| 1. |
- Andersson, Jonas
(author)
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Complement Activation Triggered by Biomaterial Surfaces : Mechanisms and Regulation
- 2003
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Doctoral thesis (other academic/artistic)abstract
- Today there are a vast number of medical devices in temporary or permanent contact with human tissues. Blood-biomaterial contact is known to trigger the complement system and results in generation of fluid phase anaphylatoxins C3a and C5a, and surface-bound C3b and iC3b. All these products together are able to attract and activate leukocytes and trigger release of inflammatory mediators leading to a systemic inflammation indirectly causing hemostatic problems and even organ failure. The aim of this study was to identify how complement is triggered on a biomaterial surface and to find ways to regulate this activation.The finding that complement activation on biomaterials can be divided into initiation and amplification will facilitate regulation of complement activation biomaterial surfaces. This concept is also compatible with the two techniques to regulate complement activation on a surface.
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| 2. |
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| 3. |
- Andersson, Marcus, 1975, et al.
(author)
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Effect of molecular mobility of polymeric implants on soft tissue reactions: An in vivo study in rats
- 2008
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In: Journal of Biomedical Materials Research Part A. - : Wiley. - 1552-4965 .- 1549-3296. ; 84A:3, s. 652-660
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Journal article (peer-reviewed)abstract
- Although numerous different polymers are used as implants or otherwise studied for many other biotechnical applications, there is a lack of basic models that correlate polymer characteristics with foreign body reactions. This study aims at developing one such model by systematically studying surface molecular mobility of polymeric implants in soft tissues in vivo. Changing the length of the alkyl side chain of poly(alkyl methacrylates) (PAMAs), provides an interesting opportunity to study the surface molecular mobility with minimal changes of the hydrophobicity of the surface. Thus, in this study three different PAMAs, with increasingly surface mobility; poly (isobutyl methacrylate) (PIBMA), poly(butyl methacrylate) (PBMA), and poly(lauryl methacralate) (PLMA) along with pure titanium (Ti) substrates were implanted in the dorsum of Sprague-Dawley rats. Inflammatory cell recruitment, cell adhesion, and cytokine release were studied after 1, 3, and 28 days of implantation. Total number of inflammatory cells in the exudate was measured but no correlation between surface mobility and cell recruitment where found. However, the number of surface associated cells where significantly lower on the surfaces with high molecular mobility (PLMA and PBMA). The histological evaluation performed after 28 days revealed thicker fibrous capsule and a higher number of blood vessels on the low molecular mobility surface (PIBMA). After 28 days the cell activity was higher on the high molecular mobility surfaces (PLMA and PBMA) compared with PIBMA, based on the cytokine release. None of the surfaces induced any significant cell-death. On the basis of the results of this study we conclude that there is a significant difference in biological response to surfaces with different in molecular mobility. This might affect the wound healing process and the biocompatibility of biomaterials. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007 -------------------------------------------------------------------------------- Received: 13 March 2006; Revised: 15 December 2006; Accepted: 29 January 2007 Digital Object Identifier (DOI) 10.1002/jbm.a.31389 About DOI
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| 4. |
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| 5. |
- Andersson, Marcus, 1975, et al.
(author)
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Quartz crystal microbalance-with dissipation monitoring (QCM-D) for real time measurements of blood coagulation density and immune complement activation on artificial surfaces
- 2005
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In: Biosensors & Bioelectronics. - : Elsevier BV. - 0956-5663. ; 21:1, s. 79-86
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Journal article (peer-reviewed)abstract
- A recently developed variant of quartz crystal microbalance (QCM) called QCM-with dissipation monitoring (QCM-D) allows simultaneous and simple measurements of changes in adsorbed mass as well as the viscoelastic property (D-factor) of deposited protein layers on the sensor surface. We have taken the QCM-D technology a step further and demonstrated its advantages in the study of protein assembly as a consequence of surface induced immune complement activation, or contact activated blood coagulation. In the present study we have continued our QCM-D investigations of surface assembly of fibrin clot formation and complement activation and incubated differently modified quartz sensor surfaces in blood plasma and sera. Polymer surfaces used were spin-coated polyethylene, poly(ethylene terephtalate), poly(methylmetacrylate) and poly(dimethylsiloxane). Also used were sputtered titanium and heparin grafted surfaces. In this investigation we found that we could describe the surface induced coagulation with four independent parameters: (1) Time of onset of coagulation, (2) fibrin deposition rate, (3) total frequency shift at stable plateau, and (4) fibrin clot density. The most important finding was that the blood plasma clot density can be assessed with the use of D determinations and that the clot density varied significantly with the chemical composition of the surface. However, the D-factor did not give any new analytical information about the possible complement activation mechanisms. Nevertheless, the QCM-D was found to be a reliable tool for the analysis of surface induced complement activation. We also compared the QCM-D technique with traditional enzyme immuno assay (EIA) measurements of soluble products from the surface activation of the complement and coagulation systems. We found that the results from EIA and QCM-D measurements corresponded well for the complement activation but not for the coagulation, probably due to the biological complexity of the coagulation system.
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| 6. |
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| 7. |
- Ausili, A., et al.
(author)
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Membrane docking mode of the C2 domain of PKCε: An infrared spectroscopy and FRET study
- 2013
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In: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736. ; 1828:2, s. 552-560
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Journal article (peer-reviewed)abstract
- The C2 domain of PKCε binds to negatively charged phospholipids but little is known so far about the docking orientation of this domain when it is bound. By using a FRET assay we have studied the binding of this domain to model membranes. We have also used ATR-Fourier transform infrared spectroscopy with polarized light (ATR-FTIR) to determine the docking mode by calculating the β-sandwich orientation when the domain is bound to different types of model membranes. The vesicle lipid compositions were: POPC/POPE/POPA (22:36:42) imitating the inner leaflet of a plasma membrane, POPC/POPA (50:50) in which POPE has been eliminated with respect to the former composition and POPC/POPE/CL (43:36:21) imitating the inner mitochondrial membrane. Results show that the β-sandwich of the PKCα-C2 domain is inclined at an angle α close to 45 to the membrane normal. Some differences were found with respect to the extent of binding as a function of phospholipid composition and small changes on secondary structure were only evident when the domain was bound to model membranes of POPC/POPA: in this case, the percentage of β-sheet of the C2 domain increases if compared with the secondary structure of the domain in the absence of vesicles. With respect to the β-sandwich orientation, when the domain is bound to POPC/POPE/CL membranes it forms an angle with the normal to the surface of the lipid bilayer (39) smaller than that one observed when the domain interacts with vesicles of POPC/POPA (49). © 2012 Elsevier B.V. All rights reserved.
