Centrifuge fractionation during purification of cellulose nanocrystals after acid hydrolysis and consequences on their chiral self-assembly.

The inherent colloidal dispersity (due to length, aspect ratio, surface charge heterogeneity) of CNCs, when produced using the typical traditional sulfuric acid hydrolysis route, presents a great challenge when interpreting colloidal properties and linking the CNC film nanostructure to the helicoidal self-assembly mechanism during drying. Indeed, further improvement of this CNC preparation route is required to yield films with better control over the CNC pitch and optical properties. Here we present a modified CNC-preparation protocol, by fractionating and harvesting CNCs with different average surface charges, rod lengths, aspect ratios, already during the centrifugation steps after hydrolysis. This enables faster CNC fractionation, because it is performed in a high ionic strength aqueous medium. By comparing dry films from the three CNC fractions, discrepancies in the CNC self-assembly and structural colors were clearly observed. Conclusively, we demonstrate a fast protocol to harvest different populations of CNCs, that enable tailored refinement of structural colors in CNC films.

Cellulose Nanofiber-Coated Perfluoropentane Droplets: Fabrication and Biocompatibility Study.

Purpose: To study the effect of cellulose nanofiber (CNF)-shelled perfluoropentane (PFP) droplets on the cell viability of 4T1 breast cancer cells with or without the addition of non-encapsulated paclitaxel. Methods: The CNF-shelled PFP droplets were produced by mixing a CNF suspension and PFP using a homogenizer. The volume size distribution and concentration of CNF-shelled PFP droplets were estimated from images taken with an optical microscope and analyzed using Fiji software and an in-house Matlab script. The thermal stability was qualitatively assessed by comparing the size distribution and concentration of CNF-shelled PFP droplets at room temperature (~22°) and 37°C. The cell viability of 4T1 cells was measured using a 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Additionally, a hemolysis assay was performed to assess blood compatibility of CNF-shelled PFP droplets. Results: The droplet diameter and concentration of CNF-shelled PFP droplets decreased after 48 hours at both room temperature and 37°C. In addition, the decrease in concentration was more significant at 37°C, from 3.50 ± 0.64×106 droplets/mL to 1.94 ± 0.10×106 droplets/mL, than at room temperature, from 3.65 ± 0.29×106 droplets/mL to 2.56 ± 0.22×106 droplets/mL. The 4T1 cell viability decreased with increased exposure time and concentration of paclitaxel, but it was not affected by the presence of CNF-shelled PFP droplets. No hemolysis was observed at any concentration of CNF-shelled PFP droplets. Conclusion: CNF-shelled PFP droplets have the potential to be applied as drug carriers in ultrasound-mediated therapy.

Measuring the Compressibility of Cellulose Nanofiber-Stabilized Microdroplets Using Acoustophoresis.

Droplets with a liquid perfluoropentane core and a cellulose nanofiber shell have the potential to be used as drug carriers in ultrasound-mediated drug delivery. However, it is necessary to understand their mechanical properties to develop ultrasound imaging sequences that enable in vivo imaging of the vaporization process to ensure optimized drug delivery. In this work, the compressibility of droplets stabilized with cellulose nanofibers was estimated using acoustophoresis at three different acoustic pressures. Polyamide particles of known size and material properties were used for calibration. The droplet compressibility was then used to estimate the cellulose nanofiber bulk modulus and compare it to experimentally determined values. The results showed that the acoustic contrast factor for these droplets was negative, as the droplets relocated to pressure antinodes during ultrasonic actuation. The droplet compressibility was 6.6-6.8 ×10-10 Pa-1, which is higher than for water (4.4×10-10 Pa-1) but lower than for pure perfluoropentane (2.7×10-9 Pa-1). The compressibility was constant across different droplet diameters, which was consistent with the idea that the shell thickness depends on the droplet size, rather than being constant.

Photoresponsive and Polarization-Sensitive Structural Colors from Cellulose/Liquid Crystal Nanophotonic Structures.

