Targeted Delivery of Atractylodin via PLGA Nanoparticles Enhances Therapeutic Efficacy in Cholangiocarcinoma
Tullayakorn Plengsuriyakarn
Center of Excellence in Molecular Biology and Pharmacology of Malaria and Cholangiocarcinoma, Chulabhorn International College of Medicine, Thammasat University, Pathum Thani 12120, Thailand.
Luxsana Panrit
Faculty of Allied Health Science, Pathum Thani University, Pathum Thani 12000, Thailand.
Kesara Na-Bangchang *
Center of Excellence in Molecular Biology and Pharmacology of Malaria and Cholangiocarcinoma, Chulabhorn International College of Medicine, Thammasat University, Pathum Thani 12120, Thailand and The Office of Advanced Science and Technology, Thammasat University, Pathum Thani 12120, Thailand.
*Author to whom correspondence should be addressed.
Abstract
Cholangiocarcinoma (CCA) is a major health challenge in the Greater Mekong subregion, particularly in north-eastern Thailand, where chronic infection with the liver fluke Opisthorchis viverrini is a principal cause. Limited treatment options and the absence of reliable early diagnostic tools impede effective CCA management. This study investigated Atractylodes lancea (Thunb.) DC., a plant traditionally used in Thai and East Asian medicine. Atractylodin (ATD)-loaded poly(lactic-co-glycolic acid) nanoparticles (ATD-PLGA NPs) were developed to improve anticancer activity against CCA. The nanoparticles were prepared using a modified nanoprecipitation method and characterised for particle size, surface charge, morphology, encapsulation efficiency, loading capacity, and in vitro drug-release profile. Anticancer activity was evaluated in CL-6 and HuCCT-1 CCA cell lines using MTT cytotoxicity, real-time migration and invasion, and caspase-3/7-mediated apoptosis assays. A CCA xenograft model was established by implanting CL-6 cells into nude mice, and tumour-related gene expression was assessed using quantitative real-time PCR. The nanoparticles had a mean diameter of 229.8 nm and an encapsulation efficiency of 83%, with a biphasic release profile reaching 92% cumulative release over seven days. They selectively reduced CL-6 and HuCCT-1 cell viability, with selectivity indices of 3.53 and 2.61, inhibited migration and invasion by up to 90% within 12 hours, and induced apoptosis in 83% of cells. Micronucleus assays indicated lower mutagenic potential than the positive control. In vivo, ATD-PLGA NPs inhibited tumour growth in a dose-dependent manner and prolonged survival. Gene-expression analysis showed downregulation of pro-tumorigenic and pro-inflammatory factors and upregulation of IL-10. These findings support further evaluation of ATD-PLGA NPs as a targeted nanotherapeutic platform for CCA.
Keywords: Atractylodin, PLGA nanoparticles, cytotoxicity, cell invasion, apoptosis, genotoxicity, anti-cholangiocarcinoma activity