Structuring gelatin methacryloyl - dextran hydrogels and microgels under shear.

Gelatin methacryloyl (GelMA) is a widely used semi-synthetic polymer for a variety of bioapplications. However, the development of versatile GelMA hydrogels requires tuning of their microstructure. Herein, we report the possibility of preparing hydrogels with various microstructures under shear from an aqueous two-phase system (ATPS) consisting of GelMA and dextran. The influence of an applied preshear on dextran/GelMA droplets and bicontinuous systems is investigated by rheology that allows the application of a constant shear and is immediately followed by in situ UV-curing of the GelMA-rich phase. The microstructure of the resulting hydrogels is examined by confocal laser scanning microscopy (CLSM). The results show that the GelMA string phase and GelMA hydrogels with aligned bands can be formed depending on the concentration of dextran and the applied preshear. The influence of the pH of the ATPS is investigated and demonstrates the formation of multiple emulsions upon decreasing the charge density of GelMA. The preshearing of multiple emulsions, following gelation, leads to the formation of porous GelMA microgels. The diversity of the formed structures highlights the application potential of preshearing ATPS in the development of functional soft materials.

On-Chip Fabrication of Colloidal Suprastructures by Assembly and Supramolecular Interlinking of Microgels.

In this report, a versatile method is demonstrated to create colloidal suprastructures by assembly and supramolecular interlinking of microgels using droplet-based microfluidics. The behavior of the microgels is systematically investigated to evaluate the influence of their concentration on their distribution between the continuous, the droplet phase, and the interface. At low concentrations, microgels are mainly localized at the water-oil interface whereas an excess of microgels results, following the complete coverage of the water-oil interface, in their distribution in the continuous phase. To stabilize the colloidal suprastructure, on-chip gelation is introduced by adding natural polyphenol tannic acid (TA) in the water phase. TA forms interparticle linking between the poly(N-vinylcaprolactam) (PVCL) microgels by supramolecular interactions. The combination of supramolecular interlinking with the variation of the microgel concentration in microfluidic droplets enables on-chip fabrication of defined colloidal suprastructures with morphologies ranging from colloidosomes to colloidal supraballs. The obtained supracolloidal structures exhibit a pH-responsive behavior with a disintegration at alkaline conditions within a scale of seconds. The destabilization process results from the deprotonation of phenolic groups and destruction of hydrogen bonds with PVCL chains at higher pH.

3D Printable Gelatin Methacryloyl (GelMA)-Dextran Aqueous Two-Phase System with Tunable Pores Structure and Size Enables Physiological Behavior of Embedded Cells In Vitro.

The restricted porosity of most hydrogels established for in vitro 3D tissue engineering applications limits embedded cells with regard to their physiological spreading, proliferation, and migration behavior. To overcome these confines, porous hydrogels derived from aqueous two-phase systems (ATPS) are an interesting alternative. However, while developing hydrogels with trapped pores is widespread, the design of bicontinuous hydrogels is still challenging. Herein, an ATPS consisting of photo-crosslinkable gelatin methacryloyl (GelMA) and dextran is presented. The phase behavior, monophasic or biphasic, is tuned via the pH and dextran concentration. This, in turn, allows the formation of hydrogels with three distinct microstructures: homogenous nonporous, regular disconnected-pores, and bicontinuous with interconnected-pores. The pore size of the latter two hydrogels can be tuned from ≈4 to 100 µm. Cytocompatibility of the generated ATPS hydrogels is confirmed by testing the viability of stromal and tumor cells. Their distribution and growth pattern are cell-type specific but are also strongly defined by the microstructure of the hydrogel. Finally, it is demonstrated that the unique porous structure is sustained when processing the bicontinuous system by inkjet and microextrusion techniques. The proposed ATPS hydrogels hold great potential for 3D tissue engineering applications due to their unique tunable interconnected porosity.

A defined heat pretreatment of gelatin enables control of hydrolytic stability, stiffness, and microstructural architecture of fibrin-gelatin hydrogel blends.

