RESUMEN
Site-specific modification is a great challenge for polysaccharide scientists. Chemo- and regioselective modification of polysaccharide chains can provide many useful natural-based materials and help us illuminate fundamental structure-property relationships of polysaccharide derivatives. The hemiacetal reducing end of a polysaccharide is in equilibrium with its ring-opened aldehyde form, making it the most uniquely reactive site on the polysaccharide molecule, ideal for regioselective decoration such as imine formation. However, all natural polysaccharides, whether they are branched or not, have only one reducing end per chain, which means that only one aldehyde-reactive substituent can be added. We introduce a new approach to selective functionalization of polysaccharides as an entrée to useful materials, appending multiple reducing ends to each polysaccharide molecule. Herein, we reduce the approach to practice using amide formation. Amine groups on monosaccharides such as glucosamine or galactosamine can react with carboxyl groups of polysaccharides, whether natural uronic acids like alginates, or derivatives with carboxyl-containing substituents such as carboxymethyl cellulose (CMC) or carboxymethyl dextran (CMD). Amide formation is assisted using the coupling agent 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM). By linking the C2 amines of monosaccharides to polysaccharides in this way, a new class of polysaccharide derivatives possessing many reducing ends can be obtained. We refer to this class of derivatives as multi-reducing-end polysaccharides (MREPs). This new family of derivatives creates the potential for designing polysaccharide-based materials with many potential applications, including in hydrogels, block copolymers, prodrugs, and as reactive intermediates for other derivatives.
Asunto(s)
Alginatos , Polisacáridos , Polisacáridos/química , Alginatos/química , Monosacáridos , Aldehídos , AmidasRESUMEN
This study was conducted to determine the effects of chitosan molecular weight and degree of deacetylation (DD) on chitosan-cellulose nanocrystal (CNC) polyelectrolyte-macroion complexes (PMCs) and their formation. Chitosan samples with three different molecular weights (81, 3 · 103, 6 · 103 kDa) and four different DDs (77, 80, 85, 89%) were used. The effects on PMC formation were determined by turbidimetric titration. An effect of the molecular weight of chitosan was not observed in turbidimetric titrations. Turbidity levels were higher for CNCs with lower sulfate group density and larger hydrodynamic diameter than for CNCs with higher sulfate group density and smaller hydrodynamic diameter. Conversely, turbidity levels were higher for chitosans with higher DD (higher charge density) than for chitosans with lower DD (lower charge density). PMC particles from chitosans with different molecular weights were characterized by scanning electron microscopy, laser Doppler electrophoresis, and dynamic light scattering. PMCs from high-molecular-weight chitosan were more spherical and those from medium-molecular-weight chitosan had a slightly larger hydrodynamic diameter than PMCs from the respective other two chitosans. The molecular weight of the chitosan was concluded to have no effect on the formation of chitosan-CNC PMC particles and only a minor effect on the shape and size of the particles. The higher turbidity levels for CNCs with lower sulfate group density and larger hydrodynamic diameter and for chitosans with higher DD were attributed to a larger number of CNCs being required for charge compensation.
RESUMEN
Elongated nanoparticles have recently been shown to have distinct advantages over spherical ones in targeted drug delivery applications. In addition to their oblong geometry, their lack of cytotoxicity and numerous surface hydroxyl groups make cellulose nanocrystals (CNCs) promising drug delivery vectors. Herein we report the synthesis of folic acid-conjugated CNCs for the targeted delivery of chemotherapeutic agents to folate receptor-positive cancer cells. Folate receptor-mediated cellular binding/uptake of the conjugate was demonstrated on human (DBTRG-05MG, H4) and rat (C6) brain tumor cells. Folate receptor expression of the cells was verified by immunofluorescence staining. Cellular binding/uptake of the conjugate by DBTRG-05MG, H4, and C6 cells was 1452, 975, and 46 times higher, respectively, than that of nontargeted CNCs. The uptake mechanism was determined by preincubation of the cells with the uptake inhibitors chlorpromazine or genistein. DBTRG-05MG and C6 cells internalized the conjugate primarily via caveolae-mediated endocytosis, whereas H4 cells internalized the conjugate primarily via clathrin-mediated endocytosis.
