Stable Cellulose Nanofibril Microcapsules from Pickering Emulsion Templates.

Electrostatic attractions are essential in any complex formation between the nanofibrils of the opposite charge for a specific application, such as microcapsule production. Here, we used cationized cellulose nanofibril (CCNF)-stabilized Pickering emulsions (PEs) as templates, and the electrostatic interactions were induced by adding oxidized cellulose nanofibrils (OCNFs) at the oil-water interface to form microcapsules (MCs). The oppositely charged cellulose nanofibrils enhanced the solidity of interfaces, allowing the encapsulation of Nile red (NR) in sunflower oil droplets. Microcapsules exhibited a low and controlled release of NR at room temperature. Furthermore, membrane emulsification was employed to scale up the preparation of microcapsules with sunflower oil (SFO) encapsulated by CCNF/OCNF complex networks.

Salt-Responsive Pickering Emulsions Stabilized by Functionalized Cellulose Nanofibrils.

Oil-in-water emulsions have been stabilized by functionalized cellulose nanofibrils bearing either a negative (oxidized cellulose nanofibrils, OCNF) or a positive (cationic cellulose nanofibrils, CCNF) surface charge. The size of the droplets was measured by laser diffraction, while the structure of the shell of the Pickering emulsion droplets was probed using small-angle neutron scattering (SANS), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), and rheology measurements. Both OCNF- and CCNF-stabilized emulsions present a very thick shell (>100 nm) comprised of densely packed CNF. OCNF-stabilized emulsions proved to be salt responsive, influencing the droplet aggregation and ultimately the gel properties of the emulsions, while CCNF emulsions, on the other hand, showed very little salt-dependent behavior.

Monovalent Salt and pH-Induced Gelation of Oxidised Cellulose Nanofibrils and Starch Networks: Combining Rheology and Small-Angle X-ray Scattering.

Water quality parameters such as salt content and various pH environments can alter the stability of gels as well as their rheological properties. Here, we investigated the effect of various concentrations of NaCl and different pH environments on the rheological properties of TEMPO-oxidised cellulose nanofibril (OCNF) and starch-based hydrogels. Addition of NaCl caused an increased stiffness of the OCNF:starch (1:1 wt%) blend gels, where salt played an important role in reducing the repulsive OCNF fibrillar interactions. The rheological properties of these hydrogels were unchanged at pH 5.0 to 9.0. However, at lower pH (4.0), the stiffness and viscosity of the OCNF and OCNF:starch gels appeared to increase due to proton-induced fibrillar interactions. In contrast, at higher pH (11.5), syneresis was observed due to the formation of denser and aggregated gel networks. Interactions as well as aggregation behaviour of these hydrogels were explored via ζ-potential measurements. Furthermore, the nanostructure of the OCNF gels was probed using small-angle X-ray scattering (SAXS), where the SAXS patterns showed an increase of slope in the low-q region with increasing salt concentration arising from aggregation due to the screening of the surface charge of the fibrils.

Effect of varying the Mg with Ca content in highly porous phosphate-based glass microspheres.

This paper reports on the role of phosphate-based glass (PBG) microspheres and their physicochemical properties including in vitro biological response to human mesenchymal stem cells (hMSCs). Solid and porous microspheres were prepared via a flame spheroidisation process. The Mg content in the PBG formulations explored was reduced from 24 to 2 mol% with a subsequent increase in Ca content. A small quantity of TiO2 (1 mol%) was added to the lower Mg-content glass (2 mol%) to avoid crystallisation. Morphological and physical characterisation of porous microspheres revealed interconnected porosity (up to 76 ± 5 %), average external pore sizes of 55 ± 5 µm with surface areas ranging from 0.38 to 0.43 m2 g-1. Degradation and ion release studies conducted compared the solid (non-porous) and porous microspheres and revealed 1.5 to 2.5 times higher degradation rate for porous microspheres. Also, in vitro bioactivity studies using simulated body fluid (SBF) revealed Ca/P ratios for porous microspheres of all three glass formulations were between 0.75 and 0.92 which were within the range suggested for precipitated amorphous calcium phosphate. Direct cell seeding and indirect cell culture studies (via incubation with microsphere degradation products) revealed hMSCs were able to grow and undergo osteogenic differentiation in vitro, confirming cytocompatibility of the formulations tested. However, the higher Mg content (24 mol%) porous microsphere showed the most potent osteogenic response and is therefore considered as a promising candidate for bone repair applications.

