Biofilms Research Center for Biointerfaces

Transcription

1 BIO FILMS INTERFACES Biofilms Research Center for Biointerfaces Biofilms Research Center for Biointerfaces A translational research programme at Malmö University Funded by the Knowledge foundation Progress report #6 January 1 st, 2010 December 31 st, 2010

2 Cover illustration Biofilms develop spontaneously on a surface in contact with a liquid phase containing biomolecules and micro-organisms. The initial phase in the development is rapid adsorption of surface active molecules, notably macromolecules such as proteins, to the surface forming an initial conditioning film. Next step is attachment of microorganisms. These organisms grow and interact with molecules from the liquid phase, cell produced matrix, and other organisms in the formation of a biological film (biofilm). Organisms and molecules within the biofilm possess unique characteristics not observed for the same species suspended or dissolved in the associated liquid phase. Contact information Biofilms Research Center for Biointerfaces Faculty of Health and Society Malmö University SE MALMÖ, Sweden Center director: Assoc. Prof. Johan Engblom (JE) Tel: +46-(0) (JE); +46-(0) (Adm. Coordinator Eva Nilsson) [email protected] Visiting adress: Skåne University Hospital, SUS (Entrance 49) MALMÖ

3 1 List of Center Members During the period of report the Center has comprised the following members (permanent staff, postdocs, PhD-students and technical and administrative staff). 1.1 Permanent staff Johan Engblom, Assoc Prof., Director Thomas Arnebrant, Prof., Director , Vice Director 2010 Tautgirdas Ruzgas, Prof. Gunilla Nordin-Fredrikson, Prof. (also part time LU) Per Ståhle von Schwerin, Prof. Ann Wennerberg, Prof. Gunnel Svensäter, Prof. Ali Massih, Prof. Håkan Ericsson, Assoc. Prof. Vitaly Kocherbitov, Assoc. Prof. Liselott Lindh, Assoc. Prof. Anette Gjörloff-Wingren, Assoc. Prof. Zoltan Blum, Assoc. Prof. Liu-Ying Wei, Assoc. Prof. Julia Davies, Assoc. Prof. Bertil Kinnby, Assoc. Prof. Christina Bjerkén, Assoc. Prof. Claes Wickström, Assoc. Prof. Lars Ohlsson, Dr Tove Sandberg, Dr. Maria Stollenwerk, Dr. Sergey Shleev, Dr. Anna Ketelsen, Dr. 1.2 Junior researchers and postdocs Olof Svensson, Dr. Olga Santos, Dr. Laura Varas, Dr. Jessica Neilands, Dr. Luis Chavez de Paz, Dr. Javier Sotres, Dr Ida Svendsen, Dr Peter Nilsson, Dr Alejandro Barrantes, Dr Jovice Bon Singh Ng, Dr Ryo Jimbo, Dr Viktor Andoralov, Dr 1.3 PhD students Jildiz Hamit Eminovski Ulf Hejman Alma Masic 3

4 Maria Pihl Sebastian Björklund (enrolled at LU) Anton Fagerström Yana Znamenskaya Adnan Safdar Christian Alfredsson Kindblom Marjan Dorkhan Kostas Bougas Magnus Falk Peter Lamberg Cathrine Albér Mariko Hayashi Lory Melin Svanborg Tuerdi Maimaitiyili 1.4 Technical and administrative staff Eva Nilsson, Administrative coordinator Ulrika Troedsson, Technician Agnethe Henriksson, Technician Madeleine Blomqvist, Technician Lina Pedersen, Technician 1.5 MSc students BMMT Master at HS Joynul Abedin Ameena Daftani Endale Asmare Hailu Rakibul Islam Lutfor Islam Abu Sayeed Khan Jabed Khandaker Shadi Movahed Bashiri Shifa Saleem Selva Kumar Subramanian Mohammed B Sunmonu Inger Anne Tveit Surendra Vutti Shaheen Mohammad Syful Islam BMMT Master at HS Haddel Ali Shoker Joy Chia Ihab Dahi Payam Delfani Susanna Tarasco Mohammad Zahir Uddin Petra Wicktor BMMT Master at HS Aseel Albayati Sheima Sultan Kadir Eleonora Dahlquist Rula Bahran Peter Lamberg Marianne Mårtensson Patrik Bauer MS Master at HS & TS Maihemutijiang Maimaiti Carl Mikaelsson Ajigul Nuermaimaiti Oyetunji Oladele Kazeem Wureguli Reheman Christian Ukoha Oji Erik Öberg Simayijiang Zhayida In addtition to these two year Master programmes, the Faculty of Odontology (OD) offers the Dentistry programme which is a continuous five year program to Master level. 4

5 1.6 Management and boards Biofilms Research Center for Biointerfaces is managed on a daily basis by the Center director and an Executive group, constituted by the heads of the individual research groups. The Steering group constitutes a link to central Malmö University, and involves the Deputy Vice-Chancellor and the Deans of the three faculties/schools involved in the research activities of the Center. The Reference group is an advisory board to the Director and contains representatives from Industry, Medicon Valley Alliance, Malmö University and other universities. Executive group Johan Engblom, Assoc. Prof., Director & Chairman for Thomas Arnebrant, Prof., Director & Chairman for Per Ståhle von Schwerin, Prof. Gunnel Svensäter, Prof. Ann Wennerberg, Prof. Gunilla Nordin-Fredrikson, Prof. Håkan Eriksson, Assoc. Prof. Steering group Eva Engquist, Deputy Vice-Chancellor, Malmö University, Chairman Margareta Östman, Prof., Dean Faculty of Health and Society Naser Eftekharian, Head School of Technology Malmö University Lars Bondemark, Prof., Dean Faculty of Odontology Reference group Martin Malmsten, Prof., Uppsala University, Chairman Ian Hamilton, Prof. em., University of Manitoba, Canada Peter Nordström, Senior Project Manager, Medicon Valley Alliance Yngve Sommarin, Dr., R&D Manager Euro-Diagnostica/Wieslab AB Magnus Christensson, Dr., R&D Manager AnoxKaldnes AB Markus Johnsson, Dr., Senior Director Pharm. Development Camurus AB Eva Engquist, Deputy Vice-Chancellor, Malmö University Zoltan Blum, Assoc. Prof., Malmö University 5

6 2 The Director s Report Biofilms - Research Center for Biointerfaces that has been established as a vital asset for Malmö University, is well reputed and carries a strong trademark within the university. Also, an increased regional awareness of the center s activities, including other academic institutions, industry and public sector, can be noted. Industry partners comment that the center has positioned Malmö University as an attractive partner and that it is an enterprise that benefits both industry and academia. During the period the center has expanded to twice its original size, now comprising 38 projects and 32 industry partners. Two strong research areas have evolved from the center activities, now established as two of the current eight research profiles at the university, i.e. Oral health and Biointerfaces. External research funding for the entire university amounts to 107 MSEK in 2010, where Biofilms Research Center for Biointerfaces contributes with 20%. Regarding output in terms of international journal publications the center share is 23% of the total university production Collaboration with external parties is a corner stone in the university strategy and the center is often put forward as a good example. Viktoria Fröjd, Helena Tassidis (enrolled at LU) and Rickard Hägglund (enrolled at LTH) successfully defended their PhD-theses in 2010, as did Ulf Hejman and Adnan Safdar defending their licentiate theses. Claes Wickström and Christina Bjerkén were accepted as Associate Professors and Sergey Shleev has pending applications as Associate Professor (Malmö) and Habil. Doctor of Science (Russia). Drs Olga Santos and Olof Svensson left their positions as researchers during the year, while five new post docs and five PhD students (three at Malmö University and two enrolled at Lund University) were recruited. Two PhD-students had research stays at partner sites, Alma Masic at University of Guelph University, Canada, and Anton Fagerström at AkzoNobel Surface Chemistry AB, Stenungsund, Sweden. Several senior researchers also payed visits to universities outside Sweden (e.g. China, Russia, Lithuania, Germany and UK). During 2010 we have produced 59(67) publications in international journals, 5 book chapters and 3 proceeding papers. Publications are in journals such as Trends in Immunology (impact factor 8.8), Atherosclerosis, Trombosis and Vascular Biology (7.5) Journal of Controlled Release (6.0), Biofouling (4.4), Langmuir (3.9), Journal of Endodontics (3.0), Clinical Oral Implant Research (2.9), International Journal of Solids and Structure (1.8) and Mathematical Biosciences (1.3). High impact factors indicate substantial scientific quality of the research carried out within the Center. In addition to this, approximately 51 oral and 21 poster presentations were made at national and international meetings. In reviewing the number citations over the past five years on papers produced by the Center members we sum up 7763 for permanent staff and 587 for junior researchers and post-docs. Five members have more than 500 citations each, Profs. Tautgirdas Ruzgas, Gunilla Nordin Fredrikson, Thomas Arnebrant, Ann Wennerberg and Dr Sergey Schleev. Center members have also refereed papers for international journals on a regular basis. 6

7 Members of the Center take part in The National Research School of Odontology (Ann Wennerberg (coordinator), Gunnel Svensäter (local coordinator) and Julia Davies, supervisors), and The Research School in Pharmaceutical Sciences, LU (Johan Engblom, supervisor). Most researchers participate in undergraduate (the BMA and TELMah programmes) and/or graduate teaching (the BMMT, MS and Dentistry programmes), as well as PhD-student supervision. We strive to further integrate education (BSc, MSc, PhD), a cornerstone for the future of the Center. Particularly, one goal is to further integrate Master-level education into our research activities also at an operational level in specific projects. Research collaboration with industry is active in all four focus areas of the Center and during 2010 we attracted Novosense AB, Biogaia AB, ACO Hud Nordic AB, YKI AB and Eviderm Institute AB as partners in new research projects. Although border lines between the focus areas are not always clear cut, the company involvement distribute according to i) Eucaryotic cell-surface interactions (Promimic, PHI), ii) Molecular transport phenomena (AkzoNobel, YKI, ACO Hud Nordic, Eviderm Institute), iii) Molecular interactions at biointerfaces (Camurus, EuroDiagnostica, Promimic, Novosense, Anordica, Arcam, Bioglan, Galenica) and iv) Microbial biofilms (ArlaFoods, AnoxKaldnes, Gambro, Arcam, Biogaia). We also have close collaboration with Medeon AB and Medicon Valley Alliance. Center members are partners in the collaborative EU FP7 funded project Threedimensional nanobiostructure-based self-contained devices for biomedical application (Dr Sergey Shleev, coordinator) and continued to be involved in the EU FP6 Marie Curie Research Training Networks (MCRTNs), Bio-interfaces: from molecular understanding to applications. We are also involved in three EU Interreg programmes; Valorisation of knowledge intensive ideas in the South Baltic area (SB-VALOR), Öresund forum for innovation within nano-, bio- and medical technology (FinNBMT) and Öresund materials innovation comunity (Ö-MIC). Center members have been responsible for arranging three workshops, Biomaterials from fundamentals to Market Application (Biofilms 6 th Annual workshop), Biofilms Members Day and together with BIOSUM, Gothenburg; Biomaterials in medicine, (within the National Research School of Odontology), one salivary symposium at IADR in Barcelona and a kick off meeting for a new research project funded by KKs. The workshop Choice of and collaboration with CRO s in pharmaceutical development (launched by the Swedish Academy of Pharmaceutical Sciences) was held at Malmö University and co-organised by the Center. Center members have been active presenting their results on numerous occasions at national and international conferences and workshops. Center activities have also been visible through e.g. articles in PS Public Service Review European Union 19, a press release highlighting the 6 th annual workshop, and Malmö University newsletters. 7

