Wound Healing, Metabolite Profiling, and In Silico Studies of Aspergillus terreus

Abstract

Burn injuries, which significantly affect global public health, require effective treatment strategies tailored to varying severity. Fungi are considered a sustainable, easily propagated source for lead therapeutic discovery. In this study, we explored the burn wound healing potential of Aspergillus terreus through a combination of in vitro, in vivo, metabolite profiling, and in silico analysis. The in vitro scratch assays performed with human skin fibroblast cells showed promising wound healing activity. Furthermore, the burn-induced rats model showed a marked improvement in cutaneous wound healing, evidenced by an accelerated rate of wound closure and better skin regeneration after A. terreus extract treatment at 14 days. The results of this study demonstrated significant enhancements in wound closure and tissue regeneration in the treated rat model, surpassing the outcomes of standard treatments. This controlled healing process, evidenced by superior collagen synthesis and angiogenesis and confirmed by histopathological studies, suggests that A. terreus has potential beyond the traditionally studied fungal metabolites. The metabolite profiling of 27 bioactive compounds was further investigated by docking analysis for the potential inhibition of the NF-κB pathway, which has an important function in inflammation and wound repair. The compounds eurobenzophenone A (7), aspernolide D (16), asperphenalenone A (23), aspergilate D (15), kodaistatin A (18), and versicolactone A (14) showed the highest binding affinity to the target protein with a pose score of -16.86, -14.65, -12.65, -12.45, -12.19, and -12.08 kcal/mol, respectively. Drug-likeness properties were also conducted. The findings suggest the potential wound healing properties of A. terreus as a source for lead therapeutic candidate discovery.

Keywords: Aspergillus terreus; LC-ESI-MS/MS; molecular docking; wound healing.

Figure 1

Wound width between edges of scratches over time using A. terreus extract treatment.

Figure 1

Wound width between edges of scratches over time using A. terreus extract treatment.

Figure 2

Photographs of burn injuries of dorsal tissues of rat groups over the experiment period.

Figure 3

Burn wound’s contracture rate for the burn groups over time intervals. ^a Significantly different from Day 0 at p ≤ 0.05; ^b Significantly varied from 7th day at p ≤ 0.05; ^α Significantly varied from corresponding 7th day of control group at p ≤ 0.05; ^β Significantly different from corresponding 14th day of mupirocin group at p ≤ 0.05.

Figure 4

Photomicrograph of skin sections of (a) the burn group, showing burned skin histology and illustrated complete destruction of superficial skin layers and inflammatory changes with signs of coagulation in dermal layers, (b) the mupirocin-treated group, showing slight regeneration of the epidermis, crust formation with separation of dermal epidermal junction, and sever granulation tissue formation, and (c) the A. terreus extract-treated group, showing mild inflammation, regeneration of the epidermis, and less granulation tissue formation and with new vascularization (H&E × 400). (d) The burn group revealed complete destruction of superficial skin layers with fewer collagen fibers, (e) the mupirocin-treated group showed marked formation of irregularly arranged collagen fibers and granulation tissues, and (f) the A. terreus extract-treated group exhibited mild granulation tissue and regularly arranged collagen fiber formation (Masson trichrome × 400).

Figure 5

ESI-TIC chromatograms of the A. terreus EtOAc extract.

Figure 6

Two- and three-dimensional interactions complex of compound (14) with the NF-κB target. (a) Two-dimensional interactions of ligand with the active site and (b) three-dimensional interactions of ligand with the active site.

Figure 7

Two- and three-dimensional interactions complex of compound (15) with the NF-κB target. (a) Two-dimensional interactions of the ligand with the active site and (b) three-dimensional interactions of the ligand with the active site.

Figure 8

Two- and three-dimensional interactions complex of compound (16) with the NF-κB target. (a) Two-dimensional interactions of the ligand with the active site and (b) three-dimensional interactions of the ligand with the active site.