Asunto(s)
Celulosa/química , Portadores de Fármacos/síntesis química , Portadores de Fármacos/metabolismo , Receptores de Folato Anclados a GPI/metabolismo , Ácido Fólico/química , Nanopartículas/química , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Animales , Caveolas/metabolismo , Clorpromazina/farmacología , Clatrina/metabolismo , Portadores de Fármacos/química , Endocitosis/efectos de los fármacos , Receptores de Folato Anclados a GPI/análisis , Genisteína/farmacología , Humanos , Ratas , Células Tumorales CultivadasRESUMEN
Pretreatment of lignocellulosic biomass with sulfuric acid may leave sulfate groups on its surface that may hinder its biochemical conversion. This study investigates the effects of sulfate groups on cellulase adsorption onto cellulose substrates and the enzymatic hydrolysis of these substrates. Substrates with different sulfate group densities were prepared from H2SO4- and HCl-hydrolyzed and partially and fully desulfated cellulose nanocrystals. Adsorption onto and hydrolysis of the substrates was analyzed by quartz crystal microbalance with dissipation monitoring (QCM-D). The surface roughness of the substrates, measured by atomic force microscopy, increased with decreasing sulfate group density, but their surface accessibilities, measured by QCM-D H2O/D2O exchange experiments, were similar. The adsorption of cellulose binding domains onto sulfated substrates decreased with increasing sulfate group density, but the adsorption of cellulases increased. The rate of hydrolysis of sulfated substrates decreased with increasing sulfate group density. The results indicated an inhibitory effect of sulfate groups on the enzymatic hydrolysis of cellulose, possibly due to nonproductive binding of the cellulases onto the substrates through electrostatic interactions instead of their cellulose binding domains.
Asunto(s)
Celulasas/metabolismo , Celulosa/química , Celulosa/metabolismo , Sulfatos/química , Adsorción , Modelos Teóricos , Especificidad por SustratoRESUMEN
Chitinases are widely distributed in nature and have wide-ranging pharmaceutical and biotechnological applications. This work highlights a real-time and label-free method to assay Chitinase activity via a quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy (AFM). The chitin substrate was prepared by spincoating a trimethylsilyl chitin solution onto a silica substrate, followed by regeneration to amorphous chitin (RChi). The QCM-D and AFM results clearly showed that the hydrolysis rate of RChi films increased as Chitinase (from Streptomyces griseus) concentrations increased, and the optimal temperature and pH for Chitinase activity were around 37 °C and 6-8, respectively. The Chitinase showed greater activity on chitin substrates, having a high degree of acetylation, than on chitosan substrates, having a low degree of acetylation.
Asunto(s)
Proteínas Bacterianas/química , Quitina/química , Quitinasas/química , Acetilación , Pruebas de Enzimas , Concentración de Iones de Hidrógeno , Hidrólisis , Cinética , Microscopía de Fuerza Atómica , Procesamiento Proteico-Postraduccional , Tecnicas de Microbalanza del Cristal de Cuarzo , Streptomyces griseus/enzimología , Propiedades de Superficie , TemperaturaRESUMEN
This work highlights a real-time and label-free method to monitor the dehydrogenative polymerization of monolignols initiated by horseradish peroxidase (HRP) physically immobilized on surfaces using a quartz crystal microbalance with dissipation monitoring (QCM-D). The dehydrogenative polymer (DHP) films are expected to provide good model substrates for studying ligninolytic enzymes. The HRP was adsorbed onto gold or silica surfaces or onto and within porous desulfated nanocrystalline cellulose films from an aqueous solution. Surface-immobilized HRP retained its activity and selectivity for monolignols as coniferyl and p-coumaryl alcohol underwent dehydrogenative polymerization in the presence of hydrogen peroxide, whereas sinapyl alcohol polymerization required the addition of a nucleophile. The morphologies of the DHP layers on the surfaces were investigated via atomic force microscopy (AFM). Data from QCM-D and AFM showed that the surface-immobilized HRP-initiated dehydrogenative polymerization of monolignols was greatly affected by the support surface, monolignol concentration, hydrogen peroxide concentration, and temperature.
