Therapeutic efficacy of hispidulin-loaded carboxymethyl cellulose nanoparticles against mammary carcinogenesis through ROS-mediated apoptosis and tissue remodelling.

OBJECTIVE: To evaluate the apoptosis-modulating and therapeutic efficacy of hispidulin-loaded carboxymethyl cellulose nanoparticles (HIS-CMC NPs) in mammary carcinogenesis through in vitro and in vivo investigations. METHODS: Apoptotic activity of HIS-CMC NPs was assessed in MCF-7 cells using cell viability, intracellular reactive oxygen species (ROS), DAPI, acridine orange/ethidium bromide (AO/EB), and Annexin-V/PI flow cytometry assays. Therapeutic efficacy was further examined in a DMBA-induced mammary carcinogenesis BALB/c mice model through haematological, biochemical, histopathological, immunohistochemical and oxidative stress analyses. RESULTS: HIS-CMC NPs significantly reduced MCF-7 cell viability and exhibited a lower IC50 value (34.36 ± 1.66 μg/mL) than free hispidulin. Enhanced ROS generation, chromatin condensation, nuclear fragmentation, membrane destabilisation, and increased late apoptotic cell populations confirmed ROS-mediated apoptosis. In vivo, HIS-CMC NPs improved haematological and biochemical parameters, restored tissue architecture, downregulated Ki67 and Nrf2 expression, restored antioxidant enzyme activities, and reduced lipid peroxidation and lactate dehydrogenase levels. CONCLUSIONS: HIS-CMC NPs effectively promoted ROS-dependent apoptosis and attenuated oxidative stress in mammary carcinogenesis. The nanoformulation demonstrated enhanced therapeutic efficacy with reduced systemic toxicity, highlighting its potential as a promising nanotherapeutic strategy for breast cancer management. HIS-CMC NPs, mammary carcinogenesis, apoptosis, oxidative stress, proliferation biomarkers, immunohistochemistry.