Cellulose nanocrystals (CNCs) possess the ability to form helical periodic structures that generate structural colors. Due to the helicity, such self-assembled cellulose structures preferentially reflect left-handed circularly polarized light of certain colors, while they remain transparent to right-handed circularly polarized light. This study shows that combination with a liquid crystal enables modulation of the optical response to obtain light reflection of both handedness but with reversed spectral profiles. As a result, the nanophotonic systems provide vibrant structural colors that are tunable via the incident light polarization. The results are attributed to the liquid crystal aligning on the CNC/glucose film, to form a birefringent layer that twists the incident light polarization before interaction with the chiral cellulose nanocomposite. Using a photoresponsive liquid crystal, this effect can further be turned off by exposure to UV light, which switches the nematic liquid crystal into a nonbirefringent isotropic phase. The study highlights the potential of hybrid cellulose systems to create self-assembled yet advanced photoresponsive and polarization-tunable nanophotonics.

A Study on the Acoustic Response of Pickering Perfluoropentane Droplets in Different Media.

Acoustic droplet vaporization (ADV) is the physical process of liquid-to-gas phase transition mediated by pressure variations in an ultrasound field. In this study, the acoustic response of novel particle-stabilized perfluoropentane droplets was studied in bulk and confined media. The oil/water interface was stabilized by cellulose nanofibers. First, their acoustic responses under idealized conditions were examined to assess their susceptibility to undergo ADV. Second, the droplets were studied in a more realistic setting and placed in a confined medium. Lastly, an imaging setup was developed and tested on the droplets. The acoustic response could be seen when the amplitude of the peak negative pressure (PNP) was above 200 kPa, suggesting that this is the vaporization pressure threshold for these droplets. Increasing the PNP resulted in a decrease in signal intensity over time, suggesting a more destructive behavior. The imaging setup was able to differentiate between the droplets and the surrounding tissue. Results obtained within this study suggest that these droplets have potential in terms of ultrasound-mediated diagnostics and therapy.

Floating Photocatalysts for Effluent Refinement Based on Stable Pickering Cellulose Foams and Graphitic Carbon Nitride (g-C3N4).

The transfer of heterogeneous photocatalysis applications from the laboratory to real-life aqueous systems is challenging due to the higher density of photocatalysts compared to water, light attenuation effects in water, complicated recovery protocols, and metal pollution from metal-based photocatalysts. In this work, we overcome these obstacles by developing a buoyant Pickering photocatalyst carrier based on green cellulose nanofibers (CNFs) derived from wood. The air bubbles in the carrier were stable because the particle surfactants provided thermodynamic stability and the derived photocatalytic foams floated on water throughout the test period (4 weeks). A metal-free semiconductor photocatalyst, g-C3N4, was facilely embedded inside the foam by mixing the photocatalyst with the air-bubble suspension followed by casting and drying to produce solid foams. When tested under mild irradiation conditions (visible light, low energy LEDs) and no agitation, almost three times more dye was removed after 6 h for the floating g-C3N4-CNF nanocomposite foam, compared to the pure g-C3N4 powder residing on the bottom of a ca. 2 cm-high water pillar. The buoyancy and physicochemical properties of the carrier material were imperative to render escalated oxygenation, high photon utilization, and faster dye degradation. The reported assembly protocol is facile, general, and provides a new strategy for assembling green floating foams that can potentially carry a number of different photocatalysts.

Unravelling the Acoustic and Thermal Responses of Perfluorocarbon Liquid Droplets Stabilized with Cellulose Nanofibers.

The attractive colloidal and physicochemical properties of cellulose nanofibers (CNFs) at interfaces have recently been exploited in the facile production of a number of environmentally benign materials, e.g. foams, emulsions, and capsules. Herein, these unique properties are exploited in a new type of CNF-stabilized perfluoropentane droplets produced via a straightforward and simple mixing protocol. Droplets with a comparatively narrow size distribution (ca. 1-5 µm in diameter) were fabricated, and their potential in the acoustic droplet vaporization process was evaluated. For this, the particle-stabilized droplets were assessed in three independent experimental examinations, namely temperature, acoustic, and ultrasonic standing wave tests. During the acoustic droplet vaporization (ADV) process, droplets were converted to gas-filled microbubbles, offering enhanced visualization by ultrasound. The acoustic pressure threshold of about 0.62 MPa was identified for the cellulose-stabilized droplets. A phase transition temperature of about 22 °C was observed, at which a significant fraction of larger droplets (above ca. 3 µm in diameter) were converted into bubbles, whereas a large part of the population of smaller droplets were stable up to higher temperatures (temperatures up to 45 °C tested). Moreover, under ultrasound standing wave conditions, droplets were relocated to antinodes demonstrating the behavior associated with the negative contrast particles. The combined results make the CNF-stabilized droplets interesting in cell-droplet interaction experiments and ultrasound imaging.

