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Nanomaterials 2020, 10, 390 8 of 16 The results indicate the safety and biocompatibility of the samples tested in this cell line. Any cytotoxic effect of these compounds may be due to their adhesion to the cell membrane, internalization and degradation of products in the cell culture medium or inside the cells. 8 of 16 Nanomaterials 2020, 10, x FOR PEER REVIEW (a) (b) (c) Figure 4. Electron micrographs of the hydrogels with silver nanoparticles at different concentrations: Nanomaterials 2020, 10, x FOR PEER REVIEW 9 of 16 Figure 4. Electron micrographs of the hydrogels with silver nanoparticles at different concentrations: 1 mM on the 200 nm scale (a) and 50 nm scale (b); 4 mM on the 200 nm (c) and 100 nm scale (d). 1 mM on the 200 nm scale (a) and 50 nm scale (b); 4 mM on the 200 nm (c) and 100 nm scale (d). cytotoxic effect of these compounds may be due to their adhesion to the cell membrane, Biological evaluation of nanoparticles-based formulations is a way of verifying the potential internalization and degradation of products in the cell culture medium or inside the cells. toxicity arising from wastes formed during the production process. To be used as a biomaterial, the 120 in vitro cytotoxicity test is the first toxic effects are assessed in norma production of silver nanopartic4l0es results are shown in Figure 5. Fro system must be biocompatible, i.e., it must interact with the physiological environment without undergoing changes or causing100tissue damage (ISO E. 10993-5, 2009). ISO 10993-5 suggests that the (d) A viability assay was carried * of cell proliferation, showing a st 0 80 alu h e m es all he om n f la mu nc am o ni d bility of a material and in this case, the 29 testing all components used for the ation of 150 μg/mL in DMSO 5%. The he silver nitrate induced the inhibition nce (p < 0.05) when compared to the 60 20 to ev l cells. out on at th m all t atistic ate t uma axi ted s y sig negative control (DMSO). None of the remaining samples showed statistically significant differences when compared to the negative control (p > 0.05). The negative control group was treated with DMSO 5% only and presented 100% cell viability. Sodium alginate, gelatin, and 4 mM silver nanoparticles treated cells resulted in 100%, 96.66%, and 96% cell viability, respectively. On the other hand, pure silver nitrate induced 47.33% of living cells, thus demonstrating its cytotoxic potential on fibroblasts. The results indicate the safety and biocompatibility of the samples tested in this cell line. Any Figure 5. Cell viability assay of gelatin, hydrogel containing 4 mM of AgNPs, sodium alginate, and Figure 5. Cell viability assay of gelatin, hydrogel containing 4 mM of AgNPs, sodium alginate, and silver nitrate of human L929 fibroblasts, determined by the methyl-thiazolyl-tetrazolium (MTT) assay Commented [1]: silver nitrate of human L929 fibroblasts, determined by the methyl-thiazolyl-tetrazolium (MTT) assay after 24 h of incubation. The vehicle used to dilute the drug (dimethyl sulfoxide, DMSO 5%) was used after 24 h of incubation. The vehicle used to dilute the drug (dimethyl sulfoxide, DMSO 5%) was used as the negative control (100% viability). The data correspond to the mean ± SEM of four independent as the negative control (100% viability). The data correspond to the mean ± SEM of four independent experiments. * p < 0.05 compared to the control group using one-way analysis of variance followed experiments. * p < 0.05 compared to the control group using one-way analysis of variance followed by Tukey’stest.The antimicrobial activity test was performed in vitro using Gram-positive (Staphylococcus by Tukey’s test. bioc ibrob m co ples, ficant aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria. MIC tests performed with gelatin, The antimicrobial activity test was performed in vitro using Gram-positive (Staphylococcus aureus) sodium alginate, silver nitrate, and hydrogel (1, 2, and 4 mM) are shown in Table 1. Gelatin and and Gram-nseogdaiutmivaelg(iPnasteuadloonme (owniathsouaterthuegiinncorspao)rabtiaocntoefrAiag.NPMs)IdCidtneosttesxhpibeirtfaonrtimiecrdobwialitahctivgietyl;atin, sodium 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 pati st L29 entr nly t iffere Viability (%) Control Gelatin Hydrogel 4 mM Sodium alginate Silver nitratePDF Image | Wound Healing Silver Nanoparticles-Composing Hydrogel
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