Fibrin-gelatin hydrogel blends exhibit high potential for tissue engineering in vitro applications. However, the means to tailor these blends in order to control their properties, thus opening up a broad range of new target applications, have been insufficiently explored. We hypothesized that a controlled heat treatment of gelatin prior to blend synthesis enables control of hydrolytic swelling and shrinking, stiffness, and microstructural architecture of fibrin-gelatin based hydrogel blends while providing tremendous long-term stability. We investigated these hydrogel blends' compressive strength, in vitro degradation stability, and microstructure in order to test this hypothesis. In addition, we examined the gel's ability to support endothelial cell proliferation and stretching of encapsulated smooth muscle cells. This research showed that a controlled heat pretreatment of the gelatin component strongly influenced the stiffness, swelling, shrinking, and microstructural architecture of the final blends regardless of identical gelatin mass fractions. All blends offered high long-term hydrolytic stability. In conclusion, the results of this study open the possibility to use this technique in order to tune low-concentrated, open-porous fibrin-based hydrogels, even in long-term tissue engineering in vitro experiments.

Physicochemical Interactions in Nanofunctionalized Alginate/GelMA IPN Hydrogels.

Polymeric hydrogels are currently at the center of research due to their particular characteristics. They have tunable physical, chemical, and biological properties making them a material of choice for a large range of applications. Polymer-composite and nanocomposite hydrogels were developed to enhance the native hydrogel's properties and to include numerous functionalities. In this work, alginate/gelatin-methacryloyl-based interpenetrating polymer network hydrogels were prepared with different alginate concentrations and investigated before and after the functionalization with nanoliposomes. The multiscale analysis was obtained through Fourier transform infrared spectroscopy and proton nuclear magnetic resonance. The results show interactions between two polymers as well as between the nanoliposomes and biopolymer.

Palmitic acid sophorolipid biosurfactant: from self-assembled fibrillar network (SAFiN) to hydrogels with fast recovery.

Nanofibres are an interesting phase into which amphiphilic molecules can self-assemble. Described for a large number of synthetic lipids, they were seldom reported for natural lipids like microbial amphiphiles, known as biosurfactants. In this work, we show that the palmitic acid congener of sophorolipids (SLC16:0), one of the most studied families of biosurfactants, spontaneously forms a self-assembled fibre network (SAFiN) at pH below 6 through a pH jump process. pH-resolved in situ small-angle X-ray scattering (SAXS) shows a continuous micelle-to-fibre transition, characterized by an enhanced core-shell contrast between pH 9 and pH 7 and micellar fusion into a flat membrane between pH 7 and pH 6, approximately. Below pH 6, homogeneous, infinitely long nanofibres form by peeling off the membranes. Eventually, the nanofibre network spontaneously forms a thixotropic hydrogel with fast recovery rates after applying an oscillatory strain amplitude out of the linear viscoelastic regime: after being submitted to strain amplitudes during 5 min, the hydrogel recovers about 80% and 100% of its initial elastic modulus after, respectively, 20 s and 10 min. Finally, the strength of the hydrogel depends on the medium's final pH, with an elastic modulus fivefold higher at pH 3 than at pH 6. This article is part of the theme issue 'Bio-derived and bioinspired sustainable advanced materials for emerging technologies (part 1)'.

pH-switchable pickering emulsions stabilized by polyelectrolyte-biosurfactant complex coacervate colloids.

HYPOTHESIS: Polyelectrolyte-surfactant complexes (PESCs) have long been employed as oil-in-water (o/w) emulsions stabilizers, but never in the structure of colloidal complex coacervates providing a Pickering effect. The complexed state of PESCs could make them unsuitable o/w Pickering emulsifiers, which instead require a balance between colloidal structure and stability, amphiphilicity and wettability. Here we hypothesize that PESCs coacervates are efficient Pickering stabilizers. Instead of classical surfactants, we employ sophorolipid (SL) biosurfactants, atypical anionic/neutral stimuli-responsive biosurfactants. Despite their tunable charge and mild amphiphilic character, they can be used in combination with cationic/neutral polyelectrolytes (chitosan, CHL, or poly-l-lysine, PLL) to form PESC coacervates for the development of biobased, but also pH-switchable, Pickering emulsions. EXPERIMENTS: Aqueous solutions of SL-CHL (or SL-PLL) complex coacervates are emulsified with dodecane. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy under cryogenic conditions (cryo-SEM) demonstrate the Pickering effect, while optical microscopy and oscillatory rheology respectively assess the emulsion formation and relative viscoelastic properties. FINDINGS: Both SL-CHL and SL-PLL PESCs stabilize o/w emulsions up to Φoil of 0.7 only in the pH region of complex coacervation (6 < pH < 9): outside this range, phase separation occurs. Rheology shows a typical solid-like response and mechanical recovery upon applying large deformations. CLSM and cryo-SEM highlight a colloidal structure, associated to the complex coacervates, of the oil/water interface and suggest a Pickering effect. These findings demonstrate the Pickering effect from PESC coacervates and the possibility to use biobased and biocompatible components, with application potential in cosmetics, food science, or oil recovery.