Using dextran of different molecular weights to achieve faster freeze-drying and improved storage stability of lactate dehydrogenase.

Freeze-drying of protein formulations is frequently used to maintain protein activity during storage. The freeze-drying process usually requires long primary drying times because the highest acceptable drying temperature to obtain acceptable products is dependent on the glass transition temperature of the maximally freeze-concentrated solution (Tg'). On the other hand, retaining protein activity during storage is related to the glass transition temperature (Tg) of the final freeze-dried product. In this study, dextrans with different molecular weight (1 and 40 kDa) and mixtures thereof at the ratio 3:1, 1:1, and 1:3 (w/w) were used as cryo-/lyoprotectant and their impact on the stability of the model protein lactate dehydrogenase (LDH) was investigated at elevated temperatures (40 °C and 60 °C). The dextran formulations were then compared to formulations containing sucrose as cryo-/lyoprotectant. Because of the higher Tg' values of the dextrans, the primary drying times could be reduced compared to freeze-drying with sucrose. Similarly, the higher Tg and Tg' of dextrans relative to sucrose led to benefits during storage which was shown through improved protection of LDH activity.

Porous Cellulose Nanofiber-Based Microcapsules for Biomolecular Sensing.

Cellulose nanofibers (CNFs) have recently attracted a lot of attention in sensing because of their multifunctional character and properties such as renewability, nontoxicity, biodegradability, printability, and optical transparency in addition to unique physicochemical, barrier, and mechanical properties. However, the focus has exclusively been devoted toward developing two-dimensional sensing platforms in the form of nanopaper or nanocellulose-based hydrogels. To improve the flexibility and sensing performance in situ, for example, to detect biomarkers in vivo for early disease diagnostics, more advanced CNF-based structures are needed. Here, we developed porous and hollow, yet robust, CNF-based microcapsules using only the primary plant cell wall components, CNF, pectin, and xyloglucan, to assemble the capsule wall. The fluorescein isothiocyanate-labeled dextrans with MW of 70 and 2000 kDa could enter the hollow capsules at a rate of 0.13 ± 0.04 and 0.014 ± 0.009 s-1, respectively. This property is very attractive because it minimizes the influence of mass transport through the capsule wall on the response time. As a proof of concept, glucose oxidase (GOx) enzyme was loaded (and cross-linked) in the microcapsule interior with an encapsulation efficiency of 68 ± 2%. The GOx-loaded microcapsules were immobilized on a variety of surfaces (here, inside a flow channel, on a carbon-coated sensor or a graphite rod) and glucose concentrations up to 10 mM could successfully be measured. The present concept offers new opportunities in the development of simple, more efficient, and disposable nanocellulose-based analytical devices for several sensing applications including environmental monitoring, healthcare, and diagnostics.

Toward Improved Understanding of the Interactions between Poorly Soluble Drugs and Cellulose Nanofibers.

Cellulose nanofibers (CNFs) have interesting physicochemical and colloidal properties that have been recently exploited in novel drug-delivery systems for tailored release of poorly soluble drugs. The morphology and release kinetics of such drug-delivery systems heavily relied on the drug-CNF interactions; however, in-depth understanding of the interactions was lacking. Herein, the interactions between a poorly soluble model drug molecule, furosemide, and cationic cellulose nanofibers with two different degrees of substitution are studied by sorption experiments, Fourier transform infrared spectroscopy, and molecular dynamics (MD) simulation. Both MD simulations and experimental results confirmed the spontaneous sorption of drug onto CNF. Simulations further showed that adsorption occurred by the flat aryl ring of furosemide. The spontaneous sorption was commensurate with large entropy gains as a result of release of surface-bound water. Association between furosemide molecules furthermore enabled surface precipitation as indicated by both simulations and experiments. Finally, sorption was also found not to be driven by charge neutralization, between positive CNF surface charges and the furosemide negative charge, so that surface area is the single most important parameter determining the amount of sorbed drug. An optimized CNF-furosemide drug-delivery vehicle thus needs to have a maximized specific surface area irrespective of the surface charge with which it is achieved. The findings also provide important insights into the design principles of CNF-based filters suitable for removal of poorly soluble drugs from wastewater.