Asunto(s)
Lignina/metabolismo , Microscopía de Fuerza Atómica , Tecnicas de Microbalanza del Cristal de Cuarzo , Adsorción , Celulosa/química , Enzimas Inmovilizadas/metabolismo , Oro/química , Peroxidasa de Rábano Silvestre/metabolismo , Hidrogenación , Lignina/biosíntesis , Lignina/química , Estructura Molecular , Tamaño de la Partícula , Polimerizacion , Porosidad , Dióxido de Silicio/química , Propiedades de SuperficieRESUMEN
The Materials Genome Initiative (MGI) seeks to accelerate the discovery and engineering of advanced materials via high-throughput experimentation (HTE), which is a challenging task, given the common trade-off between design for optimal processability vs performance. Here, we report a HTE method based on automated formulation, synthesis, and multiproperty characterization of bulk soft materials in well plate formats that enables accelerated engineering of functional composite hydrogels with optimized properties for processability and performance. The method facilitates rapid high-throughput screening of hydrogel composition-property relations for multiple properties in well plate formats. The feasibility and utility of the method were demonstrated by application to several functional composite hydrogel systems, including alginate/poly(N-isopropylacrylamide) (PNIPAM) and poly(ethylene glycol) dimethacrylate (PEGDMA)/poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) hydrogels. The HTE method was leveraged to identify formulations of conductive PEGDMA/PEDOT:PSS composite hydrogels for optimized performance and processability in three-dimensional (3D) printing. This work provides an advance in experimental methods based on automated dispensing, mixing, and sensing for the accelerated engineering of soft functional materials.
RESUMEN
We demonstrate that increasing the hydrophobic environment around the charge center of a polyelectrolyte (PE) not only decreases the water content of an adsorbed PE layer but can even dewater up to ~50% of an initially hydrated substrate. The results of this work are expected to yield new stratagies to dewater PE systems and have potential applications in mineral recovery, paper manufacturing, and biomedical materials. Adsorption of a series of cationically derivatized dextran polyelectrolytes onto sulfated nanocrystalline cellulose (SNC) has been studied using quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR). Synthesized samples of (N,N-dimethylamino)ethyldextran (DMAE-Dex), (N,N-diethylamino)ethyldextran (DEAE-Dex), and (N,N-diisopropylamino)ethyldextran (DIAE-Dex) had degrees of substitution (DS) ranging from 0.05 to 0.82. DMAE-Dex, DEAE-Dex, and DIAE-Dex all showed decreasing adsorption onto SNC and decreasing water content of the adsorbed film with increasing DS. Additionally, DEAE-Dex and DIAE-Dex films adsorbed onto SNC contained less water than DMAE-Dex films with the same DS. Interestingly, QCM-D results for high DS DIAE-Dex adsorbed onto SNC revealed mass loss, whereas SPR results clearly showed DIAE-Dex adsorbed. These observations were consistent with dehydration of the SNC substrate. This study indicates that the water content of the substrate could be tailored by controlling the DS and hydrophobic character of the adsorbed polyelectrolytes.
Asunto(s)
Nanocompuestos/química , Polímeros/química , Interacciones Hidrofóbicas e Hidrofílicas , Resonancia por Plasmón de SuperficieRESUMEN
Chitin is the second most abundant biopolymer and insight into its natural synthesis, enzymatic degradation, and chemical interactions with other biopolymers is important for bioengineering with this renewable resource. This work is the first report of smooth, homogeneous, ultrathin chitin films, opening the door to surface studies of binding interactions, adsorption kinetics, and enzymatic degradation. The chitin films were formed by spincoating trimethylsilyl chitin onto gold or silica substrates, followed by regeneration to a chitin film. Infrared and X-ray photoelectron spectroscopy, X-ray diffraction, ellipsometry, and atomic force microscopy were used to confirm the formation of smooth, homogeneous, and amorphous chitin thin films. Quartz crystal microbalance with dissipation monitoring (QCM-D) solvent exchange experiments showed these films swelled with 49% water by mass. The utility of these chitin films as biosensors was evident from QCM-D and surface plasmon resonance studies that revealed the adsorption of a bovine serum albumin monolayer.
Asunto(s)
Técnicas Biosensibles , Quitina/química , Nanocompuestos/química , Cuarzo/química , Albúmina Sérica Bovina/química , Albúmina Sérica Bovina/metabolismo , Solventes/química , Animales , Bovinos , Cristalización , Oro/química , Cinética , Microscopía de Fuerza Atómica , Espectroscopía de Fotoelectrones , Dióxido de Silicio/química , Resonancia por Plasmón de Superficie , Propiedades de Superficie , Agua/química , Difracción de Rayos XRESUMEN
This study examines a novel polyelectrolyte-macroion complex (PMC) between chitosan, a cationic polysaccharide, and cellulose nanocrystals (CNCs), anionic, cylindrical nanoparticles, for potential applications in drug delivery. CNCs were prepared by H(2)SO(4) hydrolysis of wood pulp. The formation of PMCs was monitored by turbidimetric titration. In titrations of a chitosan solution with a CNC suspension, the turbidity reached a plateau, but it had a maximum and then decreased when the direction of titration was reversed. PMC particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, dynamic light scattering, and laser Doppler electrophoresis. The particles were composed primarily of CNCs and ranged in size from a few hundred nanometers to several micrometers, depending on the cellulose/chitosan ratio. Particles formed at amino/sulfate group molar ratios >1 were nearly spherical in shape and positively charged, whereas particles formed at ratios <1 had well-defined nonspherical shapes and were negatively charged.
