In-vivo and in-vitro wound healing and tissue repair effect of Trametes versicolor polysaccharide extract

Abstract

Regarding different medical benefits of fungi, using the medical mushroom extracts as wound-healing agents is gaining popularity. This study, evaluated the wound healing characteristics of Trametes versicolor. Anti-oxidant activity addressed by employing the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay resulting 53.7% inhibitory effect. Besides, for anti-microbial ability determination, the MIC (Minimum Inhibitory Concentration) of extract measured which Escherichia coli growth was inhibited at 1.1 mg/ml, and Staphylococcus aureus did not grow at 4.38 mg/ml of extract. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method indicated dose dependence of the extract with 63 ± 3% and 28 ± 3% viability at 1250 μg/ml and 156.25 μg/ml of extract, which higher concentration caused higher cell viability. The outcome of gene expression analysis determined that overall expression of FGF2 (Fibroblast Growth Factor 2), IL-1β (Interleukin-1β), and TGF-β1 (Transforming Growth Factor-β1) was 4 times higher at 48 h than at 24 h in treated cells, suggesting a stimulating effect on cell growth. An in-vivo animal model suggested enhanced wound healing process after treatment with 0.01 g of extract. Furthermore, the number of fibroblasts, epidermal thickness, and collagen fiber was respectively 2, 3, and threefold higher in treated mice when compared to untreated mice. The treated wounds of mice showed 100% and 60% of untreated mice of healing within 14 days. The results of this research show promise for the fungus-based wound healing treatments, which may help with tissue regeneration and the healing of cutaneous wounds.

Keywords: Histopathology; Polysaccharide extract; Scratch assay; Trametes versicolor; Wound-healing.

Figure 1

MTT assay. By lowering the concentration of polysaccharide extract cell proliferation and cell growth will be decreased as well. This indicatedTrametes versicolor extract induces cell cycle and cell proliferation. IC₅₀:345.7, R squared: − 1.621, P-value < 0.05.

Figure 2

Scratch assay results. (A) Negative control of scratch assay. (B) Treated cells with extract after 24 h. Treated cells with 1250 µg/ml of polysaccharide extract express high migration and proliferation in comparison with negative control, and 100% closure after 24 h.

Figure 3

The comparison of expression of TGF-β1, IL-1β, and FGF2 at 24 and 48 h. The expression of TGF-β1 and FGF2 is higher at 24 h, which determined their significant role in early wound healing. At 48 h, IL-1β has expressed more than 24 h. TGF-β1 expression decreased after 48 h. Overall gene expression at 48 h was 4 times 24 h. P-value < 0.05.

Figure 4

Appearance changes of the wound of control and treated mice in 0 days and 14 days. (A) 0 days. (B) Negative control after 14 days. (C) Treated wound with 0.01 g of polysaccharide extract. Treated wounds showed signs of faster healing and better skin look after 14 days. This determined Trametes versicolor polysaccharide extract may have potential wound healing ability.

Figure 5

Mice skin cross-segments. (A) Untreated punched skin after 14 days. (B) Treated punched skin as negative control after 14 days. (C) Healthy control skin. Samples are captured at 10 × and 40 × magnifications.

Figure 6

Histopathology results. Fibroblasts (A), epidermal thickness (B), collagen fiber (C), and vessels (D) in treated samples observed 2-flold, threefold, threefold, and twofold more than positive control ones after 14 days, respectively. All 4 parameters have been higher in healthy skin. Neutrophil (F) was observed higher in positive control than in treated samples, indicating increased inflammation in untreated mice, and hair follicles (E) did not show significant differences in treated mice than positive control after 14 days.