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Österberg, Farooq, and Henn acknowledge Academy of Finland’s Flagship Program (project numbers 318890 and 318891, Competence Center for Materials Bioeconomy, FinnCERES), for funding. We thank Dr. Heidi Henrickson for her excellent help with proofreading, list of abbreviations, and guidance. This review presents recent advances regarding biomass-based nanomaterials, focusing on their surface interactions. Plant biomass-based nanoparticles, like nanocellulose and lignin from industry side streams, hold great potential for the development of lightweight, functional, biodegradable, or recyclable material solutions for a sustainable circular bioeconomy. However, to obtain optimal properties of the nanoparticles and materials made thereof, it is crucial to control the interactions both during particle production and in applications. Herein we focus on the current understanding of these interactions. Solvent interactions during particle formation and production, as well as interactions with water, polymers, cells and other components in applications, are addressed. We concentrate on cellulose and lignin nanomaterials and their combination. We demonstrate how the surface chemistry of the nanomaterials affects these interactions and how excellent performance is only achieved when the interactions are controlled. We furthermore introduce suitable methods for probing interactions with nanomaterials, describe their advantages and challenges, and introduce some less commonly used methods and discuss their possible applications to gain a deeper understanding of the interfacial chemistry of biobased nanomaterials. Finally, some gaps in current understanding and interesting emerging research lines are identified.

Publisher Copyright: © 2022 The Authors. Published by American Chemical Society. Keratin is a potential raw material to meet the growing demand for bio-based materials with special properties. Keratin can be obtained from feathers, a by-product from the poultry industry. One approach for keratin valorization is to use the protein to improve the properties of already existing cellulose and lignin-based materials to meet the requirements for replacing fossil-based plastics. To ensure a successful combination of keratin with lignocellulosic building blocks, keratin must have an affinity to these substrates. Hence, we used quartz crystal microbalance with a dissipation monitoring (QCM-D) technique to get a detailed understanding of the adsorption of keratin peptides onto lignocellulosic substrates and how the morphology of the substrate, pH, ionic strength, and keratin properties affected the adsorption. Keratin was fractionated from feathers with a scalable and environmentally friendly deep eutectic solvent process. The keratin fraction used in the adsorption studies consisted of different sized keratin peptides (about 1-4 kDa), which had adopted a random coil conformation as observed by circular dichroism (CD). Measuring keratin adsorption to different lignocellulosic substrates by QCM-D revealed a significant affinity of keratin peptides for lignin, both as smooth films and in the form of nanoparticles but only a weak interaction between cellulose and keratin. Systematic evaluation of the effect of surface, media, and protein properties enabled us to obtain a deeper understanding of the driving force for adsorption. Both the structure and size of the keratin peptides appeared to play an important role in its adsorption. The keratin-lignin combination is an attractive option for advanced material applications. For improved adsorption on cellulose, modifications of either keratin or cellulose would be required.

Transmembrane protein integrins play a key role in cell adhesion. Cell-biomaterial interactions are affected by integrin expression and conformation, which are actively controlled by cells. Although integrin structure and function have been studied in detail, quantitative analyses of integrin-mediated cell-biomaterial interactions are still scarce. Here, we have used atomic force spectroscopy to study how integrin distribution and activation (via intracellular mechanisms in living cells or by divalent cations) affect the interaction of human pluripotent stem cells (WA07) and human hepatocarcinoma cells (HepG2) with promising biomaterials-human recombinant laminin-521 (LN-521) and cellulose nanofibrils (CNF). Cell adhesion to LN-521-coated probes was remarkably influenced by cell viability, divalent cations, and integrin density in WA07 colonies, indicating that specific bonds between LN-521 and activated integrins play a significant role in the interactions between LN-521 and HepG2 and WA07 cells. In contrast, the interactions between CNF and cells were nonspecific and not influenced by cell viability or the presence of divalent cations. These results shed light on the underlying mechanisms of cell adhesion, with direct impact on cell culture and tissue engineering applications.

Funding Information: V.G. and P.V. acknowledge Academy of Finland’s Project No. 308931. J.J.V.D. and M.Ö. acknowledge Academy of Finland’s Flagship Programme under Project Nos. 318890 and 318891 (Competence Center for Materials Bioeconomy, FinnCERES). M.Y. acknowledges Academy of Finland’s Flagship Programme under Project No. 337430 (Gene, Cell and Nano Therapy Competence Cluster for the Treatment of Chronic Diseases, GeneCellNano). Publisher Copyright: © 2021 The Authors. Published by American Chemical Society. Bingel cyclopropanation between Buckminster fullerene and a heteroarmed malonate was utilized to produce a hexakis-functionalized C60 core, with azide and tetrazine units. This orthogonally bifunctional C60 scaffold can be selectively one-pot functionalized by two pericyclic click reactions, that is, inverse electron-demand Diels-Alder and azide-alkyne cycloaddition, which with appropriate ligands (monosaccharides, a peptide and oligonucleotides tested) allows one to control the assembly of heteroantennary bioconjugates.

