biofilms â€“research center for biointerfaces

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 #5

January 1st, 2009 – December 31st, 2009

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 in the associated liquid phase.

Contact information

Biofilms – Research Centre for Biointerfaces Faculty of Health and Society Malmö University SE-205 06 MALMÖ, Sweden Center director: Assoc. Prof. Johan Engblom (JE) Tel: +46-(0)706-08 75 25 (JE); +46-(0)40-66 57 486 (Adm. Coordinator Eva Nilsson) e-mail: [email protected] www.mah.se/biofilms Visiting adress: University Hospital MAS (Entrance 49) MALMÖ

Biofilms –Research Center for Biointerfaces Progress Report 2009

Mah: Malmö University; LU: Lund University; KTH: The Royal Institute of Technology; LuTH: Luleå Technical University; UU: Uppsala University; LiU: Linköping University; KI: Karolinska Institute; OD: Faculty of Odontology; HS: Faculty of Health and Society; CTS: School of Technology. KKs: The Knowledge Foundation; VR: The Swedish Research Council; EU: European Union; FP6: 6th Framework programme; FP7: 7th Framework programme; BMT: Biomedical Technology BMMT: Biomedical Methods and Technology; MS: Materials Science; FLÄK: The Research School in Pharmaceutical Science; BMA: Biomedical Laboratory Science; TELMah: Technology, Economy and Leadership at Mah; QCM-D: Quartz Crystal Microbalance with Dissipation Monitoring; SWAXD: Small and Wide Angle X-ray Diffraction

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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 2008- Håkan Ericsson, Assoc. Prof., Vice Director 2009 Thomas Arnebrant, Prof., Director 2005-2007 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. Vitaly Kocherbitov, Assoc. Prof. Liselott Lindh, Assoc. Prof. Anette Gjörloff-Wingren, Assoc. Prof. Zoltan Blum, Assoc. Prof. Lars Olsson, Assoc. Prof. Liu-Ying Wei, Assoc. Prof. Julia Davies, Assoc. Prof. Bertil Kinnby, Assoc. Prof. Tove Sandberg, Dr. Maria Stollenwerk, Dr. Christina Bjerkén, Dr. Sergey Shleev, Dr.

1.2 Junior researchers and postdocs

Olof Svensson, Dr. Olga Santos, Dr. Tobias Halthur, Dr. Anna Ketelsen, Dr. Laura Varas, Dr. Claes Wickström, Dr. Jessica Neilands, Dr. Luis Chavez de Paz, Dr. Javier Sotres, Dr Ida Svendsen

1.3 PhD students

Jildiz Hamit Eminovski Linda Andersson Peter Hellman Ulf Hejman Alma Masic Maria Pihl Sebastian Björklund (LU) Anton Fagerström Yana Znamenskaya



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Adnan Safdar Christian Alfredsson Kindblom Marjan Dorkhan Kostas Bougas Magnus Falk

1.4 Technical and administrative staff

Eva Nilsson, Administrative coordinator Ulrika Troedsson, Technician Agnethe Henriksson, Technician Madeleine Blomqvist, Technician

1.5 MSc students

BMMT Master at HS

2009-2011

Haddel Ali Shoker Joy Chia Ihab Dahi Payam Delfani Susanna Tarasco Mohammad Zahir Uddin Petra Wicktor

BMMT Master at HS

2008-2010

Aseel Albayati Sheima Sultan Kadir Eleonora Dahlquist Rula Bahran Peter Lamberg Marianne Mårtensson Patrik Bauer

BMMT Master at HS

2007-2009

Noor Al-Attar Fozia Elahi Tannaz Horrieh Charlotte Lerbech Jensen Shamila Khan

MS Master at HS & CTS 2008-2010

Maihemutijiang Maimaiti Carl Mikaelsson Ajigul Nuermaimaiti Oyetunji Oladele Kazeem Wureguli Reheman Christian Ukoha Oji Erik Öberg Simayijiang Zhayida

MS Master at HS & CTS 2007-2009

Adili Maimaiti Guzainuer Maimaitiyiming Huaizhi He Maimaitiyili Tuerdi Hao Qin

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.



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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 Pro-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 2008- Thomas Arnebrant, Prof., Director & Chairman for 2005-2007 Håkan Eriksson, Assoc. Prof. , Vice Director 2009 Per Ståhle von Schwerin, Prof. Gunnel Svensäter, Prof. Ann Wennerberg, Prof. Gunilla Nordin-Fredrikson, Prof.

