In vitro Evaluation of Apoptosis and Reactive Oxygen Species in Treated Cancerous and Normal Cells with Doxorubicin-Loaded Silk Fibroin Nanoparticles

Abstract

Abstract

Introduction: Breast cancer is the most common neoplastic malignancy in women in the world. The use of biodegradable and biocompatible drug nanocarriers can lead to the improvement of the most effective method of cancer treatment by increasing the drug solubility and efficacy. The aim of this study was to determine the anti-cancer effect of silk fibroin nanoparticles containing doxorubicin on cell lines, MCF7 and HFF and to determine the effective dose for cancer cell removal and to evaluate the cellular level of ROS and apoptosis in the studied cell lines.

Materials and Methods: In this study, silk fibroin nanoparticles containing doxorubicin were synthesized and characterized. Normal (HFF) and cancerous (MCF7) cells were treated with different concentrations of designed nanostructures at 24, 48 and 72 hour intervals. After the above times, MTT method was used to evaluate the vital activity of cells and V-FITC / PI Annexin kit and flow cytometry device were used to determine the rate of apoptosis. The data were analyzed by SPSS software using one-way ANOVA analysis.

Results: The inhibitory effect of fibroin-doxorubicin nanoparticles was observed on MCF7 growth by MTT test. The results of MTT assay showed that doxorubicin-coated silk fibroin nanoparticles have a dose-dependent anti-cancer effect on MCF7 cell line and low cytotoxicity on normal HFF cells. Doxorubicin-coated silk fibroin nanoparticles significantly reduced the level of reactive oxygen species (ROS) in the HFF and MCF-7 cell lines. The rate of apoptosis in MCF-7 increased significantly in the presence of doxorubicin-coated silk fibroin nanoparticles. These data suggest that the drug acts to induce cell death through the apoptotic pathway.

Conclusion: silk fibroin nanoparticles containing doxorubicin acts through the apoptotic pathway and can be considered as a candidate for new anticancer drugs.

Keywords: Breast Cancer, Silk Nanofibroin, Doxorubicin, Reactive Oxygen Species, Apoptosis