Asunto(s)
Celulosa/química , Quitosano/química , Nanoestructuras , Sistemas de Liberación de Medicamentos , Nefelometría y TurbidimetríaRESUMEN
This study examines the effects of pH and salt concentration on the formation and properties of chitosan-cellulose nanocrystal (CNC) polyelectrolyte-macroion complexes (PMCs). The components' pK values, determined by potentiometric titration, were 6.40 for chitosan and 2.46 for the CNCs. The turbidity of PMC particle suspensions was measured as a function of chitosan-CNC ratio, pH, and salt concentration. The maximum turbidity values in titrations of a chitosan solution with a CNC suspension and vice versa occurred at charge ratios of 0.47 ± 0.11 (SO(3)(-)/NH(3)(+)) and 1.16 ± 0.06 (NH(3)(+)/SO(3)(-)), respectively. A pH increase caused a turbidity decrease due to shrinking of the PMC particles upon changes in their components' degrees of ionization. An increase in salt concentration caused a decrease in turbidity due to charge-screening-related shrinking of the PMC particles. The effects of pH and salt concentration on particle size were confirmed by scanning electron microscopy.
Asunto(s)
Materiales Biocompatibles/síntesis química , Celulosa/química , Quitosano/química , Sistemas de Liberación de Medicamentos/métodos , Polímeros/química , Administración Oral , Materiales Biocompatibles/análisis , Humanos , Concentración de Iones de Hidrógeno , Iones/química , Microscopía Electrónica de Rastreo , Peso Molecular , Nanopartículas/química , Nefelometría y Turbidimetría , Tamaño de la Partícula , Polímeros/análisis , Sales (Química)/química , Cloruro de Sodio/química , SolubilidadRESUMEN
Model cellulose surfaces have attracted increasing attention for studying interactions with cell wall matrix polymers and as substrates for enzymatic degradation studies. Quartz crystal microbalance with dissipation monitoring (QCM-D) solvent exchange studies showed that the water content of regenerated cellulose (RC) films was proportional to the film thickness (d) and was consistent with about five water molecules per anhydroglucose unit. Sulfated nanocrystalline cellulose (SNC) and desulfated nanocrystalline cellulose (DNC) films had comparable water contents and contained about five times more water than RC films. A cellulase mixture served as a probe for studies of substrate accessibility and degradation. Cellulase adsorption onto RC films was independent of d, whereas degradation times increased with d. However, adsorption onto SNC and DNC films increased with d, whereas cellulase degradation times for DNC films were independent of studied d. Enhanced access to guest molecules for SNC and DNC films revealed they are more porous than RC films.
Asunto(s)
Celulosa/química , Cuarzo , Solventes/química , Agua/análisis , Cristalización , Nanopartículas , Agua/químicaRESUMEN
HYPOTHESIS: Radial capillary flow in evaporating droplets carry suspended nanoparticles to its periphery where they are deposited and form a coffee-ring. Rod-like nanoparticles seeking to minimize their capillary energy will align with their long-axis parallel to the contact line. Particles exhibiting electrostatic repulsion, such as cellulose nanocrystals (CNCs), establish a competition between capillary flow-induced impingement against a growing coffee-ring and entropic minimization leading to enhanced particle mobility. Therefore, balancing these effects by manipulating the local particle concentration in drying droplets should result in deposition with a controlled orientation of CNCs. EXPERIMENTS: The dynamic local order in aqueous suspensions of CNCs in evaporating sessile droplets was investigated through time-resolved polarized light microscopy. The spatial distribution of alignment in deposited CNCs was explored as a function of nanoparticle concentration, droplet volume, initial degree of anisotropy, and substrate hydrophobicity. Computational analysis of the rotational Péclet number during evaporation was also investigated to evaluate any effects of shear-induced alignment. FINDINGS: Multiple modes of orientation were identified suggesting local control over CNC orientation and subsequent properties can be attained via droplet-based patterning methods. Specifically, high local particle concentrations led to tangential alignment and lower local particle concentrations resulted in new evidence for radial alignment near the center of dried droplets.
