Fully synthetic, tunable poly(α-amino acids) as the base of bioinks curable by visible light.

Bioinks play a crucial role in tissue engineering, influencing mechanical and chemical properties of the printed scaffold as well as the behavior of encapsulated cells. Recently, there has been a shift from animal origin materials to their synthetic alternatives. In this context, we present here bioinks based on fully synthetic and biodegradable poly(α,L-amino acids) (PolyAA) as an alternative to animal-based gelatin methacrylate (Gel-Ma) bioinks. Additionally, we first reported the possibility of the visible light photoinitiated incorporation of the bifunctional cell adhesive RGD peptide into the PolyAA hydrogel matrix. The obtained hydrogels are shown to be cytocompatible, and their mechanical properties closely resemble those of gelatin methacrylate-based scaffolds. Moreover, combining the unique properties of PolyAA-based bioinks, the photocrosslinking strategy, and the use of droplet-based printing allows the printing of constructs with high shape fidelity and structural integrity from low-viscosity bioinks without using any sacrificial components. Overall, presented PolyAA-based materials are a promising and versatile toolbox that extends the range of bioinks for droplet bioprinting.

Synthesis and Morphology Characteristics of New Highly Branched Polycaprolactone PCL.

A simple and efficient method for the synthesis of biodegradable, highly branched polycaprolactone (PCL) is presented. The solvent-free (bulk) reaction was carried out via ring opening polymerization (ROP), catalyzed by tin octanoate Sn(Oct)2, and it employed hyperbranched polyamide (HPPA) as a macro-initiator. The core-shell structure of the obtained products (PCL-HPPA), with the hyperbranched HPPA core and linear PCL chains as shell, was in the focus of the product characterization. 1H nuclear magnetic resonance (1H NMR) and elemental analysis confirmed the covalent incorporation of the HPPA in the products, as well as a high degree of grafting conversion of its amino functional groups. Confocal Raman Micro spectroscopy, and especially Time-of-Flight Secondary Ion Mass Spectrometry, further supported the existence of a core-shell structure in the products. Direct observation of macromolecules by means of cryogenic transmission electron microscopy, as well as gel permeation chromatography (GPC), suggested the existence of a minor 'aggregated' product fraction with multiple HPPA cores, which was attributed to transesterification reactions. Differential scanning calorimetry, as well as X-ray diffraction, demonstrated that the PCL-HPPA polymers displayed a similar degree of crystallinity to linear neat PCL, but that the branched products possessed smaller and less regular crystallites.

Exceptionally Fast Temperature-Responsive, Mechanically Strong and Extensible Monolithic Non-Porous Hydrogels: Poly(N-isopropylacrylamide) Intercalated with Hydroxypropyl Methylcellulose.

Exceptionally fast temperature-responsive, mechanically strong, tough and extensible monolithic non-porous hydrogels were synthesized. They are based on divinyl-crosslinked poly(N-isopropyl-acrylamide) (PNIPAm) intercalated by hydroxypropyl methylcellulose (HPMC). HPMC was largely extracted after polymerization, thus yielding a 'template-modified' PNIPAm network intercalated with a modest residue of HPMC. High contents of divinyl crosslinker and of HPMC caused a varying degree of micro-phase-separation in some products, but without detriment to mechanical or tensile properties. After extraction of non-fixed HPMC, the micro-phase-separated products combine superior mechanical properties with ultra-fast T-response (in 30 s). Their PNIPAm network was highly regular and extensible (intercalation effect), toughened by hydrogen bonds to HPMC, and interpenetrated by a network of nano-channels (left behind by extracted HPMC), which ensured the water transport rates needed for ultra-fast deswelling. Moreover, the T-response rate could be widely tuned by the degree of heterogeneity during synthesis. The fastest-responsive among our hydrogels could be of practical interest as soft actuators with very good mechanical properties (soft robotics), while the slower ones offer applications in drug delivery systems (as tested on the example of Theophylline), or in related biomedical engineering applications.

