Identification of common textile microplastics via autofluorescence spectroscopy coupled with k-means cluster analysis.

Microplastics are an emerging anthropogenic pollutant risk with a significant body of research dedicated to understanding the implications further. To generate the databases required to characterize the impact of microplastics on our environment, and improve recovery and recycling of current plastic materials, we need rapid, in-line characterization that can distinguish individual polymer types. Here, autofluorescence spectroscopy was investigated as an alternative characterization method to the current leading techniques based on vibrational spectroscopy. It was confirmed that the autofluorescence of seven common textile polymers (acrylic, polyester, nylon, polyethylene, polypropylene, cellulose/cotton, wool) arose due to the cluster-triggered emission phenomenon. Both simulated polymer aging via photooxidation and dyeing of the polymers were found to affect the resultant autofluorescence spectra. A total of 1485 spectra from 39 unique sample groups (polymer type, colour, and degree of photooxidation) were analysed via machine learning (k-means cluster analysis). Correct identification of the polymer type was achieved in 71% of the cases from only eight input values (normalized intensity values at three autofluorescence emission wavelengths, the total autofluorescence emission intensity, the sample RGB colour values, and the sample shape). This represents a significant step towards automated polymer identification at the sub-second time scales required for the in-line characterization of microplastics.

Chemoenzymatic Synthesis of Fluorinated Cellodextrins Identifies a New Allomorph for Cellulose-Like Materials*.

Understanding the fine details of the self-assembly of building blocks into complex hierarchical structures represents a major challenge en route to the design and preparation of soft-matter materials with specific properties. Enzymatically synthesised cellodextrins are known to have limited water solubility beyond DP9, a point at which they self-assemble into particles resembling the antiparallel cellulose II crystalline packing. We have prepared and characterised a series of site-selectively fluorinated cellodextrins with different degrees of fluorination and substitution patterns by chemoenzymatic synthesis. Bearing in mind the potential disruption of the hydrogen-bond network of cellulose II, we have prepared and characterised a multiply 6-fluorinated cellodextrin. In addition, a series of single site-selectively fluorinated cellodextrins was synthesised to assess the structural impact upon the addition of one fluorine atom per chain. The structural characterisation of these materials at different length scales, combining advanced NMR spectroscopy and microscopy methods, showed that a 6-fluorinated donor substrate yielded multiply 6-fluorinated cellodextrin chains that assembled into particles presenting morphological and crystallinity features, and intermolecular interactions, that are unprecedented for cellulose-like materials.

Hydrophobization of Cellulose Nanocrystals for Aqueous Colloidal Suspensions and Gels.

Surface hydrophobization of cellulose nanomaterials has been used in the development of nanofiller-reinforced polymer composites and formulations based on Pickering emulsions. Despite the well-known effect of hydrophobic domains on self-assembly or association of water-soluble polymer amphiphiles, very few studies have addressed the behavior of hydrophobized cellulose nanomaterials in aqueous media. In this study, we investigate the properties of hydrophobized cellulose nanocrystals (CNCs) and their self-assembly and amphiphilic properties in suspensions and gels. CNCs of different hydrophobicity were synthesized from sulfated CNCs by coupling primary alkylamines of different alkyl chain lengths (6, 8, and 12 carbon atoms). The synthetic route permitted the retention of surface charge, ensuring good colloidal stability of hydrophobized CNCs in aqueous suspensions. We compare surface properties (surface charge, ζ potential), hydrophobicity (water contact angle, microenvironment probing using pyrene fluorescence emission), and surface activity (tensiometry) of different hydrophobized CNCs and hydrophilic CNCs. Association of hydrophobized CNCs driven by hydrophobic effects is confirmed by X-ray scattering (SAXS) and autofluorescent spectroscopy experiments. As a result of CNC association, CNC suspensions/gels can be produced with a wide range of rheological properties depending on the hydrophobic/hydrophilic balance. In particular, sol-gel transitions for hydrophobized CNCs occur at lower concentrations than hydrophilic CNCs, and more robust gels are formed by hydrophobized CNCs. Our work illustrates that amphiphilic CNCs can complement associative polymers as modifiers of rheological properties of water-based systems.

Employing photoluminescence to rapidly follow aggregation and dispersion of cellulose nanofibrils.

Photoluminescence of cellulose, and other polysaccharides, has long been presumed to be due to contamination of the material by other autofluorescent compounds - such as lignin, or proteins. This is attributed to the lack of known fluorescent chemical groups present in the molecular structure of polysaccharides and the weak emission intensity when compared to typical fluorophores. However, recent research suggests that the observed luminescence may actually be due to transitions involving the n orbitals containing lone electron pairs present in oxyl groups, stabilised by the molecular forces between the polysaccharide chains. Here we investigate this theory further by varying the physicochemical environment (concentration and pH) of oxidised cellulose nanofibril suspensions and observing the resultant fluorescent spectra using multi-channel confocal laser scanning spectroscopy. We confirm that both factors affect the material photoluminescence, specifically changing the intensity ratio between two localised emission maxima, supporting current theories. Furthermore, we demonstrate that this variation enables the determination of critical aggregation concentrations and the apparent pKa values of hydroxyl groups that undergo deprotonation within the examined pH range, enabling use of the technique to track rapid changes in the fibril physicochemical environment.