Superamphiphobic coatings based on liquid-core microcapsules with engineered capsule walls and functionality.

Microcapsules with specific functional properties, related to the capsule wall and core, are highly desired in a number of applications. In this study, hybrid cellulose microcapsules (1.2 ± 0.4 µm in diameter) were prepared by nanoengineering the outer walls of precursor capsules. Depending on the preparation route, capsules with different surface roughness (raspberry or broccoli-like), and thereby different wetting properties, could be obtained. The tunable surface roughness was achieved as a result of the chemical and structural properties of the outer wall of a precursor capsule, which combined with a new processing route allowed in-situ formation of silica nanoparticles (30-40 nm or 70 nm in diameter). By coating glass slides with "broccoli-like" microcapsules (30-40 nm silica nanoparticles), static contact angles above 150° and roll-off angles below 6° were obtained for both water and low surface-tension oil (hexadecane), rendering the substrate superamphiphobic. As a comparison, coatings from raspberry-like capsules were only strongly oleophobic and hydrophobic. The liquid-core of the capsules opens great opportunities to incorporate different functionalities and here hydrophobic superparamagnetic nanoparticles (SPIONs) were encapsulated. As a result, magnetic broccoli-like microcapsules formed an excellent superamphiphobic coating-layer on a curved geometry by simply applying an external magnetic field.

Cellulose nanofibers as excipient for the delivery of poorly soluble drugs.

Poor aqueous solubility of drugs is becoming an increasingly pronounced challenge in the formulation and development of drug delivery systems. To overcome the limitations associated with these problematic drugs, formulation scientists are required to use enabling strategies which often demands the use of new excipients. Cellulose nanofibers (CNFs) is such an excipient and it has only recently been described in the pharmaceutical field. In this review, the use of CNF in drug formulation with a focus on poorly soluble drugs is featured. In particular, the aim is to describe and discuss the many unique properties of CNFs, which make CNFs attractive as excipients in pharmaceutical sciences. Furthermore, the use of CNF as stabilizers for crystalline drug nanoparticles, as a matrix former to obtain a long-lasting sustained drug release over several weeks and as a film former with immediate release properties for poorly soluble drug are reported. Finally, the preparation of pharmaceutical CNF foams together with poorly soluble drugs is highlighted; foams, which offer a sustained drug delivery system with positive buoyancy.

Bioinspired Layer-by-Layer Microcapsules Based on Cellulose Nanofibers with Switchable Permeability.

Green, all-polysaccharide based microcapsules with mechanically robust capsule walls and fast, stimuli-triggered, and switchable permeability behavior show great promise in applications based on selective and timed permeability. Taking a cue from nature, the build-up and composition of plant primary cell walls inspired the capsule wall assembly, because the primary cell walls in plants exhibit high mechanical properties despite being in a highly hydrated state, primarily owing to cellulose microfibrils. The microcapsules (16 ± 4 µm in diameter) were fabricated using the layer-by-layer technique on sacrificial CaCO3 templates, using plant polysaccharides (pectin, cellulose nanofibers, and xyloglucan) only. In water, the capsule wall was permeable to labeled dextrans with a hydrodynamic diameter of ∼6.6 nm. Upon exposure to NaCl, the porosity of the capsule wall quickly changed allowing larger molecules (∼12 nm) to permeate. However, the porosity could be restored to its original state by removal of NaCl, by which permeants became trapped inside the capsule's core. The high integrity of cell wall was due to the CNF and the ON/OFF alteration of the permeability properties, and subsequent loading/unloading of molecules, could be repeated several times with the same capsule demonstrating a robust microcontainer with controllable permeability properties.

Rhamnogalacturonan-I Based Microcapsules for Targeted Drug Release.