Natural Rubber Composites Using Hydrothermally Carbonized Hardwood Waste Biomass as a Partial Reinforcing Filler-Part II: Mechanical, Thermal and Ageing (Chemical) Properties.

Natural rubber composites were reinforced by the co-fillers 'hydrochar' (HC), obtained by hydrothermal carbonization of hardwood sawdust and commercial carbon black (CB). The content of the combined fillers was kept constant while their ratio was varied. The aim was to test the suitability of HC as a partial filler in natural rubber. Due to its larger particle size and hence smaller specific surface area, large amounts of HC reduced the crosslinking density in the composites. On the other hand, due to its unsaturated organic character, HC was found to display interesting chemical effects: if it was used as the exclusive filler component, it displayed a very strong anti-oxidizing effect, which greatly stabilized the rubber composite against oxidative crosslinking (and hence embrittlement). HC also affected the vulcanization kinetics in different ways, depending on the HC/CB ratio. Composites with HC/CB ratios 20/30 and 10/40 displayed interesting chemical stabilization in combination with fairly good mechanical properties. The performed analyses included vulcanization kinetics, tensile properties, determination of density of permanent and reversible crosslinking in dry and swollen states, chemical stability tests including TGA, thermo-oxidative aging tests in air at 180 °C, simulated weathering in real use conditions ('Florida test'), and thermo-mechanical analyses of degraded samples. Generally, the results indicate that HC could be a promising filler material due to its specific reactivity.

Temperature- and pH-Responsive Super-Absorbent Hydrogel Based on Grafted Cellulose and Capable of Heavy Metal Removal from Aqueous Solutions.

In this work, we prepared highly swelling, stimuli-responsive hydrogels capable of the highly efficient adsorption of inorganic pollutants. The hydrogels were based on hydroxypropyl methyl cellulose (HPMC) grafted with acrylamide (AM) and 3-sulfopropyl acrylate (SPA) and were synthesized via the growth (radical polymerization) of the grafted copolymer chains on HPMC, which was activated by radical oxidation. These grafted structures were crosslinked to an infinite network by a small amount of di-vinyl comonomer. HPMC was chosen as a cheap hydrophilic and naturally sourced polymer backbone, while AM and SPA were employed to preferentially bond coordinating and cationic inorganic pollutants, respectively. All the gels displayed a pronounced elastic character, as well as considerably high values of stress at break (several hundred %). The gel with the highest fraction of the ionic comonomer SPA (with an AM/SPA ratio = 0.5) displayed the highest equilibrium swelling ratio (12,100%), the highest volume response to temperature and pH, and the fastest swelling kinetics, but also the lowest modulus. The other gels (with AM/SPA = 1 and 2) displayed several times higher moduli but more modest pH responses and only very modest temperature sensitivity. Cr(VI) adsorption tests indicated that the prepared hydrogels removed this species from water very efficiently: between 90 and 96% in one step. The hydrogels with AM/SPA ratios of 0.5 and 1 appeared to be promising regenerable (via pH) materials for repeated Cr(VI) adsorption.

Morphology, Micromechanical, and Macromechanical Properties of Novel Waterborne Poly(urethane-urea)/Silica Nanocomposites.

Morphology, macro-, and micromechanical properties of novel poly(urethane-urea)/silica nanocomposites were analyzed by electron microscopy, dynamic mechanical thermal analysis, and microindentation. The studied nanocomposites were based on a poly(urethane-urea) (PUU) matrix filled by nanosilica, and were prepared from waterborne dispersions of PUU (latex) and SiO2. The loading of nano-SiO2 was varied between 0 (neat matrix) and 40 wt% in the dry nanocomposite. The prepared materials were all formally in the rubbery state at room temperature, but they displayed complex elastoviscoplastic behavior, spanning from stiffer elastomeric type to semi-glassy. Because of the employed rigid and highly uniform spherical nanofiller, the materials are of great interest for model microindentation studies. Additionally, because of the polycarbonate-type elastic chains of the PUU matrix, hydrogen bonding in the studied nanocomposites was expected to be rich and diverse, ranging from very strong to weak. In micro- and macromechanical tests, all the elasticity-related properties correlated very strongly. The relations among the properties that related to energy dissipation were complex, and were highly affected by the existence of hydrogen bonding of broadly varied strength, by the distribution patterns of the fine nanofiller, as well as by the eventual locally endured larger deformations during the tests, and the tendency of the materials to cold flow.