This work made use of Aalto University Bioeconomy Facilities and OtaNano─Nanomicroscopy Center Facilities. Bin Zhao is thanked for conducting the N2 adsorption–desorption experiments. T.Z. acknowledges funding from the Novo Nordisk Foundation (SUSCELL project, Reference Number: NNF17OC0027658), M.V. acknowledges funding from Fortum and Neste Foundation (grant no. 20200079), P.P. acknowledges funding from the Academy of Finland (grant no. 315768), and M.S. acknowledges funding from the Academy of Finland (grant no. 309324). Computational resources by CSC IT Centre for Science, Finland, and RAMI─RawMatTERS Finland Infrastructure are gratefully acknowledged. The authors are also grateful for the support by the FinnCERES Materials Bioeconomy Ecosystem and Photonics Research and Innovation (PREIN) flagship. Spherical lignin nanoparticles (LNPs) fabricated via nanoprecipitation of dissolved lignin are among the most attractive biomass-derived nanomaterials. Despite various studies exploring the methods to improve the uniformity of LNPs or seeking more application opportunities for LNPs, little attention has been given to the fundamental aspects of the solvent effects on the intrinsic properties of LNPs. In this study, we employed a variety of experimental techniques and molecular dynamics (MD) simulations to investigate the solvent effects on the intrinsic properties of LNPs. The LNPs were prepared from softwood Kraft lignin (SKL) using the binary solvents of aqueous acetone or aqueous tetrahydrofuran (THF) via nanoprecipitation. The internal morphology, porosity, and mechanical properties of the LNPs were analyzed with electron tomography (ET), small-angle X-ray scattering (SAXS), atomic force microscopy (AFM), and intermodulation AFM (ImAFM). We found that aqueous acetone resulted in smaller LNPs with higher uniformity compared to aqueous THF, mainly ascribing to stronger solvent–lignin interactions as suggested by MD simulation results and confirmed with aqueous 1,4-dioxane (DXN) and aqueous dimethyl sulfoxide (DMSO). More importantly, we report that both LNPs were compact particles with relatively homogeneous density distribution and very low porosity in the internal structure. The stiffness of the particles was independent of the size, and the Young’s modulus was in the range of 0.3–4 GPa. Overall, the fundamental understandings of LNPs gained in this study are essential for the design of LNPs with optimal performance in applications.

Biomaterials of different nature have been and are widely studied for various biomedical applications. In many cases, biomaterial assemblies are designed to mimic biological systems. Although biomaterials have been thoroughly characterized in many aspects, not much quantitative information on the molecular level interactions between different biomaterials is available. That information is very important, on the one hand, to understand the properties of biological systems and, on the other hand, to develop new composite biomaterials for special applications. This work presents a systematic, quantitative analysis of self- and cross-interactions between films of collagen I (Col I), collagen IV (Col IV), laminin (LN-521), and cellulose nanofibrils (CNF), that is, biomaterials of different nature and structure that either exist in biological systems (e.g., extracellular matrices) or have shown potential for 3D cell culture and tissue engineering. Direct surface forces and adhesion between biomaterials-coated spherical micro-particles and flat substrates were measured in phosphate-buffered saline using an atomic force microscope and the colloidal probe technique. Different methods (Langmuir-Schaefer deposition, spin-coating, or adsorption) were applied to completely coat the flat substrates and the spherical micro particles with homogeneous biomaterial films. The adhesion between biomaterials films increased with the time that the films were kept in contact. The strongest adhesion was observed between Col IV films, and between Col IV and LN-521 films after 30 s contact time. In contrast, low adhesion was measured between CNF films, as well as between CNF and LN-521 films. Nevertheless, a good adhesion between CNF and collagen films (especially Col I) was observed. These results increase our understanding of the structure of biological systems and can support the design of new matrices or scaffolds where different biomaterials are combined for diverse biological or medical applications.

The transformation of a molecularly complex and irregularly shaped lignin into a nanoscale spherical architecture is anticipated to play a pivotal role in the promotion of lignin valorization. From the standpoint of using colloidal lignin particles (CLPs) as building blocks for a diverse range of applications, it has become essential to study their interactions with soluble compounds of varied origin. However, the lack of model films with well-defined surface properties similar to those of CLPs has hindered fundamental studies using surface-sensitive techniques. Here, we report well-defined and stable thin films prepared from CLPs and demonstrate their suitability for investigation of surface phenomena. Direct adsorption on substrates coated with a cationic anchoring polymer resulted in uniform distribution of CLPs as shown with atomic force microscopy (AFM). Quartz crystal microbalance with dissipation monitoring (QCM-D) experiments revealed higher adsorbed mass of cationic lignin onto the CLP-coated substrate in comparison to the film prepared from dissolved lignin, suggesting preferential adsorption via the carboxylic acid enriched surfaces of CLPs. QCM-D further enabled detection of small changes such as particle swelling or partial dissolution not detectable via bulk methods such as light scattering. The CLP thin films remained stable until pH 8 and displayed only a low degree of swelling. Increasing the pH to 10 led to some instability, but their spherical geometry remained intact until complete dissolution was observed at pH 12. Particles prepared from aqueous acetone or aqueous tetrahydrofuran solution followed similar trends regarding adsorption, pH stability, and wetting, although the particle size affected the magnitude of adsorption. Overall, our results present a practical way to prepare well-defined CLP thin films that will be useful not only for fundamental studies but also as a platform for testing stability and interactions of lignin nanoparticles with materials of technical and biomedical relevance.