Drug targeting to the colon via the oral administration route for local treatment of e.g. inflammatory bowel disease and colonic cancer has several advantages such as needle-free administration and low infection risk. A new source for delivery is plant-polysaccharide based delivery platforms such as Rhamnogalacturonan-I (RG-I). In the gastro-intestinal tract the RG-I is only degraded by the action of the colonic microflora. For assessment of potential drug delivery properties, RG-I based microcapsules (~1 µm in diameter) were prepared by an interfacial poly-addition reaction. The cross-linked capsules were loaded with a fluorescent dye (model drug). The capsules showed negligible and very little in vitro release when subjected to media simulating gastric and intestinal fluids, respectively. However, upon exposure to a cocktail of commercial RG-I cleaving enzymes, ~ 9 times higher release was observed, demonstrating that the capsules can be opened by enzymatic degradation. The combined results suggest a potential platform for targeted drug delivery in the terminal gastro-intestinal tract.

Solid cellulose nanofiber based foams - Towards facile design of sustained drug delivery systems.

Control of drug action through formulation is a vital and very challenging topic within pharmaceutical sciences. Cellulose nanofibers (CNF) are an excipient candidate in pharmaceutical formulations that could be used to easily optimize drug delivery rates. CNF has interesting physico-chemical properties that, when combined with surfactants, can be used to create very stable air bubbles and dry foams. Utilizing this inherent property, it is possible to modify the release kinetics of the model drug riboflavin in a facile way. Wet foams were prepared using cationic CNF and a pharmaceutically acceptable surfactant (lauric acid sodium salt). The drug was suspended in the wet-stable foams followed by a drying step to obtain dry foams. Flexible cellular solid materials of different thicknesses, shapes and drug loadings (up to 50wt%) could successfully be prepared. The drug was released from the solid foams in a diffusion-controlled, sustained manner due to the presence of intact air bubbles which imparted a tortuous diffusion path. The diffusion coefficient was assessed using Franz cells and shown to be more than one order of magnitude smaller for the cellular solids compared to the bubble-free films in the wet state. By changing the dimensions of dry foams while keeping drug load and total weight constant, the drug release kinetics could be modified, e.g. a rectangular box-shaped foam of 8mm thickness released only 59% of the drug after 24h whereas a thinner foam sample (0.6mm) released 78% of its drug content within 8h. In comparison, the drug release from films (0.009mm, with the same total mass and an outer surface area comparable to the thinner foam) was much faster, amounting to 72% of the drug within 1h. The entrapped air bubbles in the foam also induced positive buoyancy, which is interesting from the perspective of gastroretentive drug-delivery.

Cellulose-nanofiber/polygalacturonic acid coatings with high oxygen barrier and targeted release properties.

A bio-inspired coating consisting of pectin (polygalacturonic acid) and cationic cellulose nanofibers were successfully produced by the layer-by-layer method. The build-up and the morphology of the resulting coatings were studied with spectroscopic ellipsometry and atomic force microscopy, respectively. The coating was able to survive the exposure of a simulated gastric fluid, but was partially degraded upon exposure to pectinase enzyme, which simulate the action of the microbial symbionts present in the human colon. Prior to exposure, the oxygen permeability coefficient of the coating (0.033 ml(STP)mmm(-2)day(-1)atm(-1) at 23°C and 20% RH) was in the same order of magnitude as for ethylene vinyl alcohol films (0.001-0.01 ml(STP)mmm(-2)day(-1)atm(-1)). However, after exposure to the mimicked gastrointestinal (GI) tract conditions, the contribution of coating to the overall barrier properties was not measurable.

Photon energy upconverting nanopaper: a bioinspired oxygen protection strategy.

The development of solid materials which are able to upconvert optical radiation into photons of higher energy is attractive for many applications such as photocatalytic cells and photovoltaic devices. However, to fully exploit triplet-triplet annihilation photon energy upconversion (TTA-UC), oxygen protection is imperative because molecular oxygen is an ultimate quencher of the photon upconversion process. So far, reported solid TTA-UC materials have focused mainly on elastomeric matrices with low barrier properties because the TTA-UC efficiency generally drops significantly in glassy and semicrystalline matrices. To overcome this limit, for example, combine effective and sustainable annihilation upconversion with exhaustive oxygen protection of dyes, we prepare a sustainable solid-state-like material based on nanocellulose. Inspired by the structural buildup of leaves in Nature, we compartmentalize the dyes in the liquid core of nanocellulose-based capsules which are then further embedded in a cellulose nanofibers (NFC) matrix. Using pristine cellulose nanofibers, a sustainable and environmentally friendly functional nanomaterial with ultrahigh barrier properties is achieved. Also, an ensemble of sensitizers and emitter compounds are encapsulated, which allow harvesting of the energy of the whole deep-red sunlight region. The films demonstrate excellent lifetime in synthetic air (20.5/79.5, O2/N2)-even after 1 h operation, the intensity of the TTA-UC signal decreased only 7.8% for the film with 8.8 µm thick NFC coating. The lifetime can be further modulated by the thickness of the protective NFC coating. For comparison, the lifetime of TTA-UC in liquids exposed to air is on the level of seconds to minutes due to fast oxygen quenching.