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| 8. |
- Ausili, A., et al.
(author)
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Quartz crystal microbalance with dissipation monitoring and the real-time study of biological systems and macromolecules at interfaces
- 2012
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In: Biomedical Spectroscopy and Imaging. - 2212-8794 .- 2212-8808. ; 1:4, s. 325-338
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Journal article (peer-reviewed)abstract
- QCM-D technique is based on the physical phenomenon that generates an acoustic shear wave with an oscillating resonance in quartz resulting in an evanescent wave that arises at the interface of the quartz and the solution. The amplitude of the acoustic wave is influenced by the deposition of material onto the quartz surface and from the subsequent decrease of the frequency the bound mass can be calculated. The dissipation shift which arises inform about viscoelasticity and flexibility of the adsorbed material. QCM-D can be applied for real-time studies of several biological systems since it is a simple, fast, low-cost and sensitive technique without having to label any sample. Common applications in biological field include measurements on adsorption of lipids, proteins, DNA and cells directly onto the surface of the sensor, which generally are chemically modified by self-assembled monolayer (SAM) technique or by spin-coated polymers. QCM-D can also be used to study molecular interactions between macromolecules and adsorbed materials. Three examples of the use of this technique are presented, namely the docking orientation of the C2 domain of PKCε on phospholipid membranes, the conformational changes of fibrinogen adsorbed to model acrylic polymers and the attachment of endothelial cells to carboxylated polymers of different configuration.
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| 9. |
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| 10. |
- Berglin, Mattias, 1970, et al.
(author)
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Erosion of a model rosin-based marine antifouling paint binder as studied with quartz crystal microbalance with dissipation monitoring (QCM-D) and ellipsometry
- 2008
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In: Progress in Organic Coatings. ; 61, s. 83-88
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Journal article (peer-reviewed)abstract
- In this study two surface sensitive methods, i.e. quartz crystal microbalance with dissipation monitoring (QCM-D) and ellipsometry, were used for erosion measurements of a rosin-based marine antifouling paint binder. Thin films of the binder were applied on sensor surfaces by the means of spin-coating and the effect of water velocity over the paint film, water temperature or ionic strength on erosion was investigated. Both the acoustic QCM-D model and the optical ellipsometry model gave comparable erosion results. The initial 2-50 nm rapid erosion of the top layer was followed by steady-state erosion rate until end of experiment. For example, the steady-state erosion rate was 12 nm/24 h in artificial seawater at 23 degrees C and with a flow of 200 mu l/min over the paint surface as measured with QCM-D. The erosion rate increased with increased velocity and increased temperature. Ionic strength had no effect on the erosion rate of this model binder. At low water velocities the surface layer was highly dissipative indicating a water filled surface top layer or the formation of deposits on the surface. New characterization techniques that are able to study the erosion mechanisms on the nanometre scale are sought for as the binders get more technically complex containing, for example, nanoparticles or enzymes. Surface sensitive methods could be used to rapidly screen the effect of different binder chemistries or paint additives on the erosion during the paint development process. (C) 2007 Elsevier B.V. All rights reserved.
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| 11. |
- Berglin, Mattias, 1970, et al.
(author)
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Fibrinogen Adsorption and Conformational Change on Model Polymers: Novel Aspects of Mutual Molecular Rearrangement
- 2009
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In: Langmuir. - : American Chemical Society (ACS). - 0743-7463 .- 1520-5827. ; 25:10, s. 5602-5608
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Journal article (peer-reviewed)abstract
- By combining quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR), the organic mass, water content, and corresponding protein film structure of fibrinogen adsorbed to acrylic polymeric substrates with varying polymer chain flexibility was investigated. Albumin and immunoglobulin G were included as reference proteins. For fibrinogen, the QCM-D model resulted in decreased adsorbed mass with increased polymer chain flexibility. This stands in contrast to the SPR model, in which the adsorbed mass increased with increased polymer chain flexibility. As the QCM-D model includes the hydrodynamically coupled water, we propose that on the nonflexible polymer significant protein conformational change with water incorporation in the protein film takes place. Fibrinogen maintained a more native conformation on the flexible polymer, probably due to polymer chain rearrangement rather than protein conformational change. In comparison with immunoglobulin G and albumin, polymer chain flexibility had only minor impact on adsorbed mass and protein structure. Understanding the adsorption and corresponding conformational change of a protein together with the mutual rearrangement of the polymer chain upon adsorption not only has implications in biomaterial science but could also increase the efficacy of molecular imprinted polymers (MIPs).
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| 12. |
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| 13. |
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| 14. |
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| 15. |
- Berglin, Mattias, 1970, et al.
(author)
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Use of surface-sensitive methods for the study of adsorption and cross-linking of marine bioadhesives
- 2005
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In: Journal of Adhesion. - : Informa UK Limited. - 0021-8464 .- 1563-518X .- 1545-5823. ; 81:7-8, s. 805-822
-
Journal article (peer-reviewed)abstract
- The establishment of the bond of sessile marine organisms such as barnacles, mussels, and algae in the marine environment starts with the secretion and the adsorption of the adhesive biopolymers to the substrate. Subsequently, this is followed by the formation of cohesive interactions with the next layer of adhesive biopolymers that are deposited/adsorbed on top of the first layer. These two fundamental processes for the adhesive plaque buildup have been subjected to several investigations in recent years using model molecules, especially Mefp-1 extracted from the blue mussel Mytilus edulis. With the introduction of optical surface-sensitive methods such as ellipsometry, surface plasmon resonance (SPR), and infrared spectroscopy (IR), it has been possible to elucidate both the kinetics of adsorption and structure of the Mefp-1 film. In contrast to adsorption, the cohesive interactions or the cross-linking are not easily followed with these optical methods and new approaches and techniques are required. One such technique that has been useful is the quartz-crystal microbalance with dissipation monitoring (QCM-D), which has been used for cross-linking studies of a variety of biopolymers including bioadhesives from mussel and algae. Copyright © Taylor & Francis Inc.
