ABSTRACT:The use of a simple and reproducible model is inevitable for objective statement of the effects of external factors on wound healing. Hence, present study was conducted to establish an excisional model of skin wound healing in corticosteroid treated, and streptozotocine induced diabetic rats as well as to standardized the semi-quantitative and quantitative evaluation of selected parameters. Round full thickness skin wounds were performed on the back of male Sprague-Dawley rats. Animals were sacrificed two, six, and fourteen days after surgery. Sections were stained with hematoxylin-eosin and van Gieson. Both semi-quantitative (wound reepithelization; presence of: inflammatory cells, fibroblasts, new wessels, and collagen) and quantitative methods (polymorphonuclear leucocytes/tissue macrophages ratio, percentage of re-epithelization, area of the granulation tissue) were used to evaluate the histological changes during wound healing. As compared to the control group the wound healing process of both experimental groups was decelerated. Interestingly, wound reepithelization and angiogenesis were significantly inhibited only in the steroid rats while epithelization was accelerated in diabetic rats. In conclusion, when compared to primary sutured wound healing it can be concluded that the excisional model is more appropriate for histological assessment of the effect of various factors on wound healing. In addition, administration of corticosteroids represents simple and inexpensive model of a complex skin wound healing impairment.
Vidinsk˘ B., P. Gal, T. Toporcer, F. Longauer, L. Lenhardt, N. Bobrov, J. Sabo: Histological Study of the First Seven Days of Skin Wound Healing in Rats. Acta Vet. Brno 2006, 75: 197-202.The aim of this study was to elaborate a histological model of incisional skin wound healing in Sprague-Dawley rats. Under aseptic conditions two paravertebral full thickness skin incisions were performed on the back of 42 anesthetized male rats. Histological sections from tissue specimens were stained by hematoxylin and eosin, van Gieson, PAS + PSD, Mallory's phosphotungstic hematoxylin and azur and eosin and evaluated during the first seven days after surgery. Histological evaluation revealed that the regeneration of injured epidermis was completed five days after surgery. The inflammatory phase was recorded during the first three days of healing with the culmination of this phase between day one and day two. The beginning of the proliferative phase was dated to the first day and the peak during day five and day six. The initiation of the maturation and remodeling phase of the healing process was observed six days after wounding. At the layer of striated muscle, the centronucleated cells were described for the first time six days after surgery. The wound healing process of rat skin was histologically described during the first seven days. Results of this work can serve as an experimental model for further research using external pharmacological and physical factors (laser light, magnetic field) by which the wound healing can be favourably influenced.Sprague-Dawley rats, inflammatory phase, proliferative phase, maturation in remodeling phase, histology
The effect of Atropa belladonna L. (AB) aqueous extract on skin wound healing was studied in male Sprague-Dawley rats subjected to two parallel full-thickness skin incisions on the back. Specimens for histological evaluation were collected on days 2 and 5 whereas for biomechanical testing, they were collected on day 5. In the in vitro study, a different concentration of AB extract was used to test the differentiation of keratinocytes using a panel of selected antibodies, proliferation, and cell survival of 3T3 fibroblasts and human umbilical vein endothelial cells using the MTT-assay. Results of the in vivo experiments showed in AB-treated wounds a shortened process of inflammation and accelerated collagen formation, as well as significantly increased wound stiffness as compared with control tissues. The in vitro examination showed that control keratinocytes were cytokeratin 19 free, while samples exposed to the highest AB extract concentration expressed CK19. Moreover, all concentrations were stimulatory to human umbilical vein endothelial cell proliferation. In addition, only the AB extract at the lowest tested concentration increased fibroblast growth, but higher concentrations decreased cell survival. In conclusion, our results indicate that the AB water extract positively affects early phases of skin wound healing in rats. However, the in vitro results on the inverse relation between the concentration of the AB extract and its effects on cell proliferation may be important for future research.
The optimal parameters for low-level laser therapy (LLLT) for wound healing are still open to discussion. Hence, our study was aimed at comparing the effects of different power densities of LLLT at 670 nm in rats. Four round full-thickness skin wounds were placed on the backs of 16 rats which were divided into two groups (non-steroid and steroid-treated). Three wounds were stimulated daily with a diode laser (daily dose 5 J/cm(2)) at different power densities (5, 15 and 40 mW/cm(2), respectively), and the fourth wound served as a control. Six days after surgery all animals were killed and samples removed for histological evaluation. Significant acceleration of fibroblast proliferation and new vessel formation was observed in wounds treated at the selected power densities. No significant differences were found in corticosteroid-treated rats. In conclusion, LLLT with the methodology used improved wound healing in non-steroid rats, but was not effective after corticosteroid-treatment.
ABSTRACT:The complexity of the wound healing process, which is still poorly understood, prompted us to perform an immunohistochemical investigation using rat skin as an in vivo model. Fifteen Sprague-Dawley rats were included in the experiment. Two round full thickness wounds, 4 mm in diameter, were made on the backs of all rats. Haematoxylin and eosin basic staining as well as antibodies against wide spectrum keratin, keratin 10, keratin 14, α-smooth muscle actin, vimentin, fibronectin, collagens Type 1 and 3, and the transcription factor Sox-2 were applied to paraffin and frozen sections of skin wound specimens two, six and fourteen days after surgery, respectively. New hair follicles with Sox-2-positive cells were present after fourteen days; keratin/vimentin positivity was restricted to specimens of day two. Collagen-3 expression prevailed over collagen-1 expression at all evaluated time intervals, except in the uninjured part of the dermis. In conclusion, rat skin wound healing is a dynamic process which can serve as a model for studying phenomena such as cell-cell interactions and transitions in vivo.
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