L-Arginine-Modified Chitosan Curcumin Nanocrystals Target M1 Macrophages via CAT-2/Clathrin-Mediated Endocytosis for
Xiaowen Yang1, Shiyue Wu1, Zhiya Dou1
1School of Chinese Materia Medica, Beijing University of Chinese Medicine, Yang Guang South Street, Fang-Shan District, Beijing 102488, China.
Abstract:
Background: Acute Lung Injury/Acute Respiratory Distress Syndrome (ALI/ARDS) is a fatal inflammatory disorder driven by M1 macrophages and the associated inflammatory cascade. Targeted drug delivery to these cells is a promising therapeutic strategy. Methods: L-arginine was conjugated to chitosan of different molecular weights. The resulting curcumin nanocrystals (Arg-CS-Cur) were characterized for conjugation efficiency, zeta potential, stability, and drug release profile. Cellular uptake mechanisms and mitochondrial targeting were investigated in lipopolysaccharide (LPS)-induced M1 macrophages using specific endocytic inhibitors and confocal microscopy. Results: Low-molecular-weight chitosan (MW 50 kDa) showed the highest L-Arg conjugation efficiency (22.31%). The optimized Arg-CS-Cur nanocrystals exhibited high zeta potential (±47.5 mV), excellent stability, and a superior drug release. They were internalized by M1 macrophages more efficiently than unmodified CS-Cur or free curcumin (p < 0.05). Uptake occurred via clathrin-mediated endocytosis (p < 0.001) and was mediated by CAT-2, which was highly expressed in M1 macrophages (p < 0.001). Arg-CS-Cur specifically targeted the mitochondria, reducing ROS and NLRP3 expression, thus inhibiting the NLRP3 inflammasome pathway (p < 0.001). Conclusions: This L-arginine-modified chitosan-based nanodelivery system synergistically exploits CAT-2 and clathrin pathways to deliver curcumin to M1 macrophage mitochondria, inhibiting the NLRP3 inflammasome. This dual-targeted strategy offers a promising approach for treating ALI/ARDS.
Insights
This study developed a novel nanodelivery system using L-arginine-modified chitosan to target M1 macrophages, effectively delivering curcumin to mitochondria and inhibiting inflammation for Acute Lung Injury/Acute Respiratory Distress Syndrome (ALI/ARDS) treatment.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Pharmacology
Background:
- Acute Lung Injury/Acute Respiratory Distress Syndrome (ALI/ARDS) is a severe inflammatory condition driven by M1 macrophages.
- Targeted drug delivery to M1 macrophages presents a potential therapeutic avenue for ALI/ARDS.
Purpose of the Study:
- To develop and characterize L-arginine-modified chitosan-curcumin nanocrystals (Arg-CS-Cur) for enhanced M1 macrophage targeting.
- To investigate the cellular uptake mechanisms and mitochondrial targeting capabilities of Arg-CS-Cur in M1 macrophages.
Main Methods:
- Conjugation of L-arginine to chitosan of varying molecular weights and characterization of resulting curcumin nanocrystals.
- Assessment of cellular uptake mechanisms (clathrin-mediated endocytosis, CAT-2) and mitochondrial targeting in lipopolysaccharide (LPS)-induced M1 macrophages.
- Evaluation of reactive oxygen species (ROS) and NLRP3 inflammasome inhibition.
Main Results:
- Low-molecular-weight chitosan (50 kDa) yielded optimal L-arginine conjugation efficiency.
- Arg-CS-Cur nanocrystals demonstrated superior cellular uptake via CAT-2 and clathrin-mediated endocytosis in M1 macrophages.
- Targeted delivery to mitochondria reduced ROS and NLRP3 expression, inhibiting the NLRP3 inflammasome pathway.
Conclusions:
- L-arginine-modified chitosan nanodelivery system effectively targets M1 macrophage mitochondria via synergistic exploitation of CAT-2 and clathrin pathways.
- This dual-targeted strategy shows promise for inhibiting the NLRP3 inflammasome and treating ALI/ARDS.
