Physiological Investigation of Sap-AgNPs’ Cytotoxic and Gene-Modulatory Effects in Oral Squamous Cell Carcinoma

BACKGROUND: One of the most prevalent oral cancers, oral squamous cell carcinoma (OSCC), is distinguished by its rapid growth, invasiveness, and high metastatic potential. Green AgNPs are important because they can reduce systemic toxicity by inducing oxidative stress, cytotoxicity, and apoptosis in cancer cells. The goal of this study was to use saponins as natural stabilizers to create AgNPs, and the detrimental apoptotic effects on cancer cells were examined using high-content screening (HCS) assays such as TNI, CMP, and VCC. METHODS: The size and distribution of AgNPs were determined using saponins as natural reducing and stabilizing agents, respectively. The cytotoxic effects on OSCC-25 cells were assessed using the MTT assay, alongside real-time quantitative PCR (RT-qPCR) to identify changes in gene expression associated with apoptosis. High-content screening (HCS) was used to confirm the induction of apoptosis and to measure concentration-dependent changes in several cellular parameters. All statistical analyses were performed for each experiment. RESULTS: The results showed that the average diameter of the generated nanoparticles was 75.87 ± 15.69 nm, facilitating cellular uptake due to their narrow size distribution. Saponin-induced AgNPs significantly increased cytotoxicity and cancer cell death in OSCC-25 cells in a dose-dependent manner. Compared with the control group, treatment with 125 and 500 μg/mL resulted in a significant decrease in fluorescence intensity (p < 0.05). However, doses of 250 μg/mL and 1000 μg/mL had no significant effects. RT-qPCR analysis revealed a significant increase in the expression of IL1R, highlighting its role in apoptotic signaling. CONCLUSION: The findings suggest that the combination of the bioactive properties of saponins with the inherent cytotoxicity of AgNPs has therapeutic potential against oral squamous cell carcinoma. These results support the need for future preclinical and clinical studies and highlight the promise of integrating natural compounds with nanotechnology to develop safer and more effective anticancer therapies.