Analysing the apoptotic potential of green synthesized Nyctanthes arbor-tristis chitosan nanoparticles in MDA-MB-231 and SKOV3 cell lines

Abstract

Gynecological tumors are highly aggressive cancers in women, often treated with conventional treatments that can cause significant side effects. This study focuses on the preparation of chitosan nanoparticles from Nyctanthes arbor-tristis leaves, which possess anti-tumor properties, to address and overcome these issues. The successfully synthesized nanoparticles were characterized by UV-spectroscopy, DLS, TEM, and FTIR spectroscopy to analyze their physiochemical properties. In vitro studies, including cytotoxicity and scratch wound healing assays, along with staining and qRT-PCR, revealed the nanoparticles' anticancer efficacy against breast and ovarian cancer cells. The formation of Nat-CSNPs showed an absorbance peak at 221 nm, a particle size range of 41-56 nm with a spherical shape, polydispersity, and a positive surface charge. FTIR spectroscopy demonstrated the presence of functional groups associated with the synthesized Nat-CSNPs. It exhibited dose-dependent cytotoxicity, with IC₅₀ values of 62.40 μg/ml for MDA-MB-231 and 44.7 μg/ml for SKOV3 cells. Further assays such as wound healing assay, and DAPI/AO/EtBr staining demonstrated their antiproliferative and apoptotic effects on MDA-MB-231 and SKOV3 cells. Induction of apoptosis by the chitosan-nanoparticle via upregulation of the pro-apoptotic genes (Bax, Cas3, Cas9) and downregulation of antiapoptotic genes (Bcl2) was assessed using qRT-PCR analysis. In vivo acute toxicity assessments of Nat-CSNPs on Danio rerio revealed no significant impact on glucose levels or AST, ALT, and AChE activity, indicating low toxicity. These findings underscore the potent anticancer effects of Nat-CSNPs, particularly inducing apoptosis in MDA-MB-231 and SKOV3 cell lines. While demonstrating low toxicity in Danio rerio, Nat-CSNPs are considered a promising novel anti-cancer drug for breast and ovarian cancer treatment.

Keywords: Apoptosis; Breast cancer; Chitosan nanoparticle; Danio rerio; Nyctanthes arbor-tristis; Ovarian cancer.