The development of in vitro cell models that mimic cell behavior in organs and tissues is an approach that may have remarkable impact on drug testing and tissue engineering applications in the future. Plant-based, chemically unmodified cellulose nanofibrils (CNF) hydrogel is a natural, abundant, and biocompatible material that has attracted great attention for biomedical applications, in particular for three-dimensional cell cultures. However, the mechanisms of cell-CNF interactions and factors that affect these interactions are not yet fully understood. In this work, multi-parametric surface plasmon resonance (SPR) was used to study how the adsorption of human hepatocellular carcinoma (HepG2) cells on CNF films is affected by the different proteins and components of the cell medium. Both human recombinant laminin-521 (LN-521, a natural protein of the extracellular matrix) and poly-l-lysine (PLL) adsorbed on CNF films and enhanced the attachment of HepG2 cells. Cell medium components (glucose and amino acids) and serum proteins (fetal bovine serum, FBS) also adsorbed on both bare CNF and on protein-coated CNF substrates. However, the adsorption of FBS hindered the attachment of HepG2 cells to LN-521- and PLL-coated CNF substrates, suggesting that serum proteins blocked the formation of laminin-integrin bonds and decreased favorable PLL-cell electrostatic interactions. This work sheds light on the effect of different factors on cell attachment to CNF, paving the way for the utilization and optimization of CNF-based materials for different tissue engineering applications.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Three-dimensional (3D) printing has been an emerging technique to fabricate precise scaffolds for biomedical applications. Cellulose nanofibril (CNF) hydrogels have attracted considerable attention as a material for 3D printing because of their shear-thinning properties. Combining cellulose nanofibril hydrogels with alginate is an effective method to enable cross-linking of the printed scaffolds in the presence of Ca2+ ions. In this work, spherical colloidal lignin particles (CLPs, also known as spherical lignin nanoparticles) were used to prepare CNF-alginate-CLP nanocomposite scaffolds. High-resolution images obtained by atomic force microscopy (AFM) showed that CLPs were homogeneously mixed with the CNF hydrogel. CLPs brought antioxidant properties to the CNF-alginate-CLP scaffolds in a concentration-dependent manner and increased the viscosity of the hydrogels at a low shear rate, which correspondingly provide better shape fidelity and printing resolution to the scaffolds. Interestingly, the CLPs did not affect the viscosity at high shear rates, showing that the shear thinning behavior typical for CNF hydrogels was retained, enabling easy printing. The CNF-alginate-CLP scaffolds demonstrated shape stability after printing, cross-linking, and storage in Dulbecco's phosphate buffer solution (DPBS +) containing Ca2+ and Mg2+ ions, up to 7 days. The 3D-printed scaffolds showed relative rehydration ratio values above 80% after freeze-drying, demonstrating a high water-retaining capability. Cell viability tests using hepatocellular carcinoma cell line HepG2 showed no negative effect of CLPs on cell proliferation. Fluorescence microscopy indicated that HepG2 cells grew not only on the surfaces but also inside the porous scaffolds. Overall, our results demonstrate that nanocomposite CNF-alginate-CLP scaffolds have high potential in soft-tissue engineering and regenerative-medicine applications.

Marine ancestors of freshwater sponges had to undergo a series of physiological adaptations to colonize harsh and heterogeneous limnic environments. Besides reduced salinity, river-lake systems also have calcium concentrations far lower than seawater. Cell adhesion in sponges is mediated by calcium-dependent multivalent self-interactions of sulfated polysaccharides components of membrane-bound proteoglycans named aggregation factors. Cells of marine sponges require seawater average calcium concentration (10\xC2\xA0mM) to sustain adhesion promoted by aggregation factors. We demonstrate here that the freshwater sponge Spongilla alba can thrive in a calcium-poor aquatic environment and that their cells are able to aggregate and form primmorphs with calcium concentrations 40-fold lower than that required by marine sponges cells. We also find that their gemmules need calcium and other micronutrients to hatch and generate new sponges. The sulfated polysaccharide purified from S. alba has sulfate content and molecular size notably lower than those from marine sponges. Nuclear magnetic resonance analyses indicated that it is composed of a central backbone of non- and 2-sulfated \xCE\xB1- and \xCE\xB2-glucose units decorated with branches of \xCE\xB1-glucose. Assessments with atomic force microscopy/single-molecule force spectroscopy show that S. alba glucan requires 10-fold less calcium than sulfated polysaccharides from marine sponges to self-interact efficiently. Such an ability to retain multi-cellular morphology with low environmental calcium must have been a crucial evolutionary step for freshwater sponges to successfully colonize inland waters.

Summary At invasion, transformed mammary epithelial cells expand into the stroma through a disrupted myoepithelial (ME) cell layer and basement membrane (BM). The intact ME cell layer has thus been suggested to act as a barrier against invasion. Here, we investigate the mechanisms behind the disruption of ME cell layer. We show that the expression of basal/ME proteins CK5, CK14, and α-SMA altered along increasing grade of malignancy, and their loss affected the maintenance of organotypic 3D mammary architecture. Furthermore, our data suggests that loss of CK5 prior to invasive stage causes decreased levels of Zinc finger protein SNAI2 (SLUG), a key regulator of the mammary epithelial cell lineage determination. Consequently, a differentiation bias toward luminal epithelial cell type was detected with loss of mature, α-SMA-expressing ME cells and reduced deposition of basement membrane protein laminin-5. Therefore, our data discloses the central role of CK5 in mammary epithelial differentiation and maintenance of normal ME layer., Graphical abstract, Highlights • Cytokeratin 5 (CK5) impacts mammary epithelial cell lineage differentiation • CK5 loss at pre-invasive stage causes impaired maturation of myoepithelial cells • CK5 loss causes SLUG downregulation and differentiation bias in mammary progenitors, Cell Biology; Developmental Biology; Biophysics