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| 16. |
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| 17. |
- Dahlstrom, M., et al.
(author)
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Affinity states of biocides determine bioavailability and release rates in marine paints
- 2015
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In: Biofouling. - : Informa UK Limited. - 0892-7014 .- 1029-2454. ; 31:2, s. 201-210
-
Journal article (peer-reviewed)abstract
- A challenge for the next generation marine antifouling (AF) paints is to deliver minimum amounts of biocides to the environment. The candidate AF compound medetomidine is here shown to be released at very low concentrations, ie ng ml(-1) day(-1). Moreover, the release rate of medetomidine differs substantially depending on the formulation of the paint, while inhibition of barnacle settlement is independent of release to the ambient water, ie the paint with the lowest release rate was the most effective in impeding barnacle colonisation. This highlights the critical role of chemical interactions between biocide, paint carrier and the solid/aqueous interface for release rate and AF performance. The results are discussed in the light of differential affinity states of the biocide, predicting AF activity in terms of a high surface affinity and preserved bioavailability. This may offer a general framework for the design of low-release paint systems using biocides for protection against biofouling on marine surfaces.
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| 18. |
- Dahlström, Mia, et al.
(author)
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Evidence for different pharmacological targets for imidazoline compounds inhibiting settlement of the barnacle Balanus improvisus
- 2005
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In: Journal of Experimental Zoology. - : Wiley. - 0022-104X .- 1097-010X. ; 303A:7, s. 551-562
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Journal article (peer-reviewed)abstract
- We describe the effect of eight different imidazoline/guanidinium compounds on the settlement and metamorphosis of larvae of the barnacle Balanus improvisus. These agents were chosen on the basis of their similar pharmacological classification in vertebrates and their chemical similarity to medetomidine and clonidine, previously described as highly potent settlement inhibitors (nanomolar range). Seven of the tested compounds were found to inhibit settlement in a dose-dependent manner in concentrations ranging from 100 nM to 10 microM without any significant lethal effects. In vertebrate systems these substances have overlapping functions and interact with both alpha-adrenoceptors as well as imidazoline binding sites. Antagonizing experiments using the highly specific alpha(2)-antagonist methoxy-idazoxan or agmatine (the putative endogenous ligand at imidazoline receptors) were performed to discriminate between putative pharmacological mechanisms involved in the inhibition of cyprid settlement. Agmatine was not able to reverse the effect of any of the tested compounds. However, methoxy-idazoxan almost completely abolished the settlement inhibition mediated by guanabenz (alpha(2)-agonist, I(2) ligand), moxonidine (alpha(2)-agonist, I(1) ligand) and tetrahydrozoline (alpha-agonist, I(2) ligand). The actions of cirazoline (alpha(1)-agonist, I(2) ligand) BU 224 (I(2) ligand) and metrazoline (I(2) ligand) were not reversed by treatment with methoxy-idazoxan. These results suggest that the settlement inhibition evoked by the I(2) ligands and alpha(2)-agonists used in this study of the neurologically simple but well-organized barnacle larva is mediated through different physiological targets important in the overall settlement process.
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| 19. |
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| 20. |
- Dahlström, Mia, et al.
(author)
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Surface wettability as a determinant in the settlement of the barnacle Balanus Improvisus (DARWIN)
- 2004
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In: Journal of Experimental Marine Biology and Ecology. - : Elsevier BV. - 0022-0981. ; 305:2, s. 223-232
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Journal article (peer-reviewed)abstract
- Several studies have shown that the initial surface wettability, is of importance in the settlement of macrofouling larvae such as barnacles, bryozoans and hydroids in the field as well as in laboratory assays. In this study we present results from laboratory assays using hydrophilic and hydrophobic polystyrene (PS) and cyprid larvae of Balanus improvisus (Darwin). The results obtained differ markedly from those reported for the barnacle Balanus amphitrite (Darwin), where a high surface wettability seemed to be preferred for settlement. Our results show that a surface with intermediary wettability (hydrophilic PS) reduced settlement by 38% as compared to surfaces of low wettability (hydrophobic PS) during an 8-day period. During the experiment, the wettability in the hydrophilic PS dishes was not significantly changed as measured by advancing contact angle with mQ water. Over an 8-day period wettability of the hydrophobic PS dishes approached that of the hydrophilic PS surfaces. We further conducted experiments with highly hydrophilic and highly hydrophobic methylsilane-treated glass surfaces with known chemistry. In this experiment, the settlement of cyprid larvae was completely inhibited by the high wettability surfaces. Contact angle measurements revealed that the wettability during the length of the experiment of the hydrophilic glass surfaces was not significantly altered. We conclude by these experiments that even an intermediate wettability can significantly affect the overall settlement success of the barnacle B. improvisus. The mechanism by which the settlement is impeded might be biologically mediated through the recognition by cyprid larvae of the molecular composition of the surface when the cyprid reverts to the settlement phase, i.e. when swimming behaviour is abandoned in favour of surface exploration, or it is mediated by physicochemical forces acting between the surface and the larval body or the larval antennules. (C) 2004 Published by Elsevier B.V.
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| 21. |
- Erlandsson, Ragnar, et al.
(author)
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Scanning force microscopy - examples of applications to surface chemistry
- 1992
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In: Progress in Colloid and Polymer Science. - 0340-255X .- 1437-8027. ; 88, s. 154-161
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Journal article (peer-reviewed)abstract
- Some recent results from the scanning force microscopy activity at our laboratory are presented. A brief description of attractive mode force microscopy is followed by a discussion of the following examples: O2/H2-induced morphology changes in thin palladium films, structure of spin cast polysulfone films, fibrinogen adsorption on hydrophobic SiO2, and force measurements on hydrophobic/hydrophilic substrates.
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| 22. |
- Fant, Camilla, et al.