Steering group

Eva Engquist, Pro Vice-Chancellor, Malmö University, Chairman Margareta Östman, Prof., Dean Faculty of Health and Society Naser Eftekharian, Head School of Technology Malmö University Lars Matsson, Prof., Dean Faculty of Odontology (- June 2009) Lars Bondemark, Prof., Dean Faculty of Odontology (July 2009 -)

Reference group 2009-2011

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 Krister Thuresson, Dr., Director CMC Regulatory Affairs Camurus AB (- October 2009) Markus Johnsson, Dr., Senior Director Pharm. Development Camurus AB (November 2009 -) Eva Engquist, Pro.Vice-Chancellor, Malmö University Zoltan Blum, Assoc. Prof., Malmö University



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2 The Director´s Report During 2009 the Center project portfolio has conatained 34 projects, of which 13 are funded through our center grant from KKs. A major emphasis in 2009, apart from conducting high quality front line research, has been to secure additional funding. The efforts have been fruitful with a 94% increase in external funding compared to 2008. Some of the new projects bring new industry partners into the Center, and some also facilitate our ambition to increase focus on medical applications and to improve involvement of preclinical and clinical sciences. Linda Andersson, Ida Svendsen and Peter Hellman successfully defended their PhD-theses, as did Karolina Haberska and Marie Friberg (enrolled at LU, with supervisors at Mah). Jildiz Hamit Eminovski successfully defended her licentiate thesis. Ida Svendsen got the award for the best PhD-thesis at Malmö University 2009. Her thesis “In vitro and in vivo studies of salivary films at solid/liquid interfaces” is a good example of the potential of interfaculty collaboration within Biofilms-Research Center for Biointerfaces. Associate Professor Birgitta Rosén (Handsurgery, LU) has been part time employed as guest teacher until March 2009. Professor Anders Heyden and Drs Niels-Christian Overgaard and Tobias Halthur left their positions during the spring. Drs Javier Sortres and Olga Santos where employed in September. 10 new PhD-students where enrolled during the year, seven at Malmö University and three at Lund University, with supervision from Malmö. During 2009 we have produced 51 publications in international journals and 6 proceeding papers. Publications are in journals such as Trends in Immunology (9.910), Journal of Controlled Release (5.690), The International Journal of Cancer (4.734), Materials Science and Engineering (2.201) and FEMS Microbiology and Immunology (1.972). The impact factors span from 9.910 to 1.972 indicating a substantial scientific quality of the research carried out within the Center. In addition to this approximately 21 oral and 25 poster presentations were made at national and international meetings. In reviewing the number citations over the past six years on papers produced by the Center members we sum up 6800 for permanent staff and 900 for junior researchers and post-docs. Four members have more than 500 citations each, Profs. Tautgirdas Ruzgas, Gunilla Nordin Fredrikson, Thomas Arnebrant and Dr Sergey Schleev. Center members have also refereed more than 60 papers for international journals. Members of the Center take part in The National Research School of Odontology (Prof. Gunnel Svensäter (coordinator), Prof Ann Wennerberg and Assoc. Prof Julia Davies, supervisors). 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.



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Research collaboration with industry is active in all four focus areas of the Center and during 2009 we attracted Promimic AB, Bioglan AB and Nares AB as new partners. 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) Transport phenomena (AstraZeneca, AkzoNobel, Camurus, Galenica), iii) Molecular interactions at biointerfaces (Camurus, SinclairPharma, EuroDiagnostica, Promimic, TetraPak, Stora Enso, Anordica, Arcam, Bioglan, Nares) and iv) Microbial biofilms (ArlaFoods, AnoxKaldnes, SinclairPharma, Gambro, Arcam). We also have close collaboration with Medeon AB and Medicon Valley. Center members involve in the collaborative EU FP7 funded project “Three-dimensional nanobiostructure-based self-contained devices for biomedical application” (Dr Sergey Shleev, coordinator) and continue to be involved in the EU FP6 Marie Curie Research Training Networks (MCRTNs), “Bio-interfaces: from molecular understanding to applications”. We also involve in the EU South Baltic program “VALOR – Valorisation of knowledge intensive ideas in the South Baltic area”. A new QCM-D was purchased during the year, a valuable complement for our activities regarding thin films formation. Through a generous donation from AstraZeneca we received an X-ray diffraction camera (Kratky SWAXD) and two additional HPLC-UV set-ups, most useful in both research and education activities Center members have been responsible for arranging two workshops, “Microarrays” (Biofilms 5th Annual workshop) and “Oral Biofilm and Risk” (National Research School of Odontology), and one PhD-student day with invited mentors. The students now also meet on private initiatives, which is very encouriaging from a cross faculty collaboration perspective. Furthermore, kick-off meetings were arranged in Malmö within the two new EU-projects. Together with Medicon Valley Alliance and Trial Form Support AB we arranged a meeting to present a new education program to industry partners. 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 two press releases, Rapidus Newsletter and Malmö University newsletters.