Cross-linked and decolorized lignin nanoparticles (LNPs) were prepared enzymatically and chemically from softwood Kraft lignin. Colloidal lignin particles (CLPs, ca. 200 nm) in a non-malodorous aqueous dispersion could be dried and redispersed in tetrahydrofuran (THF) or in water retaining their stability i.e. spherical shape and size. Two fungal laccases, Trametes hirsuta (ThL) and Melanocarpus albomyces (MaL) were used in the cross-linking reactions. Reactivity of ThL and MaL on Lignoboost™ lignin and LNPs was confirmed by high performance size exclusion chromatography (HPSEC) and oxygen consumption measurements with simultaneous detection of red-brown color due to the formation of quinones. Zeta potential measurements verified oxidation of LNPs via formation of surface-oriented carboxylic acid groups. Dynamic light scattering (DLS) revealed minor changes in the particle size distributions of LNPs after laccase catalyzed radicalization, indicating preferably covalent intraparticular cross-linking over polymerization. Changes in the surface morphology of laccase treated LNPs were imaged by atomic force (AFM) and transmission emission (TEM) microscopy. Furthermore, decolorization of LNPs without degradation was obtained using ultrasonication with H2O2 in alkaline reaction conditions. The research results have high impact for the utilization of Kraft lignin as nanosized colloidal particles in advanced bionanomaterial applications in medicine, foods and cosmetics including different sectors from chemical industry.

Lignocellulosics, i.e., cellulose, lignin and hemicelluloses, are natural renewable polymers of high technological interest. The properties of products based on these polymers are largely determined by the forces at their interfaces. This review summarizes the main findings related to surface interactions relevant for papermaking and describes how the interest in novel, high performance renewable materials has changed the focus of the research to nanocellulosic materials. Areas of interest that need further work are also outlined.

Plant-based polysaccharides (cellulose and hemicellulose) are a very interesting option for the preparation of sustainable composite materials to replace fossil plastics, but the optimum bonding mechanism between the hard and soft components is still not well known. In this work, composite films made of cellulose nanofibrils (CNF) and various modified and unmodified polysaccharides (galactoglucomannan, GGM; hydrolyzed and oxidized guar gum, GGhydHox; and guar gum grafted with polyethylene glycol, GG-g-PEG) were characterized from the nano- to macroscopic level to better understand how the interactions between the composite components at nano/microscale affect macroscopic mechanical properties, like toughness and strength. All the polysaccharides studied adsorbed well on CNF, although with different adsorption rates, as measured by quartz crystal microbalance with dissipation monitoring (QCM-D). Direct surface and friction force experiments using the colloidal probe technique revealed that the adsorbed polysaccharides provided repulsive forces-well described by a polyelectrolyte brush model - and a moderate reduction in friction between cellulose surfaces, which may prevent CNF aggregates during composite formation and, consequently, enhance the strength of dry films. High affinity for cellulose and moderate hydration were found to be important requirements for polysaccharides to improve the mechanical properties of CNF-based composites in wet conditions. The results of this work provide fundamental information on hemicellulose-cellulose interactions and can support the development of polysaccharide-based materials for different packaging and medical applications.

Biomaterials for antimalarial drug transport still need to be investigated in order to attain nanocarriers that can tackle essential issues related to malaria treatment, e.g. complying with size requirements and targeting specificity for their entry into Plasmodium-infected red blood cells (pRBCs), and limiting premature drug elimination or drug resistance evolution. Two types of dendritic macromolecule that can form vehicles suitable for antimalarial drug transport are herein explored. A new hybrid dendritic-linear-dendritic block copolymer based on Pluronic® F127 and amino terminated 2,2′-bis(glycyloxymethyl)propionic acid dendrons with a poly(ester amide) skeleton (HDLDBC-bGMPA) and an amino terminated dendronized hyperbranched polymer with a polyester skeleton derived from 2,2′-bis(hydroxymethyl)propionic acid (DHP-bMPA) have provided self-assembled and unimolecular micelles. Both types of micelle carrier are biocompatible and exhibit appropriate sizes to enter into pRBCs. Targeting studies have revealed different behaviors for each nanocarrier that may open new perspectives for antimalarial therapeutic approaches. Whereas DHP-bMPA exhibits a clear targeting specificity for pRBCs, HDLDBC-bGMPA is incorporated by all erythrocytes. It has also been observed that DHP-bMPA and HDLDBC-bGMPA incorporate into human umbilical vein endothelial cells with different subcellular localization, i.e. cytosolic and nuclear, respectively. Drug loading capacity and encapsulation efficiencies for the antimalarial compounds chloroquine, primaquine and quinacrine ranging from 30% to 60% have been determined for both carriers. The resulting drug-loaded nanocarriers have been tested for their capacity to inhibit Plasmodium growth in in vitro and in vivo assays., This research was supported by grants BIO2014-52872-R, CTQ2015-70174-P, MAT2015-66208-C3-1-P (Ministerio de Ciencia, Innovación y Universidades, Spain, including FEDER funds), 2014-SGR-938 (Generalitat de Catalunya, Spain), and (Gobierno de Aragón-FSE). ISGlobal and IBEC are members of the CERCA Programme, Generalitat de Catalunya. A. L. thanks the Ministerio de Ciencia, Innovación y Universidades for his grant (FPU12/05210). L. N. B.-C. is supported by the European Commission under Horizon 2020's Marie Skłodowska-Curie Actions COFUND scheme (Grant Agreement no. 712754) and by the MINECO's Severo Ochoa programme (Grant SEV-2014-0425 (2015–2019))., The authors acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).