(author)
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Investigation of Adsorption and Cross-Linking of a Mussel Adhesive Protein Using Attenuated Total Internal Reflection Fourier Transform Infrared Spectroscopy (ATR-FTIR)
- 2010
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In: Journal of Adhesion. - : Informa UK Limited. - 0021-8464 .- 1545-5823 .- 1563-518X. ; 86:1, s. 25-38
-
Journal article (peer-reviewed)abstract
- Mytilus edulis foot protein 1 (Mefp-1) contains the redox-functional amino acid 3,4-dihydroxyphenylalanine (DOPA), which is a typical feature of most mefp proteins. We have previously shown, using combined optic (ellipsometry) and acoustic (QCM-D) measurements, that the oxidizing agent sodium periodate (NaIO4) and the transition metal ion Cu2+ promote cross-linking of Mefp-1. However, different chemical reaction mechanisms can not be distinguished using these methods. In the present study, we have complemented our previous investigations using Attenuated Total Internal Reflection Fourier Transform Infrared spectroscopy (ATR-FTIR), allowing a spectroscopic analysis of NaIO4 and Cu2+-induced cross-linking of Mefp-1 adsorbed on a ZnSe surface. In aqueous solution, adsorbed Mefp-1 displays absorption bands at 1570, 1472, 1260, and 973 cm(-1). Upon addition of NaIO4 and Cu2+, the absorptions at 1570, 1472, and 973 cm(-1) increase by approximately a factor of two. In contrast, the band at 1260 cm(-1) disappears upon cross-linking using NaIO4, but remains unchanged upon addition of Cu2+. This demonstrates that the band at 1260cm(-1) is attributed to the C O stretching vibration of the side chain hydroxyl groups in DOPA and that Cu2+ forms complexes with DOPA rather than transform it into an o-quinone. Moreover, upon addition of NaIO4 after cross-linking using Cu2+, the band at 1260cm(-1) disappears, indicating that the complex formation between DOPA and Cu2+ is reversed when DOPA is transformed into the o-quinone. These results demonstrate that NaIO4, which initiates a similar reaction to the naturally occurring enzyme catechol oxidase, contributes to the formation of di-DOPA cross-links. In contrast, the dominating contribution to the cross-linking from Cu2+, which is accumulated at high concentrations in the byssus thread of the blue mussel, is via complex formation between the metal and DOPA residues.
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| 23. |
- Fromell, Karin, et al.
(author)
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Absence of conformational change in complement factor 3 and factor XII adsorbed to acrylate polymers is related to a high degree of polymer backbone flexibility
- 2017
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In: Biointerphases. - : American Vacuum Society. - 1934-8630 .- 1559-4106. ; 12:2
-
Journal article (peer-reviewed)abstract
- In previous investigations, the authors have examined the adsorption of albumin, immunoglobulin, and fibrinogen to a series of acrylate polymers with different backbone and side-group flexibility. The authors showed that protein adsorption to acrylates with high flexibility, such as poly(lauryl methacrylate) (PLMA), tends to preserve native conformation. In the present study, the authors have continued this work by examining the conformational changes that occur during the binding of complement factor 3 (C3) and coagulation factor XII (FXII). Native C3 adsorbed readily to all solid surfaces tested, including a series of acrylate surfaces of varying backbone flexibility. However, a monoclonal antibody recognizing a "hidden" epitope of C3 (only exposed during C3 activation or denaturation) bound to the C3 on the rigid acrylate surfaces or on polystyrene (also rigid), but not to C3 on the flexible PLMA, indicating that varying degrees of conformational change had occurred with binding to different surfaces. Similarly, FXII was activated only on the rigid poly(butyl methacrylate) surface, as assessed by the formation of FXIIa-antithrombin (AT) complexes; in contrast, it remained in its native form on the flexible PLMA surface. The authors also found that water wettability hysteresis, defined as the difference between the advancing and receding contact angles, was highest for the PLMA surface, indicating that a dynamic change in the interface polymer structure may help protect the adsorbed protein from conformational changes and denaturation. (C) 2017 Author(s).
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| 24. |
- Harbecke, Olle, 1966, et al.
(author)
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Desensitization of formyl peptide receptors is abolished in calcium ionophore-primed neutrophils: an association of the ligand-receptor complex to the cytoskeleton is not required for a rapid termination of the NADPH-oxidase response.
- 1998
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In: Journal of immunology (Baltimore, Md. : 1950). - 0022-1767. ; 160:5, s. 2463-8
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Journal article (peer-reviewed)abstract
- Binding of ligands to N-formyl peptide chemoattractant receptors exposed on human neutrophils generates signals in the cells that induce an activation of the superoxide anion producing NADPH-oxidase. Ligand binding is followed by a rapid association of the ligand-receptor complex with the cytoskeleton, a process leading to desensitization of the cells with respect to NADPH-oxidase activation. We show that neutrophils that have experienced an intracellular calcium rise obtained through interaction with the calcium-specific ionophore ionomycin are "primed" with respect to the FMLP-induced production of superoxide anions. Mobilization of FMLP receptors from intracellular pools is one well-known mechanism behind the primed response. Based on our finding that ionomycin-treated neutrophils could not be desensitized, we suggest that the lack of association between the ligand-receptor complex and the cytoskeleton is an additional priming mechanism. Since in vivo-exudated neutrophils, which also had mobilized intracellular organelles, could be desensitized, we suggest that the abolished desensitization in ionomycin-treated neutrophils is not due to an inability of newly recruited receptors to couple to the cytoskeleton. We show that a rapid termination of FMLP-induced superoxide anion production is obtained in both desensitizable and nondesensitizable neutrophils, suggesting that the desensitization phenomenon is of limited importance in the oxidase termination process.
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| 25. |
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| 26. |
- Hedlund, Julia, 1975, et al.