Effects of pH, temperature and shear on the structure-property relationship of lamellar hydrogels from microbial glucolipids probed by in situ rheo-SAXS.

Lipid lamellar hydrogels are a class of soft materials composed of a defectuous lipid lamellar phase, where defects are classically stabilized by polymer or surfactant inclusions in lipid membranes. We have recently shown that bolaform microbial glucolipids, composed of a single glucose headgroup and a C18:0 fatty acid, with the carboxylic acid group located opposite to glucose, spontaneously form lamellar hydrogels at room temperature below pH 8. In this work, we combine rheology with small angle X-ray scattering (SAXS), rheo-SAXS, to correlate, in situ, the structural and mechanical properties of microbial glycolipid lamellar hydrogels upon application of three different stimuli: pH, temperature and a shear rate. In all cases we find unusual structural features of the lamellar phase if compared to classical phospholipid lamellar structures: reducing pH from alkaline to acidic induces a sol-to-gel transition during which an increasing elastic modulus is associated with an oscillatory evolution of lamellar d(100) spacing; temperature above Tm and increasing shear induce the formation of spherulitic crumpled domains, instead of a classically-expected lamellar-to-vesicle or lamellar-to-onion phase transitions.

Single-molecule lamellar hydrogels from bolaform microbial glucolipids.

Lipid lamellar hydrogels are rare soft fluids composed of a phospholipid lamellar phase instead of fibrillar networks. The mechanical properties of these materials are controlled by defects, induced by local accumulation of a polymer or surfactant in a classical lipid bilayer. Herein we report a new class of lipid lamellar hydrogels composed of one single bolaform glycosylated lipid obtained by fermentation. The lipid is self-organized into flat interdigitated membranes, stabilized by electrostatic repulsive forces and stacked in micrometer-sized lamellar domains. The defects in the membranes and the interconnection of the lamellar domains are responsible, from the nano- to the micrometer scales, for the elastic properties of the hydrogels. The lamellar structure is probed by combining small angle X-ray and neutron scattering (SAXS, SANS), the defect-rich lamellar domains are visualized by polarized light microscopy while the elastic properties are studied by oscillatory rheology. The latter show that both storage G' and loss G'' moduli scale as a weak power-law of the frequency, that can be fitted with fractional rheology models. The hydrogels possess rheo-thinning properties with second-scale recovery. We also show that ionic strength is not only necessary, as one could expect, to control the interactions in the lamellar phase but, most importantly, it directly controls the elastic properties of the lamellar gels.

Synthesis and Characterization of Nanofunctionalized Gelatin Methacrylate Hydrogels.

Given the importance of the extracellular medium during tissue formation, it was wise to develop an artificial structure that mimics the extracellular matrix while having improved physico-chemical properties. That is why the choice was focused on gelatin methacryloyl (GelMA), an inexpensive biocompatible hydrogel. Physicochemical and mechanical properties were improved by the incorporation of nanoparticles developed from two innovative fabrication processes: High shear fluid and low frequencies/high frequencies ultrasounds. Both rapeseed nanoliposomes and nanodroplets were successfully incorporated in the GelMA networks during the photo polymerization process. The impact on polymer microstructure was investigated by Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and enzymatic degradation investigations. Mechanical stability and viscoelastic tests were conducted to demonstrate the beneficial effect of the functionalization on GelMA hydrogels. Adding nanoparticles to GelMA improved the surface properties (porosity), tuned swelling, and degradability properties. In addition, we observed that nanoemulsion didn't change significantly the mechanical properties to shear and compression solicitations, whereas nanoliposome addition decreased Young's modulus under compression solicitations. Thus, these ways of functionalization allow controlling the design of the material by choosing the type of nanoparticle (nanoliposome or nanoemulsion) in function of the application.

Physico-chemical properties of alginate/shellac aqueous-core capsules: Influence of membrane architecture on riboflavin release.

