Synthesis and Cytotoxicity Studies of Wood-Based Cationic Cellulose Nanocrystals as Potential Immunomodulators

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Abstract

Polysaccharides have been shown to have immunomodulatory properties. Modulation of the immune system plays a crucial role in physiological processes as well as in the treatment and/or prevention of autoimmune and infectious diseases. Cellulose nanocrystals (CNCs) are derived from cellulose, the most abundant polysaccharide on the earth. CNCs are an emerging class of crystalline nanomaterials with exceptional physico-chemical properties for high-end applications and commercialization prospects. The aim of this study was to design, synthesize, and evaluate the cytotoxicity of a series of biocompatible, wood-based, cationic CNCs as potential immunomodulators. The anionic CNCs were rendered cationic by grafting with cationic polymers having pendant ⁺NMe ₃ and ⁺NH ₃ moieties. The success of the synthesis of the cationic CNCs was evidenced by Fourier transform infrared spectroscopy, dynamic light scattering, zeta potential, and elemental analysis. No modification in the nanocrystals rod-like shape was observed in transmission electron microscopy and atomic force microscopy analyses. Cytotoxicity studies using three different cell-based assays (MTT, Neutral Red, and LIVE/DEAD ^®) and three relevant mouse and human immune cells indicated very low cytotoxicity of the cationic CNCs in all tested experimental conditions. Overall, our results showed that cationic CNCs are suitable to be further investigated as immunomodulators and potential vaccine nanoadjuvants.

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Nanocellulose, a Versatile Green Platform: From Biosources to Materials and Their Applications

Bejoy Thomas, Midhun Raj, Athira B … (2018)

With increasing environmental and ecological concerns due to the use of petroleum-based chemicals and products, the synthesis of fine chemicals and functional materials from natural resources is of great public value. Nanocellulose may prove to be one of the most promising green materials of modern times due to its intrinsic properties, renewability, and abundance. In this review, we present nanocellulose-based materials from sourcing, synthesis, and surface modification of nanocellulose, to materials formation and applications. Nanocellulose can be sourced from biomass, plants, or bacteria, relying on fairly simple, scalable, and efficient isolation techniques. Mechanical, chemical, and enzymatic treatments, or a combination of these, can be used to extract nanocellulose from natural sources. The properties of nanocellulose are dependent on the source, the isolation technique, and potential subsequent surface transformations. Nanocellulose surface modification techniques are typically used to introduce either charged or hydrophobic moieties, and include amidation, esterification, etherification, silylation, polymerization, urethanization, sulfonation, and phosphorylation. Nanocellulose has excellent strength, high Young's modulus, biocompatibility, and tunable self-assembly, thixotropic, and photonic properties, which are essential for the applications of this material. Nanocellulose participates in the fabrication of a large range of nanomaterials and nanocomposites, including those based on polymers, metals, metal oxides, and carbon. In particular, nanocellulose complements organic-based materials, where it imparts its mechanical properties to the composite. Nanocellulose is a promising material whenever material strength, flexibility, and/or specific nanostructuration are required. Applications include functional paper, optoelectronics, and antibacterial coatings, packaging, mechanically reinforced polymer composites, tissue scaffolds, drug delivery, biosensors, energy storage, catalysis, environmental remediation, and electrochemically controlled separation. Phosphorylated nanocellulose is a particularly interesting material, spanning a surprising set of applications in various dimensions including bone scaffolds, adsorbents, and flame retardants and as a support for the heterogenization of homogeneous catalysts.

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Cell sensitivity assays: the MTT assay.

Johan van Meerloo, Gertjan Kaspers, Jacqueline Cloos (2011)

The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay is based on the conversion of MTT into formazan crystals by living cells, which determines mitochondrial activity. Since for most cell populations the total mitochondrial activity is related to the number of viable cells, this assay is broadly used to measure the in vitro cytotoxic effects of drugs on cell lines or primary patient cells. In this chapter the protocol of the assay is described including important considerations relevant for each step of the assay as well as its limitations and possible applications.

