Marine L-arginase encapsulated in poly (lactic-co-glycolic acid) nanoparticles: A novel anti-cancer strategy for effective hepatocellular carcinoma treatment in vitro and in vivo

Hepatocellular carcinoma (HCC) is one of the most challenging malignant tumors globally, owing to its intricate nature and escalating morbidity. It is often diagnosed at advanced stages, limiting treatment options. This study explores the potential of arginine deprivation therapy against HCC in vitro and in vivo using L-arginase enzyme derived from Candida labiduridarum encapsulated in poly (lactic-co-glycolic acid) (PLGA) nanoparticles. We successfully purified L-arginase from C. labiduridarum, achieving a purification fold of 5.6 and specific activity of 97.6 U/mg protein. The enzyme exhibited optimal activity at a pH of 7.5 and 45 degrees C. Encapsulation of L-arginase in PLGA nanoparticles (ARGase-PLGA) was prepared using a w/o/w emulsification technique, resulting in a nanometric size of 332.5 +/- 3.5 nm with -63.5 +/- 0.5 mV Zeta potential and excellent encapsulation efficiency reached 72.7 %. In vitro study showed that both free L-arginase and ARGase-PLGA exhibited significant cytotoxic effects on HepG-2 cells, with superior potency for ARGase-PLGA. Furthermore, in vivo studies on HCC model in Balb/C mice revealed that ARGase-PLGA treatment led to a significant reduction in the arginine level, nitric oxide content, liver function markers, tumor maker (alpha-fetoprotein), and substantial improvement in the angiogenic, apoptotic, and autophagic markers. Histopathological examinations confirmed reduced liver damage and tumor progression by ARGase-PLGA. Notably, efficacy assessment of ARGase-PLGA on arginine depletion showed sustained depletion for up to 72 h in mice. Moreover, safety evaluation of ARGase-PLGA revealed no significant adverse effects. Overall, our findings suggest that ARGase-PLGA appears promising and safe therapeutic strategy for HCC, providing a targeted approach.