To enhance physico-chemical properties of alginate liquid-core capsules, shellac was incorporated into the membrane (composite capsules) or as an additional external layer (coated capsules). The influence of pH, coating time, shellac concentration and preparation mechanism (acid or calcium precipitation) were investigated. Results showed that shellac significantly influenced the capsules properties. The feasibility of shellac incorporation was closely related to the preparation conditions as confirmed by Infrared spectroscopy. Optical, fluorescence and scanning electron microscopy, highlighted different capsules and membranes architectures. In contrast to simple and composite capsules, coated capsules showed a pH-dependent release of the entrapped vitamin especially after shellac crosslinking with calcium. Heating of coated capsules above the glass transition temperature investigated by Differential Scanning Calorimetry, led to irreversible structural change due to thermoplastic behavior of shellac and enhanced riboflavin retention under acidic conditions. This global approach is useful to control release mechanism of low molecular weight molecules from macro and micro-capsules.

Influence of internal composition on physicochemical properties of alginate aqueous-core capsules.

HYPOTHESIS: To enhance physicochemical properties of alginate aqueous-core capsules, conventional strategies were focused in literature on designing composite and coated capsules. In the present study, own effect of liquid-core composition on mechanical and release properties was investigated. EXPERIMENTS: Capsules were prepared by dripping a CaCl2 solution into an alginate gelling solution. Viscosity of CaCl2 solution was adjusted by adding cationic, anionic and non-ionic naturally derived polymers, respectively chitosan, xanthan gum and guar gum. In parallel, uniform alginate hydrogels were prepared by different methods (pouring, in situ forming and mixing). Mechanical stability of capsules and plane hydrogels were respectively evaluated by compression experiments and small amplitude oscillatory shear rheology and then correlated. Capsules permeability was evaluated by monitoring diffusion of encapsulated cochineal dye, riboflavin and BSA. The core-shell interactions were investigated by ATR-FTIR. FINDINGS: Results showed that inner polymer had an impact on membrane stability and could act as an internal coating or provide mechanical reinforcement. Mechanical properties of alginate capsules were in a good agreement with rheological behavior of plane hydrogels. Release behavior of the entrapped molecules changed considerably. This study demonstrated the importance of aqueous-core composition, and gave new insights for possible adjusting of microcapsules physicochemical properties by modulating core-shell interactions.

Physico-chemical characterization and enzymatic functionalization of Enteromorpha sp. cellulose.

Although green macro-algae represent a renewable and highly abundant biomass, they remain poorly exploited in terms of carbohydrate polymers compared to red and brown ones and other lignocellulosic materials. In this study, cellulose from the green macro-algae Enteromorpha sp. was isolated, physico-chemically characterized and enzymatically functionalized. The cellulose content was about 21.4% (w/w). FTIR analyses indicated an absence of acetyl or uronic esters confirming the absence of hemicellulose contamination. The 36% crystallinity index of the extracted cellulose revealed a high amorphous character as determined by X-ray diffraction. The moisture adsorption isotherms and specific surface measurements were respectively 42.87g/100g and 8.34m(2)/g. The Enteromorpha sp. cellulose was first enzymatically saccharified by a commercial cellulase preparation from Aspergillus niger with a hydrolysis yield of 70.4%. Besides, it was successfully functionalized based on the transglycosylation mechanism of the same enzymatic preparation, to produce highly added-value biosurfactants (butyl-glucoside) with a concentration of 8mM.

Alginate/sodium caseinate aqueous-core capsules: a pH-responsive matrix.

HYPOTHESIS: Alginate capsules have several applications. Their functionality depends considerably on their permeability, chemical and mechanical stability. Consequently, the creation of composite system by addition of further components is expected to control mechanical and release properties of alginate capsules. EXPERIMENTS: Alginate and alginate-sodium caseinate composite liquid-core capsules were prepared by a simple extrusion. The influence of the preparation pH and sodium caseinate concentration on capsules physico-chemical properties was investigated. FINDINGS: Results showed that sodium caseinate influenced significantly capsules properties. As regards to the membrane mechanical stability, composite capsules prepared at pH below the isoelectric point of sodium caseinate exhibited the highest surface Young's modulus, increasing with protein content, explained by potential electrostatic interactions between sodium caseinate amino-groups and alginate carboxylic group. The kinetic of cochineal red A release changed significantly for composite capsules and showed a pH-responsive release. Sodium caseinate-dye mixture studied by absorbance and fluorescence spectroscopy confirmed complex formation at pH 2 by electrostatic interactions between sodium caseinate tryptophan residues and cochineal red sulfonate-groups. Consequently, the release mechanism was explained by membrane adsorption process. This global approach is useful to control release mechanism from macro and micro-capsules by incorporating guest molecules which can interact with the entrapped molecule under specific conditions.