Non-volatile conductive gels made from deep eutectic solvents and oxidised cellulose nanofibrils.

Ionogels offer huge potential for a number of applications including wearable electronics and soft sensors. However, their synthesis has been limited and often relies on non-renewable or non-biocompatible components. Here we present a novel two-component ionogel made using just deep eutectic solvents (DESs) and cellulose. DESs offer a non-volatile alternative to hydrogels with highly tuneable properties including conductivity and solvation of compounds with widely varying hydrophobicity. DESs can be easily made from cheap, biodegradable and biocompatible components. This research presents the characterisation of a series of soft conductive gels made from deep eutectic solvents (DESs), specifically choline chloride-urea and choline chloride-glycerol, with the sole addition of TEMPO-oxidised cellulose nanofibrils (OCNF). A more liquid-like rather than gel-like conductive material could be made by using the DES betaine-glycerol. OCNF are prepared from sustainable sources, and are non-toxic, and mild on the skin, forming gels without the need for surfactants or other gelling agents. These DES-OCNF gels are shear thinning with conductivities up to 1.7 mS cm-1 at ∼26 °C. Given the thousands of possible DESs, this system offers unmatched tunability and customisation for properties such as viscosity, conductivity, and yield behaviour.

Recent progress in Pickering emulsions stabilised by bioderived particles.

In recent years, the demand for non-surfactant based Pickering emulsions in many industrial applications has grown significantly because of the option to select biodegradable and sustainable materials with low toxicity as emulsion stabilisers. Usually, emulsions are a dispersion system, where synthetic surfactants or macromolecules stabilise two immiscible phases (typically water and oil phases) to prevent coalescence. However, synthetic surfactants are not always a suitable choice in some applications, especially in pharmaceuticals, food and cosmetics, due to toxicity and lack of compatibility and biodegradability. Therefore, this review reports recent literature (2018-2021) on the use of comparatively safer biodegradable polysaccharide particles, proteins, lipids and combinations of these species in various Pickering emulsion formulations. Also, an overview of the various tuneable factors associated with the functionalisation or surface modification of these solid particles, that govern the stability of the Pickering emulsions is provided.

Long-Term Culture of Stem Cells on Phosphate-Based Glass Microspheres: Synergistic Role of Chemical Formulation and 3D Architecture.

Phosphate-based glasses (PBGs) are biomaterials that degrade under physiological conditions and can be modified to release various ions depending on end applications. This study utilized slow-degrading (P45:45P2O5-16CaO-24MgO-11Na2O-4Fe2O3, mol %) and comparatively faster degrading (P40:40P2O5-16CaO-24MgO-20Na2O, mol %) PBG microspheres with or without porosity, to evaluate the combined effect of chemical formulation and geometry on human mesenchymal stem cells (MSCs), a clinically relevant cell source for orthopedic applications. Scanning electron microscopy showed 2, 46, and 29% of P45 bulk (P45-B), P40 bulk (P40-B), and P40 porous (P40-P) microspheres, respectively, that had cracks or peeling off surfaces after 42 days of incubation in culture medium. Cytotoxicity assessment showed that glass debris released into the culture medium may interact with cells and affect their survival. Direct-contact cell experiments up to 42 days showed that P45-B microspheres did not sustain viable long-term cell cultures and did not facilitate extracellular matrix formation. On the other hand, P40-B microspheres enhanced alkaline phosphatase activity, calcium deposition, and collagen and osteocalcin production in MSCs. Introduction of porosity in P40 glass further enhanced these parameters and proliferation at later time points. The small pore windows (<5 µm wide) and interconnection (<10 µm wide) may have allowed limited cell penetration into the porous structures. P40-B and P40-P have potential for bone repair and reinforcement therapy based on their chemical formulation and porous geometry.