(author)
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Change of Colloidal and Surface Properties of Mytilus edulis Foot Protein 1 in the Presence of an Oxidation (NaIO4) or a Complex-Binding (Cu2+) Agent
- 2009
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In: Biomacromolecules. - : American Chemical Society (ACS). - 1525-7797 .- 1526-4602. ; 10:4, s. 845-849
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Journal article (peer-reviewed)abstract
- Quartz crystal microbalance with dissipation monitoring (QCM-D) was used to study the viscoelastic properties of the blue mussel, Mytilus edulis, foot protein 1 (Mefp-1) adsorbed on modified hydrophobic gold surfaces. The change in viscoelasticity was studied after addition of Cu2+ and Mn2+, which theoretically could induce metal complex formation with 3,4-dihydroxyphenylalanine (DOPA) moieties. We also used NaIO4, a nonmetal oxidative agent known to induce di-DOPA formation. Reduction in viscoelasticity of adsorbed Mefp-1 followed the order of NaIO4 > Cu2+ > buffer control > Mn2+. We also studied the formation of molecular aggregates of Mefp-1 in solution with the use of dynamic light scattering (DLS). We found that addition of Cu2+, but not Mn2+, induced the formation of larger DLS-detectable aggregates. Minor aggregate formation was found with NaIO4. With the analytical resolution of small angle X-ray scattering (SAXS), we could detect differences in the molecular structure between NaIO4- and Cu2+-treated Mefp-1 aggregates. We concluded from this study that Cu2+ could participate in intermolecular cross-linking of the Mefp-1 molecule via metal complex formation. Metal incorporation in the protein most likely increases the abrasion resistance of the Mefp-1 layer. NaIO4, on the other hand, resulted in mainly intramolecular formation of di-DOPA, but failed to induce larger intermolecular aggregation phenomena. The described methodological combination of surface sensitive methods, like QCM-D, and bulk sensitive methods, like DLS and SAXS, generates high resolution results and is an attractive platform to investigate intra- and intermolecular aspects of assembly and cross-linking of the Mefp proteins.
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| 27. |
- Hook, F., et al.
(author)
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Variations in coupled water, viscoelastic properties, and film thickness of a Mefp-1 protein film during adsorption and cross-linking: a quartz crystal microbalance with dissipation monitoring, ellipsometry, and surface plasmon resonance study
- 2001
-
In: Analytical Chemistry. - 0003-2700 .- 1520-6882. ; 73:24, s. 5796-804
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Journal article (peer-reviewed)abstract
- We have measured the time-resolved adsorption kinetics of the mussel adhesive protein (Mefp-1) on a nonpolar, methyl-terminated (thiolated) gold surface, using three independent techniques: quartz crystal microbalance with dissipation monitoring (QCM-D), surface plasmon resonance, and ellipsometry. The QCM-D and ellipsometry data shows that, after adsorption to saturation of Mefp-1, cross-linking of the protein layer using NaIO4 transforms it from an extended (approximately 20 nm), water-rich, and hydrogel-like state to a much thinner (approximately 5 nm), compact, and less water-rich state. Furthermore, we show how quantitative data about the thickness, shear elastic modulus, and shear viscosity of the protein film can be obtained with the QCM-D technique, even beyond the Sauerbrey regime, if frequency (f) and energy dissipation (D) measurements measured at multiple harmonics are combined with theoretical simulations using a Voight-based viscoelastic model. The modeling result was confirmed by substituting H2O for D2O. As expected, the D2O substitution does not influence the actual adsorption behavior, but resulted in expected differences in the estimated effective density and shear viscosity. These results provide new insight and understanding about the adsorption kinetics and crosslinking behavior of Mefp-1. They also demonstrate how the above three techniques complement each other for biomolecule adsorption studies.
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| 28. |
- Hulander, Mats, et al.
(author)
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Blood interactions with noble metals: coagulation and immune complement activation.
- 2009
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In: ACS applied materials & interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 1:5, s. 1053-62
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Journal article (peer-reviewed)abstract
- Noble metals are interesting biomaterials for a number of reasons, e.g., their chemical inertness and relative mechanical softness, silver's long known antimicrobial properties, and the low allergenic response shown by gold. Although important for the final outcome of biomaterials, little is reported about early events between pure noble metals and blood. In this article, we used whole blood in the "slide chamber model" to study the activation of the immune complement activation, generation of thrombin/antithrombin (TAT) complexes, and platelet depletion from blood upon contact with silver (Ag), palladium (Pd), gold (Au), titanium (Ti), and Bactiguard, a commercial nanostructured biomaterial coating comprised of Ag, Pd, and Au. The results show the highest TAT generation and platelet depletion on Ti and Au and lower on Pd, Ag, and the Bactiguard coating. The immune complement factor 3 fragment (C3a) was generated by the surfaces in the following order: Ag > Au > Pd > Bactiguard > Ti. Quartz crystal microbalance adsorption studies with human fibrinogen displayed the highest deposition to Ag and the lowest onto the Bactiguard coating. The adsorbed amounts of fibrinogen did not correlate with thrombogenicity in terms of TAT formation and platelet surface accumulation in blood. The combined results suggest, hence, that noble metal chemistry has a different impact on the protein adsorption properties and general blood compatibility. The low thrombogenic response by the Bactiguard coating cannot be explained by any of the single noble metal properties but is likely a successful combination of the nanostructure, nanogalvanic effects, or combinatory chemical and physical materials properties.
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| 29. |
- Hulander, Mats, et al.
(author)
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Gradients in surface nanotopography used to study platelet adhesion and activation
- 2013
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In: Colloids and Surfaces B-Biointerfaces. - : Elsevier BV. - 0927-7765 .- 1873-4367. ; 110, s. 261-269
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Journal article (peer-reviewed)abstract
- Gradients in surface nanotopography were prepared by adsorbing gold nanoparticles on smooth gold substrates using diffusion technique. Following a sintering procedure the particle binding chemistry was removed, and integration of the particles into the underlying gold substrate was achieved, leaving a nanostructured surface with uniform surface chemistry. After pre-adsorption of human fibrinogen, the effect of surface nanotopography on platelets was studied. The use of a gradient in nanotopography allowed for platelet adhesion and activation to be studied as a function of nanoparticle coverage on one single substrate. A peak in platelet adhesion was found at 23% nanoparticle surface coverage. The highest number of activated platelets was found on the smooth control part of the surface, and did not coincide with the number of adhered platelets. Activation correlated inversely with particle coverage, hence the lowest fraction of activated platelets was found at high particle coverage. Hydrophobization of the gradient surface lowered the total number of adhering cells, but not the ratio of activated cells. Little or no effect was seen on gradients with 36 nm particles, suggesting the existence of a lower limit for sensing of surface nano-roughness in platelets. These results demonstrate that parameters such as ratio between size and inter-particle distance can be more relevant for cell response than wettability on nanostructured surfaces. The minor effect of hydrophobicity, the generally reduced activation on nanostructured surfaces and the presence of a cut-off in activation of human platelets as a function of nanoparticle size could have implications for the design of future blood-contacting biomaterials.
