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Nanomaterials 2020, 10, 422 10 of 16 Enzyme Glutathone-S transferase Catalase Superoxide dismutase Glucose 6 phosphate dehydrogenase Glutathione reductase Glutathione peroxidase Substrate Specific Activity (U/mg Protein) Table 2. Specific activities of antioxidant enzymes. Control AgNPs/CG 0.0602 0.75 0.0675 53.15 6.46 0 3.6. Morphological and Ultrastructural Alteration Caused by AgNPs/CG CDNB 0.422 ± 0.11 H2O2 2.3 ± 0.33 Epinephrine 0.405 ± 0.06 NADP 425.22 ± 5.11 NADPH 45.23 ± 2.13 NADPH 0.00055 ± 0.0001 We further examined the effect of AgNPs/CG on the ultrastructural alteration of C. albicans using TEM. Non-treated cells showed the typical filamentous morphology of C. albicans, which was very elongated but narrow in width (0.6–1.1 μm) (Figure 6A). A septum was formed between new filamentous growth and the parent cell, and the cell wall thickness was thinner than that of the yeast-phase cells. In contrast, cells treated with AgNPs/CG showed the typical C. albicans yeast form morphology, where the ovoid cell was surrounded by a cell wall. Within the cytoplasmic area, slightly irregular organelles (such as nucleus, mitochondria, endoplasmic reticulum, and nuclei) were found to be frequently ovoid, and a double-unit membrane enclosed them. Mitochondria were elongated (Figure 6B). Figure 6. Ultrastructure alteration in C. albicans by AgNPs/CG. TEM images of C. albicans cells treated with AgNPs. (A) Yeasts without treatment showing the typical filamentous morphology of C. albicans, which was very elongated but narrow in width (0.6–1.1 μm). (B) Yeast after treatment showing the typical C. albicans yeast form morphology, where the ovoid cell was surrounded by a cell wall after 24 h of incubation and treatment with AgNPs/CG (50 μg/mL AgNPs + 200 μg/mL plant extract). 3.7. Cytotoxicity of AgNPs/CG As a preliminary step to examine the therapeutic effect of AgNPs/CG in vivo, we examined the cytotoxicity of AgNPs/CG on the human cancer cell line MCF-7 and primary mouse BMSCs using a cell viability MTT assay. As demonstrated in Figure 7A,B, AgNPs showed a cytotoxicity at concentrations above 100 μg/mL, while the plant extract was not toxic to the cells up to 300 μg/mL. Interestingly, AgNPs/CG displayed cytotoxicity at concentration above 200 μg/mL plant extract + 50 μg/mL AgNPs and started to be toxic for both cell types at 500 μg/mL plant extract + 50 μg/mL AgNPs (Figure 7C).PDF Image | Inhibition of Candidiasis Calotropis Silver Nanoparticles
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