Asunto(s)
Celulosa , Nanopartículas , Anisotropía , Desecación , SuspensionesRESUMEN
3D-printed bone scaffolds hold great promise for the individualized treatment of critical-size bone defects. Among the resorbable polymers available for use as 3D-printable scaffold materials, poly(ε-caprolactone) (PCL) has many benefits. However, its relatively low stiffness and lack of bioactivity limit its use in load-bearing bone scaffolds. This study tests the hypothesis that surface-oxidized cellulose nanocrystals (SO-CNCs), decorated with carboxyl groups, can act as multi-functional scaffold additives that (1) improve the mechanical properties of PCL and (2) induce biomineral formation upon PCL resorption. To this end, an in vitro biomineralization study was performed to assess the ability of SO-CNCs to induce the formation of calcium phosphate minerals. In addition, PCL nanocomposites containing different amounts of SO-CNCs (1, 2, 3, 5, and 10 wt%) were prepared using melt compounding extrusion and characterized in terms of Young's modulus, ultimate tensile strength, crystallinity, thermal transitions, and water contact angle. Neither sulfuric acid-hydrolyzed CNCs (SH-CNCs) nor SO-CNCs were toxic to MC3T3 preosteoblasts during a 24 h exposure at concentrations ranging from 0.25 to 3.0 mg/mL. SO-CNCs were more effective at inducing mineral formation than SH-CNCs in simulated body fluid (1x). An SO-CNC content of 10 wt% in the PCL matrix caused a more than 2-fold increase in Young's modulus (stiffness) and a more than 60% increase in ultimate tensile strength. The matrix glass transition and melting temperatures were not affected by the SO-CNCs but the crystallization temperature increased by about 5.5°C upon addition of 10 wt% SO-CNCs, the matrix crystallinity decreased from about 43 to about 40%, and the water contact angle decreased from 87 to 82.6°. The abilities of SO-CNCs to induce calcium phosphate mineral formation and increase the Young's modulus of PCL render them attractive for applications as multi-functional nanoscale additives in PCL-based bone scaffolds.
RESUMEN
Cellulose nanocrystals (CNCs) prepared by H(2)SO(4) hydrolysis have sulfate groups on their surface, which have negative implications for some CNC applications. In this study, two desulfation methods were evaluated, and the properties of desulfated CNCs were compared to those of unsulfated CNCs, prepared by HCl hydrolysis. H(2)SO(4)-hydrolyzed CNCs from softwood sulfite pulp were subjected to either a mild acid hydrolytic desulfation or a solvolytic desulfation in dimethyl sulfoxide via the pyridinium salt. Removal of the sulfate groups was confirmed by conductometric titration and X-ray photoelectron spectroscopy. The effect of the desulfation procedure on the lateral crystallite dimensions was analyzed by X-ray diffraction. The extent of particle aggregation in the samples was assessed by atomic force microscopy and dynamic light scattering. The acid hydrolytic method achieved only partial desulfation and produced gradually decreasing sulfate contents upon successive repetition. The solvolytic method achieved nearly complete desulfation in a single step. The desulfated CNCs showed similar particle aggregation as the HCl-hydrolyzed CNCs, but the extent of aggregation was slightly less.
RESUMEN
The interaction between cellulose and hemicelluloses is of fundamental importance for understanding the molecular architecture of plant cell walls. Adsorption of xyloglucan (XG) onto regenerated cellulose (RC), sulfated cellulose nanocrystal (s-CNC), and desulfated cellulose nanocrystal (d-CNC) films was studied by quartz crystal microbalance with dissipation monitoring, surface plasmon resonance, and atomic force microscopy. The amount of XG adsorbed onto different cellulose substrates increased in the order RC < s-CNC < d-CNC. The adsorption of XG onto RC films was independent of film thickness (d), whereas XG adsorption was weakly dependent on d for s-CNC films and strongly dependent on d for d-CNC films. However, approximately the same amount of XG adsorbed onto "monolayer-thin" films of RC, s-CNC, and d-CNC. These results suggest that the morphology and surface charge of the cellulose substrate played a limited role in XG adsorption and highlight the importance of film thickness of cellulose nanocrystalline films to XG adsorption.
