Abstract
Purpose: Lately, bismuth-based nanomaterials have been widely utilized in medical researches such as imaging, drug delivery and radio-sensitization. Despite their advantages, bismuth-based compounds have shown toxic effects in humans. There are few studies on cytotoxicity effects of bismuth oxide (Bi2O3) nanoparticles (NPs) in-vitro. In this study, we aimed to investigate cytotoxicity of bare and also folate and 5-aminolevulinic acid (5-ALA)-conjugated Bi2O3 NPs on nasopharyngeal carcinoma (KB) and lung cancer (A549) cell lines.Methods: Bi2O3 NPs were synthesized and conjugated with folate and 5-ALA. KB and A549 cells were cultured and incubated with 10, 20, 50 and 100 μg/ml concentrations of bare and folate-5-ALA-conjugated NPs. The survival rates were obtained after 2 and 24 hours incubation of the cells with NPs using MTT assay. Also, apoptosis and ROS generation induced by the NPs in the treated cells were obtained using Caspases-3 activity assay and flow cytometry analysis, respectively.Results: Bi2O3 NPs were successfully synthesized with average size of 19.2 ± 6.5 nm, then conjugated with 5-ALA and folate. Either naked or folate-conjugated NPs were easily taken up by the cells in a concentration-dependent manner and showed cytotoxic effects. The significant cell death was noted at the concentrations more than 50 μg/ml for both compounds.Conclusion: Results indicated low cytotoxicity of the prepared NPs at lower incubation periods, which is very important for their further applications. However, 24 hours incubation of the cells with both forms of NPs caused more cell killing and the cytotoxicity increased with increasing concentrations of the NPs.
Highlights
Recent advances in nanotechnology have led to the development of nanomaterials with potential of application in medicine, electronics, biosensors and biomaterials.[1,2,3,4] The nanomaterials have unique properties that are completely different from their bulk forms.[5]
For verifying the conjugation process, Fourier-transform infrared spectroscopy (FTIR) spectra of the synthesized Bi2O3 NPs and folic acid (FA)-conjugated NPs were done and the results are shown in Figure 2 a, b
The cytotoxicities effects of both of the synthesized NPs were studied on KB and A549 cells using three assays including MTT, caspase-3 activity, and flow cytometry
Summary
Recent advances in nanotechnology have led to the development of nanomaterials with potential of application in medicine, electronics, biosensors and biomaterials.[1,2,3,4] The nanomaterials have unique properties that are completely different from their bulk forms.[5] Due to their small size, nanomaterials have shown unique physical, optical, electronic, and chemical properties.[6,7] For example, high-atomic number nanoparticles (NPs) such as bismuth (Z=83) oxide (Bi2O3) NPs have been proven to be radioenhancer in cancer radiotherapy.[8,9,10,11,12] bismuth does not accumulate in the body and is biocompatible in-vivo.[11,12] Due to its excellent biocompatibility and low cost compared to other high-Z nanomaterials such as gold, bismuth has recently been used as a high-sensitive contrast agent in CT-scan examinations.[13,14] NPs usually induce toxicity when they enter into biological systems in medical applications.[15] Surface modification methods such as PEGylation or silica coating, allow NPs to be utilized safely in biomedical treatments;[16,17] such procedures are complex and time consuming.[18] Folate (folic acid) has been used as conjugation agent to enhance cytotoxicity of NPs for cancer treatment.[8] Folate receptors (FRs) are single-chain glycoproteins which possess high specific affinity for folic acid (FA) and are overexpressed on cell membrane of various malignant tumors,[19] such as human oral squamous carcinoma (KB cells)[20] but not in adenocarcinomic human alveolar basal epithelial cells (A549 cell line).[21] Most of human normal cells have a little expression of FR, the over-expression of FRs on membrane of the tumor cells can be exploited as a specific targeting ligand.[22] This targeting strategy can be utilized for increasing the diagnostic and therapeutic efficacy in different types of cancers
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