An Underwater Surface-Drying Peptide Inspired by a Mussel Adhesive Protein.

Water hampers the formation of strong and durable bonds between adhesive polymers and solid surfaces, in turn hindering the development of adhesives for biomedical and marine applications. Inspired by mussel adhesion, a mussel foot protein homologue (mfp3S-pep) is designed, whose primary sequence is designed to mimic the pI, polyampholyte, and hydrophobic characteristics of the native protein. Noticeably, native protein and synthetic peptide exhibit similar abilities to self-coacervate at given pH and ionic strength. 3,4-dihydroxy-l-phenylalanine (Dopa) proves necessary for irreversible peptide adsorption to both TiO2 (anatase) and hydroxyapatite (HAP) surfaces, as confirmed by quartz crystal microbalance measurements, with the coacervate showing superior adsorption. The adsorption of Dopa-containing peptides is investigated by attenuated total reflection infrared spectroscopy, revealing initially bidentate coordinative bonds on TiO2, followed by H-bonded and eventually long-ranged electrostatic and Van der Waals interactions. On HAP, mfp3s-pep-3Dopa adsorption occurs predominantly via H-bond and outer-sphere complexes of the catechol groups. Importantly, only the Dopa-bearing compounds are able to remove interfacial water from the target surfaces, with the coacervate achieving the highest water displacement arising from its superior wetting properties. These findings provide an impetus for developing coacervated Dopa-functionalized peptides/polymers adhesive formulations for a variety of applications on wet polar surfaces.

Stable Formation of Gold Nanoparticles onto Redox-Active Solid Biosubstrates Made of Squid Suckerin Proteins.

The use of biomolecules to synthesize inorganic nanomaterials, including metallic nanoparticles, offers the ability to induce controlled growth under mild environmental conditions. Here, recently discovered silk-like "suckerin" proteins are used to induce the formation of gold nanoparticles (AuNPs). Advantage is taken of the distinctive biological and physico-chemical characteristics of suckerins, namely their facile recombinant expression, their solubility in aqueous solutions, and their modular primary structure with high molar content of redox-active tyrosine (Tyr) residues to induce the formation of AuNPs not only in solution, but also from nanostructured solid substrates fabricated from suckerins.

Confocal microscopy-based goniometry of barnacle cyprid permanent adhesive.

Biological adhesives are materials of particular interest in the fields of bio-inspired technology and antifouling research. The adhesive of adult barnacles has received much attention over the years; however, the permanent adhesive of the cyprid - the colonisation stage of barnacles - is a material about which very little is presently known. We applied confocal laser-scanning microscopy to the measurement of contact angles between the permanent adhesive of barnacle cyprid larvae and self-assembled monolayers of OH- and CH3-terminated thiols. Measurement of contact angles between actual bioadhesives and surfaces has never previously been achieved and the data may provide insight into the physicochemical properties and mechanism of action of these functional materials. The adhesive is a dual-phase system post-secretion, with the behaviour of the components governed separately by the surface chemistry. The findings imply that the cyprid permanent adhesion process is more complex than previously thought, necessitating broad re-evaluation of the system. Improved understanding will have significant implications for the production of barnacle-resistant coatings as well as development of bio-inspired glues for niche applications.

A novel geometry for a laboratory-based larval settlement assay.

A novel configuration, consisting of two apposing surfaces bounding a vertical water column, is presented and evaluated for settlement assays using cyprids of Balanus amphitrite. Assays were conducted on planar surfaces, ranging from hydrophobic polystyrene to hydrophilic glass and including CH(3)- and NH(3) (+)-terminated self-assembled monolayers (SAMs). Identical apposing surfaces generated settlement rates comparable to those obtained in prior studies, while a choice assay yielded consistent results, with individual replicates each indicating the preferred surface for settlement. As gravity favours contact with the lower apposing surface, cyprids trapped at the air/water interface settled on or around the perimeter where the water column meets the lower substratum. These cyprids are capable of selecting a settlement location and are thus not lost to the assay. The assay geometry lends itself to assessing cyprid exploration and settlement on planar surfaces with chemical patterning, including relief microstructures, without using a confining material or requiring the coating of a three-dimensional well.

In situ ATR FTIR study of dextrin adsorption on anatase TiO2.

The adsorption of two dextrin-based polymers, a regular wheat dextrin (TY) and a carboxymethyl-substituted (CM) dextrin, onto an anatase TiO(2) particle film has been studied using in situ attenuated total reflection (ATR) FTIR spectroscopy. Infrared spectra of the polymer solutions and the polymer adsorbed at the anatase surface were acquired for two solution conditions: pH 3 and pH 9; below and above the isoelectric point (IEP) of anatase, respectively. Comparison of the polymer solution spectra and the adsorbed layer spectra highlighted a number of spectral differences that were attributed to involvement of the carboxyl group of CM Dextrin interacting with the anatase surface directly and the adsorption of oxidized dextrin chains in the case of regular dextrin (TY) at high pH. The adsorption/desorption kinetics were determined by monitoring spectral peaks of the pyranose ring of both polymers. Adsorption equilibrium was not established for Dextrin TY for many hours, whereas CM Dextrin reached equilibrium in its adsorption within 60 min. The extent of desorption of Dextrin TY (observed by flowing a background electrolyte dextrin-free solution) was extensive at both pH values, which reflects the poor affinity and binding of the polymer on anatase. In contrast, CM Dextrin underwent almost no desorption, indicating a high affinity between the carboxyl groups of the polymer and the anatase surface.

Effects of surface charge and Gibbs surface energy on the settlement behaviour of barnacle cyprids (Balanus amphitrite).

Gibbs surface energy has long been considered to be an important parameter in the design of fouling-resistant surfaces for marine applications. Rigorous testing of the hypothesis that settlement is related to Gibbs surface energy however has never been accomplished, due mainly to practical limitations imposed by the necessary combination of surface engineering and biological evaluation methods. In this article, the effects of surface charge and Gibbs surface energy on the settlement of cyprids of an important fouling barnacle, Balanus amphitrite, were evaluated. Settlement assays were conducted on a range of self-assembled monolayers (SAMs) (CH(3)-, OH-, COOH-, N(CH(3))(3) (+)-, NH(2)-terminated), presented in gold-coated polystyrene well plates, varying in terms of their surface charge and Gibbs surface energy. Contrary to contemporary theory, settlement was not increased by high-energy surfaces, rather the opposite was found to be the case with cyprids settling in greater numbers on a low-energy CH(3)- SAM compared to a high-energy OH- SAM. Settlement was also greater on negatively-charged SAMs, compared to neutral and positively-charged SAMs. These findings are discussed in the context of data drawn from surfaces that varied in multiple characteristics simultaneously, as have been used previously for such experiments. The finding that surface charge, rather than total surface energy, may be responsible for surface selection by cyprids, will have significant implications for the design of future fouling-resistant materials.

In situ infrared spectroscopic investigation of Perna canaliculus mussel larvae primary settlement.

The adhesion of Perna canaliculus mussel larvae on a germanium (Ge) prism in filtered seawater at 16 degrees C has been investigated by in situ attenuated total reflection infrared (ATR-IR) spectroscopy. The adhesive from the mussel larvae was spectrally monitored over 2 h and the IR spectrum showed its glycoproteinaceous nature with sulphated and carboxylated moieties. The adhesive from the mussel larvae differs from that of the adults, resembling the mucus secretion found in other benthic marine species at a larval stage. To date, this appears to be the first work describing the main chemical features of secreted adhesive associated with the primary settlement of mussel larvae. The acquired knowledge on the larval adhesive features may lead to enhanced settling methods in aquaculture or to antibiofouling strategies. The ATR-IR approach under temperature control is potentially useful for such studies on other small benthic organisms in both marine and freshwater environments.

Preventing mussel adhesion using lubricant-infused materials.

Mussels are opportunistic macrofouling organisms that can attach to most immersed solid surfaces, leading to serious economic and ecological consequences for the maritime and aquaculture industries. We demonstrate that lubricant-infused coatings exhibit very low preferential mussel attachment and ultralow adhesive strengths under both controlled laboratory conditions and in marine field studies. Detailed investigations across multiple length scales-from the molecular-scale characterization of deposited adhesive proteins to nanoscale contact mechanics to macroscale live observations-suggest that lubricant infusion considerably reduces fouling by deceiving the mechanosensing ability of mussels, deterring secretion of adhesive threads, and decreasing the molecular work of adhesion. Our study demonstrates that lubricant infusion represents an effective strategy to mitigate marine biofouling and provides insights into the physical mechanisms underlying adhesion prevention.

Chemistry-specific surface adsorption of the barnacle settlement-inducing protein complex.

Gregarious settlement in barnacle larvae (cyprids) is induced by a contact pheromone, the settlement-inducing protein complex (SIPC). The SIPC has been identified both in the cuticle of adult barnacles and in the temporary adhesive secretion (footprint) of cyprids. Besides acting as a settlement inducer, the presence of the SIPC in footprints points to its additional involvement in the adhesion process. SIPC adsorption behaviour was therefore investigated on a series of self-assembled monolayers (SAMs) by surface plasmon resonance at the pH of seawater (8.3). Fibrinogen and α2-macroglobulin (A2M) (blood complement protease inhibitors with which the SIPC shares 29% sequence homology) were used in the adsorption experiments as positive and negative standards, respectively. The mass uptake of the SIPC was comparable to that of fibrinogen, with adsorption observed even on the protein-resistant oligo(ethylene glycol) surface. Notably, on the positively charged SAM the SIPC showed a kinetic overshoot, indicating a metastable configuration causing the amount of adsorbed protein to temporarily exceed its equilibrium value. A2M adsorption was low or negligible on all SAMs tested, except for the positively charged surface, indicating that A2M adsorption is mainly driven by electrostatics. Evaluation of SIPC non-specific adsorption kinetics revealed that it adsorbed irreversibly and non-cooperatively on all surfaces tested.

Mussel adhesion is dictated by time-regulated secretion and molecular conformation of mussel adhesive proteins.

Interfacial water constitutes a formidable barrier to strong surface bonding, hampering the development of water-resistant synthetic adhesives. Notwithstanding this obstacle, the Asian green mussel Perna viridis attaches firmly to underwater surfaces via a proteinaceous secretion (byssus). Extending beyond the currently known design principles of mussel adhesion, here we elucidate the precise time-regulated secretion of P. viridis mussel adhesive proteins. The vanguard 3,4-dihydroxy-L-phenylalanine (Dopa)-rich protein Pvfp-5 acts as an adhesive primer, overcoming repulsive hydration forces by displacing surface-bound water and generating strong surface adhesion. Using homology modelling and molecular dynamics simulations, we find that all mussel adhesive proteins are largely unordered, with Pvfp-5 adopting a disordered structure and elongated conformation whereby all Dopa residues reside on the protein surface. Time-regulated secretion and structural disorder of mussel adhesive proteins appear essential for optimizing extended nonspecific surface interactions and byssus' assembly. Our findings reveal molecular-scale principles to help the development of wet-resistant adhesives.

Molecular surface chemistry in marine bioadhesion.

This review covers the in situ molecular physicochemical characterisation of bioadhesives at solid/liquid interfaces, with the aim of elucidating the adhesion strategies that lie at the root of marine biofouling. It focuses on three major foulers: mussels, algae and barnacles. The dispersal of these organisms, their colonisation of surfaces, and ultimately their survival rely critically on the ability of the organisms' larvae or spores to locate a favourable settlement site and undergo metamorphosis, thus initiating their sessile existence. Differences in the composition of adhesive secretions and the strategies employed for their temporary or permanent implementation exists between the larval and adult life stages. To date, only a few adhesive secretions from marine fouling organisms have been adequately described in terms of their chemical composition, and a survey revealed the presence of certain recurrent functional groups, specifically catechol, carboxylate, monoester-sulphate and -phosphate. This review will describe the binding modes of such functionalities to wet mineral/metal oxides surfaces. Such functionalities will be ranked based on their ability to bind to hydrophilic surfaces replacing surface-bound water (Langmuir adsorption constant) as well as other adsorbates (competitive adsorption). A plausible explanation for the propensity of the reviewed adhesive functionalities to bind to hydrous metal oxide surfaces will be given on the basis of the Hard and Soft Acids and Bases principle, Hofmeister effects and entropic considerations. From the in situ analysis of marine organism bioadhesives and adsorption studies of functionalities relevant to the bioadhesion process, insights can be gleaned for a knowledge-based innovation of antifouling strategies and the synthesis of strong, durable adhesive materials, which are suitable for implementation in wet environments.

Biotinyl moiety-selective polymer films with highly ordered macropores.

Macroporous polymer films with long-range uniformity and biotinyl-moiety selective recognition sites have been developed. A hierarchical molecular imprinting strategy afforded significant enhancements in quartz crystal microbalance (QCM) sensitivities towards biotinylated compounds.

Alginate ion adsorption on a TiO2 particle film and interactions of adsorbed alginate with calcium ions investigated by attenuated total reflection infrared (ATR-IR) spectroscopy.

The adsorption of alginic acid on a TiO(2) particle film from aqueous solution was investigated by attenuated total reflection infrared (ATR-IR) spectroscopy. ATR-IR spectra recorded at different pHs confirmed that alginate adsorption to TiO(2) is favored at pH 3.0 and no significant adsorption occurs above pH 5.0. Upon adsorption the carboxylic acid groups of alginic acid are converted to the carboxylate form and bind to surface Ti(IV) ions via bridging bidentate structures. Spectral analyses of the carboxylic acid and carboxylate stretching vibrations indicated that about three in four -COOH groups are converted to -COO(-) groups as they bind coordinately to TiO(2). Additionally, the spectral data at pH 8.0 showed specific interactions of Ca(2+) ions with the free COO(-) groups of the polysaccharide backbone of adsorbed alginic acid.