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| 30. |
- Hulander, Mats, et al.
(author)
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Immune complement activation is attenuated by surface nanotopography
- 2011
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In: International Journal of Nanomedicine. - 1176-9114 .- 1178-2013. ; 6, s. 2653-2666
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Journal article (peer-reviewed)abstract
- The immune complement (IC) is a cell-free protein cascade system, and the first part of the innate immune system to recognize foreign objects that enter the body. Elevated activation of the system from, for example, biomaterials or medical devices can result in both local and systemic adverse effects and eventually loss of function or rejection of the biomaterial. Here, the researchers have studied the effect of surface nanotopography on the activation of the IC system. By a simple nonlithographic process, gold nanoparticles with an average size of 58 nm were immobilized on a smooth gold substrate, creating surfaces where a nanostructure is introduced without changing the surface chemistry. The activation of the IC on smooth and nanostructured surfaces was viewed with fluorescence microscopy and quantified with quartz crystal microbalance with dissipation monitoring in human serum. Additionally, the ability of pre-adsorbed human immunoglobulin G (IgG) (a potent activator of the IC) to activate the IC after a change in surface hydrophobicity was studied. It was found that the activation of the IC was significantly attenuated on nanostructured surfaces with nearly a 50% reduction, even after pre-adsorption with IgG. An increase in surface hydrophobicity blunted this effect. The possible role of the curvature of the nanoparticles for the orientation of adsorbed IgG molecules, and how this can affect the subsequent activation of the IC, are discussed. The present findings are important for further understanding of how surface nanotopography affects complex protein adsorption, and for the future development of biomaterials and blood-contacting devices.
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| 31. |
- Jansson, Eva, 1972-
(author)
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Blood protein coated model biomaterials : preparation, and cell and tissue response
- 2003
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Doctoral thesis (other academic/artistic)abstract
- Solid biomaterials are widely used in bone and in soft tissue applications. Problems may then arise due to a prolonged inflammation in the proximity of the implant, resulting in the formation of a fibrous capsule and a low vascularisation near the interface.The formation of a blood plasma clot is the starting point of a normal wound healing process. One hypothesis in this thesis work was that a thin immobilised blood plasma clot may improve the integration during the early wound healing period. Model surfaces were made from titanium and silicon, and nm-μm thick blood plasma clots or protein multilayers immobilised onto the surfaces. Another hypothesis was that a submicron surface porosity further improves the integration process, and to study this some of the titanium surfaces were etched in sodium hydroxide, a treatment that resulted in 200 nm wide pores. Such porous titanium surfaces adsorbed 2 to 11 times more albumin and lgG in vitro than the corresponding smooth surfaces at varied pH and protein concentrations.The blood plasma clot coated submicron porous titanium samples were implanted subcutaneously in the back of the rat or in rabbit bone. The soft tissue response was investigated after 3 or 24 hours and the fibrous encapsulation and vessel formation after 7 or 28 days of implantation. The bone ingrowth and the implant stability in the rabbit bone were investigated after 4 weeks of implantation.The monocyte response on multilayer plasma protein coated surfaces was investigated through the analysis of tumor necrosis factor α (TNF-α) and interleukin-10 (IL-10) concentrations in the culture medium, the proportions of Annexin V and propidium iodide (PI) positive cells, and the amounts of nucleated cells. In parallel, the stability of the protein layers and the activation of the complement and coagulation cascades were investigated in vitro by ellipsometry.The results from the monocyte culture and animal experiments show that the early soft tissue inflammatory response and vascularisation can be modulated through the introduction of a surface porosity and by the immobilised protein and plasma clot coatings. However, no significant differences were observed between the different surface modifications in rabbit bone with respect to bone-to-metal contact or percentage of bone area inside the threads.
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| 32. |
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| 33. |
- Karlsson, Martin, et al.
(author)
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Reduction of irreversible protein adsorption on solid surfaces by protein engineering for increased stability
- 2005
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In: Journal of Biological Chemistry. - 0021-9258 .- 1083-351X. ; 280:27, s. 25558-25564
-
Journal article (peer-reviewed)abstract
- The influence of protein stability on the adsorption and desorption behavior to surfaces with fundamentally different properties (negatively charged, positively charged, hydrophilic, and hydrophobic) was examined by surface plasmon resonance measurements. Three engineered variants of human carbonic anhydrase II were used that have unchanged surface properties but large differences in stability. The orientation and conformational state of the adsorbed protein could be elucidated by taking all of the following properties of the protein variants into account: stability, unfolding, adsorption, and desorption behavior. Regardless of the nature of the surface, there were correlation between (i) the protein stability and kinetics of adsorption, with an increased amplitude of the first kinetic phase of adsorption with increasing stability; (ii) the protein stability and the extent of maximally adsorbed protein to the actual surface, with an increased amount of adsorbed protein with increasing stability; (iii) the protein stability and the amount of protein desorbed upon washing with buffer, with an increased elutability of the adsorbed protein with increased stability. All of the above correlations could be explained by the rate of denaturation and the conformational state of the adsorbed protein. In conclusion, protein engineering for increased stability can be used as a strategy to decrease irreversible adsorption on surfaces at a liquid-solid interface.
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| 34. |
- Liu, L, et al.
(author)
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Surface-related triggering of the neutrophil respiratory burst. Characterization of the response induced by IgG adsorbed to hydrophilic and hydrophobic glass surfaces.
- 1997
-
In: Clinical and experimental immunology. - 0009-9104. ; 109:1, s. 204-10
-
Journal article (peer-reviewed)abstract
- Hydrophilic and hydrophobic glass surfaces precoated with human albumin, fibrinogen, or IgG were investigated with respect to their ability to activate the neutrophil NADPH-oxidase. We found that IgG-coated surfaces induced a substantial and prolonged neutrophil production of reactive oxygen species (ROS). When a hydrophilic surface was used to support protein binding, a somewhat lower neutrophil response (around 35%) was obtained, compared with the response induced by IgG on a hydrophobic surface. The production of ROS was completely eliminated when cytochalasin B was added to the measuring system, suggesting the involvement of the cell cytoskeleton in the activation process. The relation between the intra- and extracellular generation of ROS was further assessed, and we found that most of the ROS produced were released from the cells, in agreement with a model in which the activating surfaces induce a 'frustrated' phagocytic response. Serum totally inhibited 'frustrated' phagocytosis provided that the IgG molecules were sticking to a hydrophilic surface.
