Comparative Evaluation of the Wound-Healing Activity of Ointments Obtained on the Basis of Some Biologically Active Complexes From Higher Basidiomycetes

Abstract

Introduction. In connection with the acquisition of antibiotic resistance of pyogenic microbiota, the problem of topical (drug) treatment of wounds has become aggravated. Aim. Comparative in vivo evaluation of the medicinal properties of ointments prepared on the basis of some biologically active complexes from higher basidiomycetes. Materials and methods. Experimental ointments were obtained on the basis of melanins from the natural raw materials of the сhaga Inonotus obliquus and from its deep-liquid culture, total polysaccharides from the oyster mushroom Pleurotus ostreatus, as well as the melanin polysaccharide complex from these mushrooms. Melanins were obtained by alkaline hydrolysis. Melanin-containing ointments were prepared using two formulations of ointment bases (the ointment base included saline, vaseline oil, emulsifiers T-2 and Tween-60, in the second case — saline, lanolin as an emulsifier and vaseline) in the first case. Total polysaccharides were obtained by 96% ethyl alcohol precipitation from an aqueous extract of a fungal homogenate heated in a boiling water bath for 12 h, followed by purification from fractions insoluble in distilled water. Melanin-polysaccharide-containing ointments were prepared on the basis of melanins from the natural raw materials of the сhaga Inonotus obliquus and from its deep culture, total polysaccharides from the oyster mushroom Pleurotus ostreatus, the ratios in the ointment formulations being (1:1 and 1:2). The ointment base of polysaccharide-containing and melanin-polysaccharide-containing ointments was similar to the specified second formulation. The wound healing efficacy of the ointments was evaluated in eight groups (20 mice each) of healthy non-inbred mice of both sexes, weighing 17–19 g, of the ICR colony of the State Scientific Center for Virology and Biotechnology Vector. The control group was a group of mice, in the treatment of which Levomicon-TFF reference ointment was used (20 mice in the group). The treatment of mice was continued until their incised skin wounds healed. The motor activity, appetite, and dynamics of wound healing were evaluated daily in mice of all groups, the change in their body weight was assessed every three days, and a histomorphological study of the incised skin wound was performed. The areas of wounds, the percentage of reduction in the area of wounds, the rate of wound contraction, and the rate of wound healing were determined. Results. It was shown that the components of the ointment bases of the two tested ointment formulations, melanin from the natural raw materials of the сhaga and from its deep-liquid culture, the total polysaccharides of the oyster mushroom, as well as the melanin polysaccharide complexes of these mushrooms, do not have toxicity for experimental animals in the concentrations of the tested ointment formulations. Ointments containing melanins from natural raw materials and deep-liquid culture of the сhaga Inonotus obliquus have similar regenerative properties, ointments with a fivefold increase in the content of these melanins, polysaccharide-containing and melanin-polysaccharide-containing ointments, have provided significantly accelerated wound healing in experimental animals. At the same time, the development of granulation tissue in the wound and the replacement of the wound defect in the skin with stratified squamous epithelium as early as on the 12th day in these groups of animals, were demonstrated in comparison with mice of the control group treated with the reference ointment Levomicon-TFF. The epidermal defect in animals of the control group was eliminated only on the 21st day. Conclusion. Ointments containing melanins from the natural raw materials of сhaga and from its deep-liquid culture, polysaccharides from the oyster mushroom, as well as a complex of these substances from these mushrooms, are superior in wound healing effectiveness, as compaired to the control ointment Levomicon-TFF, and can be considered as promising for topical treatment of wounds.