Cellulose Nanocrystals/Chitosan-Based Nanosystems: Synthesis, Characterization, and Cellular Uptake on Breast Cancer Cells

Abstract

Cellulose nanocrystals (CNCs) are elongated biobased nanostructures with unique characteristics that can be explored as nanosystems in cancer treatment. Herein, the synthesis, characterization, and cellular uptake on folate receptor (FR)-positive breast cancer cells of nanosystems based on CNCs and a chitosan (CS) derivative are investigated. The physical adsorption of the CS derivative, containing a targeting ligand (folic acid, FA) and an imaging agent (fluorescein isothiocyanate, FITC), on the surface of the CNCs was studied as an eco-friendly methodology to functionalize CNCs. The fluorescent CNCs/FA-CS-FITC nanosystems with a rod-like morphology showed good stability in simulated physiological and non-physiological conditions and non-cytotoxicity towards MDA-MB-231 breast cancer cells. These functionalized CNCs presented a concentration-dependent cellular internalization with a 5-fold increase in the fluorescence intensity for the nanosystem with the higher FA content. Furthermore, the exometabolic profile of the MDA-MB-231 cells exposed to the CNCs/FA-CS-FITC nanosystems disclosed a moderate impact on the cells’ metabolic activity, limited to decreased choline uptake and increased acetate release, which implies an anti-proliferative effect. The overall results demonstrate that the CNCs/FA-CS-FITC nanosystems, prepared by an eco-friendly approach, have a high affinity towards FR-positive cancer cells and thus might be applied as nanocarriers with imaging properties for active targeted therapy.

Natural Polymers-Based Materials: A Contribution to a Greener Future

“…Engineering natural polymers-based materials derived from polysaccharides and proteins is the main objective of the work carried out by the BioPol4fun research group. In the last decade, we have been intensively exploiting cellulose [ 63 , 64 ], nanocelluloses (e.g., bacterial nanocellulose (BNC) [ 61 , 65 , 66 ], nanofibrillated cellulose (CNFs) [ 67 , 68 ] and cellulose nanocrystals (CNCs) [ 69 ]), chitosan [ 70 , 71 , 72 , 73 ], pullulan [ 71 , 74 , 75 , 76 ], starch [ 68 ], hyaluronic acid [ 77 , 78 ], alginate [ 79 ], fucoidan [ 80 , 81 ], agar [ 82 ], lysozyme [ 83 , 84 , 85 , 86 ], and gelatin [ 87 ] ( Figure 1 ) to fabricate films [ 85 , 88 , 89 , 90 , 91 ], membranes [ 81 , 92 , 93 , 94 , 95 ], (nano)composites reference [ 64 , 96 , 97 , 98 , 99 ], coatings [ 100 , 101 , 102 , 103 ], nanosystems [ …”

“…Cellulose is considered the most abundant biopolymer on the planet, being a constituent of most green plants and algae, and naturally secreted in its pure form by some strains of non-pathogenic bacteria (e.g., Komagataeibacter ) [ 66 , 110 ]. This polysaccharide is an eminent feedstock for materials development and can be employed in its native state [ 63 , 64 ] as cellulose derivatives, or in the form of nanofibrils (CNFs) reference [ 85 , 86 , 88 , 89 , 98 , 100 , 101 , 111 ], nanorods (CNCs) [ 69 ], or three-dimensional hydrogel pellicles (BNC) [ 77 , 78 , 79 , 81 , 92 , 93 , 94 , 95 , 104 , 105 , 106 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 ] to manufacture a wide range of materials, as shown in Table 1 . Therefore, the vast majority of the works of our research group entails cellulose nanoforms, i.e., cellulose with at least one dimension in the nanoscale, for the development of nanocomposites [ 98 , ...…”

“…In a different approach, flexible electroconductive substrates with potential application in electronic devices, energy storage, or sensors were devised through the inclusion of copper nanowires [ 111 ] or organic conductive polymer solutions (e.g., poly(3,4-ethylenedioxythiophene): polystyrene sulfonate (PEDOT:PSS) [ 125 ]) in nanocellulosic substrates. The functionalization of biopolymers can also be exploited to impart luminescence or (bio)imaging functions to the ensuing materials; for instance, the blend of chitosan and pullulan films with lanthanopolyoxometalates [ 71 ], the chemical grafting of corrole macrocycles in a chitosan matrix [ 72 ], and the adsorption of a chitosan-derivative containing fluorescein isothiocyanate [ 69 ] on the surface of CNCs. In the field of water remediation, works related to the manufacture of bio-sorbent membranes of BNC/poly(2-methacryloyloxyethyl phosphorylcholine) [ 92 ] and CNFs/LNFs [ 85 ] have been suggested for the removal of organic dyes (methylene blue, methylene orange) and mercury from contaminated waters, respectively.…”

“…Similarly, lysozyme nanofibrils can provide mechanical reinforcement of polymeric matrices and impart antimicrobial properties that are desirable in active packaging and biomedical applications [ 108 ]. As a result, the proper design of functional biobased polymeric materials increases their application in high-tech fields, including active and intelligent food packaging [ 75 , 89 , 123 ], smart coatings [ 73 , 101 ], drug delivery [ 76 , 78 , 105 ], imaging [ 69 ], water remediation [ 85 , 92 ], sensors [ 111 , 125 ], and fuel cells [ 81 , 95 , 115 ].…”

Spherical Cellulose Micro and Nanoparticles: A Review of Recent Developments and Applications

“…Herein, and for clarity purposes, micro and nanoparticles solely or mostly composed of cellulose (or its derivatives) are microspheres/microcapsules with diameters at the microscale, and nanospheres/nanocapsules with diameters at the nanoscale, respectively. Therefore, cellulose nanocrystals with a rod-like morphology (elongated nanostructures), cross-sectional size of 5–70 nm and length in the range 100–250 nm [ 36 , 40 , 41 , 42 , 43 ], despite being considered cellulose nanoparticles [ 44 ], are out of the scope of this appraisal. In terms of organization, the current review includes a concise overview of the cellulose fundamentals and the fabrication methodologies of spherical particles, followed by the landscape of available examples dealing with the production of sphere-shaped micro and nanoparticles derived from the ubiquitous cellulose.…”