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respectively. The wound healing capacity of the developed hydrogels was studied over 14 days, and the results are shown in Figure 6. The area of the wound has gradually decreased over time. Since no splinted wound model has been used to prevent contraction nor induce healing by re-epithelization, the healing shown in Figure 6 has been mainly by contraction mediated by myofibroblasts. As seen Nanomaterials 2020, 10, 390 10 of 16 in the chart, the wound areas progressively decreased in both treated groups (GHP and GH) significantly. The wound size was found to be reduced considerably in GHP, and GH-coated wounds on days 3, 7, and 14 of the postoperative period, compared to uncoated injuries (GCTR, p < 0.05, wound size was found to be reduced considerably in GHP, and GH-coated wounds on days 3, 7, and 14 respectively). The effective action of the hydrogel with the incorporation of AgNPs in the group of the postoperative period, compared to uncoated injuries (GCTR, p < 0.05, respectively). The effective corresponding to GHP has been observed, specifically on the third and seventh postoperative days. action of the hydrogel with the incorporation of AgNPs in the group corresponding to GHP has been On the third day, the area of the wound was reduced by 46.03% when compared to the GCTR groups, observed, specifically on the third and seventh postoperative days. On the third day, the area of the which decreased by 17.61% and GH with a reduction of the wound area by 30.63%. On the seventh wound was reduced by 46.03% when compared to the GCTR groups, which decreased by 17.61% and day, the GHP was also effective in reducing the total area of the wound by 81.14%, while the GH GH with a reduction of the wound area by 30.63%. On the seventh day, the GHP was also effective in decreased by 65.11% higher than the GCTR with 45.66% reduction of the injured area 45.66%. On the reducing the total area of the wound by 81.14%, while the GH decreased by 65.11% higher than the fourteenth day, the effective action of the GHP and GCTR hydrogels are possibly associated with the GCTR with 45.66% reduction of the injured area 45.66%. On the fourteenth day, the effective action formation of the granulation tissue, characteristic of the last cicatricial phase, and because it is no of the GHP and GCTR hydrogels are possibly associated with the formation of the granulation tissue, longer corresponding to the inflammatory period, which is more sensitive to the action of characteristic of the last cicatricial phase, and because it is no longer corresponding to the inflammatory microorganisms. It corroborates with the resolution of the use of AgNPs incorporated in the period, which is more sensitive to the action of microorganisms. It corroborates with the resolution of hydrogel, their antimicrobial activity made possible the reduction in the area of the wound mainly the use of AgNPs incorporated in the hydrogel, their antimicrobial activity made possible the reduction on the third and seventh day (Figure 6, left). in the area of the wound mainly on the third and seventh day (Figure 6, left). 100 80 60 40 20 0 GCTR GH GHP * 0 3 7 14 Days * * ** Figure 6. Non-splinted model showing the percentage of the non-epithelialized surface of the wound of the groups: GCTR (Control Group), GH (Group with hydrogel sodium alginate/gelatin (80:20), and GHP (Group hydrogel with AgNP 4 mM AgNO3). All values are mean ± S.E. Statistical analysis comprised ANOVA followed by Tukey’s test. * P < 0.05 in relation to GCTR, GH, and GHP groups, respectively (n = 21/group). The histological wound healing occurred without intercurrence over the time-course of the experiment in all the studied groups (Figure 7). On day 3, an intense inflammatory infiltrate composed by polymorphonuclear neutrophils (PMN), and macrophages were observed throughout the wounded area of all groups. Interstitial edema was remarkable, particularly in GCTR and GH. Interestingly, there was a more conspicuous chronic lymphocytic infiltrate in the bottom of GH wounds, whereas GHP presented the formation of immature granulation tissue, rich in hyperemic capillary vessels and proliferative endothelial-like spindle cells, in the bottom of the wounds. On day 7, granulation tissue containing plump, active fibroblasts forms was observed in all groups but at different maturation grades. The inflammation remained intense in GCTR, but there was a balance in the content of PMN, macrophages, and lymphocytes, which characterized a “persistent” subacute inflammatory infiltrate. Narrowed (slit-shaped) capillary vessels were concentrated on the edges and bottom of the wounds. In GH, the inflammatory response was essentially lymphocytic (chronic inflammation). The stromal spindle cells, interpreted as fibroblasts and endothelial cells, were densely dispersed throughout the wound area and arranged parallel to the wound surface. Most vessels were widely dilated and hyperemic. In GHP, the granulation tissue had more mature morphological features, such as reduction of the inflammatory response, higher content of hyperemic blood vessels, and more intense proliferation of fibroblast-like spindle cells. On day 14, there was the persistence of vascular granulation tissue in GCTR, whereas GH and GHP presented a cellular fibrous scar. All groups exhibited full epithelization, but only GH and GHP showed Wound area (%)PDF Image | Wound Healing Silver Nanoparticles-Composing Hydrogel
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