In vitro cell culture or tissue models that mimic in vivo cellular response have potential in tissue engineering and regenerative medicine, and are a more economical and accurate option for drug toxicity tests than animal experimentation. The design of in vivo-like cell culture models should take into account how the cells interact with the surrounding materials and how these interactions affect the cell behavior. Cell-material interactions are furthermore important in cancer metastasis and tumor progression, so deeper understanding of them can support the development of new cancer treatments. Herein, the colloidal probe microscopy technique was used to quantify the interactions of two cell lines (human pluripotent stem cell line WA07 and human hepatocellular carcinoma cell line HepG2) with natural, xeno-free biomaterials of different chemistry, morphology, and origin. Key components of extracellular matrices –human collagens I and IV, and human recombinant laminin-521−, as well as wood-derived, cellulose nanofibrils –with evidenced potential for 3D cell culture and tissue engineering– were analysed. Both strength of adhesion and force curve profiles depended on biomaterial nature and cell characteristics. The successful growth of the cells on a particular biomaterial required cell-biomaterial adhesion energies above 0.23 nJ/m. The information obtained in this work supports the development of new materials or hybrid scaffolds with tuned cell adhesion properties for tissue engineering, and provides a better understanding of the interactions of normal and cancerous cells with biomaterials in the human body.

Lignin has interesting functionalities to be exploited in adhesives for medicine, foods and textiles. Nanoparticles (NPs) &lt, 100 nm coated with poly (L-lysine), PL and poly(L-glutamic acid) PGA were prepared from the laccase treated lignin to coat nanocellulose fibrils (CNF) with heat. NPs ca. 300 nm were prepared, &beta, casein coated and cross-linked with transglutaminase (Tgase) to agglutinate chamois. Size exclusion chromatography (SEC) and Fourier-transform infrared (FTIR) spectroscopy were used to characterize polymerized lignin, while zeta potential and dynamic light scattering (DLS) to ensure coating of colloidal lignin particles (CLPs). Protein adsorption on lignin was studied by quartz crystal microbalance (QCM). Atomic force microscopy (AFM) was exploited to examine interactions between different polymers and to image NPs with transmission electron microscopy (TEM). Tensile testing showed, when using CLPs for the adhesion, the stress improved ca. 10 and strain ca. 6 times compared to unmodified Kraft. For the &beta, casein NPs, the values were 20 and 8, respectively, and for the &beta, casein coated CLPs between these two cases. When NPs were dispersed in adhesive formulation, the increased Young&rsquo, s moduli confirmed significant improvement in the stiffness of the joints over the adhesive alone. Exploitation of lignin in nanoparticulate morphology is a potential method to prepare bionanomaterials for advanced applications.

This paper provides evidence for the adsorption of suberin onto cellulose and illustrates the effect of suberin and suberin fatty acid (SFA) treatment on the physicochemical surface properties of the modified cellulose substrate. Suberin and SFA were extracted from the outer bark of birch (Betula pendula Roth) and characterized by Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry (GC–MS). The results confirmed the extracts to comprise primarily of depolymerised suberin and SFA. The adsorption of suberin and SFA on cellulose was studied by treating the filter paper with solutions of suberin and SFA. Changes in the surface topography of the filter paper following treatment were analysed with atomic force microscopy and the effect on wettability assessed through contact angle measurement. X-ray photoelectron spectroscopy analysis showed that the cellulose substrate was evenly covered. GC–MS analysis showed which suberin monomers were preferentially adsorbed on the cellulose surfaces. Based on these results, it can be inferred that suberin and SFA have the potential for cellulose surface modification. This finding supports the view that suberin and SFA could be used to improve the interfacial properties of cellulose fibre reinforced composites. The novelty in this study is that it provides detailed information, derived though advanced experimental techniques, about the manner and mechanisms behind changes in surface physiochemical properties of cellulose obtained through suberin and SFA adsorption.

Prompt and accurate diagnosis of malaria is part of an effective disease management (1) , because if not treated malaria can quickly become life-threatening, whereas false positives increase treatment costs and drug-induced resistance, giving a wrong idea of therapeutic efficacy. Since the symptoms of malaria are nonspecific, the observation of clinical features alone might not be enough and should be confirmed with a parasitological analysis. Microscopic examination of Giemsa-stained thin and/or thick blood smears remains the conventional approach for diagnosis (2). The sensitivity of this relatively inexpensive method is excellent, allowing the detection of as few as 5 parasites per µL of blood, and permitting also the determination of the infecting species and of the developmental stage of circulating parasites. In addition, smears provide a permanent record for quality assessment of the diagnosis. However, microscopy requires considerable expertise learned through extended training, the procedure is labor-intensive and time-consuming, and the variability in stains and in techniques used to collect and process blood affects slide interpretation (3). Finally, routine clinical microscopy cannot reliably detect very low parasitemias (

openaire: EC/H2020/720303/EU//ZELCOR Coating of colloidal lignin particles (CLPs), or lignin nanoparticles (LNPs), with proteins was evaluated in order to establish a safe, self-assembly mediated modification technique to tune their surface chemistry. Gelatin and poly- l-lysine formed the most pronounced protein corona on the CLP surface, as determined by dynamic light scattering (DLS) and zeta potential measurements. Spherical morphology of individual protein coated CLPs was confirmed by transmission electron (TEM) and atomic force (AFM) microscopy. A mechanistic adsorption study with several random coiled and globular model proteins was carried out using quartz crystal microbalance with dissipation monitoring (QCM-D). The three-dimensional (3D) protein fold structure and certain amino acid interactions were decisive for the protein adsorption on the lignin surface. The main driving forces for protein adsorption were electrostatic, hydrophobic, and van der Waals interactions, and hydrogen bonding. The relative contributions of these interactions were highly dependent on the ionic strength of the surrounding medium. Capillary electrophoresis (CE) and Fourier transform infrared spectroscopy (FTIR) provided further evidence of the adsorption-enhancing role of specific amino acid residues such as serine and proline. These results have high impact on the utilization of lignin as colloidal particles in biomedicine and biodegradable materials, as the protein corona enables tailoring of the CLP surface chemistry for intended applications.