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Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique.

Richard Murdock, Laura K Braydich-Stolle, Amanda Schrand … (2008)

The need to characterize nanoparticles in solution before assessing the in vitro toxicity is a high priority. Particle size, size distribution, particle morphology, particle composition, surface area, surface chemistry, and particle reactivity in solution are important factors which need to be defined to accurately assess nanoparticle toxicity. Currently, there are no well-defined techniques for characterization of wet nanomaterials in aqueous or biological solutions. Previously reported nanoparticle characterization techniques in aqueous or biological solutions have consisted of the use of ultra-high illumination light microscopy and disc centrifuge sedimentation; however, these techniques are limited by the measurement size range. The current study focuses on characterizing a wide range of nanomaterials using dynamic light scattering (DLS) and transmission electron microscopy, including metals, metal oxides, and carbon-based materials, in water and cell culture media, with and without serum. Cell viability and cell morphology studies were conducted in conjunction with DLS experiments to evaluate toxicological effects from observed agglomeration changes in the presence or absence of serum in cell culture media. Observations of material-specific surface properties were also recorded. It was also necessary to characterize the impact of sonication, which is implemented to aid in particle dispersion and solution mixture. Additionally, a stock solution of nanomaterials used for toxicology studies was analyzed for changes in agglomeration and zeta potential of the material over time. In summary, our results demonstrate that many metal and metal oxide nanomaterials agglomerate in solution and that depending upon the solution particle agglomeration is either agitated or mitigated. Corresponding toxicity data revealed that the addition of serum to cell culture media can, in some cases, have a significant effect on particle toxicity possibly due to changes in agglomeration or surface chemistry. It was also observed that sonication slightly reduces agglomeration and has minimal effect on particle surface charge. Finally, the stock solution experienced significant changes in particle agglomeration and surface charge over time.

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Author and article information

Journal

Journal ID (nlm-ta): Nanomaterials (Basel)

Journal ID (iso-abbrev): Nanomaterials (Basel)

Journal ID (publisher-id): nanomaterials

Title: Nanomaterials

Publisher: MDPI

ISSN (Electronic): 2079-4991

Publication date (Electronic): 15 August 2020

Publication date Collection: August 2020

Volume: 10

Issue: 8

Electronic Location Identifier: 1603

Affiliations

[1 ]Department of Chemistry, State University of New York at Plattsburgh, Plattsburgh, New York, NY 12901, USA; yimti001@ 123456plattsburgh.edu (Y.I.); btuga001@ 123456plattsburgh.edu (B.T.); c.w.smith022@ 123456gmail.com (C.W.S.); arabi005@ 123456plattsburgh.edu (A.R.); lnguy007@ 123456plattsburgh.edu (L.N.)

[2 ]Aquatic and Crop Resource Development Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, Canada; yali.liu@ 123456cnrc-nrc.gc.ca (Y.L.); sabahudin.hrapovic@ 123456cnrc-nrc.gc.ca (S.H.)

Author notes

[* ]Correspondence: kckle001@ 123456plattsburgh.edu (K.C.); rajesh.sunasee@ 123456plattsburgh.edu (R.S.)

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Christopher W. Smith https://orcid.org/0000-0002-0828-3370

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Publisher ID: nanomaterials-10-01603

DOI: 10.3390/nano10081603

PMC ID: 7466698

PubMed ID: 32824129

SO-VID: 09379068-5ed3-427d-8d8d-23b56291ae7f

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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

Synthesis and Cytotoxicity Studies of Wood-Based Cationic Cellulose Nanocrystals as Potential Immunomodulators

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Most cited references 31

Nanocellulose, a Versatile Green Platform: From Biosources to Materials and Their Applications

Cell sensitivity assays: the MTT assay.

Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique.

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