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Journal of Nanostructure in Chemistry Fig. 6 DPPH free radical scavenging assay of S-AgNPs and arbutin (values are mean ± SD of three determinations) 65.2%, respectively. The DPPH inhibition by S-AgNPs can be due to the fact that silver that acts as a good oxidant can easily lose electrons [29]. There is a possibility that the interaction of plant metabolites with metal ions during nanoparticle formation may result in free radical scavenging compounds. Toxicity analysis of S‐AgNPs Pharmacological research indicates that solar UV irradia- tion plays a key role in skin carcinogenesis and photoaging [30]. Chronic solar UV irradiation results in oxidative stress, inflammation, direct and ROS-mediated DNA damage, and dysregulation of cellular signaling pathways, all of which promote skin cancer. Recently, nanoparticles synthesized from the medicinal plant have been widely reported for their anti-inflammatory, anti-cancer, anti-oxidant and antimicro- bial effects [11, 31, 32]. There are very few studies showing the effect of nanoparticles on photoaging for e.g., Lee et al. [33] reported the effect of maghemite nanoparticles in pro- tection against ultraviolet-induced photo damage in human skin fibroblasts. Solid lipid nanoparticles of N-6-furfuryl adenine has been reported for the protective effect against photoaging [34]. Here, we hypothesized that S-AgNPs syn- thesized has photoprotective ability against UVB-induced photoaging. First, MTT analysis was performed to inves- tigate the cytotoxic effects of S-AgNPs on UVB-irradiated and non-UVB-irradiated HaCaT cells. Exposure of cells to UVB caused damage, but this was not statistically signifi- cant. S-AgNPs was non-cytotoxic to cells treated with 1, 10 and 100 μg mL−1 concentrations (Fig. 7). But in the case of UVB-irradiated cells, the cytotoxicity was observed at the concentration of 100 μg mL−1, the cell viability was reduced to 62%. The cell death was due to the UVB expo- sure. S-AgNPs were found to be non-cytotoxic to HaCaT. Fig.7 Effect of S-AgNPs on cell viability of HaCaT cells. HaCaT cells were either irradiated with 100 mJ cm−2 or nonirradiated, and then all cells were treated with the indicated concentration of the syn- thesized S-AgNPs (1, 10 and 100 μg mL−1) for 24 h. Data are pre- sented as the mean ± SD of three determination Inhibitory effect of S‐AgNPs on the production of MMP‐1 and IL‐6 UV stimulated the expression of pro-inflammatory cytokines and epidermal growth factors, which in turn resulted in an increase in the level of MMPs [35]. MMP is a major col- lagenolytic enzyme responsible for collagen damage in the UVB-irradiated human skin [27]. Reports suggested that collagen damage due to MMP is a major contributor to the photoaging of human skin [36–39]. Therefore, inhibition of MMPs is an important target of treatment in photoag- ing. We evaluated the effect of S-AgNPs on the secretion of MMP-1 in cultured HaCaT cells. As shown in Fig. 8a, b, UVB exposure led to an abnormal increase in MMP-1 secretion. When treated with S-AgNPs, the level of MMP-1 production started decreasing in a dose-dependent manner. HaCaT cells treated with 10 and 100 μg mL−1 of S-AgNPs had 47.8% and 53.3% levels of MMP-1 secretion, respec- tively. Our result is in line with the previous research which showed the inhibitory effect of maghemite nanoparticles on MMP-1 secretion [33]. Interleukin (IL)-6 is a pro-inflammatory cytokine, which results in overexpression of MMPs and eventually photo- aging [15]. We next tested the effect of S-AgNPs on the secretion of IL-6. Our data suggested that UVB irradiation also resulted in a severe increase of the pro-inflammatory cytokine IL-6. After, the treatment with S-AgNPs signifi- cantly calmed the activation of the IL-6 protein, which showed an inhibition rate of 17.8% and 54.4% at 10 μg mL−1 and 100 μg mL−1 (Fig. 8b). UV irradiation alters the physical properties of skin including loss of type 1 collagen in dermal ECM. Because type I collagen is the main structural protein of dermal ECM, 13PDF Image | green silver nanoparticles from leaf used as UVb protection
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