essential oil
Figure 8.
Figure 8.
In vivo animal experiments with positive drug, CBM/CMC/EEO hydrogel and saline on days 3, 7, 14 and 21 respectively. (a) Representative visual wound appearance (the scale presents the dimensions in mm). (b) Wound healing rate comparison for treatment. GEL indicated CBM/CMC/EEO hydrogel. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, PC positive control, NC negative control
Figure 9.
Figure 9.
Histology of the wound healing site in the mice with positive drug, CBM/CMC/EEO hydrogel and saline on days 3, 7, 14 and 21 respectively (a) The H&E-stained images of the mice wound tissue for treatment. (b) Masson-stained images of the mice wound tissue for treatment. GEL indicated CBM/CMC/EEO hydrogel. CBM carbomer 940, CMC carboxymethyl chitosan, EEO eucalyptus essential oil, PC positive control, NC negative control
Figure 10.
Figure 10.
Quantitative analysis of various cytokine content in the wound tissue by employing the ELISA method. The values have been expressed as mean ± SD. One-way ANOVA was used to evaluate the differences. *p < 0.05 and **p < 0.01 vs NC (a) Quantitative determination of IL-6 content. (b) Quantitative determination of TNF-α content. (c) Quantitative determination of TGF-β1 content. (d) Quantitative determination of VEGF content. (e) Quantitative determination of EGF content. GEL indicated CBM/CMC/EEO hydrogel. PC positive control, NC negative control, VEGF vascular epidermal growth factor, EGF epidermal growth factor
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