Rapid Determination of the Distribution of Cellulose Nanomaterial Aggregates in Composites Enabled by Multi-Channel Spectral Confocal Microscopy.

There is increased interest in the use of cellulose nanomaterials for the mechanical reinforcement of composites due to their high stiffness and strength. However, challenges remain in accurately determining their distribution within composite microstructures. We report the use of a range of techniques used to image aggregates of cellulose nanocrystals (CNCs) greater than 10 µm2 within a model thermoplastic polymer. While Raman imaging accurately determines CNC aggregate size, it requires extended periods of analysis and the limited observable area results in poor reproducibility. In contrast, staining the CNCs with a fluorophore enables rapid acquisition with high reproducibility, but overestimates the aggregate size as CNC content increases. Multi-channel spectral confocal laser scanning microscopy is presented as an alternative technique that combines the accuracy of Raman imaging with the speed and reproducibility of conventional confocal laser scanning microscopy, enabling the rapid determination of CNC aggregate distribution within composites.

Cellulose ionics: switching ionic diode responses by surface charge in reconstituted cellulose films.

Cellulose films as well as chitosan-modified cellulose films of approximately 5 µm thickness, reconstituted from ionic liquid media onto a poly(ethylene-terephthalate) (PET, 6 µm thickness) film with a 5, 10, 20, or 40 µm diameter laser-drilled microhole, show significant current rectification in aqueous NaCl. Reconstituted α-cellulose films provide "cationic diodes" (due to predominant cation conductivity) whereas chitosan-doped cellulose shows "anionic diode" effects (due to predominant anion conductivity). The current rectification, or "ionic diode" behaviour, is investigated as a function of NaCl concentration, pH, microhole diameter, and molecular weight of the chitosan dopant. Future applications are envisaged exploiting the surface charge induced switching of diode currents for signal amplification in sensing.

Combining random walk and regression models to understand solvation in multi-component solvent systems.

Polysaccharides, such as cellulose, are often processed by dissolution in solvent mixtures, e.g. an ionic liquid (IL) combined with a dipolar aprotic co-solvent (CS) that the polymer does not dissolve in. A multi-walker, discrete-time, discrete-space 1-dimensional random walk can be applied to model solvation of a polymer in a multi-component solvent mixture. The number of IL pairs in a solvent mixture and the number of solvent shells formable, x, is associated with n, the model time-step, and N, the number of random walkers. The mean number of distinct sites visited is proportional to the amount of polymer soluble in a solution. By also fitting a polynomial regression model to the data, we can associate the random walk terms with chemical interactions between components and probe where the system deviates from a 1-D random walk. The 'frustration' between solvents shells is given as ln x in the random walk model and as a negative IL:IL interaction term in the regression model. This frustration appears in regime II of the random walk model (high volume fractions of IL) where walkers interfere with each other, and the system tends to its limiting behaviour. In the low concentration regime, (regime I) the solvent shells do not interact, and the system depends only on IL and CS terms. In both models (and both regimes), the system is almost entirely controlled by the volume available to solvation shells, and thus is a counting/space-filling problem, where the molar volume of the CS is important. Small deviations are observed when there is an IL-CS interaction. The use of two models, built on separate approaches, confirm these findings, demonstrating that this is a real effect and offering a route to identifying such systems. Specifically, the majority of CSs - such as dimethylformide - follow the random walk model, whilst 1-methylimidazole, dimethyl sulfoxide, 1,3-dimethyl-2-imidazolidinone and tetramethylurea offer a CS-mediated improvement and propylene carbonate results in a CS-mediated hindrance. It is shown here that systems, which are very complex at a molecular level, may, nonetheless, be effectively modelled as a simple random walk in phase-space. The 1-D random walk model allows prediction of the ability of solvent mixtures to dissolve cellulose based on only two dissolution measurements (one in neat IL) and molar volume.

Surface modified cellulose scaffolds for tissue engineering.

We report the ability of cellulose to support cells without the use of matrix ligands on the surface of the material, thus creating a two-component system for tissue engineering of cells and materials. Sheets of bacterial cellulose, grown from a culture medium containing Acetobacter organism were chemically modified with glycidyltrimethylammonium chloride or by oxidation with sodium hypochlorite in the presence of sodium bromide and 2,2,6,6-tetramethylpipiridine 1-oxyl radical to introduce a positive, or negative, charge, respectively. This modification process did not degrade the mechanical properties of the bulk material, but grafting of a positively charged moiety to the cellulose surface (cationic cellulose) increased cell attachment by 70% compared to unmodified cellulose, while negatively charged, oxidised cellulose films (anionic cellulose), showed low levels of cell attachment comparable to those seen for unmodified cellulose. Only a minimal level of cationic surface derivitisation (ca 3% degree of substitution) was required for increased cell attachment and no mediating proteins were required. Cell adhesion studies exhibited the same trends as the attachment studies, while the mean cell area and aspect ratio was highest on the cationic surfaces. Overall, we demonstrated the utility of positively charged bacterial cellulose in tissue engineering in the absence of proteins for cell attachment.