8 3 List of Research Activities The research activities of the Center during 2010 may be described by the following headlines. Projects are listed with partners and funding in paranthesis. Projects in bold are funded by the center grant from KKs. In addition, we are also involved in three EU Interreg programmes; Valorisation of knowledge intensive ideas in the South Baltic area (SB-VALOR), Öresund forum for innovation within nano-, bio- and medical technology (FinNBMT) and Öresund materials innovation community (Ö-MIC). 3.1 Eucaryotic cell-surface interactions 1. Biomaterial 1(2), The influence of biochemical coat for implant bone incorporation, The in vivo part (Promimic AB, funded by KKs-Biofilms) 2. Cell-to-bio-mimetic interface interactions (funded by EU Marie Curie Research Training Networks) 3. Digital holography for cell studies (Phase Holographic Imaging AB, funded by the Crafoord foundation, the Magnus Bergwall foundation and Mah) 4. Biological responses to photo-reactive hydrophilic nano-size structures (funded by VR) 5. Hydrophilic and hydrophobic implant surfaces (funded by Vilhelm and Martina Lundgren Foundation) 6. Histological and 3-dimensional analysis of laminin coated polished ceramic implants (funded by Hjalmar Svensson Research Foundation) 7. The use of CaO as luting material and bone substitute (funded by VR) 8. Facilitation of soft tissue healing upon implant treatment in patients with supressed healing ability (funded by Mah) 3.2 Molecular transport phenomena 9. Adjuvants for products used in agriculture (AkzoNobel Surface Chemistry AB, funded by KKs-Biofilms) 10. Water a crucial factor in regulating biomembrane permeability (Physical Chemistry 1, LU, funded by FLÄK, LU) 11. Humectants and their mechanisms in skin (YKI AB, ACO Hud Nordic AB, Eviderm Institute AB, funded by KKs) 12. Miniature biofuel cells for self-contained bio-devices: electron transfer in threedimensional nanobiostructures (funded by VR & EU) 13. Grinfeld surface instabilities (funded by VR) 14. Effect of a gradient in water chemical potential on buccal drug delivery (Food technology, LU, funded by Mah) 3.3 Molecular interactions at biointerfaces 15. Mucoadhesion: Drug carrier interactions at biologically relevant interfaces (Camurus AB, funded by KKs-Biofilms) 16. Bioassay: New concept for lipid-based surface coatings in bioassays (EuroDiagnostica AB, funded by KKs-Biofilms) 8

9 17. Biomaterial 2(2), The influence of biochemical coat for implant bone incorporation, The in vitro part (Promimic AB, funded by KKs-Biofilms) 18. Development of biofuel cells for powering wireless transmission (Novosense AB, funded by KKs-Biofilms) 19. Attachable Diagnostic Devices with Individualised Referencing (ADDIR) (Galenica AB, funded by KKs-biofilms) 20. Symptomatic vs. asymptomatic atherosclerotic plaques (CRC, LU, funded in part by Mah) 21. Biocompatibility of metals (Anordica AB & Arcam AB, funded by KKs) 22. Hydration of mucous gel (funded by Mah) 23. Topical hydrogen peroxide in wound healing (Bioglan AB, pilot project) 24. Membrane impedance spectroscopy as a tool to study skin barrier function and variability (Galenica AB & Dermatology (LU), pilot project) 25. Development of novel multi-functional salivary substitutes for dry mouth syndrome patients (funded by the Swedish Laryng Foundation) 26. Screening of phase behavior in the DOPS/DOPE/water system and effects on lipid morphology from a decrease in water chemical potential (funded by CLRF) 3.4 Microbial biofilms 27. Milk Protein: Investigation of interactions between osteopontin and oral biofilm bacteria (Arla Foods AB, funded by KKs-Biofilms) 28. Carrier: Investigation and modelling of convection in biofilms for different carriers (AnoxKaldnes AB, funded KKs-Biofilms) 29. Probiotics: Effects of probiotic lactobacilli on biofilm formation and acid tolerance (Biogaia AB, funded by KKs-Biofilms) 30. Catheters: Biofilm formation on Peritoneal Dialysis catheters (Gambro Lundia AB, funded by KKs-Biofilms) 31. Biologically induced stress corrosion crack growth (Arcam AB, funded by KKs-Biofilms) 32. Biofilm activity as a marker to identify patients at risk of caries mechanisms underlying microbial stress tolerance (funded by KKs-Biofilms) 33. Caries prevention with fluoridated milk a prospective clinical and microbiological study of root caries (funded by Swedish Patent Revenue Foundation) 34. Biofilms on oral mucosal surfaces (funded by Swedish Dental Society) 35. Mucins and microbial biofilms a symbiotic relationship for health (funded by Mah and Crafoord Foundation) 36. Mucosal interactions as inducers of acid tolerance in oral microorganisms (funded by Crafoord Foundation and Swedish Patent Revenue Foundation) 37. Activities of microbial biofilms on bioactive implant surfaces (funded by Mah) 38. The plasminogen activating system interaction with microorganisms and a potential risk marker (funded by Swedish Dental Society) 9

10 4 Research highlights 4.1 Friction force spectroscopy for the study of the strength of protein layers Protein layers can confer diverse properties on surfaces such as molecule-binding, biocompatibility, or simply act as protective barriers against the surrounding medium. Considering the vast number of applications where they are involved, these layers can be subjected to multiple damage sources, including those of mechanical origin. Thus, the study of the mechanical properties of protein layers can provide with useful information for i) a better understanding of how the proteins interact both between themselves and with the underlying substrate, but also for ii) their design so that their resistance against mechanical damage is increased. During the last decade, the Atomic Force Microscope (AFM), where a sharp nm-sized tip is used to probe surfaces, has emerged as a powerful tool to study mechanical properties of protein layers. We have approached the study of these properties by a new methodology based on the Friction Force Spectroscopy (FFS) operation mode of the AFM, which allows studying the sample response to both normal and shear applied stresses. This methodology is based on the continuous two-dimensional scanning of a surface while ramping the applied load force. For each load force, a topography image of the scanned sample area and the average applied friction force during the scan are recorded. Therefore, FFS allows the characterization of the layers with friction-load curves, and with the characteristic topographies (like the initial rupture or the total removal) that correspond to the different regimes of the scratching process. This technique has been tested on β- and κ-casein monolayers. Caseins do not only act as natural emulsifiers in milk, but are also used as emulsifiers or dispersants in many technological and industrial applications. Our experiments showed that they can support pressures up to hundred of MPa before being removed while still exhibiting a high frictional behaviour, supporting their good performance as emulsifiers. Moreover, the technique was also proved to be able to study the dependence of the cohesion of the layers with properties of the surrounding liquid medium such as ph and ionic strength. Recent experiments on systems such as serum-proteins and saliva layers support the wide applicability of the technique. Figure 4.1. a) Schematic draw of a FFS measurement. b) and c) Topography images and frictionload curve corresponding to the scratching of a β- -casein monolayer. Sotres J, Svensson O and Arnebrant T., Friction force spectroscopy of β- and κ-casein monolayers. Langmuir 2011, 27(3),

11 4.2 Bioelectrocatalytic interfaces: Redox Enzymes Electronically Connected at Three-Dimensional Materials for Extracting Electrical Energy from Biofuels Bioelectrocatalytic devices, such as biosensors, have proven to be useful in different areas of applications including biomedicine. Current research in bioelectrochemistry is massively focused on the improvement of biofuel cells. The last year we have aimed to design three-dimensional (3D) conducting materials which, when loaded with redox enzymes, can constitute high power biofuel cells. Conducting 3D materials were based on redox hydrogels [1], carbon [2] or gold [3] nanoparticles, and nanostructured silica [4]. It was found that all these materials provide a possibility to control the procedures of assembly of 3D electrodes. The work was devoted to tune or adjust selectivity and optimize catalytic activity of the redox enzymes incorporated into the 3D structures of these electrodes. Recently developed 3D electrodes of biofuel cells show at least 10 times higher current densities if compare with 2D electrodes. 3D cathodes developed in our laboratory provide µa/cm 2 current densities. This means that a few micrometer thick biofuel cell of approximately 1 cm 2 area already now might power wireless biomedical devices. Figure 4.2. An example of 3D electrode based on micro-/ nanostructured silica layer. Relevant publications: 1. Design of a bioelectrocatalytic electrode interface for oxygen reduction in biofuel cells based on a specifically adapted Oscomplex containing redox polymer with entrapped Trametes hirsuta laccase. Ackermann, Y.; Guschin, D.A.; Eckhard, K.; Shleev, S.; Schuhmann, W. Electrochemistry Communications, 2010, 12(5), Stable floating air diffusion biocathode based on direct electron transfer reactions between carbon particles and high redox potential laccase. Shleev S.; Shumakovich G.; Morozova O.; Yaropolov A. Fuel Cells, 2010, 10(4), Laccase-gold nanoparticle assisted bioelectrocatalytic reduction of oxygen. Dagys, Marius; Haberska, Karolina; Shleev, Sergey; Arnebrant, Thomas; Kulys, Juozas; Ruzgas, Tautgirdas. Electrochemistry Communications, 2010, 12(7), Bioelectrochemical studies of azurin and laccase confined in three-dimensional chips based on gold-modified nano- /microstructured silicon. Ressine A., Vaz-Dominguez C., Fernandez V.M., De Lacey A.L., Laurell T., Ruzgas T., Shleev S. Biosensors and Bioelectronics, 2010, 25(5), The following projects support the development of 3D biofuel cells: 1. Wireless self-powered biodevices: Function of nanowired multicentre redox enzymes and living cells", Main applicant: Sergey Shleev. The Swedish Research Council, project number: Bioelectrochemical 3D nanobiostructures in physiological liquids and cell based in-vitro platforms Main applicant: Tautgirdas Ruzgas. The Swedish Research Council, project number: Three-dimensional nanobiostructure-based self-contained devices for biomedical application, Coordinator: Sergey Shleev. EU FP7-NMP-2008-SMALL-2. Grant Agreement Number:

12 4.3 Factors affecting transport of tebuconazole over leaf cuticle The complex structure of plant cuticles constitutes the main barrier for fungicide uptake in leaves. Consequently, the detailed underlying mechanism of action of adjuvants, often used to facilitate fungicide permeation in leaves, is also complex and remains to be resolved. The overall aim of this project is thus to obtain a better understanding of the mechanisms of action of specific surfactants, in order to facilitate the design of better adjuvants for agricultural fungicides. We have combined in vitro diffusion methodology (Franz cells) with sorption isotherms and membrane impedance spectroscopy to evaluate the effect of two specific adjuvants (C 10 EO 7 and C 8 G 1.6 ) on the bioavailability of tebuconazole as model active ingredient, using the adaxial side of leafs from the model plant Clivia Miniata Regel as the membrane. The barrier properties of plant leaves may respond to external factors, like changes in ambient relative humidity or gradients imposed by individual formulations applied and therefore, inert silicone membranes were employed in parallel to plant membranes to distinguish between factors affecting the diffusion coefficient in the membrane (D i ) and the gradient in chemical potential (d i /dz) over the membrane. From the data given below it is evident that fungicide permeation over silicone membrane is strictly dependent on the gradient in tebuconazole chemical potential (Figure 4.3.1), while the prescence of surfactants has a strong impact on the diffusion coefficient of tebuconazole in Clivia cuticle, resulting in up to four times higher permeability (Figure 4.3.2). C 10 EO 7 is more effective than C 8 G 1.6 in promoting tebuconazole permeation through the Clivia cuticle, and C 10 EO 7 also has a more pronounced ability to decrease the extremely long lag-time. Moreover, the Clivia cuticle can accommodate large amounts of tebuconazole and thereby act as a depot over time. We have also shown using impedance spectroscopy that the barrier properties of Clivia cuticle improves with maturation of the leaf. Figure Tebuconazole permeability over a silicone membrane. Carrier: water, and watersurfactant (4%) mixtures. a teb = Figure Tebuconazole permeability over a Clivia membrane. Carrier: water, and watersurfactant (4%) mixtures. a teb = Factors Affecting Transport of Tebuconazole over Silicone Membrane and Leaf Cuticle, Fagerström A, Kocherbitov V, Lamberg P, Bergström K, Westbye P, Ruzgas T and Engblom J. in 9th International Symposium on Adjuvants for Agrochemicals, ISAA Society; Baur P and Bonnet M Eds. August 2010, Pages ISBN Effects of adjuvants on Tebuconazole leaf cuticle penetration, Fagerström et al, Manuscript to be submitted Surfactant induced fluidization of plant leaf cuticle, Fagerström et al, Manuscript to be submitted

