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B: Biophysical Chemistry and Biomolecules
Investigating the Interaction of Silicon Dioxide Nanoparticles with Human Hemoglobin and Lymphocyte Cells by Biophysical, Computational and Cellular Studies Negin Sabziparvar, Yosra Saeedi, Mina Nouri, Atefeh Sadat Najafi Bozorgi, Elahe Alizadeh, Farnoosh Attar, Keivan Akhtari, Seyyedeh Elaheh Mousavi, and Mojtaba Falahati J. Phys. Chem. B, Just Accepted Manuscript • DOI: 10.1021/acs.jpcb.8b00193 • Publication Date (Web): 14 Mar 2018 Downloaded from http://pubs.acs.org on March 15, 2018
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The Journal of Physical Chemistry
Investigating the Interaction of Silicon Dioxide Nanoparticles with Human Hemoglobin and Lymphocyte Cells by Biophysical, Computational and Cellular Studies Negin Sabziparvar1#, Yosra Saeedi1#, Mina Nouri1, Atefeh Sadat Najafi Bozorgi2, Elahe Alizadeh2,Farnoosh Attar3, Keivan Akhtari4, Seyyedeh Elaheh Mousavi5*, Mojtaba Falahati6* 1
Department of Cellular and Molecular Biology, Faculty of Advance Science and Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran (IAUPS). 2 Department of Toxicology–Pharmacology, Faculty of Pharmacy, Pharmaceutical Science Branch, Islamic Azad University , Tehran, Iran (IAUPS). 3 Department of Biology, Faculty of Food Industry & Agriculture, Standard Research Institute (SRI), Karaj, Iran; 4 Department of Physics, University of Kurdistan, P.O. Box 416, Sanandaj, Iran. 5 Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran 6 Department of Nanotechnology, Faculty of Advance Science and Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran (IAUPS).
*Corresponding author: Mojtaba Falahati: Department of Nanotechnology, Faculty of Advance Science and Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran (IAUPS).
[email protected]. *Co-corresponding author: Seyyedeh Elaheh Mousavi: Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, P.O. Box: 13145784, Tehran, Iran.
[email protected].
#These authors contributed equally to this work
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Abstract NPs have received a great attention in biological and medical applications due to their unique features. However, their induced adverse effects on the biological system are not well explored. Herein, the interaction of silicon dioxide nanoparticle (SiO2 NP) with human hemoglobin (Hb) and lymphocyte cell line was evaluated under physiological conditions by multi spectroscopic (intrinsic and synchronous fluorescence spectroscopy, and circular dichrosim (CD)), molecular docking, and cellular (3-[4,5dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) and acridine orange/ethidium bromide (AO/EB) staining) methods. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) revealed the nanosized and spherical shaped SiO2 particle. The fluorescence and lifetime decay results indicated that SiO2 NP quenched the intrinsic intensity of Hb through a static quenching mechanism. Binding affinity of SiO2 NP towards Hb was directly correlated with temperature. The sign of thermodynamic parameters demonstrated that hydrophobic forces played a pivotal role in the interaction of SiO2 NP with Hb. Results of synchronous fluorescence experiments displayed that Tyr residues are moved to a more hydrophilic microenvironment. Molecular docking studies exhibited that SiO2 and Si NPs were bound to Hb primarily by hydrophobic residues. The findings from CD data verified no alteration in secondary structure of Hb upon binding to SiO2 NP. The human lymphocyte cell line was treated with SiO2 NP at varying concentrations and time intervals and the cytotoxicity assays by MTT and AO/EB staining showed that cells viability was reduced by SiO2 NP-induced apoptosis mechanism in a dose and time-dependent manner. Therefore, it may be suggested comprehensive details regarding the interaction of NPs and biological systems such as cells and proteins can provide useful information in development of NP-based systems.