Natural Rubber Composites Using Hydrothermally Carbonized Hardwood Waste Biomass as a Partial Reinforcing Filler- Part I: Structure, Morphology, and Rheological Effects during Vulcanization.

A new generation biomass-based filler for natural rubber, 'hydrochar' (HC), was obtained by hydrothermal carbonization of hardwood waste (sawdust). It was intended as a potential partial replacement for the traditional carbon black (CB) filler. The HC particles were found (TEM) to be much larger (and less regular) than CB: 0.5-3 µm vs. 30-60 nm, but the specific surface areas were relatively close to each other (HC: 21.4 m2/g vs. CB: 77.8 m2/g), indicating a considerable porosity of HC. The carbon content of HC was 71%, up from 46% in sawdust feed. FTIR and 13C-NMR analyses indicated that HC preserved its organic character, but it strongly differs from both lignin and cellulose. Experimental rubber nanocomposites were prepared, in which the content of the combined fillers was set at 50 phr (31 wt.%), while the HC/CB ratios were varied between 40/10 and 0/50. Morphology investigations proved a fairly even distribution of HC and CB, as well as the disappearance of bubbles after vulcanization. Vulcanization rheology tests demonstrated that the HC filler does not hinder the process, but it significantly influences vulcanization chemistry, canceling scorch time on one hand and slowing down the reaction on the other. Generally, the results suggest that rubber composites in which 10-20 phr of CB are replaced by HC might be promising materials. The use of HC in the rubber industry would represent a high-tonnage application for hardwood waste.

Self-Healing and Super-Elastomeric PolyMEA-co-SMA Nanocomposites Crosslinked by Clay Platelets.

Novel solvent-free ultra-extensible, tough, and self-healing nanocomposite elastomers were synthesized. The self-assembled materials were based on the copolymer matrix poly(methoxyethyl acrylate-co-sodium methacrylate) physically crosslinked by clay nano-platelets ('poly[MEA-co-SMA]/clay'). Depending on the content of SMA, the super-elastomers were predominantly hydrophobic, water-swelling, or fully water-soluble, and hence repeatedly processible. The SMA co-monomer introduces a tremendous increase in tensile strength, an increase in toughness, while ultra-extensibility is preserved. By tuning the contents of nano-clay and SMA co-monomer, a very wide range of product properties was achieved, including extreme ultra-extensibility, or high stiffness combined with more moderate super-extensibility, or very different values of tensile strength. There was very attractive, great improvement in autonomous self-healing ability induced by SMA, combined with tremendously enhanced self-recovery of internal mechanical damage: even complete self-recovery could be achieved. The ionic SMA repeat units were found to assemble to multiplets, which are phase-separated in the hydrophobic polyMEA matrix. The dynamics of SMA-units-hopping between these aggregates was of key importance for the mechanical, visco-elastic, tensile, and self-healing properties. The studied super-elastomers are attractive as advanced self-healing materials in engineering, soft robotics, and in medical or implant applications.

Biodegradable Thermoplastic Starch/Polycaprolactone Blends with Co-Continuous Morphology Suitable for Local Release of Antibiotics.

We report a reproducible preparation and characterization of highly homogeneous thermoplastic starch/pol(ε-caprolactone) blends (TPS/PCL) with a minimal thermomechanical degradation and co-continuous morphology. These materials would be suitable for biomedical applications, specifically for the local release of antibiotics (ATB) from the TPS phase. The TPS/PCL blends were prepared in the whole concentration range. In agreement with theoretical predictions based on component viscosities, the co-continuous morphology was found for TPS/PCL blends with a composition of 70/30 wt.%. The minimal thermomechanical degradation of the blends was achieved by an optimization of the processing conditions and by keeping processing temperatures as low as possible, because higher temperatures might damage ATB in the final application. The blends' homogeneity was verified by scanning electron microscopy. The co-continuous morphology was confirmed by submicron-computed tomography. The mechanical performance of the blends was characterized in both microscale (by an instrumented microindentation hardness testing; MHI) and macroscale (by dynamic thermomechanical analysis; DMTA). The elastic moduli of TPS increased ca four times in the TPS/PCL (70/30) blend. The correlations between elastic moduli measured by MHI and DMTA were very strong, which implied that, in the future studies, it would be possible to use just micromechanical testing that does not require large specimens.