Self-assembly of amphiphilic polyoxometalates for the preparation of mesoporous polyoxometalate-titania catalysts.

Amphiphilic polyoxometalate (POM) surfactants were prepared by covalently grafting double hydrophobic tails with chain lengths C12H25, C14H29, C16H33 or C18H37 onto the lacunary Wells-Dawson {P2W17O61} headgroup. The critical micelle concentrations (CMCs) of these novel surfactants in aqueous solutions were determined by conductivity, and micelle formation was studied by small angle neutron scattering (SANS). Surprisingly, the amphiphiles with longer hydrophobic tails tend to form less elongated and more globular micelles in water. The self-assembled amphiphilic polyoxometalates were used as templates in the hydrothermal synthesis of mesoporous TiO2 containing dispersed, immobilised {P2W17O61} units, which showed enhanced activity for the photodegradation of rhodamine B (RhB). The catalyst was recycled eight times with no loss of efficiency, demonstrating the stability of the hybrid structure. The amphiphilic polyoxometalates, therefore have excellent potential for the synthesis of various types of catalytically active porous materials.

Impact of wormlike micelles on nano and macroscopic structure of TEMPO-oxidized cellulose nanofibril hydrogels.

In this work, we investigated the effect of adding surfactant mixtures on the rheological properties of TEMPO-oxidized cellulose nanofibril (OCNF) saline dispersions. Three surfactant mixtures were studied: cocamidopropyl betaine (CAPB)/sodium dodecyl sulfate (SDS), which forms wormlike micelles (WLMs); cocamidopropylamine oxide (CAPOx)/SDS, which forms long rods; and CAPB/sodium lauroyl sarcosinate (SLS), which forms spherical micelles. The presence of micelles in these surfactant mixtures, independent of their morphology, leads to an increase of tan δ, making the gels less solid-like, therefore acting as a plasticizer. WLMs were able to suppress strain stiffening normally observed in OCNF gels at large strains. OCNF/WLM gels have lower G' values than OCNF gels while the other micellar morphologies have a reduced impact on G'. The presence of unconnected micelles leads to increased dissipative deformation in OCNF gels without affecting the connectivity of the fibrils, while the presence of entangled micelles interferes with the OCNF network.

Core-Shell Spheroidal Hydrogels Produced via Charge-Driven Interfacial Complexation.

Through charge-driven interfacial complexation, we produced millimeter-sized spheroidal hydrogels (SH) with a core-shell structure allowing long-term stability in aqueous media. The SH were fabricated by extruding, dropwise, a cationic cellulose nanofibril (CCNF) dispersion into an oppositely charged poly(acrylic acid) (PAA) bath. The SH have a solid-like CCNF-PAA shell, acting as a semipermeable membrane, and a liquid-like CCNF suspension in the core. Swelling behavior of the SH was dependent on the osmotic pressure of the aging media. Swelling could be suppressed by increasing the ionic strength of the media as this enhanced interfibrillar interactions and thus strengthened the outer gel membrane. We further validated a potential application of SH as reusable matrixes for glucose oxidase (GOx) entrapment, where the SH work as microreactors from which substrate and product are freely able to migrate through the SH shell while avoiding enzyme leakage.

Filler size effect in an attractive fibrillated network: a structural and rheological perspective.

The effect of the filler size on the structural and mechanical properties of an attractive fibrillated network composed of oxidised cellulose nanofibrils (OCNF) in water was investigated. Silica nanoparticles with a diameter of ca. 5 nm (SiNp5) and and ca. 158 nm (SiNp158) were chosen as non-interacting fillers of the OCNF network. These filler sizes were chosen, respectively, to have a particle size which was either similar to that of the network mesh size or much larger than it. Contrast matched small angle neutron scattering (SANS) experiments revealed that the presence of the fillers (SiNp5 and SiNp158) did not perturb the structural properties of the OCNF network at the nanometer scale. However, the filler size difference strongly affected the mechanical properties of the hydrogel upon large amplitude oscillatory shear. The presence of the smaller filler, SiNp5, preserved the mechanical properties of the hydrogels, while the larger filler, SiNp158, allowed a smoother breakage of the network and low network recoverability after breakage. This study showed that the filler-to-mesh size ratio, for non-interacting fillers, is pivotal for tailoring the non-linear mechanical properties of the gel, such as yielding and flow.