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| 35. |
- Lundgren, Anders, 1978, et al.
(author)
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Gold-nanoparticle-assisted self-assembly of chemical gradients with tunable sub-50 nm molecular domains
- 2014
-
In: Particle & particle systems characterization. - : Wiley. - 0934-0866 .- 1521-4117. ; 31:2, s. 209-218
-
Journal article (peer-reviewed)abstract
- A simple and efficient principle for nanopatterning with wide applicability in the sub-50 nanometer regime is chemisorption of nanoparticles; at homogeneous substrates, particles carrying surface charge may spontaneously self-organize due to the electrostatic repulsion between adjacent particles. Guided by this principle, a method is presented to design, self-assemble, and chemically functionalize gradient nanopatterns where the size of molecular domains can be tuned to match the level corresponding to single protein binding events. To modulate the binding of negatively charged gold nanoparticles both locally (<100 nm) and globally (>100 μm) onto a single modified gold substrate, ion diffusion is used to achieve spatial control of the particles' mutual electrostatic interactions. By subsequent tailoring of different molecules to surface-immobilized particles and the void areas surrounding them, nanopatterns are obtained with variable chemical domains along the gradient surface. Fimbriated Escherichia coli bacteria are bound to gradient nanopatterns with similar molecular composition and macroscopic contact angle, but different sizes of nanoscopic presentation of adhesive (hydrophobic) and repellent poly(ethylene) glycol (PEG) domains. It is shown that small hydrophobic domains, similar in size to the diameter of the bacterial fimbriae, supported firmly attached bacteria resembling catch-bond binding, whereas a high number of loosely adhered bacteria are observed on larger hydrophobic domains. Chemical gradients with the resolution needed to address complex biological binding events at the single protein level are prepared using surface-deposited gold nanoparticles as a versatile template for orthogonal chemical modifications. The effect of hydrophobic domain arrangement on the sub-50 nm scale is shown to influence binding of fimbriae carrying E. coli bacteria. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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| 36. |
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| 37. |
- Nilsson Ekdahl, Kristina, et al.
(author)
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Innate immunity activation on biomaterial surfaces: A mechanistic model and coping strategies
- 2011
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In: Advanced Drug Delivery Reviews. - : Elsevier BV. - 0169-409X .- 1872-8294. ; 63:12, s. 1042-1050
-
Research review (peer-reviewed)abstract
- When an artificial biomaterial (e.g., a stent or implantable pump) is exposed to blood, plasma proteins immediately adhere to the surface, creating a new interface between the biomaterial and the blood. The recognition proteins within the complement and contact activation/coagulation cascade systems of the blood will be bound to, or inserted into, this protein film and generate different mediators that will activate polymorphonuclear leukocytes and monocytes, as well as platelets. Under clinical conditions, the ultimate outcome of these processes may be thrombotic and inflammatory reactions, and consequently the composition and conformation of the proteins in the initial layer formed on the surface will to a large extent determine the outcome of a treatment involving the biomaterial, affecting both the functionality of the material and the patient's life quality. This review presents models of biomaterial-induced activation processes and describes various strategies to attenuate potential adverse reactions by conjugating bioactive molecules to surfaces or by introducing nanostructures.
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| 38. |
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| 39. |
- Olofsson, Ann-Cathrin, 1970, et al.
(author)
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Use of a Quartz Crystal Microbalance To Investigate the Antiadhesive Potential of N-Acetyl-L-Cysteine
- 2005
-
In: Applied and Environmental Microbiology. ; 71:5, s. 2705-2712
-
Journal article (peer-reviewed)abstract
- The reduction of bacterial biofilm formation on stainless steel surfaces by N-acetyl-L-cysteine (NAC) is attributed to effects on bacterial growth and polysaccharide production, as well as an increase in the wettability of steel surfaces. In this report, we show that NAC-coated stainless steel and polystyrene surfaces affect both the initial adhesion of Bacillus cereus and Bacillus subtilis and the viscoelastic properties of the interaction between the adhered bacteria and the surface. A quartz crystal microbalance with dissipation was shown to be a powerful and sensitive technique for investigating changes in the applied NAC coating for initial cell surface interactions of bacteria. The kinetics of frequency and dissipation shifts were dependent on the bacteria, the life cycle stage of the bacteria, and the surface. We found that exponentially grown cells gave rise to a positive frequency shift as long as their cell surface hydrophobicity was zero. Furthermore, when the characteristics of binding between the cell and the surface for different growth phases were compared, the rigidity increased from exponentially grown cells to starved cells. There was a trend in which an increase in the viscoelastic properties of the interaction, caused by the NAC coating on stainless steel, resulted in a reduction in irreversibly adhered cells. Interestingly, for B. cereus that adhered to polystyrene, the viscoelastic properties decreased, while there was a reduction in adhered cells, regardless of the life cycle stage. Altogether, NAC coating on surfaces was often effective and could both decrease the initial adhesion and increase the detachment of adhered cells and spores. The most effective reduction was found for B. cereus spores, for which the decrease was caused by a combination of these two parameters
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| 40. |
- Pinori, Emiliano, et al.
(author)
-
Multi-seasonal barnacle (Balanus improvisus) protection achieved by trace amounts of a macrocyclic lactone (ivermectin) included in rosin-based coatings.
