In VitroUltrastructural Changes of MCF-7 for Metastasise Bone Cancer and Induction of Apoptosis via Mitochondrial Cytochrome C Released by CaCO₃/Dox Nanocrystals

Abstract

Bones are the most frequent site for breast cancer cells to settle and spread (metastasise); bone metastasis is considered to have a substantial impact on the quality of patients with common cancers. However, majority of breast cancers develop insensitivity to conventional chemotherapy which provides only palliation and can induce systemic side effects. In this study we evaluated the effect of free Dox and CaCO3/Dox nanocrystal on MCF-7 breast cancer using MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide), neural red, and lactate dehydrogenase colorimetric assays while DNA fragmentation and BrdU genotoxicity were also examined. Apoptogenic protein Bax, cytochrome C, and caspase-3 protein were analysed. Morphological changes of MCF-7 were determined using contrast light microscope and scanning and transmission electron microscope (SEM and TEM). The findings of the analysis revealed higher toxicity of CaCO3/Dox nanocrystal and effective cells killing compared to free Dox, morphological changes such as formation of apoptotic bodies, membrane blebbing, and absent of microvilli as indicated by the SEM analysis while TEM revealed the presence of chromatin condensation, chromosomal DNA fragmentation, cell shrinkage, and nuclear fragmentation. Results of TUNEL assay verified that most of the cells undergoes apoptosis by internucleosomal fragmentation of genomic DNA whereas the extent of apoptotic cells was calculated using the apoptotic index (AI). Therefore, the biobased calcium carbonate nanocrystals such as Dox carriers may serve as an alternative to conventional delivery system.

Doxorubicin-loaded quaternary ammonium palmitoyl glycol chitosan polymeric nanoformulation: uptake by cells and organs

“…55 Cytotoxicity experiments were conducted at 24, 48, and 72 h of incubation of cells with all concentrations of DOX and DOX-GCPQ. 56,57 Since the best cytotoxic results were obtained after 48 h, this time was selected to calculate IC 50 . The in vitro interaction of DOX-GCPQ nanoformulation with RD cell line showed significant cytotoxic effect as compared to free DOX and GCPQ polymer alone (P,0.05) due to more internal localization of DOX-GCPQ.…”

Ultrastructural Changes and Antitumor Effects of Doxorubicin/Thymoquinone-Loaded CaCO3 Nanoparticles on Breast Cancer Cell Line

“…The cell samples were mounted onto an aluminum stub, point dried, sputtered gold coated (E5100 Polaron, UK) and they were examined under SEM (JOEL-64000, Japan). Surface and morphological changes in the cells are then observed and recorded (19).…”

Bioprospecting the Curculigoside-Cinnamic Acid-Rich Fraction from Molineria latifolia Rhizome as a Potential Antioxidant Therapeutic Agent

“…The NRU assay was performed based on the modified protocol described by Kamba et al [54]. After 24 h of exposure to the test samples, the cells were incubated with 100 μg/mL neutral red dye dissolved in serum-free medium for 2 h at 37 °C.…”