Cellulose nanofiber/nanocrystal reinforced capsules: a fast and facile approach toward assembly of liquid-core capsules with high mechanical stability.

Liquid-core capsules of high mechanical stability open up for many solid state-like applications where functionality depending on liquid mobility is vital. Herein, a novel concept for fast and facile improvement of the mechanical properties of walls of liquid-core capsules is reported. By imitating nature's own way of enhancing the mechanical properties in liquid-core capsules, the parenchyma plant cells found in fruits and vegetables, a blend of short cellulose nanofibers (<1 µm, NFC) and nanocrystals (CNC) was exploited in the creation of the capsule walls. The NFC/CNC blend was prepared from a new version of the classical wood pulp hydrolysis. The capsule shell consisted of a covalently (by aromatic diisocyanate) cross-linked NFC/CNC structure at the outer capsule wall and an inner layer dominated by aromatic polyurea. The mechanical properties revealed an effective capsule elastic modulus of 4.8 GPa at 17 wt % NFC/CNC loading, about six times higher compared to a neat aromatic polyurea capsule (0.79 GPa) and 3 orders of magnitude higher than previously reported capsules from regenerated cellulose (0.0074 GPa). The outstanding mechanical properties are ascribed to the dense nanofiber structure, present in the outer part of the capsule wall, that is formed by oriented NFC/CNC of high average aspect ratio (L/d ∼ 70) and held together by both covalent (urethane bonds) and physical bonds (hydrogen bonds).

Transparent films based on PLA and montmorillonite with tunable oxygen barrier properties.

Polylactide (PLA) is viewed as a potential material to replace synthetic plastics (e.g., poly(ethylene terephthalate) (PET)) in food packaging, and there have been a number of developments in this direction. However, for PLA to be competitive in more demanding uses such as the packaging of oxygen-sensitive foods, the oxygen permeability coefficient (OP) needs to be reduced by a factor of ~10. To achieve this, a layer-by-layer (Lbl) approach was used to assemble alternating layers of montmorillonite clay and chitosan on extruded PLA film surfaces. When 70 bilayers were applied, the OP was reduced by 99 and 96%, respectively, at 20 and 50% RH. These are, to our knowledge, the best improvements in oxygen barrier properties ever reported for a PLA/clay-based film. The process of assembling such multilayer structures was characterized using a quartz crystal microbalance with dissipation monitoring. Transmission electron microscopy revealed a well-ordered laminar structure in the deposited multilayer coatings, and light transmittance results demonstrated the high optical clarity of the coated PLA films.

Cellulose nanocomposite biopolymer foam--hierarchical structure effects on energy absorption.

Starch is an attractive biofoam candidate as replacement of expanded polystyrene (EPS) in packaging materials. The main technical problems with starch foam include its hygroscopic nature, sensitivity of its mechanical properties to moisture content, and much lower energy absorption than EPS. In the present study, a starch-based biofoam is for the first time able to reach comparable mechanical properties (E = 32 MPa, compressive yield strength, 630 kPa) to EPS at 50% relative humidity and similar relative density. The reason is the nanocomposite concept in the form of a cellulose nanofiber network reinforcing the hygroscopic amylopectin starch matrix in the cell wall. The biofoams are prepared by the freezing/freeze-drying technique and subjected to compressive loading. Cell structure is characterized by FE-SEM of cross sections. Mechanical properties are related to cell structure and cell wall nanocomposite composition. Hierarchically structured biofoams are demonstrated to be interesting materials with potential for strongly improved mechanical properties.