Autofluorescence spectroscopy for quantitative analysis of cellulose nanocrystals.

The ability to determine the physicochemical properties of nanoparticles, such as cellulose nanocrystals, in suspension is critically important to maximize their potential. Currently, various techniques are required to ascertain different properties, which results in a laborious analysis procedure. Here, autofluorescence arising from the cluster-triggered emission (CTE) photoluminescence mechanism is utilized as an analytical spectroscopic tool to determine multiple properties from one data acquisition sequence. This study confirms that key properties - including the nanoparticle concentration in suspension, the critical concentration for liquid crystal formation, and the surface charge content - can be obtained simultaneously. Measured values are accurate to within 10% of conventional techniques with average residual errors of 0.4 wt% for the critical concentration, and 11 mmol kg-1 CNC for the surface charge content. This charge-coupled device (CCD) sensor-based methodology is rapid and does not require the addition of further chemicals. These results support the theory behind CTE and represent a new opportunity for quantitatively analysing non-aromatic, heteroatom-containing nanoparticles in flow based on understanding their inter- and intra-particle interactions.

Enhanced ligand-free attachment of osteoblast to poly(3-hydroxybutyrate-co-3-hydroxyvalerate) nanoparticles.

Polymeric nanoparticles have previously been used as substrates for cell attachment and proliferation due to their ability to mimic the extracellular matrix, but in general, they require surface chemical modifications to achieve this purpose. In this study, polymeric nanoparticles were developed and used without any matrix ligands functionalized on their surface to promote cell attachment and proliferation of human osteoblasts (MG63s). First, telechelic, reduced molar mass and diol-functionalized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was prepared by transesterification using ethylene glycol. Then, PHBV-diol was used to prepare biodegradable nanoparticles via the solvent evaporation technique. MG63s were cultured in the presence of PHBV nanoparticles and growth kinetics were compared to that on tissue culture polystyrene (TCPS). Cell attachment on non-tissue culture polystyrene pre-coated with nanoparticles was assessed and compared to attachment on TCPS. The cell attachment study demonstrated that cells readily attached and were well spread onto the nanoparticle surfaces compared to non-tissue culture polystyrene. These findings reveal the potential of PHBV nanoparticles for cell attachment and growth to be used in tissue engineering.

Hydrophobized cellulose nanocrystals enhance xanthan and locust bean gum network properties in gels and emulsions.

Locust bean/xanthan gum (LBG/XG) synergistic networks have previously been well studied, with evidence that junction zones between the two polymers result in hydrophobic domains. Here we report on the effect of both hydrophilic and hydrophobic cellulose nanocrystals (CNCs) on the rheological properties of the individual gums, the gum networks, and emulsion gels consisting of the gum network and corn oil. We also take advantage of differences in the autofluorescent spectra for each of the components to map their distribution within the gel and emulsion gel systems. Whilst both types of CNC confer thermal stability to the systems, hydrophilic CNCs induce minor changes in rheological properties of synergistic gels and prove to be detrimental to the stability of the emulsion gels. In contrast, hydrophobic CNCs associate with the LBG/XG network, affecting the rheological response. Their inclusion in the emulsion gel system results in smaller, more homogeneously distributed oil droplets with a resultant increase in the storage modulus by an order of magnitude compared to the CNC-free and hydrophilic CNC systems. We conclude that hydrophobic CNCs play a critical role in stabilising LBG/XG network gels and emulsions.

Predicting Ligand-Free Cell Attachment on Next-Generation Cellulose-Chitosan Hydrogels.

There is a growing appreciation that engineered biointerfaces can regulate cell behaviors, or functions. Most systems aim to mimic the cell-friendly extracellular matrix environment and incorporate protein ligands; however, the understanding of how a ligand-free system can achieve this is limited. Cell scaffold materials comprised of interfused chitosan-cellulose hydrogels promote cell attachment in ligand-free systems, and we demonstrate the role of cellulose molecular weight, MW, and chitosan content and MW in controlling material properties and thus regulating cell attachment. Semi-interpenetrating network (SIPN) gels, generated from cellulose/ionic liquid/cosolvent solutions, using chitosan solutions as phase inversion solvents, were stable and obviated the need for chemical coupling. Interface properties, including surface zeta-potential, dielectric constant, surface roughness, and shear modulus, were modified by varying the chitosan degree of polymerization and solution concentration, as well as the source of cellulose, creating a family of cellulose-chitosan SIPN materials. These features, in turn, affect cell attachment onto the hydrogels and the utility of this ligand-free approach is extended by forecasting cell attachment using regression modeling to isolate the effects of individual parameters in an initially complex system. We demonstrate that increasing the charge density, and/or shear modulus, of the hydrogel results in increased cell attachment.