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Nanomaterials 2020, 10, 422 6 of 16 MTT to metabolize to formazan. The optical density was determined at 550 nm using an ELISA plate reader [37]. Cell viability was represented as a percentage of the control non-treated cells. 2.13. Statistical Analysis All values are represented as mean ± SD, of at least three independent experiments. A power calculation was performed for two-samples using an unpaired Student’s t-test (two-tailed) assuming equal variation in the two groups. Differences were considered statistically significant at * p < 0.05, and ** p < 0.005. 3. Results 3.1. Preparation and Characterization of AgNPs In the present study, aqueous leaf extract of C. gigantea, a traditional medicinal plant, was used as a reducing and stabilizing agent for the green synthesis of AgNPs. The ability of the bioactive components of C. gigantea to act as biocatalysts for the reduction of Ag+ to Ag0 was evaluated. The change of color of the biomass filtrate after adding AgNO3 as a precursor from colorless to yellowish brown was observed (Figure 1B). The UV-visible spectra of biosynthesized AgNPs showed an absorption band peaking at 450 nm (Figure 2A). The color of the solution supports the absorption wavelength in the visible region. The XRD pattern showed clear diffraction line at low angles (30◦–80◦). The Bragg reflections at angles 2θ of 38.18◦, 44.35◦, 64.4◦, and 77.3◦ corresponded to the 111, 200, 220, and 311 bands, respectively (Figure 2B). This pattern verified the structure of AgNPs as a face-centered cubic structure. XRD results confirmed the crystal structure of the silver in the C. gigantean extract. The FTIR spectrum of biosynthesized AgNPs displayed seven different peaks: 1116, 1457, 1629, 2361, 2853, 2923, and 3421 cm−1 (Figure 3A). The peaks at 3421cm−1 and 2923 cm−1 were due to the NH stretch vibration of the primary and secondary amides of protein. The peak at 2853 cm−1 was due to the C–H symmetrical stretch vibration of alkanes. The peak at 2361 cm−1 was due to the primary amine group of proteins. The peaks at 1457 cm−1 was due to the amino and amino-methyl stretching groups of proteins. The peak at 1116 cm−1 was due to the C–O stretching vibrations mode. Finally, TEM gave additional information about the morphology, size, and distribution profile of the AgNPs. The results obtained from the TEM demonstrated that the biosynthesized AgNPs were spherical in shape and the size was in the range of ≈10–70 nm (Figure 3B). In addition, AgNPs were distributed uniformly without significant agglomeration. 3.2. Anti-Candidal Activity of AgNPs We studied the anti-candidal activity of AgNPs, C. gigantea plant extract, and AgNPs/CG using a disk diffusion method assay. In this assay, the commercially available amphotericin b showed a higher inhibitory effect against C. albicans with an inhibition zone diameter of 19 mm. The AgNPs showed a moderate anti-candidal activity with inhibition zone diameter of 11.33 mm, while the plant extract displayed a lower anti-candidal activity with an inhibition zone diameter of 6.1 mm (Table 1 and Figure 4A). The MIC values of AgNPs and plant extract were 50 and 200 μg/mL, respectively (Table 1).PDF Image | Inhibition of Candidiasis Calotropis Silver Nanoparticles
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