Novel Tough and Transparent Ultra-Extensible Nanocomposite Elastomers Based on Poly(2-methoxyethylacrylate) and Their Switching between Plasto-Elasticity and Viscoelasticity.

Novel stiff, tough, highly transparent and ultra-extensible self-assembled nanocomposite elastomers based on poly(2-methoxyethylacrylate) (polyMEA) were synthesized. The materials are physically crosslinked by small in-situ-formed silica nanospheres, sized 3-5 nm, which proved to be a very efficient macro-crosslinker in the self-assembled network architecture. Very high values of yield stress (2.3 MPa), tensile strength (3.0 MPa), and modulus (typically 10 MPa), were achieved in combination with ultra-extensibility: the stiffest sample was breaking at 1610% of elongation. Related nanocomposites doubly filled with nano-silica and clay nano-platelets were also prepared, which displayed interesting synergy effects of the fillers at some compositions. All the nanocomposites exhibit 'plasto-elastic' tensile behaviour in the 'as prepared' state: they display considerable energy absorption (and also 'necking' like plastics), but at the same time a large but not complete (50%) retraction of deformation. However, after the first large tensile deformation, the materials irreversibly switch to 'real elastomeric' tensile behaviour (with some creep). The initial 'plasto-elastic' stretching thus causes an internal rearrangement. The studied materials, which additionally are valuable due to their high transparency, could be of application interest as advanced structural materials in soft robotics, in implant technology, or in regenerative medicine. The presented study focuses on structure-property relationships, and on their effects on physical properties, especially on the complex tensile, elastic and viscoelastic behaviour of the polyMEA nanocomposites.

Macro-, Micro- and Nanomechanical Characterization of Crosslinked Polymers with Very Broad Range of Mechanical Properties.

This work is focused on the comparison of macro-, micro- and nanomechanical properties of a series of eleven highly homogeneous and chemically very similar polymer networks, consisting of diglycidyl ether of bisphenol A cured with diamine terminated polypropylene oxide. The main objective was to correlate the mechanical properties at multiple length scales, while using very well-defined polymeric materials. By means of synthesis parameters, the glass transition temperature (Tg) of the polymer networks was deliberately varied in a broad range and, as a result, the samples changed their mechanical behavior from very hard and stiff (elastic moduli 4 GPa), through semi-hard and ductile, to very soft and elastic (elastic moduli 0.006 GPa). The mechanical properties were characterized in macroscale (dynamic mechanical analysis; DMA), microscale (quasi-static microindentation hardness testing; MHI) and nanoscale (quasi-static and dynamic nanoindentation hardness testing; NHI). The stiffness-related properties (i.e., storage moduli, indentation moduli and indentation hardness at all length scales) showed strong and statistically significant mutual correlations (all Pearson's correlation coefficients r > 0.9 and corresponding p-values < 0.001). Moreover, the relations among the stiffness-related properties were approximately linear, in agreement with the theoretical prediction. The viscosity-related properties (i.e., loss moduli, damping factors, indentation creep and elastic work of indentation at all length scales) reflected the stiff-ductile-elastic transitions. The fact that the macro-, micro- and nanomechanical properties exhibited the same trends and similar values indicated that not only dynamic, but also quasi-static indentation can be employed as an alternative to well-established DMA characterization of polymer networks.

Low-Temperature-Meltable Elastomers Based on Linear Polydimethylsiloxane Chains Alpha, Omega-Terminated with Mesogenic Groups as Physical Crosslinker: A Passive Smart Material with Potential as Viscoelastic Coupling. Part II-Viscoelastic and Rheological Properties.