RESUMEN
The study evaluates cellulose nanocrystals (CNCs) as nanocarriers for targeted, intracellular delivery of molecular agents. CNCs were labeled with fluorescein-5'-isothiocyanate as an imaging agent and conjugated to folic acid (FA) as a targeting ligand. The CNC conjugates were characterized by UV-vis spectroscopy, ζ-potential analysis, dynamic light scattering, and atomic force microscopy. Cellular binding/uptake of the FA-conjugated CNCs by KB and MDA-MB-468 cells was quantified with cellular uptake assays. Internalization of the particles was confirmed by confocal microscopy. Uptake mechanisms were determined by inhibition studies with chlorpromazine and genistein. Binding affinity was qualitatively assessed with a free folate inhibition assay. Both KB and MDA-MB-468 cells exhibited significant and folate-receptor specific binding/uptake of FA-conjugated CNCs. Clathrin-mediated endocytosis was a significant uptake mechanism in both cell types, whereas caveolae-mediated endocytosis only played a significant role in MDA-MB-468 cells. Uptake inhibition of FA-conjugated CNCs by KB cells required high concentrations (>1 mM) of free FA. The observed FR-specific internalization of FA-conjugated CNCs by FR-positive cancer cells and tumors and their remarkable high affinity for the FR demonstrate the great potential of CNCs as novel nanocarriers for imaging agents and chemotherapeutics in the early detection and treatment of cancer.
RESUMEN
Residual lignin in pretreated biomass impedes enzymatic hydrolysis. Nonionic surfactants are known to enhance the enzymatic hydrolysis of lignocellulosic biomass but their mechanisms of action are incompletely understood. This study investigates the effect of a nonionic surfactant, Tween 80, on the adsorption of cellulases onto model lignin substrates. Lignin substrates were prepared by spin coating of flat substrates with three different types of lignin: organosolv lignin, kraft lignin, and milled wood lignin. The functional group distributions in the lignins were quantitatively analyzed by 31P NMR spectroscopy. The surface energies and surface roughnesses of the substrates were determined by contact angle measurements and atomic force microscopy, respectively. Tween 80 and cellulase adsorption onto the lignin substrates was analyzed with a quartz crystal microbalance with dissipation monitoring. Tween 80 adsorbed rapidly and primarily (≥85%) via dispersion interactions onto the lignin substrates and effected solubilization of lignin molecules, most notably with organosolv lignin, having the largest dispersive surface energy component and smallest molar mass. Cellulase adsorption onto the lignin substrates was mostly irreversible and had both a rapid and a gradual adsorption stage. Rapid cellulase adsorption was primarily (≥64%) mediated by dispersion interactions. The subsequent gradual mass increase is postulated to involve swelling of the lignin substrates. Adsorbed Tween 80 rendered lignin surfaces more hydrophilic by increasing their polar surface energy component and reduced both the extent of rapid cellulase adsorption as well as the rate of the subsequent gradual mass increase. The effect of Tween 80 on the rate and extent of the gradual mass increase depended strongly on the chemical properties of the lignin.
Asunto(s)
Celulasas/química , Lignina/química , Polisorbatos/química , Tensoactivos/química , Trichoderma/enzimología , Adsorción , Propiedades de Superficie , TermodinámicaRESUMEN
Cellulose nanocrystals are rod-shaped, crystalline nanoparticles that have shown prom-ise in a number of industrial applications for their unique chemical and physical properties. However, investigations of their abilities in the biomedical field are limited. The goal of this study is to show the potential use of folic acid-conjugated cellulose nanocrystals in the potentiation of irreversible electroporation-induced cell death in folate receptor (FR)-positive cancers. We optimized key pulse parameters including pulse duration, intensity, and incubation time with nanoparticles prior to electroporation. FR-positive cancer cells, KB and MDA-MB-468, were preincubated with cellulose nanocrystals (CNCs) conjugated with the targeting molecule folic acid (FA), 10 and 20 min respectively, prior to application of the optimized pulse electric field (PEF), 600 and 500 V/cm respectively. We have shown cellulose nanocrystals' ability to potentiate a new technique for tumor ablation, irreversible electroporation. Pre-incubation with FA-conjugated CNCs (CNC-FA) has shown a significant increase in cytotoxicity induced by irreversible electroporation in FR-positive cancer cells, KB and MDA-MB-468. Non-targeted CNCs (CNC-COOH) did not potentiate IRE when preincubated at the same parameters as previously stated in these cell types. In addition, CNC-FA did not potentiate irreversible electroporation-induced cytotoxicity in a FR-negative cancer cell type, A549. Without changing irreversible electroporation parameters it is possible to increase the cytotoxic effect on FR-positive cancer cells by exploiting the specific binding of FA to the FR, while not causing further damage to FR-negative tissue.