br Thermal gravimetric analysis TGA revealed that
Thermal gravimetric analysis (TGA) revealed that there is a higher weight loss of protein for the cAgNPs as compared to AgNPs. The
Fig. 7. Anticancer effect of AgNPs and cAgNPs against various MCF-7, HCT-116, MG-63 cancer Necrostatin-1 and normal fibroblast 3T3 cells where (C) control normal cells (A) Treated cells with AgNPs (B) Treated cells with cAgNPs. Treated cells showing membrane blabbing and morphological changes.
Fig. 9. Graphical representation of release of LDH (%) when treated with AgNPs and cAgNPs against various cell lines (A) MCF-7 cell line (B) HCT-116 cell line (C) MG-63 cell line.
Fig. 10. Apoptotic activity of cAgNPs against (A) MCF-7 cell line (B) HCT-116 cell line (C) MG-63 where (c) control living green cells (A) showing apoptotic bodies orange (D) dead cells using the IC50 con-centration showing chromatin condensation and membrane blebbing. (For interpreta-tion of the references to color in this figure legend, the reader is referred to the web version of this article.)
cAgNPs showed 10.9% weight loss as compared to AgNPs which showed the weight loss of 8.9% (Fig. 6). The TGA graph also showed the mass profile as compared to non cAgNPs this is because of bovine
serum. The cAgNPs contain higher mass profile due to the highly re-active and binding affinity of AgNPS to bind with amino and thiol group of bovine serum albumin as shown in Fig. 6 .
3.1. Cytotoxic study of AgNPs and cAgNPs
control cells as reported in the previous studies . Nosrati. H (2018) reported the enhanced anticancer activity of MCF-7 cell line when treated with Chrysin-BSA NP as compared with chrysin alone . Nosrati. H et al. (2018) also reported BSA is an excellent nanocarrier for the therapeutic drugs to the target site .
Alongside, these AgNPS and cAgNPs are tested against normal skin fibroblast 3 T3 cell line. To examine the effect on normal cells the concentration of the drugs were used from 60, 80, 100, 120, 140, until 160 μg/mL. The results showed that the IC50 value at 140 μg/mL and 120 μg/mL for cAgNPs and AgNPs, respectively (as shown in Fig. 7 and Fig. 8(D)). It is quite obvious that an increase in the nanoparticles concentration increases the cytotoxic effect towards cancer cells on the dose-dependent manner as reported by previous researchers [25,26]. Lin w et al. 2006 also reported the toxicity of silica nanoparticles to-wards the lung cancer cells by using different concentrations of nano-particles . The above results clearly shown that cAgNPs had a good anticancer effect but less toxicity towards normal cell line as shown in Fig. 7 and Fig. 8(D).
The release of LDH increases when the MCF-7, HCT-116, and MG-63 cells were treated with different concentrations of the AgNPs and the cAgNPs on doses manner (shown in Fig. 9). After the cells were exposed to drugs for 24 h of incubation, the results revealed that the cAgNPs was capable of more release of LDH compared to AgNPs. The cAgNPs en-hances the cytotoxicity as compared to AgNPs by the release of LDH enzymes when using the concentration ranges from 20, 40, 60,80 and 100 μg/mL for MCF -7, HCT-116, and MG-63 cell lines (shown in Fig. 9). The previous research report showed that there is a significant increase in the LDH when the rat lung epithelial Type-I cell line R3/1 showing toxic effect of silver nanoparticles .
In normal cells, glycolysis is the process for the conversion of glu-cose into pyruvate to provide the energy in the form of ATPs. But in the case of cancer cells, these glycolysis process increases tremendously because there is high demand for energy, so that cancer cells actively divide. it causes uncontrolled proliferation of the cells. In glycolysis, the pyruvate is converted into lactate in the presence of enzyme lactate dehydrogenase (LDH) that takes the H ions from NADH and converts it into NAD. This NAD makes the cell more prone for programmed cell death (PCD). The decrease of the cell viability was the major cause for the number of cell reduction. The cell reduction increased the amount of LDH enzyme that causes the cell membrane damage. Oberdorster G et al. 2005 reported viability of the A549 cell decreased when treated
with 15 and 46 nm of amorphous silica nanoparticles at the con-centration of 50 mg/mL for 48 h incubation. Hence, smaller size of the nanoparticles increases the toxicity towards the cells .
The cancer cells of MCF-7, HCT-116, and MG-63 were further stu-died for apoptotic activity by AO/EB dual staining method. These all were treated with IC50 value concentration and incubated for 24 h. The results showed significant decrease in the cell size, membrane blabbing, orange and red bodies, all these changes indicated that the DNA was condensed and observed under fluoresce microscope. But in control normal live green cells with full cell volume means cells are alive and viable confirms the DNA was intact inside the cell as shown Fig. 10 [29,30]. The percentage of apoptotic bodies produced by the cancer cells upon treatment with cAgNPs can be seen in Fig. 11. Hence the above results showed the number of viable cells decreased when the cells were treated with the cAgNPs.