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3 List of Research Activities The research activities of the Center during the period of report 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 also involve in the EU South Baltic program “VALOR – Valorisation of knowledge intensive ideas in the South Baltic area”.

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)

3.2 Molecular transport phenomena

4. Hydration. Interactions between pharmaceutical materials and water (AstraZeneca, funded by KKs-Biofilms)

5. Adjuvants for products used in agriculture (AkzoNobel Surface Chemistry AB, funded by KKs-Biofilms)

6. Stress driven diffusion (pilot project funded by Mah ) 7. Water – a crucial factor in regulating biomembrane permeability (Physical

Chemistry 1, LU and Camurus AB, funded by FLÄK, LU) 8. Miniature biofuel cells for self-contained bio-devices: electron transfer in three-

dimensional nanobiostructures (funded by VR & EU) 9. Grinfeld surface instabilities (funded by VR) 10. Stress corrosion (funded by VR) 11. Set up and optimisation of an in vitro mucosa permeation model (Food

technology, LU, pilot project)

3.3 Molecular interactions at biointerfaces

12. Mucoadhesion: Drug carrier interactions at biologically relevant interfaces (Camurus AB, funded by KKs-Biofilms)

13. Antiplaque 1(2): Adsorption and biofilm formation at oral interfaces (SinclairPharma AB, funded by KKs-Biofilms)

14. Bioassay 1(2): New concept for lipid-based surface coatings in bioassays (EuroDiagnostica AB, funded by KKs-Biofilms)

15. Bioassay 2(2): New concept for lipid-based surface coatings in bioassays (EuroDiagnostica AB, funded by KKs-Biofilms)

16. Biomaterial 2(2), The influence of biochemical coat for implant bone incorporation, The in vitro part (Promimic AB, funded by KKs-Biofilms)



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17. Symptomatic vs. asymptomatic atherosclerotic plaques (CRC, LU, funded in part

by Mah) 18. Fracture of biofibre-, polymer- and metal composites (TetraPak AB & Stora Enso

AB, funded by KKs) 19. Biocompatibility of metals (Anordica AB & Arcam AB, funded by KKs) 20. Hydration of mucous gel (funded by Mah) 21. Topical microemulsion as nasal drug delivery system (Bioglan AB & Nares AB,

pilot project)

3.4 Microbial biofilms

22. Milk Protein: Investigation of interactions between osteopontin and oral biofilm bacteria (Arla Foods AB, funded KKs-Biofilms)

23. Carrier: Investigation and modelling of convection in biofilms for different carriers (AnoxKaldnes AB, funded KKs-Biofilms)

24. Antiplaque 2(2): The effect of surface coatings on bacterial biofilm formation (SinclairPharma AB, funded KKs-Biofilms)

25. Biofilm formation on Peritoneal Dialysis catheters (Gambro Lundia AB, funded by KKs-Biofilms)

26. Biologically induced stress corrosion crack growth (Arcam AB, funded by KKs-Biofilms)

27. Caries prevention with fluoridated probiotic milk – a prospective clinical and microbiological study of root caries (funded by Swedish Patent Revenue Foundation)

28. Survival strategies of bacteria in oral biofilms (funded by Swedish Research Council)

29. Mucins and microbial biofilms – a symbiotic relationship for health (funded by Mah and Crafoord Foundation)

30. Mucosal interactions as inducers of acid tolerance in oral microorganisms (funded by Crafoord Foundation and Swedish Dental Society)

31. Biofilm activity as a marker to identify patients at risk of caries – mechanisms underlying microbial stress tolerance (funded by Mah)

32. Activities of microbial biofilms on bioactive implant surfacrs (funded by Mah) 33. The plasminogen activating system – interaction with microorganisms and a

potential risk marker (funded by Crafoord foundation and Swedish Dental Society)



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4 Research highlights

4.1 A water gradient can be used to regulate drug transport across skin

At normal conditions there is a substantial water gradient over the skin as it separates the water-rich inside of the body from the dry outside. This leads to a variation in the degree of hydration from the inside to the outside of skin and changes in this gradient may affect the structure and function of skin. Sebastian Björklund et al have shown that a water gradient can be used to regulate transport of drugs with different lipophilic characteristics across the skin barrier. The transport of metronidazole (log Po/w=0.0) and methyl salicylate (log Po/w=2.5) across skin increases abruptly at low water gradients, corresponding to high degrees of skin hydration, and that this effect is reversible.This phenomenon is highly relevant to drug delivery applications. Further, the results contribute to the understanding of the occlusion effect and indicate the boundary conditions of the water gradient needed to make use of this effect. Björklund S, Engblom J, Thuresson K and Sparr E, A water gradient can be used to regulate drug transport across skin, J Control Release, in press