- 2011
-
In: Biofouling. - : Informa UK Limited. - 1029-2454 .- 0892-7014. ; 27:9, s. 941-53
-
Journal article (peer-reviewed)abstract
- Rosin-based coatings loaded with 0.1% (w/v) ivermectin were found to be effective in preventing colonization by barnacles (Balanus improvisus) both on test panels as well as on yachts for at least two fouling seasons. The leaching rate of ivermectin was determined by mass-spectroscopy (LC/MS-MS) to be 0.7ng cm(-2) day(-1). This low leaching rate, as deduced from the Higuchi model, is a result of the low loading, low water solubility, high affinity to the matrix and high molar volume of the model biocide. Comparison of ivermectin and control areas of panels immersed in the field showed undisturbed colonisation of barnacles after immersion for 35 days. After 73 days the mean barnacle base plate area on the controls was 13mm(2), while on the ivermectin coating it was 3mm(2). After 388 days, no barnacles were observed on the ivermectin coating while the barnacles on the control coating had reached a mean of 60mm(2). In another series of coated panels, ivermectin was dissolved in a cosolvent mixture of propylene glycol and glycerol formal prior to the addition to the paint base. This method further improved the anti-barnacle performance of the coatings. An increased release rate (3ng cm(-2) day(-1)) and dispersion of ivermectin, determined by fluorescence microscopy, and decreased hardness of the coatings were the consequences of the cosolvent mixture in the paint. The antifouling mechanism of macrocyclic lactones, such as avermectins, needs to be clarified in further studies. Beside chronic intoxication as ivermectin is slowly released from the paint film even contact intoxication occurring inside the coatings, triggered by penetration of the coating by barnacles, is a possible explanation for the mode of action and this is under investigation.
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| 41. |
- Pinori, Emiliano, et al.
(author)
-
The impact of coating hardness on the anti-barnacle efficacy of an embedded antifouling biocide
- 2013
-
In: Biofouling. - : Informa UK Limited. - 0892-7014 .- 1029-2454. ; 29:7, s. 763-773
-
Journal article (peer-reviewed)abstract
- The efficacy of antifouling coatings designed to minimise the release of biocide, either by embedded (non-covalent) or tethered (covalently bonded) biocides, relies on sufficient bioavailability of the active compound upon contact between the organism and the coating. This investigation is focused on whether coating hardness affects the efficacy of embedded coating systems. Two experimental, non-eroding and waterborne latex paint formulations composed mainly of polystyrene (PS) or polyvinyl versatate (PV) were chosen for their difference in mechanical properties measured in terms of Buchholz indentation resistance. Ivermectin was added to both formulations to a final concentration of 0.1% (w/v) and the steady state release rate was measured according to ISO 15181 at between 34 and 70ngcm(-2)day(-1) for both formulations. Field trials conducted over 3months showed significant differences in anti-barnacle efficacy between the formulations despite their similar release profiles. The softer PV coating showed complete anti-barnacle efficacy, ie no barnacles were detected, while the harder PS coating showed no efficacy against barnacle colonisation during the same time period. The results indicate a new antifouling strategy whereby a route of intoxication is triggered by the organism itself upon interaction with the coating and its embedded biocide. This finding opens new possibilities in controlling macrofouling by low emission antifouling coatings.
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| 42. |
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| 43. |
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| 44. |
- Sott, Kristin, 1974, et al.
(author)
-
Adsorption behavior and enzymatically or chemically induced cross-linking of a mussel adhesive protein
- 2000
-
In: Biofouling. - : Informa UK Limited. - 0892-7014 .- 1029-2454. ; 16:2-4, s. 119-132
-
Journal article (peer-reviewed)abstract
- The adsorption behavior of the mussel adhesive protein Mytilus edulis foot protein-1 (Mefp-1) has been investigated on a negatively charged polar SiO2 surface and an electrically inert non-polar CH3-terminated thiolated gold surface. How the structure of adsorbed Mefp-1 is changed upon chemically and enzymatically induced cross-linking using sodium periodate (NaIO4) and catechol oxidase, both of which transform DOPA residues in Mefp-1 into highly reactive o-quinones, was also investigated. The results are compared with those resulting from addition of Cu2+ to adsorbed Mefp-1, which forms complexes with and catalyses oxidation of DOPA residues, previously suggested to participate in the cohesive and adhesive properties of the byssus thread of M. edulis. By combining surface plasmon resonance (SPR) and quartz crystal microbalance/dissipation (QCM-D) measurements, the effects of these agents were investigated with respect to changes in the amount of coupled water, the viscoelastic properties (rigidity) and the hydrodynamic thickness of the protein adlayers. The layer of Mefp-1 formed on the bare CH3-terminated surface was elongated, flexible and coupled hydrodynamically a substantial amount of water, whereas Mefp-1 formed a rigidly attached adlayer on the SiO2 surface. Upon enzymatically and chemically induced cross-linking of Mefp-1 formed on the CH3 surface, the rigidity of the adlayer(s) increased significantly. A similar increase in the rigidity was observed also upon addition of Cu2+, suggesting that the high level of metal ions present in the byssus thread might be essential for the cohesive and adhesive properties of this protein. For the mass-uptake kinetics of enzymatically induced cross-linking, three different phases were observed and are interpreted as competition between binding of protein and release of coupled water. For the reaction with NaIO4 and Cu2+ only release of water affected the coupled mass. The importance of this type of information for an improved understanding of the strong adhesion and cohesive properties in marine environments is discussed.
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| 45. |
- Ytreberg, Erik, 1980, et al.
(author)
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A novel XRF method to measure environmental release of copper and zinc from antifouling paints
- 2017
-
In: Environmental Pollution. - : Elsevier BV. - 0269-7491 .- 1873-6424. ; 225, s. 490-496
-
Journal article (peer-reviewed)abstract
- The release of copper (Cu) and zinc (Zn) from vessels and leisure crafts coated with antifouling paints can pose a threat to water quality in semi-enclosed areas such as harbors and marinas as well as to coastal archipelagos. However, no reliable, practical and low-cost method exists to measure the direct release of metals from antifouling paints. Therefore, the paint industry and regulatory authorities are obliged to use release rate measurements derived from either mathematical models or from laboratory studies. To bridge this gap, we have developed a novel method using a handheld X-Ray Fluorescence spectrometer (XRF) to determine the cumulative release of Cu and Zn from antifouling paints. The results showed a strong linear relationship between XRF K-alpha net intensities and metal concentrations, as determined by ICP-MS. The release of Cu and Zn were determined for coated panels exposed in harbors located in the Baltic Sea and in Kattegat. The field study showed salinity to have a strong impact on the release of Cu, i.e. the release increased with salinity. Contrary, the effect of salinity on Zn was not as evident. As exemplified in this work, the XRF method also makes it possible to identify the governing parameters to the release of Cu and Zn, e.g. salinity and type of paint formulation. Thus, the XRF method can be used to measure environmentally relevant releases of metallic compounds to design more efficient and optimized antifouling coatings. (C) 2017 The Authors. Published by Elsevier Ltd.
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