Biocompatible madecassoside encapsulated alginate chitosan nanoparticles, their anti-proliferative activity on C6 glioma cells

Abstract

Madecassoside (MAD), an ursane type pentacyclic triterpenoid saponin, isolated from Centella asiatica (CA) possesses various pharmacological activities. The aim of this study was to evaluate therapeutic potential of MAD encapsulated alginate chitosan nanoparticles (MACNPs) on C6 glioma cells. Synthesized nanoparticles were characterized by Thermogravimetric analysis (TGA), Differential thermal analysis (DTA), Differential scanning calorimetry (DSC), Scanning electron microscopy (SEM), Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR). Cell viability and compatibility of both MAD and MACNP were observed on primary astrocytes. Cytotoxicity and anti-proliferative effect of MACNPs were studied on C6 glioma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5′-ethynyl-2′-deoxyuridine (Edu) assays. MACNPs showed 90% cytotoxicity, and depicted anti-proliferative activity against C6 cells. Annexin V-Alexa flour-488/propidium iodide (PI) staining followed by flow cytometry revealed that MACNPs induced necrosis in C6 glioma cells. Cell death was also observed using acridine orange/ethidium bromide (AO/EtBr) and Hoechst staining. 2′,7′-Dichlorodihydrofluorescein diacetate (DCFDA) assay by flow cytometry showed an increase in reactive oxygen species (ROS) production in MACNP treated C6 glioma cells. MACNPs showed a greater uptake and distribution in C6 glioma cells and inhibited the proliferation of C6 cells via increased intracellular ROS production. Our findings suggest MACNP as a promising drug delivery carrier for the management of glioma and shed new insights to brain malaises.

Keywords

• Madecassoside
• Nanoparticle
• C6 glioma cells
• ROS production
• Anti-proliferative activity