Cationic surfactants as a non-covalent linker for oxidised cellulose nanofibrils and starch-based hydrogels.

Rheological properties of hydrogels composed of TEMPO-oxidised cellulose nanofibrils (OCNF)-starch in the presence of cationic surfactants were investigated. The cationic surfactants dodecyltrimethylammonium bromide (DTAB) and cetyltrimethylammonium bromide (CTAB) were used to trigger gelation of OCNF at around 5 mM surfactant. As OCNF and DTAB/CTAB are oppositely charged, an electrostatic attraction is suggested to explain the gelation mechanism. OCNF (1 wt%) and soluble starch (0.5 and 1 wt%) were blended to prepare hydrogels, where the addition of starch to the OCNF resulted in a higher storage modulus. Starch polymers were suggested to form networks with cellulose nanofibrils. The stiffness and viscosity of OCNF-Starch hydrogels were enhanced further by the addition of cationic surfactants (5 mM of DTAB/CTAB). ζ -potential and amylose-iodine complex analyses were also conducted to confirm surface charge and interaction of OCNF-starch-surfactant in order to provide an in-depth understanding of the surfactant-induced gel networks.

Deep eutectic solvent in water pickering emulsions stabilised by cellulose nanofibrils.

Deep eutectic solvent (menthol : dodecanoic acid) in water (30 : 70) emulsions stabilised with partially oxidised cellulose nanoparticles remained stable for 200 days at room temperature. Deep eutectic-based emulsions offer potential for non-aqueous reaction systems, chemical extraction, and controlled release. Pickering emulsions using polysaccharides are less toxic and more stable than surfactant-stabilised emulsions.

The effect of MgO/TiO2 on structural and crystallization behavior of near invert phosphate-based glasses.

Varying formulations in the glass system of 40P2 O5 ─(24 - x)MgO─(16 + x)CaO─(20 - y)Na2 O─yTiO2 (where 0 ≤ x ≤ 22 and y = 0 or 1) were prepared via melt-quenching. The structure of the glasses was confirmed by X-ray diffraction (XRD), Fourier transform infrared (FTIR), micro Raman and solid-state nuclear magnetic resonance (NMR) spectroscopies. The thermal properties and the activation energy of crystallization (Ec ) were measured using thermal analysis and the Kissinger equation, respectively. The glass forming ability of the formulations investigated was seen to decrease with reducing MgO content down to 8 mol% and the glass stability region also decreased from 106 to 90°C with decreasing MgO content. The activation energy of crystallization (Ec ) values also decreased from 248 (for 24 mol% MgO glass) to 229 kJ/mol (for the 8 mol% MgO content) with the replacement of MgO by CaO for glasses with no TiO2 . The formulations containing less than 8 mol% MgO without TiO2 showed a strong tendency toward crystallization. However, the addition of 1 mol% TiO2 in place of Na2 O for these glasses with less than 8 mol% MgO content, inhibited their crystallization tendency. Glasses containing 8 mol% MgO with 1 mol% TiO2 revealed a 12°C higher glass transition temperature, a 14°C increase in glass stability against crystallization and a 38 kJ/mol increase in Ec in comparison to their non TiO2 containing counterpart. NMR spectroscopy revealed that all of the formulations contained almost equal percentages of Q1 and Q2 species. However, FTIR and Raman spectroscopies showed that the local structure of the glasses had been altered with addition of 1 mol% TiO2 , which acted as a network modifier, impeding crystallization by increasing the cross-linking between phosphate chains and consequently leading to increased Ec as well as their glass forming ability.

Formulating injectable pastes of porous calcium phosphate glass microspheres for bone regeneration applications.