A lack of renewable resources and their inefficient use is a major challenge facing the society. Lignin is a natural biopolymer obtained mainly as a by-product from the pulp- and paper-making industries, and is primarily burned to produce energy. However, interest for using lignin in more advanced applications has increased rapidly. In particular, lignin based nanoparticles could find potential use in functional surface coatings, nanoglue, drug delivery, and microfluidic devices. In this work, a straightforward method to produce lignin nanoparticles from waste lignin obtained from kraft pulping is introduced. Spherical lignin nanoparticles were obtained by dissolving softwood kraft lignin in tetrahydrofuran (THF) and subsequently introducing water into the system through dialysis. No chemical modification of lignin was needed. Water acts as a non-solvent reducing lignin's degrees of freedom causing the segregation of hydrophobic regions to compartments within the forming nanoparticles. The final size of the nanoparticles depended on the pre-dialysis concentration of dissolved lignin. The stability of the nanoparticle dispersion as a function of time, salt concentration and pH was studied. In pure water and at room temperature the lignin nanoparticle dispersion was stable for over two months, but a very low pH or high salt concentration induced aggregation. It was further demonstrated that the surface charge of the particles could be reversed and stable cationic lignin nanoparticles were produced by adsorption of poly(diallyldimethylammonium chloride) (PDADMAC).

The development of materials that combine the excellent mechanical strength of cellulose nanofibrils (CNF) with the lubricating properties of hyaluronic acid (HA) is a new, promising approach to cartilage implants not explored so far. A simple, solvent-free method to produce a very lubricating, strong cellulosic material by covalently attaching HA to the surface of CNF films is described in this work. A detailed analysis of the tribological properties of the CNF films with and without HA is also presented. Surface and friction forces at micro/nanoscale between model hard surfaces (glass microspheres) and the CNF thin films were measured using an atomic force microscope and the colloid probe technique. The effect of HA attachment, the pH and the ionic strength of the aqueous medium on the forces was examined. Excellent lubrication was observed for CNF films with HA attached in conditions where the HA layer was highly hydrated. These results pave the way for the development of new nanocellulose-based materials with good lubrication properties that could be used in biomedical applications. (C) 2015 Elsevier B.V. All rights reserved.

Early metazoans had to evolve the first cell adhesion mechanism addressed to maintain a distinctive multicellular morphology. As the oldest extant animals, sponges are good candidates for possessing remnants of the molecules responsible for this crucial evolutionary innovation. Cell adhesion in sponges is mediated by the calcium-dependent multivalent self-interactions of sulfated polysaccharides components of extracellular membrane-bound proteoglycans, namely aggregation factors. Here, we used atomic force microscopy to demonstrate that the aggregation factor of the sponge Desmapsamma anchorata has a circular supramolecular structure and that it thus belongs to the spongican family. Its sulfated polysaccharide units, which were characterized via nuclear magnetic resonance analysis, consist preponderantly of a central backbone composed of 3-alpha-Glc1 units partially sulfated at 2-and 4-positions and branches of Pyr(4,6)alpha-Gal1 -> 3-alpha-Fuc2(SO3)1 -> 3-alpha-Glc4(SO3) 1 -> 3-alpha-Glc -> 4-linked to the central alpha-Glc units. Single-molecule force measurements of self-binding forces of this sulfated polysaccharide and their chemically desulfated and carboxyl-reduced derivatives revealed that the sulfate epitopes and extracellular calcium are essential for providing the strength and stability necessary to sustain cell adhesion in sponges. We further discuss these findings within the framework of the role of molecular structures in the early evolution of metazoans.

The self-assembly of peptides and proteins into amyloid fibrils of nanometric thickness and up to several micrometres in length, a phenomenon widely observed in biological systems, has recently aroused a growing interest in nanotechnology and nanomedicine. Here we have applied atomic force microscopy and single molecule force spectroscopy to study the amyloidogenesis of a peptide derived from human amylin and of its reverse sequence. The spontaneous formation of protofibrils and their orientation along well-defined directions on graphite and DMSO-coated graphite substrates make the studied peptides interesting candidates for nanotechnological applications. The measured binding forces between peptides correlate with the number of hydrogen bonds between individual peptides inside the fibril structure according to molecular dynamics simulations.

