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. 2022 May 17:2022:1646687.
doi: 10.1155/2022/1646687. eCollection 2022.

In Vitro Evaluation of Antibacterial, Antioxidant, and Antidiabetic Activities and Glucose Uptake through 2-NBDG by Hep-2 Liver Cancer Cells Treated with Green Synthesized Silver Nanoparticles

Shahnaz Majeed  1 , Mohammed Danish  2 , Norul Aini Zakariya  1 , Rokiah Hashim  2 , Mohammed Tahir Ansari  3 , Saad Alkahtani  4 , Md Saquib Hasnain  5
Affiliations

In Vitro Evaluation of Antibacterial, Antioxidant, and Antidiabetic Activities and Glucose Uptake through 2-NBDG by Hep-2 Liver Cancer Cells Treated with Green Synthesized Silver Nanoparticles

Shahnaz Majeed et al. Oxid Med Cell Longev. .

Abstract

The alarming rise in diabetes owing to drug resistance necessitates the implementation of prompt countermeasures in the treatment module of diabetes. Due to their unique physicochemical features, silver nanoparticles may have potential applications in the medical and pharmaceutical industries. Silver nanoparticles (AgNPs) were synthesized from the culture filtrate of Salmonella enterica (ATCC-14028). UV-Vis spectrophotometry, FTIR, SEM, and energy dispersive X-rays were used in the characterization of the nanoparticles. Transmission electron microscopy (TEM) revealed that AgNPs are spherical and highly scattered and vary in size from 7.18 nm to 13.24 nm. AgNP stability and protein loss were confirmed by thermogravimetric analysis (TGA) at different temperatures. The AgNPs had excellent antibacterial activity and a strong synergistic effect against methicillin-resistant bacteria Staphylococcus aureus (MRSA) ATCC-4330 and Streptococcus epidermis (MRSE) ATCC-51625. The DPPH experiment revealed that the AgNPs had high antioxidant activity. The antidiabetic assay revealed that these AgNPs had an IC50 for alpha-amylase of 428.60 μg/ml and an IC50 for alpha-glucosidase of 562.02 μg/ml. Flow cytometry analysis of Hep-2 cells treated with AgNPs (40 μg/ml) revealed higher expression of 2-NBDG glucose absorption (uptake) compared to control metformin. These AgNPs have promising antidiabetic properties and could be used in pharmaceuticals and biomedical industries.

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Conflict of interest statement

There is no potential conflict of interest among authors.

Figures

Figure 1
Figure 1
Biosynthesis of AgNP: colour change into dark brown confirms AgNP formation.
Figure 2
Figure 2
UV spectral analysis of biosynthesized AgNPs showed absorption peak at 410 nm while no peak revealed for the bacterial extract.
Figure 3
Figure 3
FTIR analysis of synthesized AgNPs showing that various functional groups are associated with AgNPs where (a) is FTIR of AgNPs and (b) is the FTIR for the cell-free extract.
Figure 4
Figure 4
SEM analysis of AgNPs while EDX confirms the presence of element Ag.
Figure 5
Figure 5
TEM analysis of synthesized AgNPs showing that particles are spherical in shape and well dispersed and their size ranges from 7.18 nm to 13.24 nm.
Figure 6
Figure 6
TGA of synthesized AgNPs showed the thermostability of AgNPs.
Figure 7
Figure 7
Antibacterial activity of AgNPs against MRSA and MRSE showed that AgNPs possess good antibacterial activity as well as synergistic effect against various antibiotics. Van: vancomycin; VanNP: Van+AgNPs; Caf: ceftriaxone; CafNP: ceftriaxone+AgNPs; Gen: gentamycin; GenNP: gentamycin+AgNPs.
Figure 8
Figure 8
Antioxidant activity of synthesized AgNPs showed excellent antioxidant property.
Figure 9
Figure 9
Alpha-amylase inhibition of AgNPs in a dose-dependent manner along positive control acarbose (a) while (b) showed IC50 of AgNPs and acarbose showing AgNPs able to inhibit the alpha-amylase efficiently. The readings are shown as triplicates.
Figure 10
Figure 10
Alpha-glucosidase inhibition of AgNPs in a dose-dependent manner along with positive control voglibose (a) while (b) showed IC50 of AgNPs and voglibose. The AgNPs were able to inhibit the alpha-glucosidase efficiently. The readings are shown as triplicates.
Figure 11
Figure 11
Graphical representation showing the anticancer effect of AgNPs against Hep-2 liver cancer cells in a dose-dependent manner. The readings are shown as triplicates.
Figure 12
Figure 12
Anticancer activity of AgNPs on Hep-2 cells in a dose-dependent manner: (a) metformin; (b) cells at 10 μg/ml; (c) cells at 20 μg/ml and 40 μg/ml; (d) cells at 40 μg/ml; (e) cells at 60 μg/ml; (f) cells at 80 μg/ml; (g) untreated cells.
Figure 13
Figure 13
Comparative glucose uptake of the Hep-2 cells treated with AgNPs and the expression of FSC-H signal through fluorescence microscopy showing that the treated cells with AgNPs increase the uptake of glucose significantly as compared with nontreated cells.
Figure 14
Figure 14
Glucose uptake study of AgNPs against HepG2 cells using BD FACSCalibur. 2-NBDG histogram of the gated HepG2 singlets distinguishes cells at the M1 and M2 phases (here M1 refers to the negative expression/region and M2 refers to the positive expression/region). Gating of M1 and M2 phases is approximate and can be refined using software (CellQuest Pro Software, version 6.0) analysis.
Figure 15
Figure 15
(a) Graphical representation of 2-NBDG expression. (b) Overlaid 2-NBDG expression of HepG2 cells in treated and untreated conditions.
See this image and copyright information in PMC

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