An investigation of antibiofilm and cytotoxic property of MgO nanoparticles

https://doi.org/10.1016/j.bcab.2019.101069Get rights and content

Highlights

  • Magnesium oxide nanoparticles (MgONPs) synthesis and characterization.

  • Reduction of the biofilm forming uropathogens upon treatment with MgONPs.

  • MgONPs induce apoptosis in MCF-7 cell line.

  • It can be used as a coating material for medical implants.

Abstract

MgO nanoparticles (MgONPs) have been widely used as antibacterial agents with the advantages of them being nontoxic and their unique biological properties. In this study, we synthesized MgONPs by co-precipitation method and characterized them by XRD, SEM and EDS analysis. The antibiofilm activity of MgONPs as observed quantitatively by crystal violet assay and their action on biofilm architecture were assessed in both Gram-positive and Gram negative uropathogenic bacteria. In addition, the cytotoxic effect of synthesized MgONPs was evaluated against human breast cancer, MCF-7 cells. The morphological changes of apoptosis were observed by Acridine Orange/Ethidium Bromide (AO/EB) staining using a fluorescent microscope. MgONPs inhibited more than 50% of the biofilms in most of the tested uropathogenic bacteria; notably 80% inhibition in the case of K. pneumoniae. MgONPs successfully inhibited the viability of MCF-7 cells at a concentration of 50 μg.mL−l. Given their antibiofilm properties; MgONPs could be used as a potential nanomaterial for in vivo applications such as coating for a medical implant.

Introduction

Nanotechnology is an area of emerging interest in the field of science and technology due to its wide variety of applications in the field of biomedicine, optics, and electronics; especially for purpose of developing new nanoscale materials (Albrecht et al., 2006). Various physical, chemical, biological, and hybrid methods are currently being employed for nanoparticle synthesis (MubarakAli et al., 2015a, MubarakAli et al., 2015b). Nanoparticles can be made by using various biological substrates such as bacteria, algae, diatom, actinomycetes, plants, and biomolecules (Priyadarshini et al., 2013; MubarakAli et al., 2011a, 2011b; MubarakAli et al., 2011a, MubarakAli et al., 2011b, MubarakAli et al., 2011c, MubarakAli et al., 2013a, MubarakAli et al., 2013b).

Urolithiasis (kidney stone disease) is one of the most common urological diseases with high prevalence globally (Manzoor et al., 2017 and 2018a). Urinary tract infections (UTIs) and urolithiasis are inevitably linked and studies suggested that patients with kidney stone are more likely to have UTIs than the normal population (Barr-Beare et al., 2015). Bacterial biofilms play an important role in urolithiasis and many bacteria causing UTIs are associated with biofilm formation (Shabeena et al., 2018, Manzoor et al., 2018b, Manzoor et al., 2018c, Manzoor et al., 2018d). The biofilms are multi-cellular, surface-attached microbial communities with particular physiologic and architecture characteristics, which can sometimes confer resistance to different classes of antibiotics (Vahedi et al., 2017). This biofilm formation is an important virulence factor for a wide range of microbes that cause chronic infections, and are responsible for 75% of human microbial infections (Koo et al., 2017). Most of the Gram positive and Gram negative bacteria have the ability to form biofilms and main ones include Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Staphylococcus aureus, Streptococcus viridans, Proteus mirabilis and Pseudomonas aeruginosa (Donlan, 2001; MubarakAli et al., 2015).

The control of the bacterial biofilm formation is of importance for public health and economy, especially in the case nosocomial infections which leads to various life-threatening diseases (Naiyf, and Jamal, 2017). Several synthesized nanoparticles have shown their effectiveness for treating infectious microbes, including antibiotic-resistant strains (Huh et al., 2011; Ghosh et al., 2015). Several studies have demonstrated the in vitro inhibition of bacterial biofilm by various nanoparticles (Markowska et al., 2013). Most of the nanoparticles such as silver, gold, copper, and zinc have the ability to inhibit the biofilm growth (Gopinath et al., 2015; Chari et al., 2017; LewisOscar et al., 2015). Furthermore, certain inorganic nanoparticles have important implications for the treatment of microbial infections and post-surgical complication as wound dressing materials (Gopinath et al., 2012; MubarakAli et al., 2013a, MubarakAli et al., 2013b). Breast cancer is one of the most common cancers worldwide. The global burden of breast cancer exceeds all other cancers and it is a leading cause of cancer death in women and the incidence rates of breast cancer are increasing globally (Jemal et al., 2010). One of the main advantages of these nanoparticles is their low cytotoxicity and antioxidant property (MubarakAli et al., 2018). Importantly, some of the synthesized nanoparticles have been shown to be effective against a broad range of human breast cancer cells (Jeyaraj et al., 2013). MCF-7 is a widely used epithelial cancer cell line, derived from breast adenocarcinoma and used for in vitro breast cancer studies (Lee et al., 2015).

Magnesium oxide (MgO) is a versatile biomaterial, with wide applications in materials science and biomedical diagnostics. However, the toxicity of nanoparticles of MgO to bacterial/human cells and organs remains fairly unknown. The role of MgONPs as efficient biofilm inhibitors has not yet been given considerable attention (Hayat et al., 2018). Therefore, the present study was designed to synthesize and characterize MgONPs, and the antibiofilm potential was assessed against urolithiasis associated uropathogenic bacteria. The MgONPs were characterized using X-ray diffraction (XRD), fourier transform infrared (FTIR) spectscopy, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The biofilm architecture was analyzed by CLSM. In addition, the anti-cancer activity was studied using MCF-7 breast cancer cell line and the relative rate of proliferation was studied by Acridine Orange/Ethidium Bromide (AO/EB) apoptotic staining method using a fluorescent microscope.

Section snippets

Materials

Chemicals such as Acridine orange (C17H19N3), ethidium bromide (C21H20BrN3), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (RM1131, Hi-media Mumbai), and Dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St Louis, MO, USA).

Collection of clinical samples, cell culture, and maintenance

Ten clinical strains of bacteria including Gram positive and Gram negative strains were obtained from the Yenepoya Research Centre in Mangalore, India and sub-cultured on Luria Bertani Agar. The morphological and physiological

Results and discussion

The SEM, XRD and EDS pattern of MgONPs obtained from coprecipitation synthesis were as shown in Fig. 1(a–d). The synthesized MgONPs showed sharp plane showing at 42 °C by XRD analysis. A cluster of fine spherical nanoparticles was observed with the size range from 100 to 200 nm by SEM analysis. EDS spectrum showed Mg and O elemental signals at a high percentage. The antibiofilm efficacy of the MgONPs was evaluated against both Gram-positive and Gram-negative uropathogenic bacteria. Biofilm

Conclusion

The study highlighted the capacity of MgONPs as an antibiofilm against uropathogens and potential cytotoxicity against MCF-7 cells. The results clearly revealed a significant reduction of the biofilm formation upon treatment of MgONPs, and the wide spectrum of morphological differences in biofilm architectures. We believe that the present study will encourage further studies on the applications of MgONPs as potential biocompatible and nanomaterial for in vitro and in vivo application. Further

Acknowledgment

The authors gratefully acknowledge the financial support from the DBT (Govt. of India), for the establishment of National Repository for Microalgae and Cyanobacteria – Freshwater (BT/PR7005/PBD26/357/2012). This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean Government (No 2016R1D1A1A09918072).

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    Present Address: ICAR–Indian Institute of Spices Research, Kozhikode, 673012, Kerala, India.

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