Wound Healing Silver Nanoparticles-Composing Hydrogel

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Wound Healing Silver Nanoparticles-Composing Hydrogel ( wound-healing-silver-nanoparticles-composing-hydrogel )

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Nanomaterials 2020, 10, 390 9 of 16 alginate, silver nitrate, and hydrogel (1, 2, and 4 mM) are shown in Table 1. Gelatin and sodium alginate alone (without the incorporation of AgNPs) did not exhibit antimicrobial activity; the bacterial growth was therefore expected. These natural polymers contribute both to achieve the adequate consistency of the hydrogel through hydrogen bonds interaction, formed between the functional carboxylic and hydroxyl group. All tested concentration ratios of AgNPs incorporated in the hydrogel showed bactericidal activity. The Minimum Inhibitory Concentration (MIC) values recorded for the treatment with Hydrogels at 1.0 mM and 2.0 mM remained constant for gram-negative and gram-positive bacteria. The increased hydrogel concentration up to 4.0 mM induced the reduction of the MIC values in both strains. A significant bactericidal action was observed both against Pseudomonas aeruginosa with a minimum bacterial increase of 0.50 μg/mL and against Staphylococcus aureus with a minimum bacterial increase of 53 μg/mL (Table 1). This result corroborates the study of Rescignano et al. [44] in which all hydrogels incorporating AgNPs showed inhibition of bacterial growth, suggesting that the antimicrobial activity is associated with the direct contact of the AgNPs with bacteria. Table 1. MIC of the gelatin, sodium alginate, and silver nitrate solutions, hydrogels with silver nanoparticles at different concentrations (1 mM, 2 mM, and 4 mM). Bacteria Gram-negative Pseudomonas aeruginosa ATCC 27853 Gram-positive Staphylococcus aureus ATCC 25923 Gelatin Bacterial Growth Bacterial Growth Sodium Alginate Bacterial Growth Bacterial Growth AgNO3 No bacterial growth No bacterial growth Hydrogel 1 mM 1.050 μg.mL−1 130 μg.mL−1 Hydrogel 2 mM 1.050 μg.mL−1 130 μg.mL−1 Hydrogel 4 mM 00.50 μg.mL−1 53.0 μg.mL−1 As AgNO3 is toxic to microorganisms, it already exhibits bactericidal action. Therefore, the use of AgNO3 was also evaluated in this study, obtaining an expected result with the absence of bacterial growth when tested against Pseudomonas aeruginosa and Staphylococcus aureus. Thus, the antimicrobial activity related to the use of AgNPs occurs in Gram-positive and Gram-negative bacteria, which determines that this formulation has a bactericidal action of broad-spectrum, offering potential antimicrobial activity [45]. Our results show that AgNPs synthesized from AgNO3 require concentration around micrograms for bacterial growth inhibition. The results of the present study were similar to those reported by Kanmani et al. [46], which gelatin/AgNPs nanocomposite films (30 and 40 mg) showed potent antimicrobial activity against Gram-positive and Gram-negative foodborne pathogens. AgNPs are capable of interacting physically with the cell surface of several bacteria. It is particularly important in the case of Gram-negative bacteria, where numerous studies report the adhesion and accumulation of AgNPs on the bacterial surface. Many studies have reported that AgNPs may impair cell membranes leading to structural changes, which make bacteria more permeable [47,48]. Mekkawy et al. [49] developed AgNPs stabilized with polymer, for which the MIC values were in the range of 0.93–7.5 and 3.75–15 μg/mL, respectively, Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli). Similar studies were also done by Ashmore et al. [50] showing that AgNP inhibited the growth of E. coli only at 0.621 mg/mL; and by Rath et al. [51] who obtained MIC of AgNPs against S. aureus and P. aeruginosa 5.8 ± 0.3 mg/ml and 7.4 ± 0.2 mg/mL, 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 in the chart, the wound areas progressively decreased in both treated groups (GHP and GH) significantly. The

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