Efeitos do D-limoneno incorporado em membranas bioativas de colágeno sobre a cicatrização de feridas cutâneas

Abstract

Chronic wounds are responsible for significantly impacting the health and quality of life of patients worldwide. Treatment of these is considered expensive and difficult to access. Knowing the need for new proposals for its treatment, the objective was to evaluate the effect of D-limonene incorporated into bioactive collagen membranes on the healing of cutaneous wounds. Analysis of the effect of D-limonene incorporated into bioactive collagen membranes was performed by skin excision in female Wistar rats, 250-300 g, divided into three groups and each submitted to treatment with COL / HP-bioactive membrane control collagen containing hydroxypropyl-β-cyclodextrin (HP-β-CD); COL / HPLim - bioactive collagen membrane containing D-limonene complexed to hydroxypropyl-β-cyclodextrin; LP (Clean) - 0.9% saline solution. The wounds were evaluated in the period of 7 and 14 days. In order to evaluate the final area of the wound, they were measured and photographed on the day of preparation and on the day of removal of the lesion. For the evaluation of the reepithelialization index of the wound surface, the formation of stratified squamous epithelium in the region corresponding to the surface of the surgical wound was observed. For the morphological and histopathological evaluation of the healing tissue, slides were made by histochemical techniques (stained by the HE-Hematoxylin-Eosin method). The Western Blot technique was used to evaluate the immunocontent of VEGF (Vascular Endothelial Growth Factor) and NFB (Nuclear Transcription Factor kappa b). It was observed in the 7-day wound evaluation that the bioactive membrane containing D-limonene complexed to HP-β-CD (COL / HPLim) reduced the final area of the wounds. The rate of re-epithelialization in the COL / HPLim group was greater when compared to the other groups. The inflammatory response was considered severe in all groups and the leukocyte infiltrate was predominantly subacute in the LP and COL / HP groups and predominantly non-specific chronic in the COL / HPLim group. It was also observed that the COL / HPLim group increased the levels of VEGF concentration and decreased the concentration of NFB, favoring the cicatrization process, inducing the formation of new vessels and, consequently, tissue nutrition, besides proving its anti- inflammatory in this model of healing. In the evaluation of 14 days the wounds were practically epithelialized, evidenced by the histological aspect with presence of lymphocytes and rudiments of cutaneous appendages such as the presence of hair follicles in formation and glands and a substantial decrease in the increase of vascularization, possibly indicating a rapid overcoming of the inflammatory phase and anticipation of the remodeling phase. Thus, this work showed that D-limonene complexed with HP-β-CD incorporated in bioactive membrane of collagen promoted the acceleration in the healing process in this animal model, becoming a product with potential to be evaluated for the healing of cutaneous wounds.