The role of amyloid β (Aβ) peptide in the onset and progression of Alzheimer's disease is linked to the presence of soluble Aβ species. Sulfated glycosaminoglycans (GAGs) promote Aβ fibrillogenesis and reduce the toxicity of the peptide in neuronal cell cultures, but a satisfactory rationale to explain these effects at the molecular level has not been provided yet. We have used circular dichroism, Fourier transform infrared spectroscopy, fluorescence microscopy and spectroscopy, protease digestion, atomic force microscopy (AFM), and molecular dynamics simulations to characterize the association of the 42-residue fragment Aβ(42) with sulfated GAGs, hyaluronan, chitosan, and poly(vinyl sulfate) (PVS). Our results indicate that the formation of stable Aβ(42) fibrils is promoted by polymeric GAGs with negative charges placed in-frame with the 4.8-Å separating Aβ(42) monomers within protofibrillar β-sheets. Incubation of Aβ(42) with excess sulfated GAGs and hyaluronan increased amyloid fibril content and resistance to proteolysis 2- to 5-fold, whereas in the presence of the cationic polysaccharide chitosan, Aβ(42) fibrillar species were reduced by 25% and sensitivity to protease degradation increased ∼3-fold. Fibrils of intermediate stability were obtained in the presence of PVS, an anionic polymer with more tightly packed charges than GAGs. Important structural differences between Aβ(42) fibrils induced by PVS and Aβ(42) fibrils obtained in the presence of GAGs and hyaluronan were observed by AFM, whereas mainly precursor protofibrillar forms were detected after incubation with chitosan. Computed binding energies per peptide from -11.2 to -13.5 kcal/mol were calculated for GAGs and PVS, whereas a significantly lower value of -7.4 kcal/mol was obtained for chitosan. Taken together, our data suggest a simple and straightforward mechanism to explain the role of GAGs as enhancers of the formation of insoluble Aβ(42) fibrils trapping soluble toxic forms.

The environmental cues received by the cells from synthetic substrates in vitro are very different from those they receive in vivo. In this study, we applied the LangmuireSchaefer (LS) deposition, a variant of LangmuireBlodgett technique, to fabricate a biomimetic microenvironment mimicking the structure and organization of native Bruch's membrane for the production of the functional human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells. Surface pressure-area isotherms were measured simultaneously with Brewster angle microscopy to investigate the self-assembly of human collagens type I and IV on air-subphase interface. Furthermore, the structure of the prepared collagen LS films was characterized with scanning electron microscopy, atomic force microscopy, surface plasmon resonance measurements and immunofluorescent staining. The integrity of hESC-RPE on double layer LS films was investigated by measuring transepithelial resistance and permeability of small molecular weight substance. Maturation and functionality of hESC-RPE cells on double layer collagen LS films was further assessed by RPE-specific gene and protein expression, growth factor secretion, and phagocytic activity. Here, we demonstrated that the prepared collagen LS films have layered structure with oriented fibers corresponding to architecture of the uppermost layers of Bruch's membrane and result in increased barrier properties and functionality of hESC-RPE cells as compared to the commonly used dip-coated controls.

Hydrogen peroxide is the product of various enzymatic reactions, and is thus typically utilized as the analyte in biosensors. However, its detection with conventional materials, such as noble metals or glassy carbon, is often hindered by slow kinetics and biofouling of the electrode. In this study electrochemical properties and suitability to peroxide detection as well as ability to resist biofouling of Pt-doped ta-C samples were evaluated. Pure ta-C and pure Pt were used as references. According to the results presented here it is proposed that combining ta-C with Pt results in good electrocatalytic activity towards H2O2 oxidation with better tolerance towards aqueous environment mimicking physiological conditions compared to pure Pt. In biofouling experiments, however, both the hybrid material and Pt were almost completely, blocked after immersion in protein-containing solutions and did not produce any peaks for ferrocenemethanol oxidation or reduction. On the contrary, it was still possible to obtain clearpeaks for H2O2 oxidation with them after similar treatment Moreover, quartz crystal microbalance experiment showed less protein adsorption on the hybrid sample compared to Pt which is also supported by the electrochemical biofouling experiments for H2O2 detection. (C) 2015 Elsevier B.V. All rights reserved.

Current malaria therapeutics demands strategies able to selectively deliver drugs to Plasmodium-infected red blood cells (pRBCs) in order to limit the appearance of parasite resistance. Here, the poly(amidoamines) AGMA1 and ISA23 have been explored for the delivery of antimalarial drugs to pRBCs. AGMA1 has antimalarial activity per se as shown by its inhibition of the in vitro growth of Plasmodium falciparum, with an IC50 of 13.7 μM. Fluorescence-assisted cell sorting data and confocal fluorescence microscopy and transmission electron microscopy images indicate that both polymers exhibit preferential binding to and internalization into pRBCs versus RBCs, and subcellular targeting to the parasite itself in widely diverging species such as P. falciparum and Plasmodium yoelii, infecting humans and mice, respectively. AGMA1 and ISA23 polymers with hydrodynamic radii around 7 nm show a high loading capacity for the antimalarial drugs primaquine and chloroquine, with the final conjugate containing from 14.2% to 32.9% (w/w) active principle. Intraperitoneal administration of 0.8 mg/kg chloroquine as either AGMA1 or ISA23 salts cured P. yoelii-infected mice, whereas control animals treated with twice as much free drug did not survive. These polymers combining into a single chemical structure drug carrying capacity, low unspecific toxicity, high biodegradability and selective internalization into pRBCs, but not in healthy erythrocytes for human and rodent malarias, may be regarded as promising candidates deserving to enter the antimalarial therapeutic arena. © 2014 Elsevier B.V.

Glycosaminoglycans (GAGs) play an important role in the sequestration of Plasmodium falciparum-infected red blood cells (pRBCs) in the microvascular endothelium of different tissues, as well as in the formation of small clusters (rosettes) between infected and non-infected red blood cells (RBCs). Both sequestration and rosetting have been recognized as characteristic events in severe malaria. Here we have used heparin and pRBCs infected by the 3D7 strain of P. falciparum as a model to study GAG–pRBC interactions. Fluorescence microscopy and fluorescence-assisted cell sorting assays have shown that exogenously added heparin has binding specificity for pRBCs (preferentially for those infected with late forms of the parasite) vs. RBCs. Heparin–pRBC adhesion has been probed by single-molecule force spectroscopy, obtaining an average binding force ranging between 28 and 46 pN depending on the loading rate. No significant binding of heparin to non-infected RBCs has been observed in control experiments. This work represents the first approach to quantitatively evaluate GAG–pRBC molecular interactions at the individual molecule level.

