Abstract
This study deals with the preparation and characterization of selenium incorporated guar gum nanoparticle (SGG), and its effect on H9c2 cardiomyoblast. Herein, nanoprecipitation techniques had been employed for the preparation of SGG nanoparticle. The prepared nanoparticle had been subjected to various types of analytical techniques like transmission electron microscopy (TEM), X-ray diffraction (XRD) and particle size analysis to confirm the characteristics of nanoparticle as well as for selenium incorporation. Physical characterization of nanoparticle showed that the size of nanoparticles increase upto ∼69–173 nm upon selenium incorporation from ∼41–132 nm. Then the prepared nanoparticles were evaluated for its effect on H9c2 cells. In this regard, the effect of nanoparticle on various vital parameters of H9c2 cells was studied. Parameters like cell viability, uptake of selenium incorporated guar gum nanoparticle by the cells, effect of SGG on DNA integrity, apoptosis, reactive oxygen species generation, alteration in transmembrane potential of mitochondria and cytoskeletal integrity had been investigated. Viability results showed that up to 25 nM of SGG was safe (10.31%) but beyond that it induces cytotoxicity. Cellular uptake of selenium showed that cell permeability for SGG is significantly high compared to normal selenium (7.2 nM of selenium for 25 nM SGG compared with 5.2 nM selenium for 25 nM sodium selenite). There was no apoptosis with SGG and also it protects DNA from hydroxyl radical induced breakage. Likewise no adverse effect on mitochondria and cytoskeleton was observed for 25 nM of SGG. Overall results reveal that SGG is highly suitable for biomedical research application.
Highlights
Nanoscience has become an important area of research in biomedical sciences
In order to see the interaction of selenium incorporated guar gum nanoparticle (SGG) with cells we systematically investigated the effect of nanoparticle on H9c2 cells by analyzing various parameters like cell viability, apoptosis, DNA protection, reactive oxygen species (ROS) generation, mitochondrial transmembrane potential change and alteration in cytoskeleton
Materials and Reagents Guar gum powder, mannanase enzyme from Helix pomatia, sodium selenite, triton X-100, isopropanol, 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyl tetrazolium bromide (MTT), sodium tripolyphosphate, dimethyl sulfoxide (DMSO), 29,79 dichlorodihydrofluorescein diacetate (DCFH-DA), acridine orange (AO), ethidium bromide (EtBr), 2, 3 diaminonaphthalene, JC-1 (5,59,6,69-tetrachloro-1,19,3,39tetraethylbenzimidazolyl carbocyanine iodide), 49,6-diamidino-2-phenylindole (DAPI), phallodin and pUC-18 plasmid DNA were purchased from Sigma Chemicals, USA
Summary
Nanoscience has become an important area of research in biomedical sciences. Nanoparticles deliver a wide range of drugs (hydrophilic drugs, hydrophobic drugs, proteins, vaccines, biological macromolecules) to target areas of the body (lymphatic system, brain, arterial walls, lungs, liver, spleen, or made for long-term systemic circulation) for sustained periods of time [1]. A large number of studies have been conducted on polysaccharides and their derivatives for their potential application as drug delivery systems [2,3,4]. Nanoprecipitation is a general route to prepare polymeric nanoparticles under mild conditions and is well suitable for biological applications [5,6]. These techniques have many advantages as it is a straightforward technique, rapid and easy to perform [7,8,9]. Nanoparticles made of biodegradable polymers like proteins and polysaccharides can act as efficient drug delivery vehicles for sustained, controlled and targeted release, aiming to improve the therapeutic effects and to reduce the side effects of the formulated drugs [10]
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