Rheological and viscoelastic properties of physically crosslinked low-temperature elastomers were studied. The supramolecularly assembling copolymers consist of linear polydimethylsiloxane (PDMS) elastic chains terminated on both ends with mesogenic building blocks (LC) of azobenzene type. They are generally and also structurally highly different from the well-studied LC polymer networks or LC elastomers: The LC units make up only a small volume fraction in our materials and act as fairly efficient physical crosslinkers with thermotropic properties. The aggregation (nano-phase separation) of the relatively rare, small and spatially separated terminal LC units generates temperature-switched viscoelasticity in the molten copolymers. Their rheological behavior was found to be controlled by an interplay of nano-phase separation of the LC units (growth and splitting of their aggregates) and of the thermotropic transitions in these aggregates (which change their stiffness). As a consequence, multiple gel points (up to three) are observed in temperature scans of the copolymers. The physical crosslinks also can be reversibly disconnected by large mechanical strain in the 'warm' rubbery state, as well as in melt (thixotropy). The kinetics of crosslink formation was found to be fast if induced by temperature and extremely fast in case of internal self-healing after strain damage. Thixotropic loop tests hence display only very small hysteresis in the LC-melt-state, although the melts show very distinct shear thinning. Our study evaluates structure-property relationships in three homologous systems with elastic PDMS segments of different length (8.6, 16.3 and 64.4 repeat units). The studied copolymers might be of interest as passive smart materials, especially as temperature-controlled elastic/viscoelastic mechanical coupling.

Low-Temperature Meltable Elastomers Based on Linear Polydimethylsiloxane Chains Alpha, Omega-Terminated with Mesogenic Groups as Physical Crosslinkers: A Passive Smart Material with Potential as Viscoelastic Coupling. Part I: Synthesis and Phase Behavior.

Physically crosslinked low-temperature elastomers were prepared based on linear polydimethylsiloxane (PDMS) elastic chains terminated on both ends with mesogenic building blocks (LC) of azobenzene type. They are generally (and also structurally) highly different from the well-studied LC polymer networks (light-sensitive actuators). The LC units also make up only a small volume fraction in our materials and they do not generate elastic energy upon irradiation, but they act as physical crosslinkers with thermotropic properties. Our elastomers lack permanent chemical crosslinks-their structure is fully linear. The aggregation of the relatively rare, small, and spatially separated terminal LC units nevertheless proved to be a considerably strong crosslinking mechanism. The most attractive product displays a rubber plateau extending over 100 °C, melts near 8 °C, and is soluble in organic solvents. The self-assembly (via LC aggregation) of the copolymer molecules leads to a distinctly lamellar structure indicated by X-ray diffraction (XRD). This structure persists also in melt (polarized light microscopy, XRD), where 1-2 thermotropic transitions occur. The interesting effects of the properties of this lamellar structure on viscoelastic and rheological properties in the rubbery and in the melt state are discussed in a follow-up paper ("Part II"). The copolymers might be of interest as passive smart materials, especially as temperature-controlled elastic/viscoelastic mechanical coupling. Our study focuses on the comparison of physical properties and structure-property relationships in three systems with elastic PDMS segments of different length (8.6, 16.3, and 64.4 repeat units).

Control of Gelation and Properties of Reversible Diels-Alder Networks: Design of a Self-Healing Network.

Reversible Diels-Alder (DA) type networks were prepared from furan and maleimide monomers of different structure and functionality. The factors controlling the dynamic network formation and their properties were discussed. Evolution of structure during both dynamic nonequilibrium and isothermal equilibrium network formation/breaking was followed by monitoring the modulus and conversion of the monomer. The gelation, postgel growth, and properties of the thermoreversible networks from tetrafunctional furan (F4) and different bismaleimides (M2) were controlled by the structure of the maleimide monomer. The substitution of maleimides with alkyl (hexamethylene bismaleimide), aromatic (diphenyl bismaleimide), and polyether substituents affects differently the kinetics and thermodynamics of the thermoreversible DA reaction, and thereby the formation of dynamic networks. The gel-point temperature was tuned in the range Tgel = 97-122 °C in the networks of the same functionality (F4-M2) with different maleimide structure. Theory of branching processes was used to predict the structure development during formation of the dynamic networks and the reasonable agreement with the experiment was achieved. The experimentally inaccessible information on the sol fraction in the reversible network was received by applying the theory. Based on the acquired results, the proper structure of a self-healing network was designed.