Current trends in regenerative medicine treatments for bone repair applications focus on cell-based therapies. These aim to deliver the treatment via a minimally invasive injection to reduce patient trauma and to improve efficacy. This paper describes the injectability of porous calcium phosphate glass microspheres to be used for bone repair based on their formulation, rheology and flow behavior. The use of excipients (xanthan gum, methyl cellulose and carboxyl methyl cellulose) were investigated to improve flow performance. Based on our results, the flow characteristics of the glass microsphere pastes vary according to particle size, surface area, and solid to liquid ratio, as well as the concentration of viscosity modifiers used. The optimal flow characteristics of calcium phosphate glass microsphere pastes was found to contain 40 mg/mL of xanthan gum which increased viscosity whilst providing elastic properties (∼29,000 Pa) at shear rates that mirror the injection process and the resting period post injection, preventing the glass microspheres from both damage and dispersion. It was established that a base formulation must contain 1 g of glass microspheres (60-125 µm in size) per 1 mL of cell culture media, or 0.48 g of glass microspheres of sizes between 125 and 200 µm. Furthermore, the glass microsphere formulations with xanthan gum were readily injectable via a syringe-needle system (3-20 mL, 18G and 14G needles), and have the potential to be utilized as a cell (or other biologics) delivery vehicle for bone regeneration applications.

Charge-driven interfacial gelation of cellulose nanofibrils across the water/oil interface.

Interfacial gels, obtained by the interaction of water-dispersible oxidised cellulose nanofibrils (OCNF) and oil-soluble oleylamine (OA), were produced across water/oil (W/O) interfaces. Surface rheology experiments showed that the complexation relies on the charge coupling between the negatively-charged OCNF and OA. Complexation across the W/O interface was found to be dependent on the ζ-potential of the OCNF (modulated by electrolyte addition), leading to different interfacial properties. Spontaneous OCNF adsorption at the W/O interface occurred for particles with ζ-potential more negative than -30 mV, resulting in the formation of interfacial gels; whilst for particles with ζ-potential of ca. -30 mV, spontaneous adsorption occurred, coupled with augmented interfibrillar interactions, yielding stronger and tougher interfacial gels. On the contrary, charge neutralisation of OCNF (ζ-potential values more positive than -30 mV) did not allow spontaneous adsorption of OCNF at the W/O interface. In the case of favourable OCNF adsorption, the interfacial gel was found to embed oil-rich droplets - a spontaneous emulsification process.

Porous Phosphate-Based Glass Microspheres Show Biocompatibility, Tissue Infiltration, and Osteogenic Onset in an Ovine Bone Defect Model.

Phosphate-based glasses (PBGs) are bioactive and fully degradable materials with tailorable degradation rates. PBGs can be produced as porous microspheres through a single-step process, using changes in their formulation and geometry to produce varying pore sizes and interconnectivity for use in a range of applications, including biomedical use. Calcium phosphate PBGs have recently been proposed as orthobiologics, based on their in vitro cytocompatibility and ion release profile. In this study, porous microspheres made of two PBG formulations either containing TiO2 (P40Ti) or without (P40) were implanted in vivo in a large animal model of bone defect. The biocompatibility and osteogenic potential of these porous materials were assessed 13 weeks postimplantation in sheep and compared to empty defects and autologous bone grafts used as negative and positive controls. Histological analysis showed marked differences between the two formulations, as lower trabeculae-like interconnection and higher fatty bone marrow content were observed in the faster degrading P40-implanted defects, while the slower degrading P40Ti material promoted dense interconnected tissue. Autologous bone marrow concentrate (BMC) was also incorporated within the P40 and P40Ti microspheres in some defects; however, no significant differences were observed in comparison to microspheres implanted alone. Both formulations induced the formation of a collagen-enriched matrix, from 20 to 40% for P40 and P40Ti2.5 groups, suggesting commitment toward the bone lineage. With the faster degrading P40 formulation, mineralization of the tissue matrix was observed both with and without BMC. Some lymphocyte-like cells and foreign body multinucleated giant cells were observed with P40Ti2.5, suggesting that this more durable formulation might be linked to an inflammatory response. In conclusion, these first in vivo results indicate that PBG microspheres could be useful candidates for bone repair and regenerative medicine strategies and highlight the role of material degradation in the process of tissue formation and maturation.

Flame-Spheroidized Phosphate-Based Glass Particles with Improved Characteristics for Applications in Mesenchymal Stem Cell Culture Therapy and Tissue Engineering.

