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Bio-fabrication of silver nanoparticles by phycocyanin, characterization, in vitro anticancer activity against breast cancer cell line and in vivo cytotxicity.
El-Naggar NE
,
Hussein MH
,
El-Sawah AA
.
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In recent decades, researchers were attracted towards cyanobacterial components which are potential low-cost biological reagents for silver nanoparticle biosynthesis. This article describes the biological synthesis of silver nanoparticles using a proteinaceous pigment phycocyanin extracted from Nostoc linckia as reducing agent. The synthesized silver nanoparticles have a surface plasmon resonance band centered at 425 nm. Face-centered central composite design used for optimization of silver nanoparticles (AgNPs) biosynthesis using phycocyanin. The maximum AgNPs biosynthesis obtained using the optimized four variables, initial pH level (10), AgNO3 concentration (5 mM), phycocyanin pigment concentration (1 mg/mL) and incubation period (24 h) was 1100.025 µg/mL. The TEM analysis of AgNPs showed spherical nanoparticles with mean size between 9.39 to 25.89 nm. FTIR spectra showed major peaks of proteins involved in AgNPs biosynthesis by identifying different functional groups involved in effective capping of AgNPs. The biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic Gram-positive (Staphylococcus aureus), Gram-negative bacteria (Pseudomonas aeruginosa, E. coli and Klebsiella pneumonia). The synthesized AgNPs exhibited effective cytotoxic activity against MCF-7 and the inhibitory concentration (IC50) was recorded at 27.79 ± 2.3 µg/mL. The in vivo studies clearly indicated that AgNPs has a capacity to inhibit the growth of tumor in Ehrlich ascites carcinoma bearing mice.
Figure 1. UV–Vis absorption spectrum of phycocyanin pigment with λA max = 614 nm.
Figure 2. (A) Phycocyanin pigment (B) visible observation of AgNPs biosynthesis by phycocyanin pigment after exposure to AgNO3 solution (5 mM). (C) UV–Vis absorption spectra of silver nanoparticles synthesized by phycocyanin pigment.
Figure 3. (A) The normal probability plot of the residuals. (B) Correlation between the experimented and predicted values for silver nanoparticles biosynthesis using phycocyanin pigment determined by the second-order polynomial equation. (C) Box-Cox plot of model transformations.
Figure 4. Three-dimensional response surface plots (A–F) showing the interactive effects of independent variables: initial pH level, AgNO3 concentration, phycocyanin pigment concentration and incubation period on biosynthesis of silver nanoparticles.
Figure 5. EDX spectrum recorded showing peak approximately near 3 keV confirming the presence of silver.
Figure 6. X-Ray Diffraction for silver nanoparticles synthesized by using phycocyanin pigment.
Figure 7. Transmition electron microscopy images of produced silver nanoparticles using phycocyanin pigment. Size-controlled silver nanoparticles synthesized over the range 9.39 to 25.89 nm.
Figure 8. FTIR spectrum recorded by making KBr disc with synthesized silver nanoparticles by using phycocyanin pigment.
Figure 9. The zeta potential distribution graph showing negative zeta potential value for silver nanoparticles synthesized by using phycocyanin pigment.
Figure 10. Antibacterial activity of silver nanoparticles produced by using phycocyanin pigment against bacterial species.Inhibition zones were 9, 10, 10, 9 mm against Staphylococcus aureus, Pseudomonas aeruginosa, E. coli and Klebsiella pneumonia; respectively.
Figure 11.
In vitro anti-cancer activity of various concentrations of AgNPs on mammary gland breast cancer cell line (MCF-7), human lung fibroblast (WI38) and human amnion (WISH) the cell lines . 5-fluorouracil was used as a standard anticancer drug for comparison.
Figure 12. Schematic representation of the proposed mode of action of AgNPs on MCF-7 cells.
Figure 13. Microscopy images demonstrate the cytotoxic effect of AgNPs and 5-FU on EAC cells, cell damage by AgNPs due to loss of cell membrane integrity and apoptosis.
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