Resveratrol-loaded Silver Nanoparticles: A Novel Nanocarrier Approach for the Treatment of Hepatocellular Carcinoma: In-Vitro and In-Vivo

Md Abdur Rashid¹Rohini Kharwade^2*Purushottam Gangane²Sanket Meshram²Naushad Javed³Shirish Jain²Mohammed Kaleem^4,5*Madhav Chakolkar⁶Turky Omar Asar⁷Yahya Alhamhoom¹

• ¹King Abdulaziz University Department of Pharmacy Practice, Jeddah, Saudi Arabia
• ²Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur, India
• ³The University of Texas Rio Grande Valley, Brownsville, United States
• ⁴King Abdulaziz University, Jeddah, Saudi Arabia
• ⁵Savitribai Phule Pune University, Pune, India
• ⁶Rajarshi Shahu College of Pharmacy, Buldana, India
• ⁷University of Jeddah College of Science, Jeddah, Saudi Arabia

Abstract-Hepatocellular carcinoma (HCC) is the most common liver cancer and is associated with poor clinical outcomes. Its high mortality rate is due to the limited effectiveness of conventional chemotherapy, along with issues of drug resistance and systemic toxicity. To address these challenges, naturally derived bioactive compounds are increasingly being explored as safer and more effective treatments. In this context, the present study focused on developing resveratrol-loaded silver nanoparticles (R-AgNPs) using a chemical reduction method. The synthesized nanocarrier was thoroughly characterized for its physicochemical properties, solid-state behavior, and in vitro drug release profile to assess its potential as a treatment for HCC. The R-AgNPs produced were nanometric in size (130.56±2.36 nm) with a low PDI (0.136 ± 0.02) and high entrapment efficiency of 94.65 ± 1.38%. The cumulative resveratrol release reached 93.05±1.23% and showed sustained release over 32 hours at pH 6.8, following Higuchi drug release kinetics. Cell studies involving resveratrol-loaded AgNPs were conducted using the HepG2 human hepatocellular carcinoma cell line and compared to standard Camptothecin and pure resveratrol. These included cytotoxicity assays, percentage apoptosis, drug uptake via confocal microscopy, and cell cycle analysis through flow cytometry. The IC50 values for pure resveratrol and R-AgNPs were 46 µg/mL and 23 µg/mL, respectively. Confocal microscopy and flow cytometry results demonstrated that R-AgNPs significantly increased resveratrol uptake in HepG2 cells, with notable indications of apoptosis and arrest in the S phase of cell proliferation. Therefore, R-AgNPs confirmed the potential therapeutic efficacy of resveratrol nanoparticles for managing hepatocellular carcinoma.