The chemical formulation of phosphate-based glasses (PBGs) can be tailored to fit particular end applications such as bone tissue engineering. While most reports to date have evaluated the effect of PBG chemical formulation on bone cells, this study specifically explored the manufacturing process, the changes in physical and chemical properties of PBG particles after flame spheroidization, and subsequent effects on human mesenchymal stem cells (hMSCs), a prime cell type for regenerative medicine applications. Flame spheroidization involves feeding irregular PBG particles (microparticles, MP) into a hot flame, causing them to melt and mold into solid spherical PBG particles (microspheres, MS). The laser diffraction analysis showed an increase in the volume-weighted mean diameter of particles from 48 to 139 µm after spheroidization and also revealed changes in the chemical composition of smaller MS (< 45 µm in size), whereas MS in other size ranges did not show significantly different chemical composition compared to MP. Additionally, some air bubbles were entrapped inside particles during spheroidization, causing a 2% drop in relative density of MS. However, the packing density of MS was 30% higher than that of MP. Culture of hMSCs on the particles showed significant improvement in cell spreading on MS compared to that on MP and nearly 2 times higher cell metabolic activity after 7 days of culture, suggesting that MS provided a more favorable support and geometry for hMSC attachment and growth for tissue engineering.

Flexible and transparent films produced from cellulose nanowhisker reinforced agarose.

Transparent and flexible nanocomposite films with a range of Agarose to Cellulose Nano-Whisker (CNW) ratios were produced using never-dried CNWs. The incorporation of never-dried CNWs within Agarose played an important role in the surface roughness (Ra 7-15 nm) and light transparency of the films (from 84 to 90%). Surface induced crystallisation of Agarose by CNWs was also found with increasing percentage of crystallinity (up to 79%) for the nanocomposite films, where CNW acted as nucleating sites. The enhanced tensile strength (ca. 30% increase) and modulus (ca. 90% increase) properties of the nanocomposite films compared to the control Agarose film indicated the effectiveness of the nanowhiskers incorporation. The storage modulus of the nanocomposite films increased also to be tripled Agarose alone as the CNWs content reached 43%. The swelling kinetics of the nanocomposites revealed that addition of CNWs reduced the long-term swelling capacity and swelling rate of the nanocomposite.

Porous calcium phosphate glass microspheres for orthobiologic applications.

Orthobiologics is a rapidly advancing field utilising cell-based therapies and biomaterials to enable the body to repair and regenerate musculoskeletal tissues. This paper reports on a cost-effective flame spheroidisation process for production of novel porous glass microspheres from calcium phosphate-based glasses to encapsulate and deliver stem cells. Careful selection of the glass and pore-forming agent, along with a manufacturing method with the required processing window enabled the production of porous glass microspheres via a single-stage manufacturing process. The morphological and physical characterisation revealed porous microspheres with tailored surface and interconnected porosity (up to 76 ±â€¯5%) with average pore size of 55 ±â€¯8 µm and surface areas ranging from 0.34 to 0.9 m2 g-1. Furthermore, simple alteration of the processing parameters produced microspheres with alternate unique morphologies, such as with solid cores and surface porosity only. The tuneable porosity enabled control over their surface area, degradation profiles and hence ion release rates. Furthermore, cytocompatibility of the microspheres was assessed using human mesenchymal stem cells via direct cell culture experiments and analysis confirmed that they had migrated to within the centre of the microspheres. The novel microspheres developed have huge potential for tissue engineering and regenerative medicine applications. STATEMENT OF SIGNIFICANCE: This manuscript highlights a simple cost-effective one-step process for manufacturing porous calcium phosphate-based glass microspheres with varying control over surface pores and fully interconnected porosity via a flame spheroidisation process. Moreover, a simple alteration of the processing parameters can produce microspheres which have a solid core with surface pores only. The tuneable porosity enabled control over their surface area, degradation profiles and hence ion release rates. The paper also shows that stem cells not only attach and proliferate but more importantly migrate to within the core of the porous microspheres, highlighting applications for bone tissue engineering and regenerative medicine.