13 4.4 Low levels of fluoride inhibit acid tolerance of plaque bacteria in vivo Fluoride is used for prevention of dental caries mainly due to its potential to decrease solubility and promote remineralisation of enamel and root dentin. In vitro studies show that fluoride also affects the physiology of oral streptococci. However, data on the effect of fluoride on acid tolerance in plaque in vivo is limited. The aim of this investigation was to study the effect of fluoride on plaque acid tolerance and composition, lactic acid production, and the clinical effect on re-mineralization of root caries lesions. The test group (F-group) consumed 200 ml of cow s milk supplemented with 5 mg/l NaF as a single dose once per day, the milk control group (Mgroup) drank 200 ml of unsupplemented cow s milk and a no-milk control group (C-group) did not consume milk in this manner. The length of the study was 15 months. Dental plaque samples were taken at baseline and after 15 months. The proportion of acid tolerant bacteria in plaque was estimated using LIVE/DEAD BacLight TM staining after exposure to ph 3.5 for 2 hours. Lactic acid production after glucose pulsing was measured enzymatically. The Electronic Caries Monitor (ECM) was used to measure the electrical resistance of root surface lesions. Plaque from subjects in the F-group showed a statistically significant decrease in plaque acid tolerance and a significant increase in ECM values were found after 15 months compared to baseline indicating re-mineralization. Lactic acid production from glucose was also lower in the F-group although not statistically significant. This experimental clinical study shows that daily intake of fluoride reduces plaque acid tolerance and lactic acid production in vivo and promote remineralization of root caries lesions. BASELINE AFTER 15 MONTHS DECREASED PLAQUE ACID TOLERANCE INCREASED PLAQUE ACID TOLERANCE Figure 4.5. Bacteria in plaque stained with LIVE/DEAD BacLight TM staining. Green cells are acid tolerant while red cells are not. Neilands J., L.G. Petersson, D. Beighton and G. Svensäter. Fluoride inhibits acid tolerance of root surface biofilms. Manuscript 13

14 4.5 Peritoneal dialysis catheters show presence of bacteria without clinical signs of infection Fifteen peritoneal dialysis catheters extracted from patients undergoing renal transplantation (i.e. from patients with no clinical signs of infection) were investigated for the presence of bacteria using microbiological culture and confocal laser scanning microscopy (CLSM). For reference, two catheters from patients with infections were also investigated. The results showed 82% of the catheters to be colonised by bacteria, although in a rather low numbers. The bacteria were heterogeneously spread all over the catheter surface and several species often colonised the same area (Figure 1). The major species found were Staphylococus epidermidis and Propionibacterium acnes, but several others were also detected, including Micrococcus spp, Staphylococcus lugdunensis, Staphylococcus warneri, Corynebacterium spp, Proteus mirabilis, Rothia mucilaginosa, Streptococcus sanguis and Staphylococus aureus. Figure 4.6. Species found on peritoneal dialysis catheters positive in the microbiological cultures. Pihl M, Davies JR, Johansson A-C and Svensäter G. Occurrence of bacteria on catheters in patients undergoing peritoneal dialysis Submitted to Journal of Medical Microbiology. 14

15 4.6 Adhesion of Streptococcus oralis to titanium surfaces - effects of surface roughness and conditioning films Initial healing is a critical phase in dental implant therapy and optimum surface roughness is one of the key factors of importance for successful osseo-integration. Surfaces designed to promote osteoblast activity may however also enhance bacterial adhesion and stimulate microbial activity thus increasing the risk of peri-implant infections in the longer-term. The aim of this investigation was to determine how surface roughness affects the adhesion of Streptococcus oralis (a species frequently isolated from peri-implant infections) to titanium surfaces used in dental implants in the presence of conditioning films derived from saliva and serum. Titanium plates with average surface roughness (Sa) of 0,5 or 1,5 µm, uncoated or coated with 25% whole saliva or 5% human serum were exposed to exponential growth phase cells of Streptococcus oralis (LA 11) in a flow-cell system for 2 h. After washing for 1h, the numbers of adhered bacteria were assessed using confocal laser scanning microscopy (CLSM) after staining with the Baclight Live/Dead kit (Figure 1). The mean percentage bacterial coverage on the uncoated smooth surface (Sa = 0.5 µm) was 2±1% while that on the moderately rough surface (Sa = 1.5 µm) was significantly greater (mean percentage coverage = ± 0.88%, p < 0.01). Figure 4.7. CLSM images showing biofilms formed on moderately rough titanium surfaces in (a) the absence of a conditioning film or in the presence of (b) a saliva- or (c) and serum-derived conditioning film. For both the moderately rough and smooth surfaces, a conditioning film of saliva significantly increased the adhesion of bacteria (p< 0.01) whereas bacterial binding in the presence of a serumderived film showed no differences to that on the control (uncoated) surface. These data suggest that increased surface roughness, as well as the presence of salivary proteins, are important determinants of the level of colonization by streptococci on oral implant surfaces. Dorkhan M, Chávez de Paz M, Svensäter G and Davies JR. Adhesion of Streptococcus oralis to turned and blasted titanium surfaces - effects of saliva- and serum-derived pellicles. Manuscript 15

16 4.7 How to detect protease activity of dental biofilms in situ Proteolytic bacteria export proteases to their immediate surroundings. The extracellular proteolytic activity of these bacteria can thus be monitored through the addition of a protease substrate to the culture medium. Casein is a substrate for the four major types of proteases (serine, aspartic, cysteine and metalloproteases) and is thus a suitable substrate for screening protease activity. Using fluorescein-labeled casein, protease activity can be detected. Proteolytic activity of bacteria grown in biofilms has been visualized by confocal microscopy after addition of fluorescein-labeled casein. Figure 4.8. Proteolytic activity of bacteria in a clinical subgingival biofilm sample visualized as green fluorescence. Kinnby B, Wickström C & Svensäter G, Method development

17 4.8 Selective adhesion and phenotypic changes in oral streptococci revealed through interaction with mucinconditioned surfaces The resident microflora of dental plaque changes as it matures over time. Different streptococcal species are suggested to adhere and colonize at different time points in the maturation process, where early colonizers have the ability to adhere and grow, using the host proteins as substrates and late colonizers require other bacteria or their products to be able to adhere and grow. We show here, that two streptococcal species, Streptococcus mitis biovar 2 and Streptococcus mutans, display very different behaviors when introduced to surfaces conditioned with human salivary mucin MUC5B, the major glycoprotein found in the mucus film covering all oral surfaces. Using test surfaces conditioned with the salivary MUC5B mucin, S. mitis biovar 2 showed avid adherence as well as a phenotypic shift towards more protease active cells. When introduced to surfaces conditioned with other salivary proteins, this effect was not seen, suggesting a specific interaction between S. mitis biovar 2 and the MUC5B molecule. However, when S. mutans was studied under the same conditions, very few cells adhered to the conditioned surfaces, MUC5B or the other salivary proteins, suggesting an inability of S. mutans to adhere to the natural occurring salivary film components found in vivo. The same increase in protease active cells observed for S. mitis biovar 2 was found in the S. mutans cells, although there was no difference between the two substrates. This work clearly shows a specific difference between two streptococcal species in the way they adapt to different environments, S. mitis biovar 2 being able to adhere and colonize a salivary conditioning film, whereas S. mutans cannot. Figure 4.9. Adhesion of S. mitis biovar 2 (A) and S. mutans UA159 (B) to surfaces coated with salivary proteins, MUC5B or low-density salivary proteins. Bacteria were allowed to adhere to coated or uncoated surfaces for 2 hours and attached cells were visualized in a CLSM after BacLight LIVE/DEAD staining. * Differences are statistically significant (p < 0,05) using the Mann-Whitney t-test. Christian Kindblom, Gunnel Svensäter & Claes Wickström Salivary proteins influence phenotypic changes in Streptococcus mutans and Streptococcus mitis biovar 2 biofilm cells differences in adhesion and protease activity. To be submitted. 17

18 4.9 Spatial distribution of multiple species in complex biofilm communities In nature, bacterial biofilm communities are highly organized structures composed of multiple species that are believed to interact with each other in order to coexist. Synergistic coexistence of bacteria in mixed biofilm communities is suggested to play a crucial role in development of chronic infections as many species can become extremely resistant against antimicrobials and host defences. To better understand the spatial structure of multiple species communities it is necessary to visually characterize the distribution of individual species. We have developed an automated in vitro system to study spatial distribution of bacteria in multi-species biofilm communities. Our system is based in a combination of fluorescence in situ hybridization (FISH), confocal laser scanning microscopy (CLSM), and digital image analysis. FISH probes targeting the 16S rrna gene are designed to verify the abundance and spatial location of microbial community members. Overall, the knowledge gained by this method about distribution and interactions in multi-species biofilms will facilitate the rapid analysis of microbial communities in the sense of assessing changes in microbial populations as a function of time or environmental conditions. Prospectively, this methodology can be applied in combination with specific fluorescent markers targeting metabolic processes to allow the investigation of in situ structure/function analysis of complex microbial communities. A B Figure Spatial distribution of two oral biofilm communities. (A) A four-species biofilm community composed of clinical root canal isolates of Lactobacillus salivarius (red), Streptococcus gordonii (green/yellow), Actinomyces naeslundii (blue) and Enterococcus faecalis (violet). (B) A three-species biofilm community composed of supra-gingival isolates of Lactobacillus fermentum (red), Streptococcus gordonii (green/yellow) and Actinomyces naeslundii (blue). Bar = 10 µm. Chavez de Paz L,

19 4.10 On Ca2+ incorporation and nanoporosity of titanium surfaces and the effect on implant performance Implants need to perform in three biological arenas: in relation to bone-tissue, soft-tissue, and microbial biofilms. An implant should be properly osseointegrated and have a tight adaptation of surrounding soft-tissues but it should at the same time not be prone for extensive biofilm formation or be difficult to clean. More specifically we aimed for: Bone: To evaluate the importance of anodic oxidation and Ca2+ incorporation/surface chemistry of commercially pure titanium implants regarding osseointegration, and whereas the chemical modification may compensate for a minimal surface roughness. Oral mucosa: To evaluate the effect of sol-gel derived nanoporous TiO2 coating of commercially pure titanium for the adaptation of oral mucosa. Bacterial adhesion and biofilm formation: To investigate bacterial adhesion, as well as biofilm formation and retention of oral bacteria in vitro on smooth and moderately rough, anodized and Ca2+ incorporated, as well as, nanoporous surfaces. Figure Bone adhesion towards Ca ion implanted surfaces, in trabecular and cortical bone Figure Soft tissue adhesion demonstrated with TEM We have shown that surface chemistry/anodic oxidation and Ca2+ incorporation of titanium surfaces may enhance the osseointegration and could possibly compensate for a minimal surface roughness. Nanoporous TiO 2 coating indicates some advantages in relation to unmodified titanium regarding the sealing of oral mucosa. A tendency of increased biofilm accumulation of oral bacteria in vitro was found for moderately rough (Sa 1-2 μm) blasted surfaces compared to smooth ones (Sa <0.5 μm). Moderately rough surfaces, in addition, retained more bacteria after mechanical removal of adhered biofilms compared to smooth. Nanoporosity or Ca2+ incorporation did not affect the bacterial adhesion or biofilm formation compared to turned surfaces. Increased bone contact to a Ca2+ incorporated oxidized c.p. titanium implant: an in vivo study in rabbit. Fröjd V, Franke-Stenport V, Meirelles L, Wennerberg A. Int J Oral Maxillofac Surg 37:6 (2010) Importance of Ca2+ modifications for osseointegration of smooth and moderately rough anodized titanium implants a removal torque and histological evaluation in rabbit. Fröjd V, Wennerberg A, Franke-Stenport V. Clin Impl Dent Rel Res Nanoporous TiO2 thin film on titanium oral implants for enhanced human soft tissue adhesion - a histological evaluation in three different levels of resolution. In situ analysis of biofilm formation on titanium surfaces Fröjd V, Chávez de Paz L, Andersson M, Wennerberg A, Davies J, Svensäter G. Submitted. Microbial biofilm formation on smooth nanoporous TiO2 coated and anodized Ca2+ modified surfaces. Fröjd V, Linderbäck P, Wennerberg A, Chávez de Paz L, Svensäter G, Davies J. Submitted. 19