Monolithic intercalated PNIPAm/starch hydrogels with very fast and extensive one-way volume and swelling responses to temperature and pH: prospective actuators and drug release systems.

Remarkable monolithic (non-porous) hydrogels based on poly(NIPAm-co-sodium methacrylate) intercalated by starch were prepared, and were found to display very fast and extensive one-way solvent (water) release, induced by both pH and temperature. With centimeter-sized 3D specimens, the achieved response times were as short as 4 min (for 70% water release), in combination with very large volume responses (shrinking ratios up to 15). The response time can be tuned from minutes, over tens of minutes, up to hours. The pH-induced deswelling is always slower than the temperature-induced one, but at the highest starch content, ca. 5.5 min are needed for 70% completion of the pH-triggered process. Simultaneous temperature- and pH-stimuli expectedly also lead to very fast water release. The unique intercalated structure and the temperature-dependent hydrogen bridging between the intercalated phases, as well as between these phases and water, were found to play the key role in the ability of the gels to rapidly release water and shrink, which was deeper elucidated in this work. The hydrogels are of interest as soft actuators, but also for chemical release systems or for drug release applications. The latter was successfully tested with theophylline as the drug.

Insight into the cryopolymerization to form a poly(N-isopropylacrylamide)/clay macroporous gel: structure and phase evolution.

The cryopolymerization and formation of a macroporous poly(N-isopropylacrylamide) (PNIPA)/clay cryogel were investigated. The mechanism of the cryopolymerization and cryogel formation was elucidated. Two processes, cryostructuration and cryopolymerization, proceed simultaneously and their relative rates determine the structure evolution and the cryogel morphology - porosity. The cryostructuration in the PNIPA/clay system during freezing, controlled by the freezing temperature and the rate of cooling, includes both water and NIPA crystallization, formation of a highly concentrated non-frozen liquid phase (NFLP) and clay aggregation. The rate of cryopolymerization and gelation is governed by the following effects: by a low polymerization temperature and after freezing, by the high cryoconcentration and a steric confinement, manifested by a reduced reagent mobility. Moreover, it depends on the cooling rate and the evolution of cryostructuration. The progress of cryostructuration and cryopolymerization during freezing was described and experimentally proved step by step. Both the phase development during freezing and the progress of cryopolymerization including gelation were monitored in situ by NMR, DSC, chemorheology and SAXS. The morphology and porosity of the cryogels were characterized by SEM and TEM.

Poly(N-isopropylacrylamide)-clay based hydrogels controlled by the initiating conditions: evolution of structure and gel formation.

The formation of the hydrogel poly(N-isopropylacrylamide)-clay (LAPONITE®) by redox polymerization was investigated, and the main factors governing the gel build-up were determined. The significant effect of the redox initiating system ammonium peroxodisulfate (APS) and tetramethylethylenediamine (TEMED) on gel formation and structure was established, making it possible to control the structure of the gel. Moreover, the pre-reaction stage involving the quality of the clay exfoliation in an aqueous suspension and the interaction of reaction components with the clay play a role in controlling the polymerization and gel structure. The molecular and phase structure evolution during polymerization was followed in situ by the following independent techniques: Fourier transform infrared spectroscopy (FTIR), chemorheology, small-angle X-ray scattering (SAXS) and ultraviolet-visible spectroscopy (UV/Vis). The combination of these methods enabled us to describe in detail particular progress stages during the gel formation and determine the correlation of the corresponding processes on a time and conversion scale. The mechanism of gel formation was refined based on these experimental results.