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The Journal of Physical Chemistry
1. Introduction With the invention of nanomaterials came the achievement of their extensive implementation to many interdisciplinary researches, especially nano-medicine1-3. The distinctive features of nanomaterials have demonstrated to be predominantly connected with their dimension, morphology, chemical type, and functional groups4, 5. Of all the nanoparticles (NPs) available today, SiO2 NPs has gained growing interest owing to its plasmonic characteristics in conjunction with low or no immediate adverse effects against biological systems6, 7. Likewise, the simplicity of fabrication and chemical stability adjoins to the general interest of SiO2 NPs. Recently; the leading edge of attention has been given to explore the material features of NPs, which led to the expansion of fabrication routes and characterization methods8. However, with the ever-growing increase in the nano-bio implementation of SiO2 NPs, there is a necessity to explore their interaction with biological systems9. Serum proteins like hemoglobin (Hb) and blood components such as lymphocytes circulate through the blood and, therefore are possibly biological targets to interact with SiO2 NPs. Also, the non-inert essence of these NPs has been identified in both in vitro and in vivo investigations exhibiting adverse effects or altered cellular function following NPs exposure10-12. Actually, SiO2 NPs can interact with biomolecules and show the ability to alter the outcome of chemical reactions and modulate cellular pathways. Currently; exploring the interaction of NPs with proteins and protein corona formation has been an active area of study. Understanding mechanisms of protein corona formation at the nao/bio interface is critical in elucidating the biological effects and functions of NPs in the body13. NP-protein corona interaction induces strong effects on pharmacokinetic aspect of bound NPs as it influences the absorption, distribution, metabolism, and excretion of NP in the body14, 15. These factors have been documented to play important roles in assessing the safety of NPs, because they influence physicochemical properties and kinetics of NPs in vivo16, 17. Various NPs bind reversibly to the serum protein such as hemoglobin with different binding affinities. Low binding affinity of a NP to protein 3 ACS Paragon Plus Environment
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causes short life-span and poor distribution of the NP in the body, whereas strong binding lead to the NP-mediated conformational changes and cytotoxicity18. Therefore, exploring the NP-biological system interactions may provide useful information in advancement of therapeutic efficiency of the NPs. It has been well reported that proteins adsorb on the NP surface and mortality of the cells depending on the chemical type of NPs. For example Sun et al., (2012) reported that zwitterionic sulfobetaine copolymer‐coated mesoporous Si NPs can be utilized for temperature‐responsive drug delivey19. Pisani et al., (2017) revealed the biocompatibility of modified magnetic mesoporous Si NPs in human HepaRG cells20. Sahoo et al., (2014) suggested that mesoporous Si-coated superparamagnetic manganese ferrite NP exhibits enhanced biocompatibility for selective drug delivery and MR imaging implementations21. In other side, Yang et al., (2016) reported the toxicity and DNA-damage in mouse macrophage incubated with SiO2 NPs22. Gong et al., (2017) explored that PAPR-1 can protect human HaCaT cells against SiO2 NPs-initiated adverse effects23. Moreover, Shamsi et al., (2017) revealed the conformational changes of kidney cystatin mediated by SiO2 NPs24. Givens et al., (2017) also reported that the secondary structure of protein was altered by SiO2 NPs25. However, Falahati et al., (2011) suggested that mesoporous SiO2 NPs can be employed as a potential host for immobilization of superoxide dismutase26. Therefore, conflicting reports have been dedicated to the effects of SiO2 NPs on biological systems such as cells and proteins. Hence, the main aim of ongoing research was to explore the influence of SiO2 NPs on Hb and lymphocyte as potential models of blood system by means of biophysical, docking and cellular approaches.
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The Journal of Physical Chemistry
2. Materials and methods 2.1. Materials Hb was purchased from Sigma Company (St. Louis, MO, USA). SiO2 nanopowder (Purity: 99.5%, APS: 20-30nm, SSA: 180-600m2/g, Color: white, Bulk Density: 0 and ∆S>0 display the existence of hydrophobic interactions (b) ∆H