Figure 4.1. Effect of a water gradient on metronidazole permeability in vitro over corresponding excised skin (left) and silicone membrane (right)

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4.2 Adsorption and elution of salivary films from solid surfaces

The salivary pellicle is formed by selective adsorption of salivary proteins on tooth enamel. It has lubricating and protective functions, which may be altered during plaque (biofilm) control treatments. In order to develop products aimed at controlling e.g. plaque growth or to develop new or improved salivary substitutes it is important to get a better understanding of protein behaviour at the interface. We have therefore investigated the interfacial properties of human salivary films formed on silica and hydroxyapatite (HA) surfaces in different adsorption media (PBS and water) by the use of quartz crystal microbalance with dissipation (QCM-D) and ellipsometry. We observed that saliva adsorbs at a higher extent onto the HA surfaces compared to the silica surfaces, regardless of the adsorption media. For both surfaces a higher adsorption of saliva was obtained when the adsorption media was PBS compared to water. Since Sodium dodecyl sulphate (SDS) is a common surfactant used in many types of oral care products we have also investigated how SDS solutions interact with the saliva film at different SDS concentrations. We observed that SDS was able to remove the saliva film completely from the silica surfaces but only partially from the HA surfaces. As expected, the highest pellicle removal from both surfaces was observed at the concentration corresponding to the cmc. Our results also implied that a softer saliva layer was build up in PBS for both surfaces. In water a softer saliva layer was formed onto the HA surfaces, while in PBS the saliva layer formed at both surfaces seemed to have similar structure. Olga Santos, Liselott Lindh, Tobias Halthur, and Thomas Arnebrant, Adsorption behaviour and elution of saliva from silica and hydroxyapatite surfaces with SDS and delmopinol studied by quartz crystal microbalance. Manuscript.

Figure 4.2. Adsorption of saliva pellicle on silica and HA surfaces in PBS and elution with SDS

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4.3 Bioelectrocatalytic interfaces: Redox Enzymes Electronically Connected at Three-Dimensional Materials for Extracting Electrical Energy from Biofuels

Bioelectrochemical devices, such as biosensors, have proven to be useful in different areas of applications including biomedicine. Current research in bioelectrochemistry is massively focused on development of biofuel cells and defining principals of biocomputing devices. During the last year at our department we have focused on the studies of bioelectrocatalytic function of enzymes in three-dimensional (3D) conducting materials with the purpose to increase the power density of biofuel cells. Two major approaches were pursued, i.e. layer by layer deposition of bioelectrocatalytic enzyme –nanoparticle conjugates [1] and making conducting 3D nanostructures from silicon [2]. Direct and mediated electron transfer reactions between different enzymes, viz. blue multicopper oxidases (laccase [2] and ceruloplasmin [3]) and cellobiose dehydrogenase [4] on planar and 3D electrodes were compared to understand the mechanism of functioning of both cathodic (oxygen reducing enzymes) and anodic (glucose oxidizing enzymes) bioelements. As a final outcome a biofuel cell operating in human serum (see figure below) was constructed. The cell exploits the principal of direct electron transfer for bioelectrocatalysis on both anode and cathode. As a consequence of this the biofuel cell is simple and does not contain leaking and toxic redox mediators [5]. 1. Haberska K. Protein and polyelectrolyte layer-by-layer films: Assembly and electron transfer. Doctoral dissertation, Lund University, Lund, 2009, pp 116. 2. Ressine A., Vaz-Dominguez C., Fernandez V.M., De Lacey A.L., 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, 2010, 25(5), 1001-1007 3. Haberska K., Vaz-Domínguez C., De Lacey A.L., Dagys M., Reimann C.T., Shleev S. Direct electron transfer reactions between human ceruloplasmin and electrodes. Bioelectrochemistry, 2009, 76(1-2), 34-41. 4. Stoica L., Ruzgas T., Gorton L. Electrochemical evidence of self-substrate inhibition as functions regulation for cellobiose dehydrogenase from Phanerochaete chrysosporium. Bioelectrochemistry, 2009, 76(1-2), 42-52. 5. 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, 2009, DOI: 10.1002/fuce200900121.

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2 H2O

cellobiose dehydrogenaseblue multicopper oxidase

Ired Iox

+

0.17-0.5 V

IredIox



11

4.4 Hydration of proteins: water sorption isotherm of lysozyme

Investigations the hydration of proteins is an important part of protein science. The hydration of proteins can be studied by two different approaches. In one approach, protein-water interactions are studied in dilute solutions of proteins in water. In the other approach, changes in structural, dynamic, and thermodynamic properties of proteins are monitored as functions of water content at relatively low hydration levels. Water sorption isotherms of proteins are usually interpreted with such models as BET or GAB that imply the formation of multilayers at solid-gas interface. However, this approach is not applicable to globular proteins such as humid lysozyme where a solid-gas interface does not exist. We proposed to interpret the water sorption isotherms of proteins considering adsorption of water at protein-protein interface. Thermodynamic analysis based on experimental data on enthalpy and entropy of hydration of lysozyme showed that the Langmuir sorption isotherm can be used to describe hydration of protein-protein interface. The result of the fitting of the sorption isotherm of lysozyme with the Langmuir model shows good agreement with the structural data on lysozyme. Hydration of lysozyme: the protein-protein interface and the enthalpy-entropy compensation. Langmuir, 2010. DOI: 10.1021/la903210e

Figure 4.4. Schematic structures of lysozyme in solution (left) and at a low hydration level (right). The structure of lysozyme in solution is from the protein database (193 L); water molecules are not shown. The low-hydration-level structure is for illustration purposes only and does not correspond to any experimentally obtained structure



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4.5 Effect of Hydration on Structural Properties of Mucous Gel

The mucus barrier and its transport properties are essential for proper functioning of the digestive, respiratory and reproductive systems of vertebrates, including humans. From an engineering point of view, mucus is an outstanding water-based lubricant. The principal components of mucus are the glycoprotein mucin and water. Mucin forms the macromolecular matrix of mucus and dominates its rheological properties.

Mucin molecules obtained from different sources have different structures ranging from bottle brush to dumbbell type structures. It is known from literature that mucin can form liquid crystalline phases but exact phase behaviour of mucin at different temperatures and hydration levels has yet to be determined. The phase behaviour of a particular type of mucin is dependent on its molecular structure.

In this work we visualised the molecular structure of porcine gastric mucin (PGM) in dehydrated state using the atomic force microscopy (AFM). Samples deposited on mica were studied in air by AFM operated in the tapping mode. Atomic force microscopy indicates the presence of a dumbbell structure (b, c, d) as well as fiberlike structure (a) at higher concentrations. Znamenskaya Y, Engblom J, Sotres J, Arnebrant T and Kocherbitov K. (To be published)

Figure 4.5. Images of mucin structures obtained after evaporation of solutions. Initial mucin concentrations are: a) 10

-3 %; b) 10

-5 %; c) 10

-5 %; d) 10

-5 %, 3-D image of dumbbell mucin

molecule

a) b)

c) d)



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4.6 Differential effects of Pseudomonas aeruginosa on biofilm formation by fresh isolates and laboratory strains of Staphylococcus epidermidis

Natural biofilms, such as plaque on teeth and the biofilms in the intestines consists of hundreds of different species. However, biofilms on medical implants are deprived of the rich flora and usually consist only of one or a few species. Pseudomonas aeruginosa and Staphylococcus epidermidis are common species found in infections related to medical implants. Studying dual species biofilms of P. aeruginosa and S. epidermidis, using 16S rRNA FISH probes and confocal microscopy, we showed presence of P. aeruginosa to reduce biofilm formation of S. epidermidis. This inhibition was due to extracellular products of P. aeruginosa, possibly polysaccharides. Interestingly, different strains of S.

epidermidis varied in their ability to withstand the inhibition of P. aeruginosa, with fresh isolates far exceeding laboratory strains. Maria Pihl, Julia R. Davies, Luis E. Chávez de Paz, Gunnel Svensäter. Differential effects of Pseudomonas aeruginosa on biofilm formation by fresh isolates and laboratory strains of Staphylococcus epidermidis. Submitted to FEMS Immunol Microbiol

5 µm Figure 4.6. The figure show 9 hour biofilms of two different S. epidermidis strains (Mia and C103) alone (a, c) and in dual-species biofilms (b, d) with P. aeruginosa NCTC 6750, clearly showing P. aeruginosa to be able to reduce biofilm formation of S. epidermidis.



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4.7 The effects of antimicrobials in endodontic biofilm bacteria

In the present study, confocal microscopy, a mini flow-cell system and image analysis were combined to test in situ the effect of antimicrobials and alkali on biofilms of Enterococcus

faecalis, