Dendritic saccharide surfactant polymers as antifouling interface materials to reduce platelet adhesion

Here, we report on the synthesis of dendritic saccharide surfactant polymers as antifouling interface materials to reduce platelet adhesion. An acetal-protected poly(amidoamine) (PAMAM) dendron (5, G = 2) was first synthesized by using aminoacetaldehyde dimethyl acetal (1) as the starting material to provide a monovalent focal structure with dimethyl acetal-protected aldehyde functionality. Maltose dendron (M4, 6) was obtained by reacting the peripheral amine groups of acetal-dendron (5) with maltonolactone. The dendritic surfactant polymers (9) were then synthesized via a two-step method by sequential addition of maltose dendron and hexanal to react with the amine groups on the poly(vinylamine) (PVAm) backbone. Surface activity of the amphiphilic glycopolymers at the air/water interface was demonstrated by reduction in water surface tension. Adsorption of the amphiphilic glycopolymers at the solid/water interface was examined on octadecyltrichlorosilane (OTS)-coated coverslips by water contact angle measurements. A nanoscale understanding of surface-induced self-assembly of the dendritic surfactant polymer on highly oriented pyrolytic graphite (HOPG) was gained using AFM operated in fluid tapping mode. A lateral ordering of adsorbing surfactant polymer was visualized with a pattern in strands 60 degrees out of alignment. The static platelet adhesion tests show that the hexyl side chains can facilitate adsorption of the surfactant polymers onto hydrophobic substrates, while the maltose dendron side chains can provide a dense canopy of protective glycocalyx-like layer as an antifouling interface to reduce platelet adhesion.     

Polygodial: a contact active antifouling biocide

Ongoing investigation of the candidate antifouling (AF) biocide polygodial (PG) has revealed that this compound may be contact active, whereby it can confer effect while remaining bound within a stable matrix. To test this hypothesis, the AF activity of PG-laced coatings was compared to that of seawater in which PG-laced coatings had been soaked. Four coating types spanning high to low affinity for PG were examined and AF activity was assessed based on inhibition of settlement and metamorphosis of larvae of three fouling organisms: Ciona savignyi Herdman, Mytilus galloprovincialis Lamarck and Spirobranchus caraniferus Gray. Direct exposure to the coatings had a significantly greater impact on larval metamorphosis than indirect exposure to seawater in which the coatings had been soaked. In particular, metamorphosis was almost completely inhibited by high-affinity coatings containing ≥ 200 ng of PG per replicate, while corresponding soaking waters had no detectable effect. These findings support the assertion that PG is contact active.     

Effect of secochiliolide acid isolated from the Patagonian shrub Nardophyllum bryoides as active component in antifouling paints

Environmental concerns about the use of toxic antifoulants have led to an increased interest in the development of new alternatives. So far, most of the antifouling natural products have been obtained from marine organisms. However, some secondary metabolites from terrestrial plants could be promising antifoulant candidates. The antifouling performance of secochiliolide acid, the main component isolated from Nardophyllum bryoides ethanolic extract, was evaluated for inclusion in rosin-based coatings.Field testing was conducted during the summer months at Mar del Plata harbor, Argentina. The results indicated that secochiliolide acid-based paints completely inhibited the settlement of Bugula neritina colonies, Polydora sp., Hydroides elegans, Corophium sp. and solitary ascidians, and also reduced the attachment of some algae as Enteromorpha intestinalis and Ectocarpus sp. In addition, a lower density and diversity of microfouling species was registered.These results highlighted the importance of terrestrial plants as a sustainable source of potential environmentally friendly antifoulants.     

Bio-inspired antifouling approaches: the quest towards non-toxic and non-biocidal materials

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Shogaols from Zingiber officinale as promising antifouling agents

We isolated the highly potent attachment-inhibitors (three times more active than standard CuSO4 in the blue mussel assay), trans-6-, 8-, and 10-shogaols, from a hexane extract of the roots of ginger, Zingiber officinale Roscoe. Trans-8-shogaol showed the highest antifouling activity comparable with that of tributyltin fluoride (TBTF), which is recognized as one of the most effective antifouling agents, in the conventional submerged assay.     

Screening of bromotyramine analogues as antifouling compounds against marine bacteria

Rapid and efficient synthesis of 23 analogues inspired by bromotyramine derivatives, marine natural products, by means of CuSO₄-catalysed [3+2] alkyne–azide cycloaddition is described. The final target was then assayed for anti-biofilm activity against three Gram-negative marine bacteria, Pseudoalteromonas ulvae (TC14), Pseudoalteromonas lipolytica (TC8) and Paracoccus sp. (4M6). Most of the synthesised bromotyramine/triazole derivatives are more active than the parent natural products Moloka’iamine (A) and 3,5-dibromo-4-methoxy-β-phenethylamine (B) against biofilm formation by the three bacterial strains. Some of these compounds were shown to act as non-toxic inhibitors of biofilm development with EC₅₀ < 200 μM without any effect on bacterial growth even at high concentrations (200 μM).     

Nontoxic piperamides and their synthetic analogues as novel antifouling reagents

Bioassay-guided isolation of an acetone extract from a terrestrial plant Piper betle produced four known piperamides with potent antifouling (AF) activities, as evidenced by inhibition of settlement of barnacle cypris larvae. The AF activities of the four piperamides and 15 synthesized analogues were compared and their structure–activity relationships were probed. Among the compounds, piperoleine B and 1-[1-oxo-7-(3′,4′-methylenedioxyphenyl)-6E-heptenyl]-piperidine (MPHP) showed strong activity against settlement of cyprids of the barnacle Balanus amphitrite, having EC₅₀ values of 1.1?±?0.3 and 0.5?±?0.2?μg?ml^?1, respectively. No toxicity against zebra fish was observed following incubation with these two compounds. Besides being non-toxic, 91% of piperoleine B-treated cyprids and 84% of MPHP-treated cyprids at a concentration of 100?μM completed normal metamorphosis in recovery bioassays, indicating that the anti-settlement effect of these two compounds was reversible. Hydrolysis and photolysis experiments indicated that MPHP could be decomposed in the marine environment. It is concluded that piperamides are promising compounds for use in marine AF coatings.     

Evaluation of dihydrooroidin as an antifouling additive in marine paint

Methods used to deter biofouling of underwater structures and marine vessels present a serious environmental issue and are both problematic and costly for government and commercial marine vessels worldwide. Current antifouling methods include compounds that are toxic to aquatic wildlife and marine ecosystems. Dihydrooroidin (DHO) was shown to completely inhibit Halomonas pacifica biofilms at 100 μM in a static biofilm inhibition assay giving precedence for the inhibition of other marine biofilm-forming organisms. Herein we present DHO as an effective paint-based, non-cytotoxic, antifouling agent against marine biofouling processes in a marine mesocosm.     

Natural products as antifouling compounds: recent progress and future perspectives

Since early 2008, an increasing number of countries have ratified an international treaty to ban the application of antifouling (AF) coatings based on organotin compounds (eg tributyltin (TBT) and triphenyltin). As a result, the demand for environmentally friendly, non-toxic or low-toxicity AF compounds and technologies (green AF agents) has become an urgent reality. Marine coatings based on Cu₂O and various other biocides have a negative impact on the environment and they must eventually be replaced by new, effective, and environmentally friendly AF compounds. This mini-review describes important AF compounds discovered from a variety of organisms from 2004 until mid 2009, and discusses recent and general trends in the discovery of AF compounds. Finally, a perspective on the future of AF compound development is presented. The discussion is aimed at updating scientists and engineers on the current challenges facing AF research.     

Preparation and characterization of amphiphilic triblock terpolymer-based nanofibers as antifouling biomaterials

Antifouling surfaces are critical for the good performance of functional materials in various applications including water filtration, medical implants, and biosensors. In this study, we synthesized amphiphilic triblock terpolymers (tri-BCPs, coded as KB) and fabricated amphiphilic nanofibers by electrospinning of solutions prepared by mixing the KB with poly(lactic acid) (PLA) polymer. The resulting fibers with amphiphilic polymer groups exhibited superior antifouling performance to the fibers without such groups. The adsorption of bovine serum albumin (BSA) on the amphiphilic fibers was about 10-fold less than that on the control surfaces from PLA and PET fibers. With the increase of the KB content in the amphiphilic fibers, the resistance to adsorption of BSA was increased. BSA was released more easily from the surface of the amphiphilic fibers than from the surface of hydrophobic PLA or PET fibers. We have also investigated the structural conformation of KB in fibers before and after annealing by contact angle measurements, transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDX), and coarse-grained molecular dynamics (CGMD) simulation to probe the effect of amphiphilic chain conformation on antifouling. The results reveal that the amphiphilic KB was evenly distributed within as-spun hybrid fibers, while migrated toward the core from the fiber surface during thermal treatment, leading to the reduction in antifouling. This suggests that the antifouling effect of the amphiphilic fibers is greatly influenced by the arrangement of amphiphilic groups in the fibers.     

Hybrid xerogel films as novel coatings for antifouling and fouling release

Hybrid sol-gel-derived xerogel films prepared from 45/55 (mol ratio) n-propyltrimethoxysilane (C3-TMOS)/tetramethylorthosilane (TMOS), 2/98 (mol ratio) bis[3-(trimethoxysilyl)propyl]-ethylenediamine (enTMOS)/tetraethylorthosilane (TEOS), 50/50 (mol ratio) n-octyltriethoxysilane (C8-TEOS)/TMOS, and 50/50 (mol ratio) 3,3,3-trifluoropropyltrimethoxysilane (TFP-TMOS)/TMOS were found to inhibit settlement of zoospores of the marine fouling alga Ulva (syn. Enteromorpha) relative to settlement on acid-washed glass and give greater release of settled zoospores relative to glass upon exposure to pressure from a water jet. The more hydrophobic 50/50 C8-TEOS/TMOS xerogel films had the lowest critical surface tension by comprehensive contact angle analysis and gave significantly greater release of 8-day Ulva sporeling biomass after exposure to turbulent flow generated by a flow channel than the other xerogel surfaces or glass. The 50/50 C8-TEOS/TMOS xerogel was also a fouling release surface for juveniles of the tropical barnacle Balanus amphitrite. X-ray photon electron data indicated that the alkylsilyl residues of the C3-TMOS-, C8-TEOS-, and TFP-TMOS-containing xerogels were located on the surface of the xerogel films (in a vacuum), which contributes to the film hydrophobicity. Similarly, the amine-containing silyl residues of the enTMOS/TEOS films were located at the surface of the xerogel films, which contributes to the more hydrophilic character and increased critical surface tension of these films.     

Viscoelasticity of living materials: mechanics and chemistry of muscle as an active macromolecular system

At the molecular and cellular level, mechanics and chemistry are two aspects of the same macromolecular system. We present a bottom-up approach to such systems based on Kramers' diffusion theory of chemical reactions, the theory of polymer dynamics, and the recently developed models for molecular motors. Using muscle as an example, we develop a viscoelastic theory of muscle in terms of an simple equation for single motor protein movement. Both A.V. Hill's contractile component and A.F. Huxley's equation of sliding-filament motion are shown to be special cases of the general viscoelastic theory of the active material. Some disparity between the mechanical and the chemical views of cross-bridges and motor proteins are noted, and a duality between force and energy in discrete states and transitions of macromolecular systems is discussed.     

Bacterial assay for the rapid assessment of antifouling and fouling release properties of coatings and materials

An assay has been developed to accurately quantify the growth and release behaviour of bacterial biofilms on several test reference materials and coatings, using the marine bacterium Cobetia marina as a model organism. The assay can be used to investigate the inhibition of bacterial growth and release properties of many surfaces when compared to a reference. The method is based upon the staining of attached bacterial cells with the nucleic acid-binding, green fluorescent SYTO 13 stain. A strong linear correlation exists between the fluorescence of the bacterial suspension measured (RFU) using a plate reader and the total bacterial count measured with epifluorescence microscopy. This relationship allows the fluorescent technique to be used for the quantification of bacterial cells attached to surfaces. As the bacteria proliferate on the surface over a period of time, the relative fluorescence unit (RFU) measured using the plate reader also shows an increase with time. This was observed on all three test surfaces (glass, Epikote and Silastic T2) over a period of 4 h of bacterial growth, followed by a release assay, which was carried out by the application of hydrodynamic shear forces using a custom-made rotary device. Different fixed rotor speeds were tested, and based on the release analysis, 12 knots was used to provide standard shear force. The assay developed was then applied for assessing three different antifouling coatings of different surface roughness. The novel assay allows the rapid and sensitive enumeration of attached bacteria directly on the coated surface. This is the first plate reader assay technique that allows estimation of irreversibly attached bacterial cells directly on the coated surface without their removal from the surface or extraction of a stain into solution.     

Degradation of antifouling biocides

The relative biodegradability in seawater of a number of compounds in current use in antifouling paints viz. Sea-Nine? 211 antifoulant (4,5-dichloro-2-n-octyl-4-isothiazolin-3-one), Irgarol(R) 1051 (2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine), diuron (3-(3,4-dichlorophenyl)l-l-dimethylurea), chlorothalonil (tetrachloroisophthalonitrile) and TBTO (tributyltin oxide) was investigated. The disappearance of the each compound from seawater was monitored over 8 w by bioassay using the ship-fouling diatom Amphora coffeaeformis. The results show, that under the test conditions employed, biodegradability ranges from very readily biodegradable (Sea-Nine 211) to non-biodegradable (diuron and Irgarol 1051). The results are discussed in relation to published data on biocide degradation.     

Acyl silicates and acyl aluminates as activated intermediates in peptide formation on clays

Glycine reacts with heating on dried clays and other minerals to give peptides in much better yield than in the absence of mineral. This reaction was proposed to occur by way of an activated intermediate such as an acyl silicate or acyl aluminate (i.e., the anhydride of a carboxylic acid with Si–OH or Al–OH), analogous to acyl phosphates involved in several biochemical reactions including peptide bond synthesis. We confirmed the proposed mechanism by trapping the intermediate, as well as by direct spectroscopic observation of a related intermediate. The reaction of amino acids on periodically dried mineral surfaces represents a widespread, geologically realistic setting for prebiotic peptide formation viain situ activation.     

Acyl silicates and acyl aluminates as activated intermediates in peptide formation on clays

Glycine reacts with heating on dried clays and other minerals to give peptides in much better yield than in the absence of mineral. This reaction was proposed to occur by way of an activated intermediate such as an acyl silicate or acyl aluminate (i.e., the anhydride of a carboxylic acid with Si-OH or Al-OH), analogous to acyl phosphates involved in several biochemical reactions including peptide bond synthesis. We confirmed the proposed mechanism by trapping the intermediate, as well as by direct spectroscopic observation of a related intermediate. The reaction of amino acids on periodically dried mineral surfaces represents a widespread, geologically realistic setting for prebiotic peptide formation via in situ activation.     

The performance of hybrid titania/silica-derived xerogels as active antifouling/fouling-release surfaces against the marine alga Ulva linza: in situ generation of hypohalous acids

Mixed titania/silica xerogels were prepared using titanium tetraisopropoxide (TTIP) and tetraethoxy orthosilicate (TEOS). Xerogel properties were modified by incorporating n-octyltriethoxysilane (C8). The xerogels catalyze the oxidation of bromide and chloride with hydrogen peroxide (H₂O₂) to produce hypohalous acids at pH 7 and pH 8. The antifouling/ fouling-release performance of a TTIP/C8/TEOS xerogel in the presence and absence of H₂O₂ was evaluated for the settlement of zoospores of the marine alga Ulva linza and for the removal of sporelings (young plants). In the absence of H₂O₂, differences in the settlement of zoospores and removal of sporelings were not significant relative to a titanium-free C8/TEOS xerogel. Addition of H₂O₂ gave a significant reduction in zoospore settlement and sporeling removal relative to the C8/TEOS xerogel and relative to peroxide-free conditions. The impact of TTIP on xerogel characteristics was evaluated by comprehensive contact angle analysis, scanning electron microscopy, and X-ray photoelectron spectroscopy.