20 4.11 Biologically induced stress corrosion as an indicator of residual stress. In general, surface instabilities play an important role in industry. The applications are initial stages of biologically induced corrosion. The examples from nuclear, petroleum and offshore industry are numerous. Problems arise during manufacturing and in operation because of high stresses and exposure to aggressive environment. As much as 25% of all accidents reported to the Swedish Plant Inspectorate are claimed caused by stress corrosion. A large part of these can be attributed to biological induced corrosion. A severe circumstance is that the cracks propagate at very small loads. Occasionally the general status of the environment is sufficient to cause stress corrosion, but in general the extreme local micro-environment under microbial colonies are responsible for so called pitting and initiation of edge cracks that subsequently grow into the structure. Apart from the immediate advantage of more knowledge, several applications taking advantage of the possibilities for non destructive testing of stresses could have been identified. The most striking example is that inspection of a developing surface roughness has been proven to work as a tool to discover high mechanical stresses and risk zones for corrosion cracks. Theoretical studies of an amorphous material explain the mechanism behind the repeated branching of cracks that is observed for cracks in aggressive environment. Numerically simulated crack growth was performed using a moving boundary finite element formulation. The results show great agreement with the experiments. An observed scatter in results in both experiments and in the numerical simulations reflects an inherent perturbation sensitivity of environmentally assisted cracking. Figure Left) Typical initiation pattern of small pits resulting from an aggressive environment. The pit density of pits reveal the mechanical stress in the structure. Right) Maximal principal strain field surrounding a newly branched crack. The arrows indicate where the surface straining equals the threshold strain. On initiation of chemically assisted crack growth and crack propagation paths of branching cracks in polycarbonate, Hejman, Ulf, 2010, Licentiate Thesis, 77 pages, Media-Tryck AB, Lund Sweden Dissolution driven crack branching in polycarbonate, Hejman, Ulf, Bjerkén, Christina, Fatigue and Fracture of Engineering Materials and Structures, 2010 Environmentally assisted initiation and growth of multiple surface cracks, Hejman, Ulf, Bjerkén, Christina, International Journal of Solids and Structures, 14-15, vol. 47, p ,

21 5 PhD Theses Supervised by Center Members Theses defended: 1. Rickard Hägglund, LTH (Supervisors Per Ståhle, P Isaksson): Damage of paper materials ( ) 2. Helena Tassids, LU. (Supervisors: Anette Gjörloff Wingren, HS and Pirkko Härkönen, LU): Expression and function of the protein tyrosine phosphatases SHP-1a nd SHP-2 in prostate cancer ( ). 3. Victoria Fröjd, Malmö University (Supervisors Ann Wennerberg, Gunnel Svensäter, Julia Davies, Victoria Franke Stenport (GU) On Ca 2+ incorporation and nanoporosity of titanium surfaces and the effect on implant performance ( ) Theses in progress: 4. Jildiz Hamit Eminovski, Malmö University (Supervisors Thomas Arnebrant and Krister Eskilsson (Kemira AB)) "Interactions of adsorbed layers of carbohydrate containing polymers - Saliva, mucins and bacterial surfaces" (2005-, Lic thesis defended 2009, doctoral thesis will be defended sept 2011) 5. Ulf Hejman, Malmö University (Supervisors Per Ståhle, Christina Bjerkén): "Biologically induced stress corrosion" (2005-, Lic thesis defended 2010, doctoral thesis will be defended autumn 2011) 6. Adnan Safdar, Lunds University (Supervisors Liu-Ying Wei, Per Ståhle) "Biocompatibility of ion beam melted materials" (2008-, Lic thesis defended Dec 2010, doctoral thesis will be defended autumn of Maria Pihl, Malmö University (Supervisors Gunnel Svensäter, Bertil Kinnby, Thomas Arnebrant) Biofilms on Peritoneal Dialysis Catheters (2005-) 8. Alma Masic, Malmö University (Supervisors Per Ståhle, Johan Helsing) "Mathematical modeling of biofilms" (2007-) 9. Sebastian Björklund, Lund University (Supervisors Emma Sparr (LU), Johan Engblom (Mah) and Krister Thuresson (Camurus AB)): "Water - a crucial factor in regulating biomembrane permeability" (2008-) 10. Anton Fagerström, Malmö University (Supervisors Johan Engblom, Vitaly Kocherbitov and Karin Bergström (AkzoNobel)): Bioavailability of active ingredients used in agriculture (2009-) 11. Yana Znamenskaya, Malmö University (Supervisors Vitaly Kocherbitov and Johan Engblom): Hydration of mucous gel (2009-) 12. Christian Alfredsson Kindblom, Malmö University (Supervisors Gunnel Svensäter, Claes Wickström, Madeleine Rohlin): Biofilm activity as a marker to identify patients at risk mechanisms underlying microbial stress tolerance (2009-) 13. Marjan Dorkhan, Malmö University (Supervisors Julia Davies, Gunnel Svensäter, Ann Wennerberg): Activities of microbial biofilms on bioactive implant surfaces (2009-) 14. Kostas Bougas, Malmö University (Supervisors Ann Wennerberg, Pentti Tengvall GU), Victoria Franke Stenport (GU)): Protein coat and bone incorporation (2009-) 15. Lory Melin, Malmö University (Supervisors Ann Wennerberg, Martin Andersson (Promimic AB): On the importance of nanometer structures for implant incorporation in bone tissue (2009-) 16. Magnus Falk, Malmö University (Supervisors Tautgirdas Ruzgas, Sergey Shleev): Three-dimensional nanobiostructure-based self-contained devices for biomedical application (2009-) 21

22 17. Jon Lind, LTH (Supervisors Ali Massih, Christina Bjerkén): Methods for evaluation of evaluation of the hydride embrittlement of Ni-based super alloys (2009-) 18. Tuerdi Maymaytilli, LTH (Supervisors Christina Bjerkén, Per Ståhle): Influence of plastic deformation on the formation and growth of embritteling metal hydride's" (2009-) 19. Liselott Ellmarker-Löfquist, Malmö University (supervisors: Liselott Lindh, Björn Söderfeldt): A Clinical and subjective evaluation of single implant treatments. A retroscpective study. (2009-) 20. Mariko Hayashi, Malmö University (Supervisors: Liselott Lindh, Ann Wennerberg, Ryo Jimbo): Biological Responses to Amphiphilic Nano-size Structures (2010-) 21. Peter Lamberg, Malmö University (Supervisors: Tautgirdas Ruzgas, Thomas Arnebrant): Three-dimensional (3D) bioelectrochemical systems assembled from nanoparticles (2010-) 22. Cathrine Albér, Malmö University (Supervisors: Johan Engblom, Vitaly Kocherbitov, Marie Lodén, Robert Corkery): Humectants and their effectson skin barrier function a mechanistic approach (2010-) 23. Caitriona Ericsson, Lund University (Supervisor: Gunilla Nordin Fredrikson): The role of IL-16 in atherosclerosis development (2010-) 24. Polyxeni Mantani, Lund University (Supervisor: Gunilla Nordin Fredrikson): Atheroprotective humoral and T cell immune responses against oxidized LDL (2010-) 6 Collaborative Partners of the Center 6.1 Industry collaborators (* Partners within KKs-Biofilm grant) 1. ACO Hud Nordic AB 2. Akzo Nobel AB* 3. Anordica AB 4. AnoxKaldnes AB* 5. Arcam AB* 6. Arla Foods AB* 7. Biogaia AB* 8. Bioglan AB 9. Bioinvent International AB 10. Camurus AB* 11. Euro-Diagnostica AB* 12. Eviderm AB 13. Galenica AB 14. Gambro Lundia AB* 15. Genovis AB 16. InnoScandinavia AB 17. Medeon AB 18. Medicon Valley Alliance 19. Nares AB 20. Nobel Biocare AB 21. Novosense AB* 22. Novozymes A/S, Denmark 23. Phase Holographic Imaging AB 24. Promimic AB* 25. PVA-MV AG, Germany 26. QuNano AB 27. Quantumwise A/S, Denmark 28. Stora Enso AB 29. Studsvik Nuclear AB 30. TetraPak AB 31. Volvo Aero AB 32. YKI AB 22

23 6.2 Academic collaborators Collaborations with other universities and research institutions in Sweden 1. Prof. em. Kåre Larsson, Camurus Lipid Research Foundation,, Lund 2. Prof. Gunnar Bergenholtz, Microbiology/Endodontics University of Gothenburgh 3. Prof. Per Claesson, The Royal Institute of Technology (KTH) and Surface Chemistry Institute (YKI), Stockholm 4. Prof Gunnar Dahlén, Microbiology/Endodontics University of Gothenburgh 5. Prof. Lo Gorton, Biochemistry, Lund University 6. Prof. Christer Hansson, Dermatology, Lund University 7. Prof. Martin Malmsten, Pharmacy, Uppsala University 8. Prof. Jan Nilsson, CRC UMAS, Lund University 9. Prof. Tommy Nylander, Physical Chemistry 1, Lund University 10. Prof. Adrian Rennie, Physics, Uppsala University 11. Prof. Mark Rutland, The Royal Institute of Technology (KTH) and Surface Chemistry Institute (YKI), Stockholm 12. Prof. Olle Söderman, Physical Chemistry 1, Lund University 13. Prof. Per Uvdal, MAX lab, Lund University 14. Prof. Martin Andersson, Dept Applied Chemistry, Chalmers university of Technology. 15. Prof. Pentti Tengvall, Dept Biomaterials, Sahlgrenska Academy, Göteborg University 16. Prof. Tomas Albrektsson, Dept Biomaterials, Sahlgrenska Academy, Göteborg University 17. Prof Marie Wahlgren, Food Technology, LTH, Lund University 18. Prof. Ingegerd Johansson, Cariology, Umeå University 19. Prof. Artur Schmidtchen, Department of Clinical Sciences, Lund University 20. Prof. Swante Twetman, Specialist Clinic for Oral Health Care, Hallands Läns Landsting, Halmstad 21. Prof Bengt-Göran Rosén, Dept. of Business and Engineering Halmstad University 22. Assoc. Prof. Lars Norlén, Cell- and Molecular biology, Karolinska Institute 23. Assoc. Prof. Viveka Alfredsson, Physical Chemistry 1, Lund University 24. Assoc. Prof. Ola Bergendorff, Dermatology, Lund University 25. Assoc. Prof. Emma Sparr, Physical Chemistry 1, Lund University 26. Assoc.Prof. Peter Siesjö, Department of Clinical Sciences, BMC, Lund University 27. Assoc. Prof.Eva Blomberg, The Royal Institute of Technology (KTH) and Surface Chemistry Institute (YKI), Stockholm 28. Associate Professor Victoria Franke-Stenport, Dept Prosthodontics, Sahlgrenska Academy, Göteborg University 29. Dr Yngve Cerenius, MAX lab, Lund University 30. Dr Adam Feiler, The Royal Institute of Technology (KTH) and Surface Chemistry Institute (YKI), Stockholm 31. Dr Isabel Goncalves, CRC UMAS, Lund University 32. Dr Robert Corkery, The Royal Institute of Technology (KTH) and Surface Chemistry Institute (YKI), Stockholm 33. Dr Anna Westerlund, Odontology, Gothenburg University 34. Dr. Ivan Maximov, Solid State Physics, Lund University 35. Dr Valera Veryazov, Theoretical Chemistry, Lund University 36. Dr Karina Persson, Odontology, Umeå University 37. Dr Jenny Liao Persson, Center for Molecular Pathology, Lund University 23

24 38. Per Rabe, ÖTL, Specialist Clinic for Oral Health Care, Hallands Läns Landsting, Halmstad 39. Björn Svensson, ÖTL, Specialist docent, Clinic fororal Health Care, Örebro Läns Landsting, Örebro International collaborations 1. Prof. em. Ian Hamilton, University of Manitoba, Dept of Oral Biology, Winnipeg, Canada 2. Prof. Robert Baier, University of Buffalo, Industry/University Cooperative Research Center for Biosurfaces, Buffalo, USA 3. Prof. F. C. Shih, King Fei Sals University, Dubai 4. Prof. Leslie Banks-Sills, Cornell University, Ithaca, NY, USA and University of Tel Aviv, Israel 5. Prof. Iwona Beech, University of Portsmouth, UK 6. Prof. Jan Sunner, University of Portsmouth, UK 7. Prof. David Beighton, Guy's Kings and St Thomas' Dental Institute, Joint Microbiology Research Unit, London, UK 8. Prof. R. Singh, Indian Institute of Technology, Mumbai, India 9. Prof. Bent Sörensen, Denmark Technical University, Riso Labs, Denmark 10. Prof. R. K. Thomas, Oxford University, Dept of Physical Chemistry, and ISIS at RAL (Neutron reflection facility), Oxford 11. Prof. Regine Willumeit, GKSS Research Centre, Geesthacht, Germany 12. Prof. Mark Hertzberg, University of Minnesota, Department of Microbiology, Minneapolis, USA 13. Prof. A. Needleman, Brown university, Providence, USA 14. Prof. Jukka Meurman, Helsinki University Central Hospital, Dept of Oral and Maxillofacial Diseases, Helsinki, Finland 15. Prof. Gordon Proctor, Kings College, London, UK 16. Prof. Christopher Exley, Keele University, UK 17. Prof. Takashe Sawase, Dept Prosthodontics, University of Nagasaki, Japan. 18. Prof. Wolfgang Schuhmann, Ruhr-Universität Bochum, Germany 19. Prof. Edmond Magner, University of Limerick, Ireland 20. Prof. Dietmar Haltrich, Universität für Bodenkultur Wien, Austria 21. Prof. Phil Bartlett, The University of Southampton, UK 22. Prof. Dennis Cvitkovitch, University of Toronto, Dept of Microbiology, Toronto, Canada 23. Prof. Jeannine Brady, University of Florida, Dept of Oral Biology, Gainesville, USA 24. Prof Kamal Mustafa, University of Bergen, Norway 25. ProfAlexander Toikka, Faculty of Chemistry, Saint Petersburg State University, Russia 26. Prof. Hermann Eberl, University of Guelph,Canada 27. Assoc. Prof. Marie Ranson, School of Biological Sciences, Scientific Director Cancer, Illawarra Health and Medical Research Institute, University of Wollongong, Wollongong, Australia 28. Assoc. Prof. Duncan Sutherland, Aarhus University, Denmark 29. Assoc. Prof. Pablo Galindo, Department of Oral surgery and Implant Dentistry, Granada University 30. Dr Justas Barauskas, Institute of Biochemistry, Vilnius, Lithuania 31. Dr Michael Ortize, Caltech, USA 32. Dr Andrey Jivkov, Manchester University, UMIST, UK 24

25 33. Dr. Isaac Klapper, Montana State University, Center of Biofilms Research, Montana, USA 34. Dr Juozas Kulys, Inst. of Biochemistry, Vilnius, Lithuania 35. Dr. Anne Meyer, University of Buffalo, Industry/University Cooperative Research Center for Biosurfaces, Buffalo, USA 36. Dr Srikumar Banerjee, Indian Atomic Research Centre, India 37. Dr. G. Fragneto, ILL (Neutron reflection facility), Grenoble 38. Dr Adam Heller, University of Texas at Austin, TX, USA 39. Dr Kenneth Holmberg, Tekniska högskolan i Helsinki, Finland 40. Dr Rafael Taborisky, Denmark Technical University, Riso Labs, Denmark 41. Dr Alexander Yaropolov, Inst. of Biochemistry, Moscow 42. Dr. Sergei Lobov, School of Biological Sciences, University of Wollongong, Wollongong, Australia 43. Dr. Miguel Alcalde, Consejo Superior de Investigaciones Científicas, Applied Biocatalysis group, Spain 44. Dr. Antonio L. De Lacey,, Consejo Superior de Investigaciones Científicas, Bioelectrocatalysis group, Spain 45. Dr. Donald Leech, National University of Ireland, Galway, Ireland 46. Dr. Paulo G. Coelho, Department of Biomimetics, New York University 47. Dr. Yuusuke Kozai, Department of Radiology, Kanagawa Dental University 48. Dr. Jelena Kosoric, Centre for Oral Growth and Development, Queen Mary s School of Medicine and Dentistry, University of London, UK 7 List of Publications of the Center from 2010 and Onwards All titles listed under journal articles, review papers, and book chapters have been or are subjected to peer review. Center publications for the period can be found at Journal articles and invited review papers in journals 1. Ackermann, Y.; Guschin, D.A.; Eckhard, K.; Shleev, S.; Schuhmann, W., Design of a bioelectrocatalytic electrode interface for oxygen reduction in biofuel cells based on a specifically adapted Os-complex containing redox polymer with entrapped Trametes hirsuta laccase. Electrochemistry Communications, 2010, 12(5), Bjerken C, Ortiz M (2010) "Evolution of anodic stress corrosion cracking in a coated material" INTERNATIONAL JOURNAL OF FRACTURE, 165, 2, p Bjerkén, Christina (2010) "The influence of biaxial loading on branching of a dissolution driven stress corrosion crack" Engineering Fracture Mechanics 11, vol. 77, p Björkbacka H, Lavant EH, Fredrikson GN, Melander O, Berglund G, Carlson JA and Nilsson J. Weak associations between human leukocyte antigen (HLA) genotype and acute myocardial infarction. Journal of Internal Medicine, 268:1 (2010) Björklund S, Engblom J, Thuresson K and Sparr E, A water gradient can be used to regulate drug transport across skin, J Control Release, 143:2 (2010) Chávez de Paz LE, Bergenholtz G, Svensäter G. The effects of antimicrobials on endodontic biofilm bacteria. J Endod 2010;36:

26 7. Coelho PG, Granato R, Marin C, Jimbo R, Lin S, Witek L, Suzuki M, Bonfante EA. Effect of Si Addition on Ca- and P- Impregnated Implant Surfaces with Nanometer Scale Roughness. An Experimental Study in Dogs. Clin Oral Impl Res; in press 8. Coman, V.; Ludwig, R.; Harreither, W.; Haltrich, D.; Gorton, L.; Ruzgas, T.; Shleev, S., A direct electron transfer-based glucose/oxygen biofuel cell operating in human serum. Fuel Cells, 2010, 10(1), Dagys, Marius; Haberska, Karolina; Shleev, Sergey; Arnebrant, Thomas; Kulys, Juozas; Ruzgas, Tautgirdas., Laccase-gold nanoparticle assisted bioelectrocatalytic reduction of oxygen. Electrochemistry Communications, 2010, 12(7), Dohan Ehrenfest DM, Lemo N, Jimbo R, Sammartino G. Selecting a relevant animal model for testing thein vivo effects of Choukroun s Platelet-Rich Fibrin (PRF) : rabbit tricks and traps. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 110(4): Dunér P; To F, Berg K, Alm R, Björkbacka H, Engelbertsen D, Fredrikson GN, Nilsson J, Bengtsson E. Immune responses against aldehyde-modified laminin accelerate atherosclerosis in Apoe-/- mice. Atherosclerosis, Oct;212(2):457-65, Eliasson A, Wennerberg A, Johansson A, Ortorp A, Jemt T. The Precision of Fit of Milled Titanium Implant Frameworks (I-Bridge(R)) in the Edentulous Jaw. Clin Implant Dent Relat Res. 2010;12: Exley C, Siesjö P and Eriksson H. The Immunobiology of Aluminium Adjuvants: How do they really work? Trends in Immunology, 31 (2010) Fransson H, Petersson K., Davies JR. Dentine sialoprotein and Collagen I expression after experimental pulp capping in humans using Emdogain Gel. Int Endod J 2011, 44, Fredrikson GN, Nilsson J, Björkbacka H. Autoimmunitet mot blodfett kan ge både skydd och sjukdom. Incitament, 19(7): , Fröjd V, Wennerberg A, Franke-Stenport V. Importance of Ca2+ modifications for osseointegration of smooth and moderately rough anodized titanium implants-a removal torque and histological evaluation in rabbits. Clin Implant Dent Relat Res 2010;26 E pub ahead of print. 17. Halthur TJ, Arnebrant T, Macakova L, Feiler A. Sequential Adsorption of Bovine Mucin (BSM) and Lactoperoxidase (LPO) to various substrates studied with Quartz Crystal Microbalance with Dissipation (QCM-D). Langmuir 26:7 (2010) Hamit-Eminovski J, Eskilsson K and Arnebrant T, An ellipsometry study on the effect of aluminium chloride and ferric chloride formulations on mucin layers adsorbed at hydrophobic surfaces. Biofouling, 26 (2010), Hamit-Eminovski J, Eskilsson K and Arnebrant T, Change in surface properties of Microthrix parvicella upon addition of polyaluminium chloride as characterized by atomic force microscopy, Biofouling, 26 (2010), Hejman, Ulf, Bjerkén, Christina (2010) "Dissolution driven crack branching in polycarbonate" Fatigue and Fracture of Engineering Materials and Structures, In Press 21. Hejman, Ulf, Bjerkén, Christina (2010) "Environmentally assisted initiation and growth of multiple surface cracks" International Journal of Solids and Structures 14-15, vol. 47, p Hjalmarsson L, Smedberg JI, Wennerberg A. Material degradation in implant-retained cobalt-chrome and titanium framworks. J Oral Rehabilitation 2010;29 E-PUB ahead of print. 23. Jalali, Y., Lindh, L. A randomized prospective clinical evaluation of two desensitizing agents on cervical dentine sensitivity: A pilot study. Swed Dent J. 34: 79-86,

27 24. Jimbo R, Ivarsson M, Koskela A, Sul YT, Johansson CB. Protein adsorption to surface chemistry and crystal structure modification of titanium surfaces. J Oral Maxillofac Res 2010; 1(3):e Johansson CB, Jimbo R, Steffenson P. Ex Vivo and In Vivo Biomechanical Test of Implant Attachment to Various Materials. Introduction of a New User-friendly Removal Torque Equipment. Clin Implant Dent Relat Res 2010; In press 26. Karlsson C, Ahrné S, Molin G, Berggren A, Palmquist I, Fredrikson GN, and Jeppsson B. Probiotic therapy to men with incipient arteriosclerosis initiates increased bacterial diversity in colon: a randomized controlled trial. Atherosclerosis, 208 (2010) Klingenberg R, Lebens M, Hermansson A, Fredrikson GN, Strodthoff D, Rudling M, Ketelhuth D, Gerdes N, Holmgren J, Nilsson J and Hansson GK. Intranasal immunization with an apob-100 fusion protein induces antigen-specific regulatory T cells and reduces atherosclerosis. Arterioscler Thromb Vasc Biol, 30(5):946-52, Kocherbitov V and Alfredsson V. Assessment of porosities of SBA-15 and MCM-41 using water sorption calorimetry. Accepted for publication in Langmuir 29. Kocherbitov V, Ulvenlund S, Briggner L-E, Kober M, Arnebrant T. Hydration of a natural polyelectrolyte Xanthan Gum: comparison with non-ionic carbohydrates, Carbohydrate Polymers 82 (2010) Kocherbitov, V.; Arnebrant, T. Hydration of lysozyme: the protein-protein interface and the enthalpy-entropy compensation. Langmuir (2010), 26(6), Kolbus D, Ramos O, Olofsson KE, Persson J, Wigren M, Björkbacka H, Fredrikson GN and Nilsson J. CD8+ T cell activation predominate early immune responses to hypercholesterolemia in Apoe-/- mice. BMC Immunol, 2010 Dec 2;11(1):58. [Epub ahead of print]. 32. Kolbus D, Wigren M, Ljungcrantz I, Söderberg I, Björkbacka H, Nilsson J and Fredrikson GN. Immunization with cationized BSA inhibits progression of disease in Apobec-1/LDL receptor deficient mice with manifest atherosclerosis. Immunobiology, in press. 33. Masic A, Bengtsson J, Overgaard NC, Christensson M, Heyden A. Measuring and modeling the oxygen profile in a nitrifying Moving Bed Biofilm Reactor. Mathematical Biosciences, 227:1 (2010) Melin Svanborg L, Hoffman M, Andersson M, Currie F, Kjellin P, Wennerberg A. The effect of hydroxyapatite nanocrystals on early bone formation surrounding dental implants. Int J Oral Maxillofac Surg. In press 35. Nilsson J and Fredrikson GN. The B cell in atherosclerosis - teaming up with the bad guys? Clin Chem, Dec;56(12): , Nilsson-Berglund LM, Zetterqvist AV, Nilsson-Ohman J, Sigvardsson M, González Bosc LV, Smith ML, Salehi A, Agardh E, Fredrikson GN, Agardh CD, Nilsson J, Wamhoff BR, Hultgårdh-Nilsson A, Gomez MF. Nuclear factor of activated T cells regulates osteopontin expression in arterial smooth muscle in response to diabetes-induced hyperglycemia. Arterioscler Thromb Vasc Biol, 30(2):218-24, Ono D, Jimbo R, Kawachi G, Ioku K, Ikeda T, Sawase T. Lateral bone augmentation with a newly developed beta-tri-calcium phosphate block: an experimental study in the rabbit mandible. Clin Oral Impl Res; In press 38. Pedersen TX, Binder CJ, Fredrikson GN, Nilsson J, Bro S, Nielsen LB. The proinflammatory effect of uremia overrules the anti-atherogenic potential of immunization with oxidized LDL in apoe-/- mice. Nephrol Dial Transplant, Aug;25(8): ,

28 39. Pihl M, Chávez de Paz LE, Schmidtchen A, Svensäter G, Davies JR. Effects of clinical isolates of Pseudomonas aeruginosa on Staphylococcus epidermidis biofilm formation. FEMS Immunol Med Microbiol 2010;59: Pihl M, Davies JR, Chávez de Paz LE, Svensäter G. Differential effects of Pseudomonas aeruginosa on biofilm formation by different strains of Staphylococcus epidermidis. FEMS Immunol Med Microbiol 2010;59: Ressine A, Vaz-Dominguez C, Fernandez VM, De Lacey AL, Laurell T, Ruzgas T, Shleev S. Bioelectrochemical studies of azurin and laccase confined in three-dimensional chips based on gold-modified nano-/microstructured silicon. Biosensors and Bioelectronics, 25:5 (2010) Santos O, Lindh L, Halthur T and Arnebrant T, Adsorption from saliva to silica and hydroxyapatite surfaces and elution of salivary films by SDS and delmopinol, Biofouling, 26 (2010), Shleev S.; Shumakovich G.; Morozova O.; Yaropolov A., Stable floating air diffusion biocathode based on direct electron transfer reactions between carbon particles and high redox potential laccase. Fuel Cells, 2010, 10(4), Sotres J, Svensson O and Arnebrant T., Friction force spectroscopy of β- and κ-casein monolayers. Langmuir 2011, 27(3), Stahle, P. (2010) "Foreword of the KB Broberg symposium issue" International Journal of Fracture, 165(2): p Stahle, P., Barenblatt, G.I., and Rice, J.R. (2010) Biography of Professor Knut Bertram Broberg" International Journal of Fracture, 165(2): p Stahle, P., Singh, R.N., and Banerjee, S. (2010) "Spontaneous fracture of growing precipitates with large misfit strain" International Journal of Fracture, 165(2): p Stollenwerk MM, Svensson O, Schiopu A, Jansson B, Arnebrant T, and Fredrikson GN. Adsorption of low density lipoprotein, its oxidation and subsequent binding of specific recombinant antibodies an in situ ellipsometric study. Biochimica et Biophysica Acta (BBA) -General Subjects 2011, 1810(2), Strietzel FP, Lafaurie GI, Bautista Mendoza GR, Alajbeg I, Pejda S, Vuletić L, Mantilla R, Falcão DP, Leal SC, Barreto Bezerra AC, Tran SD, Ménard HA, Kimoto S, Pan S, Martín-Granizo RA, Maniegas Lozano ML, Zunt SL, Krushinski CA, Melilli D, Campisi G, Paderni C, Dolce S, Yepes JF, Lindh L, Koray M, Mumcu G, Elad S, Zeevi I, Aldape Barrios BC, López Sánchez RM, Beiski BZ, Wolff A, Konttinen YT. Efficacy and safety of an intraoral electrostimulation device for xerostomia relief: a multicenter randomized trial. Arthritis & Reumatism. 63 (2010) Svanborg LM, Andersson M, Wennerberg A. Surface Characterization of Commercial Oral Implants on the nanometer level. J Biomed Mater Res B Appl Biomater. 2010; 92: Svensson O and Arnebrant T, Adsorption of serum albumin on silica The influence of surface cleaning procedures. Journal of Colloid and Interface Science 2010, 344(1), Svensson O and Arnebrant T, Mucin layers and Multilayers. Current Opinion in Colloid & Interface Science 2010, 15(6), Svensson O, Halthur T, Sjödin T and Arnebrant T, The adsorption of delmopinol at the solid liquid interface the role of the acid base equilibrium. Journal of Colloid and Interface Science 2010, 350, Tassidis, H., Culig Z., Gjörloff Wingren, A, Härkönen, P. Role of protein tyrosine phosphatase SHP-1 in interleukin-6 regulation of prostate cancer. The Prostate 70 (2010)

29 55. Wennerberg A, Albrektsson T. Current Challenges in Successful Rehabilitation with Oral Implants. J Oral Rehabil 2010 Oct 25 Epub ahead of print 56. Wennerberg A, Albrektsson T. On implant surfaces: a review of current knowledge and opinions. Int J Oral Maxillofac Implants 25 (2010) Wigren M, Kolbus D, Dunér P, Ljungcrantz I, Söderberg I, Björkbacka H, Fredrikson GN, Nilsson J. Evidence for a role of regulatory T cells in mediating the athero-protective effect of apolipoprotein B peptide vaccine. J Int Med, 2010 Nov 3. doi: /j x. [Epub ahead of print] 58. Zhao M, Wigren M, Dunér P, Bengtsson D, Olofsson K, Björkbacka H, Nilsson J, Fredrikson GN. Journal of Immunology, 184:5 (2010) Östman PO, Wennerberg A, Albrektsson T. Immediate occlusal loading of NanoTite PREVAIL implants: a prospective 1-year clinical and radiographic study. Clin Implant Dent Relat Res. 12:1 (2010) Submitted: 60. Chávez de Paz LE, Davies J, Svensäter G. Fluorescence in situ hybridization for intracellular mrna detection in adherent bacteria. Appl Environ Microbiol, submitted 61. Fröjd V, Chàvez de Paz L, Andersson M, Wennerberg A, Davies JR, Svensäter G. In situ analysis of multi-species biofilm formation on customized titanium surfaces. Submitted 62. Fröjd V, Linderbäck P, Wennerberg A, Svensäter G, Davies JR. Microbial biofilm formation on smooth nanoporous titanium dioxide coated and anodized calcium ion modified titanium surfaces. (Submitted) 63. Jimbo R, Sotres J, Johansson C, Breding K, Currie F, Wennerberg A. The biological responses to three different nanostructures applied on smooth implant surfaces. Clinical Oral Implant Res. Submitted 64. Leisnert L, Karlsson M, Franklin I, Lindh L, Wretlind K., Improving teamwork between students from two professional centers in dental education. Eur J Dent Educ. Submitted 65. Lindh L, Santos O, Svendsen IE, Sotres J, Breding K, Wennerberg A, Arnebrant T. Adsorption of plasma proteins onto hydroxyapatite surfaces. Submitted 66. Pihl M, Arvidsson A, Skepö M, Svensäter G, Davies JR. Biofilm formation by Staphylococcus epidermidis on peritoneal dialysis catheters and the effects of extracellular products from Pseudomonas aeruginosa. Submitted 67. Pihl M, Davies JR, Johansson AC, Svensäter G. Microbial biofilms in patients with peritoneal dialysis catheters. (submitted) Proceeding papers: 68. Fagerström A, Kocherbitov V, Lamberg P, Bergström K, Westbye P, Ruzgas T and Engblom J. Factors Affecting Transport of Tebuconazole over Silicone Membrane and Leaf Cuticle; in 9th International Symposium on Adjuvants for Agrochemicals, ISAA Society; Baur P and Bonnet M Eds.; August 2010, Pages ISBN Safdar A, Wei Liu-Ying, Snis A, Lai Z, Evaluations of microstructural development in electron beam melted Ti-6Al-4V, Proc. MSE Conference 2010, Aug, 2010, Darmstadt Germany. 70. Sparr, Emma; Björklund, Sebastian; Engblom, Johan; Thuresson, Krister, A water gradient can be used to regulate drug transport across skin - A responding membrane, Biophysical Journal, Volume 98, Issue 3, Supplement 1, January 2010, Page 627a 29

30 7.2 Books and book chapters 1. Albrektsson T, Wennerberg A. The science of osseointegration. In; Prosthodontic Treatment for Edentulous Patients, ed 13. Submitted Alm K, Cirenajwis H, Gisselsson L, Gjörloff Wingren A, Janicke B, Mölder A, Oredsson S, Persson J. Digital holography and cell studies. In press, Gjörloff Wingren A, et al. Digital holographic microscopy - innovative and nondestructive analysis of living cells. Microscopy Book Series - Volume # 4: Microscopy: Science, Technology, Applications and Education Release date: Autumn Julia R. Davies, Claes Wickström and David J. Thornton. Gel-forming and cellassociated mucins preparation for structural and functional studies in Methods in Molecular Biology Edited by McGuckin and Thornton. In press Winning TA, Skinner VJ, Kinnell A, Townsend, GC, Svensäter G, Rohlin M, Davies, JR. The influence of two PBL curricula contexts on students understandings of PBL, approaches to learning and outcome in Researching Problem-based Learning in Clinical Education: The Next Generation. Edited by Bridges S, McGrath C, Whitehill T. In press Popular articles 1. El-Schish Z, Mölder A, Sebesta M, Gjörloff Wingren A. Det digitala holografiska mikroskopet innovativ teknik för analys av levande celler. 2010, Bioingenjören (Norge). 2. Engblom J, Interfaces in biomedicine PS Public service review European Union 19, PSCA Int. Ltd 2010, Engblom J, Interfacing the future, PS Public service review European Union 19, PSCA Int. Ltd 2010, Fridberg M, Tassidis H, Gjörloff Wingren A. PTPN7 (protein tyrosine phosphatase, nonreceptor type 7). Atlas Genet Cytogenet Oncol Haematol. January Reportage i Mun & Hals (tidning för mun- och halscancerförbundets medlemmar), 2010, nr 2:8 om forskningsprojekt om saliversättningsmedel. 8 Visitors and Seminars at the Center 1. Prof Mark Herzberg, University of Minneapolis, USA, Seminar: Characterization of hydrogen peroxide-induced DNA release by streptococci Prof Charles Shuler, University of British Columbia, Canada, Seminar: Visions and strategies in dental education Prof Arunas Ramanavicius, Faculty of Chemistry, Vilnius University, Vilnius, Lithuania 4. Dr Almyra Ramanaviciene, Faculty of Chemistry, Vilnius University, Vilnius, Lithuania 5. Domhnall Mac Aodha, PhD student at National University of Ireland, Galway, Ireland, Marius Dagys, PhD student at the Institute of Biochemistry, Vilnius, Lithuania, Vida Krikstolaityte, PhD student at the Faculty of Chemistry, Vilnius University, Vilnius, Lithuania, Diana Mate, PhD student from Instituto de Catálisis y Petroleoquímica, CSIC, Madrid, Spain

31 9. Asta Makaraviciute, MSc-student at the Faculty of Chemistry, Vilnius University, Vilnius, Lithuania. 10. Alexandra Pulyalina, PhD student at Faculty of chemistry, St Petersburg University, Russia, Marija Jankunec, PhD student at Institue of Biochmistry, Vilnius, Lithuania, & Prof Per-Olof Glantz, Faculty of Odontology, Malmö University:Seminar: From forbidden to high priority Prof Göran Lundborg, Hand Surgery, Lund University, Seminar: Hand to brain in research and development Prof Tomas Albrektsson, Biomaterial Research, University of Gothenburg, Seminar: On survival and success of oral implants Prof Tommy Nylander, Physical Chemistry, Lund University, Seminar: On the interaction of drug delivery vehicles with model biomembranes Carin Daal, Region Skåne, Seminar: The innovation system in Skåne what use can a researcher have of it? Klementina Österberg, GU Holding, Seminar: Investing in making a profitable university business Dr Evy Lundgren-Åkerlund, Ideon Bioincubator, Seminar: From adhesion to business Dr Jeanette Sundberg, PULS AB, Seminar: What makes an innovation successful? Drs Karin Bryskhe & Anna Stenstam, Colloidal Resource AB, Seminar: Business windows at the intersection of academia and business Jonas Gulliksson, Advokatbyrån Gulliksson, Seminar: IP strategies Ashkan Pouya, Serendipity Innovations AB, Seminar: The Creative Dance Eva Jensen, Advokatbyrån Lindahls, Seminar: Agreeing on intellectual property rights 24. Takashe Sawase, Nagasaki University, Japan 25. Humbeto Osvaldo Schwartz Filho, PhD-student at UNESP, Sao Paulo State University, Brazil, (11 months) 26. Dr Lana Karlmark, Läkemedelsverket, Uppsala, Seminar: Myndighetens syn på CROverksamhet Dr Torbjörn Larsson, Medivir, Huddinge, Seminar: Samarbetet med CRO - Ur uppdragsgivarens synvinkel Dr Lars Wannerberger, Galenica AB, Malmö, Seminar: Samarbetet med CRO - Ur uppdragstagarens synvinkel 29. Odd Swarting, Setterwalls advokatbyrå, Stockholm, Seminar: Juridiska aspekter och avtal vid anlitande av CRO Maria Stenbäck, Awapatent, Malmö, Seminar: Patentperspektivet Vad bör man tänka på? Prof Per Jönsson, School of Technology, Malmö University, Assoc. Prof Yuanji Cheng, School of Technology, Malmö University, Internal seminars at the Center 33. Vitaly Kocherbitov Hydration of proteins Yana Znamenskaya Effect of hydration on the structural properties of mucous gel Anna Gustafsson "Some aspects on immunomodulation by cationic peptides on bacterial toxins"

32 36. Varvara Petrova Chemical composition of the epicuticular wax layers of Clivia miniata leaves Helena Tassidis "Expression and function of the protein tyrosine phosphatases SHP-1 and SHP-2 in prostate cancer" Emmy Nilsson and Anette Gjörloff Wingren "Interfacing antibody-based microarrays and digital holography enables detection of cell death-induced lymphocytic cell lines" Javier Sotres Friction and wear of adsorbed casein layers studied by AFM Vida Krikstolaityte Assembly and catalytic properties of nanoparticles and enzymes Gabriella Sinkiewicz Påverkan av munhålans bakterieflora i en frisk population efter tillförsel av Lactobacillus reuteri via tuggummi" Magnus Falk Three-dimensional nanobiostructure-based biofule cells for biomedical applications Maria Stollenwerk och Anette Gjörloff Wingren Pedagogiskt seminarium om bedömning Vida Krikstolaityte Evaluation of (1) gold nanoparticle/laccase assembly at different ionic strengths by using QCM-D and (2) bioelectrocatalysis of oxygen using LSV Diana Maté Laboratory Evolution of a High Redox Potential Laccase, Alejandro Barrantes "Interactions between DNA and Alzheimer's disease amyloid peptides, and MAP tau, detected by Surface Plasmon Resonance" Lina Pedersen Effect of a gradient in water chemical potential on buccal drug delivery Jovice Boon Sing Ng Mesoporous spheres as model materials for molecular transport studies Julia Hedlund z-lab, an new and compact impedance measurement instrument for sensitive surface analyses Yana Znamenskaya Effect of hydration on the structural properties of mucin Pontus Dunér Immune responses against aldehyde-modified laminin accelerate atherosclerosis in ApoE deficient mice Maria Wigren MHC class II deficiency increases atherosclerosis in apoe-/- mice Anton Fagerström Factors affecting transport of tebuconazole over silicone and leaf cuticle Viktor Andoralov ''Bioelectrocatalytic bechaviour of Octaheme Nitride Reductase from Thioalkalivibrio paradoxus'' Workshops and Conferences Organized by the Center During 2010 the Center has launched the 6 th Annual workshop as a joint venture with the EU project Valorisation of knowledge intensive ideas in the South Baltic area (SB- VALOR).,Focus this year was Biomaterials - From Fundamentals to Market Application and the event attracted 110 participants from different universities as well as industries, innovation agencies, solicitor s offices and risk capitalists The workshop was 32

33 preceeded by a PhD-student day with Prof. Per-Olof Glantz, former Vice-Chancellor of Malmö University, and Prof. Tommy Nylander, Lund University as mentors. The Center has also launched a Members day for sharing progress in the different projects with all partners, and coorganized a workshop on Choice of and collaboration with CRO s in pharmaceutical development at Malmö University, launched by the Swedish Academy of Pharmaceutical Sciences. Ann Wennerberg has been responsible for arranging a 2-week workshop within the National Research School of Odontology; Biomaterials in medicine, together with BIOSUM, Gothenburg. Liselott Lindh arranged a salivary symposium together with Guy Carpenter (London) at IADR in Barcelona. Johan Engblom arranged a kick-off meeting for the new project Humectants and their effects on skin barrier function a mechanistic approach, funded by KKs Our internal seminars were organised by Tautgirdas Ruzgas and Anette Gjörloff Wingren, and Gunilla Nordin Fredrikson was responsible for the seminars at Clinical Research Center, Lund University 10 Contributions to and Participation in Conferences and Workshops 10.1 Oral presentations 2010 International 1. Fagerström A, Kocherbitov V, Lamberg P, Bergström K, Westbye P, Ruzgas T, Engblom J. Factors affecting transport of tebuconazole over silicone membrane and leaf cuticle, 9th international symposium on Adjuvants for Agrochemicals (ISAA 2010) August 16-20, Fröjd V,Svensäter G, Andersson M, Wennerberg A, Chàvez de Paz L. In situ analysisof biofilm formationon titanium surfaces. IADR, 88th general session, Barcelona, Spain, July14-17, Kocherbitov V and Wadsö L. Application of Sorption Calorimetry for Studies of Hydration of Polymers, POLYSOLVAT-8, 8th International IUPAC Conference Polymer-Solvent Complexes & Intercalates, 5-8 July 2010, Strasbourg, France 4. Krikstolaityte V, Dagys M, Kulys J, Ramanavicius A, Arnebrant T, Shleev S, Ruzgas T. Reduction of O2 at laccase modified gold nanoparticles. 4th NanoSchool. Centre of Nanotechnology and Materials Science NanoTechnas, Faculty of Chemistry, Vilnius University, Lithuania, Krikstolaityte V, Dagys M, Kulys J, Ramanavicius A, Arnebrant T, Shleev S, Ruzgas T. Reduction of O2 at laccase modified gold nanoparticles. The 61st Annual Meeting of the International Society of Electrochemistry. Nice, France, Lindh L. "Salivary films formed on different materials future applications", IADR, Barcelona, Lindh L. Invited speaker: Oral Mouthfeel. IADR, Barcelona. J Dent Res, 89: Sp Iss

34 8. Shleev S, Falk M, Sotres J, Blum Z, Ruzgas T. Semiconductor-like behaviour of a fungal laccase. 4th NanoSchool. Centre of Nanotechnology and Materials Science NanoTechnas, Faculty of Chemistry, Vilnius University, Lithuania, Shleev S. Implantable glucose-oxygen biofuel cell. 217th Meeting of the Electrochemical Society. Vancouver, Canada, Shleev S. Semiconductor-like behaviour of a fungal laccase. The 61st Annual Meeting of the International Society of Electrochemistry. Nice, France, Sotres J. AFM a versatile tool for mapping surface properties with nm resolution. 4th NanoSchool. Centre of Nanotechnology and Materials Science NanoTechnas, Faculty of Chemistry, Vilnius University, Lithuania, Wennerberg A. Invited speaker: Implant treatment. Seminar Gent University, Belgium February 26th 2010, 8h 13. Wickström C. Invited speaker at the IADR Annual meeting, Barcelona July 17 th, Wickström C. Mucin interaction with microorganisms. IADR, 88th general session, Barcelona, Spain, July14-17, National 15. Arnebrant T. Invited speaker: "Properties of salivary and mucin films on different materials and possibilities for multicomponent surface assemblies", 24 March 2010, Chalmers, Materials and Health Platform in the Strategic Materials Initiative at Chalmers and GU Biomaterials arranged by Peter Thomsen, GU and Martin Andersson, Chalmers 16. Davies J. Invited speaker at 'The Pufendorf Institute, Lund, March 29 th, Davies J.-Invited speaker at 'The Department of Oral Biology, KI, Stockholm, March 5 th, Engblom J. Invited speaker: Utan vatten inget upptag Medeon AB, 25th anniversary) May 15th, Lindh L. Biofilm formation in oral environment saliva and salivary films, Nationella forskarskolan i Odontologi, Göteborg 20. Lindh L. Invited speaker: Saliv och saliversättningsmedel, vid Mun- & Halscancerförbundets riksstämma, Knivsta, Lindh L. Invited speaker: Tandvård nya tandvårdsstödet. Vad gäller för dig som strålbehandlats, vid Mun- & Halscancerförbundets riksstämma, Knivsta, Lindh L. Invited speaker: Den nya tandvårdstaxan, samt hjälpmedel för mun- och halscancerdrabbade vid lokalföreningen Mun- och Halscancerföreningens rehab-möte, Ronneby Brunn, Lindh L. Invited speaker: Hjälpmedel för mun- och halscancerdrabbade, vid lokalföreningen Mun- och Halscancerföreningens rehab-möte, Ronneby Brunn, Svensäter G. Invited speaker at 'Cross-disciplinary course in Biomaterials', Göteborg, February 3 rd, Svensäter G. Invited speaker at 'Diplomkurs I Odontologisk Teknologi', Göteborg, May 27th, Svensäter G. Invited speaker at 'The Pufendorf Institute, Lund, March 29 th, Wennerberg A. Invited speaker: Forskning och framtid för dentala implantat. Tylösandsdagarna September Wennerberg A. Invited speaker: Surfaces: Where are we today and where are we going? The Gothenburg Research and Technology Forum. Gothenburg, Sweden October Wennerberg A. Invited speaker: Teknologi inom tandvården. Medicin för tekniker, Lunds universitet, 16/

35 30. Wennerberg A. Invited speaker: Topographical measurements- techniques and applications h. In research course Models and Methods in Dental Implant Research. Sahlgrenska Academy, Göteborg. Local 31. Arnebrant T, Kocherbitov V. Drug carrier interactions at biologically relevant interfaces. Members Day of Biofilms Feb10, 2010, Malmö 32. Arnebrant T, Kocherbitov V. Interactions between pharmaceutical materials and water. Members Day of Biofilms Feb10, 2010, Malmö 33. Arnebrant T. Adsorption and biofilm formation at oral interfaces, Members Day of Biofilms Feb10, 2010, Malmö 34. Davies J, Investigation of interactions between osteopontin and oral biofilm bacteria, Members Day of Biofilms Feb10, 2010, Malmö 35. Eriksson H, Arnebrant T. New concept for lipid based surface coatings in bioassays. Members Day of Biofilms Feb10, 2010, Malmö 36. Fagerström A, Adjuvants for products used in agriculture, Members Day of Biofilms Feb10, 2010, Malmö 37. Fagerström A, Factors affecting transport of tebuconazole over silicone and leaf cuticle. 6th Annual Workshop of Biofilms, Malmö. 38. Maimaitiyili T, The misfit strain analysis on hydrides with synchrotron radiation. 6th Annual Workshop of Biofilms, Malmö. 39. Ng JBS, Using mesophorous silica spheres as biomolecule supports. 6th Annual Workshop of Biofilms, Malmö. 40. Pihl M. Halvtidsseminarie Feb12, 2010., Biofilms on peritoneal dialysis catheters, Opponent: Prof Pentti Tengvall, Sahlgrenska academy, Göteborg. 41. Pihl M. Microbial biofilms in patients with peritoneal dialysis catheters. 6th Annual Workshop of Biofilms, Malmö. 42. Pihl M. Microbial biofilms in patients with peritoneal dialysis catheters. PhD day Oct ,, 6th Annual Workshop of Biofilms, Malmö 43. Pihl M. Microbial biofilms on peritoneal dialysis catheters. Members Day of Biofilms Feb10, 2010, Malmö 44. Ståhle P, Biologically induced stress corrosion crack growth. Members Day of Biofilms Feb10, 2010, Malmö 45. Ståhle P, Investigation and modeling of convection in biofilms for different carriers, Members Day of Biofilms Feb10, 2010, Malmö 46. Svanborg L, On the importance of nanometer structures for implant incorporation in bone soft tissue. 6th Annual Workshop of Biofilms, Malmö. 47. Svensäter G, Sjödin T. Adsorption and biofilm formation at oral interfaces, Members Day of Biofilms Feb10, 2010, Malmö 48. Wennerberg A, Arnebrant T. Implants and their interface towards bone tissue. Members Day of Biofilms Feb10, 2010, Malmö 49. Wennerberg A. An example of a PhD program that influenced the development of oral implants. 6th Annual Workshop of Biofilms, Malmö. 50. Wennerberg A. Invited speaker: Development of oral implants. Examples of research contribution. The 6th annual workshop of Biofilms. Research Center for Biointerfaces, October 13-15th, Znamenskaya Y, Effect of hydration on structural and thermodynamic properties of mucin. 6th Annual Workshop of Biofilms, Malmö. 35

36 10.2 Posters 2010 International 1. Dorkhan M, Chàvez de Paz L, Davies JR. Effect of titanium surface roughness on adhesion of Streptococcus oralis. IADR, 88th general session, Barcelona, Spain, July14-17, Kindblom C, Wickström C, Svensäter G. Binding of Streptococcus mutans to salivary components. IADR, 88th general session, Barcelona, Spain, July14-17, Kocherbitov V, Arnebrant T. Adsorption of water at solid-vapour and solid-solid interfaces. 24th Conference of the European Colloid and Interface Society (ECIS 2010), Prague, September 5-10, Neilands J, Svensäter G. Effect of fluoride and probiotics on plaque acid tolerance. IADR, 88th general session, Barcelona, Spain, July14-17, Svanborg L, Hoffman M, Andersson M, Wennerberg A. The effect of hydroxyapatitenanocrystals on early bone formation surrounding dental implants. IADR, Barcelona, 15th of July ID Svensson O and Arnebrant T. Adsorption of serum albumin on silica: Influence of surface cleaning procedures. 24th Conference of the European Colloid and Interface Society (ECIS 2010), Prague, September 5-10, Znamenskaya Y, Engblom J, Sotres J, Arnebrant T and Kocherbitov V. Effect of hydration on structural and thermodynamic properties of mucin. 24th Conference of the European Colloid and Interface Society (ECIS 2010), Prague, September 5-10, 2010 National 8. Björklund S, Sparr E, Engblom J, Thuresson K. The effect of water and other small polar molecules on skin permeability, Swedish Neutron Scattering Society Meeting and Biointerfaces, August 24-27, Engblom J, Pedersen L, Nilsson P, Kocherbitov V. Cubic phases in the DOPS/DOPE/water system. MAX-lab Annual user meeting, Lund, November 8-10, Fagerström A., Kocherbitov V., Lamberg P., Bergström K., Westbye P., Ruzgas T., Engblom J., Factors affecting transport of tebuconazole over silicone membrane and leaf cuticle, 10th ASCS Molecular Processes at Solid Surfaces, Lund, November 24-26, Local 11. Barauskas J, Cervin C, Jankunec M, Špandyreva M, Ribokaitė K, Tiberg F, Johnsson M. Interactions of lipid-based liquid crystalline nanoparticles with model and cell. 6th Annual Workshop of Biofilms, Malmö. 12. Björklund S, Dahi I, Engblom J, Sparr E, Ruzgas T. Skin response to hydration confirmed by impedance spectroscopy. 6th Annual Workshop of Biofilms, Malmö. 13. Björklund S, Sparr E, Engblom J, Thuresson K. The effect of water and other small polar molecules on skin permeability. 6th Annual Workshop of Biofilms, Malmö. 14. Dagys M, Shleev S, Arnebrant T, Niaura G, Kulys J, Ruzgas T. Bioelectrocatalysis of oxygen reduction with Trametes hirsuta laccase immobilized on gold nanoparticles. 6th Annual Workshop of Biofilms, Malmö. 15. Engblom J, Pedersen L, Nilsson P, Kocherbitov V. Cubic phases in the DOPS/DOPE/water system. 6th Annual Workshop of Biofilms, Malmö. 16. Fagerström A., Kocherbitov V., Lamberg P., Bergström K., Westbye P., Ruzgas T., Engblom J. Factors affecting transport of tebuconazole over silicone membrane and leaf cuticle. 6th Annual Workshop of Biofilms, Malmö. 36

37 17. Hamit-Eminovski J, Eskilsson K, Arnebrant T. An ellipsometry study on the effect of aluminium chloride and ferric chloride formulations on mucin layers adsorbed at hydrophobic surfaces. 6th Annual Workshop of Biofilms, Malmö. 18. Kindblom C., Wickström C., Svensäter G. Binding of Streptococcus mutans to salivary components. 6th Annual Workshop of Biofilms, Malmö. 19. Neilands J, Petersson L, Beighton D, Svensäter G. Fluoride inhibits acid tolerance of root surface biofilms. 6th Annual Workshop of Biofilms, Malmö. 20. Pedersen L, Wahlgren M, Engblom J. Effect of a gradient in water chemical potential on buccal drug delivery. 6th Annual Workshop of Biofilms, Malmö. 21. Znamenskaya Y, Engblom J, Sotres J, Arnebrant T and Kocherbitov V. Effect of hydration on structural and thermodynamic properties of mucin. 6th Annual Workshop of Biofilms-Research Center for Biointerfaces, Malmö, October 13-15, BSc and MSc education 11.1 BSc-level 1. Biomedical laboratory science (HS) 2. TELMah-Biomedical Technology (TS & HS) 3. Dentistry program (OD) 11.2 MSc-level 1. Biomedical Methods and Technology (HS) 2. Materials science (TS & HS) 3. Dentistry program (OD) 12 Other Activities by Center Members Gunnel Svensäter and Tautgirdas Ruzgas have been opponents at dissertations for doctors degree at Faculty of Health Sciences, Copenhagen University, and at Dept of Physics, Linköping University, respectively. Gunilla Nordin Fredrikson has been opponent at a half-time control (Faculty of Health Sciences, Linköping University). Håkan Eriksson, Bertil Kinnby, Julia Davies, Anette Gjörloff-Wingren, Ann Wennerberg, Liselott Lindh, Tautgirdas Ruzgas, Thomas Arnebrant and Per Såhle have been board members at dissertations at 12 occasions (Lnu, LU, GU, LuTH, Mah) and Julia Davies and Gunnel Svensäter have been external examiners at Leeds Dental Institute, UK. Julia Davies has also been external examiner at Trinity College Dublin, Ireland. Gunilla Nordin Fredrikson, Liselott Lindh, Per Ståhle, Tautgirdas Ruzgas and Thomas Arnebrant have been external reviewers for promotions to professor (2), associate professor (1) and employment as senior lecturer (3). Center members have been referees for international journals on a regular basis. The list of journals include; Acta Biomaterialia, Acta Oncologica, Analytical Chemistry, Angewandte Chemie International Edition, Arteriosclerosis, Thrombosis and Vascular, Biology (ATVB), Atherosclerosis, Biofouling, Biomacromolecules, Biosensors and 37

38 Bioelectronics, Biotechnology Progress, Bioelectrochemistry, Caries research, Chemistry Communications, Clinical Immunology, Colloid and Polymer Science, Colloids and Surfaces B: Biointerfaces, Colloids and Surfaces A, Current Opinion in Colloid and Interface Science, Electroanalysis, Electrochimica Acta, Electrochemical Communications, European Heart Journal, European Journal of Oral Sciences, European Journal of Histochemistry, Future Cardiology, Industrial & Engineering Chemistry Research, International Journal of Biomedical Science (IJBS), Journal of Clinical Pathology, Journal of Proteome Research, Journal of the American Chemical Society (JACS), Journal of Biomedical Materials Research: Part A, Journal of Internal Medicine, Journal of Molecular Signaling, Journal of Physical Chemistry B, Journal of Chemical Physics, Journal of Colloid and Interface Science, Journal of Biomedical Biochemistry (JBB), Journal of Food Engineering, Langmuir, Macromolecules, Scandinavian Journal of Immunology, Sensors and actuators, Talanta, Water Environment Research, JJOM, IJSS, JAM, European Journal of Dental Education, Proteome Science, Journal of Periodontal Research, Thrombosis and Haemostasis, PLoS, Cancer Letters, International Journal of Endodontics, Caries Research, Journal of Medical Microbiology, Microbiology, Archives Oral Biology, Archives of Oral Biology, Journal of Oral Rehabilitation, Journal of Biomedical Materials Research, Biofouling, Journal of Colloid and Interface Science, Microbiology, Journal of Colloids and Surfaces B: Biointerfaces, Biomacromolecules, Journal of Clinical Periodontology, Acta Biomaterialia, Oral Diseases, European Journal of Oral Sciences, Journal of Dental research. Ann Wennerberg is on the editorial board for Acta Biomaterialia, International Journal of Prosthodontics and Journal of Oral Rehabilitation. Tautgirdas Ruzgas is on the editorial board for Nonlinear analysis: Modelling and control, and Per Ståhle was guest editor of International Journal of Fracture. Ann Wennerberg was coordinator, and Gunnel Svensäter was the local coordinator at Mah, of the Swedish National Graduate School in Odontological Science. Svensäter was also member of the scientific advisory board at the Swedish Council on Technology Assessment in Health Care (SBU) and the Research committee at Faculty of Odontology (OD, Mah). Julia Davies was Director of Postgraduate Research Education, delegate on Faculty Working Group for Quality Development, delegate on Faculty Library Working Group and faculty representative on University group for Research Education. Ann Wennerberg was member of the scientific advisory board at the European Association for Osseointegration and board member at Swedish research counsil (NT-A). Anette Gjörloff Wingren was chairman of the Education board at Faculty of Health and society. Tautgirdas Ruzgas was member of the docent board at Malmö University. Johan Engblom was member of the Quality assurance group and the Education board at Faculty of Health and society, and member of the Faculty Board at School of Technology (TS, Mah). Gunilla Nordin Fredrikson was convener of the Evaluation committee of DPLU (Diabetes Program Lund University) and consulted as a judgement expert of Biomedical Scientist educations in Sweden by the National Agency of Higher Education. Thomas Arnebrant was at the faculty board of Health and society, and also assistant Dean. 38

39 Cover story Biofilms annual workshop this year attracted 110 guests from different universities as well as industries, innovation agencies, solicitor s offices and risk capitalists. During the three day workshop a wide range of interesting lectures were held on the topic Biomaterials - From Fundamentals to Market Application. - We are very happy that we this year could combine the annual workshop with the workshop of an EU-project as it allowed us to offer lectures on research results as well as on the commercialization of scientific innovations, says Johan Engblom, Director of Biofilms - Research Center for Biointerfaces - It has been a very interesting workshop! I have got an up date on what is going on, and at the same time I have had the opportunity to meet many interesting people in the area, says Christy Whiddon, AkzoNobel Surface Chemistry AB.. [Malmö University News Letter, ]

40 Center Mission Statement Biofilms Research Center for Biointerfaces is a translational research programme covering six research groups within three faculties/schools (HS, OD & TS) at Malmö University. The core strengths of the Center is i) our broad expertise, spanning the range from theoretical modeling to clinical sciences, and ii) our long experience in working with industry and relating to their needs. The general aim of the research activities at the Center is to understand, predict and control material/cell/tissue interactions with medical, dental, food and environmental applications. We strive to further integrate education (BSc, MSC, PhD), a cornerstone for the future of the Center. Particularly, one goal is to further integrate Master-level education into our research activities also at an operational level in specific projects. Biofilms Research Center for Biointerfaces has a goal to become a regional venue for a creative environment of biomedical technology in the Öresund region, facilitating collaboration between academic research, higher education and industry, based on pharmaceutical technology, biotechnology and medical technology, which together constitutes a key area of significant commercial growth potential. We strive to become a prime entry point for regional industry seeking translational academic expertise in the Biomedical technology field. Center Location Biofilms Research Center for Biointerfaces Malmö University SE MALMÖ, Sweden Center director: Assoc. Prof. Johan Engblom (JE) Tel: +46-(0) (JE); +46-(0) (Adm. Coordinator Eva Nilsson) [email protected] Visiting adress: Skåne University Hospital, SUS (Entrance 49) MALMÖ