ABSTRACT
Analysis of factors that play a role on the healing process in exudates from skin wounds might shed light on the effect that grafted artificial tissue has in wound regeneration and repair. The first objective of this work was to standardize an optic surface plasmon resonance method based on self-assembled monolayers to quantify healing mediator factors (angiopoietin-2, epidermal growth factor, tumour necrosis factor-α, transforming growth factor-ß1, and vascular endothelial growth factor) in wound exudates. Optimal conditions for self-assembling of alkanethiol monolayers, immobilization of antibodies antifactors, and regeneration of sensor surfaces were established. A second objective was to compare healing of wounds grafted with artificial dermis with wounds left to heal by secondary intention (control) in a lagomorph model of full-thickness skin wound. Each animal included in this study had a control wound and an identical contralateral wound grafted with artificial dermis that was made by seeding autologous skin fibroblasts into unidirectional or multidirectional collagen type I scaffolds. Histological and histomorphometric analyses were carried out when animals were sacrificed, in addition to quantifying the factors in the exudates of wounds sampled 3 days after surgery. There were significant differences between the concentrations of evaluated factors in the exudates from grafted and control wounds. This finding coincides with differences observed in the histological and histomorphometric analyses of repaired tissue formed in treated and control wounds.
Subject(s)
Dermis , Fibroblasts , Skin, Artificial , Wound Healing , Animals , Autografts , Fibroblasts/metabolism , Fibroblasts/pathology , Fibroblasts/transplantation , Male , Rabbits , Wounds and Injuries/metabolism , Wounds and Injuries/pathology , Wounds and Injuries/therapyABSTRACT
Impaired wound healing is an important clinical problem in diabetes mellitus and results in failure to completely heal diabetic foot ulcers (DFUs), which may lead to lower extremity amputations. In the present study, collagen based dressings were prepared to be applied as support for the delivery of neurotensin (NT), a neuropeptide that acts as an inflammatory modulator in wound healing. The performance of NT alone and NT-loaded collagen matrices to treat wounds in streptozotocin (STZ) diabetic induced mice was evaluated. Results showed that the prepared dressings were not-cytotoxic up to 72h after contact with macrophages (Raw 264.7) and human keratinocyte (HaCaT) cell lines. Moreover, those cells were shown to adhere to the collagen matrices without noticeable change in their morphology. NT-loaded collagen dressings induced faster healing (17% wound area reduction) in the early phases of wound healing in diabetic wounded mice. In addition, they also significantly reduced inflammatory cytokine expression namely, TNF-α (p<0.01) and IL-1ß (p<0.01) and decreased the inflammatory infiltrate at day 3 post-wounding (inflammatory phase). After complete healing, metalloproteinase 9 (MMP-9) is reduced in diabetic skin (p<0.05) which significantly increased fibroblast migration and collagen (collagen type I, alpha 2 (COL1A2) and collagen type III, alpha 1 (COL3A1)) expression and deposition. These results suggest that collagen-based dressings can be an effective support for NT release into diabetic wound enhancing the healing process. Nevertheless, a more prominent scar is observed in diabetic wounds treated with collagen when compared to the treatment with NT alone.