VK: T10413 50 %; T10402 50 % There is a growing interest to design biomimetic self-assembled composite films from renewable resources aimed at a combination of high toughness, strength and stiffness. However, the relationship between interfacial interactions of the components and the mechanical performance of the composite is still poorly understood. In this work we present evidence of the link between mechanical performance of carbohydrate-based composites with nanolubrication and with direct surface forces between the hard and soft domain in the system. Our approach was to use nanofibrillated cellulose (NFC) as the major reinforcing domain and to modify it by adsorption of a small amount of soft polyethylene glycol grafted carboxymethyl cellulose (CMC-g-PEG). The effect of the soft polymer on direct normal and friction forces in air between cellulose surfaces was evaluated using colloidal probe microscopy. The fibrillar structure of the NFC thin film affected the frictional behaviour; when decreasing load, the friction between pure cellulose surfaces increased, suggesting partial pull-out of fibrils, a phenomenon not observed for non-fibrillar cellulose substrates. Adsorption of CMC-g-PEG on both surfaces decreased the friction considerably but adhesion was still high. The symmetric system, having both cellulose substrates covered with polymer, was compared to asymmetric systems where only one surface was covered with polymer. Furthermore, a free standing composite film was prepared by non-ionic self-assembly of NFC and CMC-g-PEG with 99:1 weight-ratio; the mechanical properties of the macroscopic films were related to the nanoscaled interactions between the components. The composition studied showed excellent mechanical properties which do not follow the simple rule of mixture. Thus, a synergy in the direct surface forces and mechanical properties was found. This approach offers a robust path to aid in the efficient design of next generation biomimetic composites.

Antimicrobial peptide drugs are increasingly attractive therapeutic agents as their roles in physiopathological processes are being unraveled and because the development of recombinant DNA technology has made them economically affordable in large amounts and high purity. However, due to lack of specificity regarding the target cells, difficulty in attaining them, or reduced half-lives, most current administration methods require high doses. On the other hand, reduced specificity of toxic drugs demands low concentrations to minimize undesirable side-effects, thus incurring the risk of having sublethal amounts which favour the appearance of resistant microbial strains. In this scenario, targeted delivery can fulfill the objective of achieving the intake of total quantities sufficiently low to be innocuous for the patient but that locally are high enough to be lethal for the infectious agent. One of the major advances in recent years has been the size reduction of drug carriers that have dimensions in the nanometer scale and thus are much smaller than -and capable of being internalized by- many types of cells. Among the different types of potential antimicrobial peptide-encapsulating structures reviewed here are liposomes, dendritic polymers, solid core nanoparticles, carbon nanotubes, and DNA cages. These nanoparticulate systems can be functionalized with a plethora of biomolecules providing specificity of binding to particular cell types or locations; as examples of these targeting elements we will present antibodies, DNA aptamers, cell-penetrating peptides, and carbohydrates. Multifunctional Trojan horse-like nanovessels can be engineered by choosing the adequate peptide content, encapsulating structure, and targeting moiety for each particular application.

The specific functional structure of natural proteins is determined by the way in which amino acids are sequentially connected in the polypeptide. The tight sequence/structure relationship governing protein folding does not seem to apply to amyloid fibril formation because many proteins without any sequence relationship have been shown to assemble into very similar β-sheet-enriched structures. Here, we have characterized the aggregation kinetics, seeding ability, morphology, conformation, stability, and toxicity of amyloid fibrils formed by a 20-residue domain of the islet amyloid polypeptide (IAPP), as well as of a backward and scrambled version of this peptide. The three IAPP peptides readily aggregate into ordered, β-sheet-enriched, amyloid-like fibrils. However, the mechanism of formation and the structural and functional properties of aggregates formed from these three peptides are different in such a way that they do not cross-seed each other despite sharing a common amino acid composition. The results confirm that, as for globular proteins, highly specific polypeptide sequential traits govern the assembly pathway, final fine structure, and cytotoxic properties of amyloid conformations.

The histopathological hallmarks of Alzheimer disease are the self-aggregation of the amyloid beta peptide (Abeta) in extracellular amyloid fibrils and the formation of intraneuronal Tau filaments, but a convincing mechanism connecting both processes has yet to be provided. Here we show that the endogenous polysaccharide chondroitin sulfate B (CSB) promotes the formation of fibrillar structures of the 42-residue fragment, Abeta(1-42). Atomic force microscopy visualization, thioflavin T fluorescence, CD measurements, and cell viability assays indicate that CSB-induced fibrils are highly stable entities with abundant beta-sheet structure that have little toxicity for neuroblastoma cells. We propose a wedged cylinder model for Abeta(1-42) fibrils that is consistent with the majority of available data, it is an energetically favorable assembly that minimizes the exposure of hydrophobic areas, and it explains why fibrils do not grow in thickness. Fluorescence measurements of the effect of different Abeta(1-42) species on Ca(2+) homeostasis show that weakly structured nodular fibrils, but not CSB-induced smooth fibrils, trigger a rise in cytosolic Ca(2+) that depends on the presence of both extracellular and intracellular stocks. In vitro assays indicate that such transient, local Ca(2+) increases can have a direct effect in promoting the formation of Tau filaments similar to those isolated from Alzheimer disease brains.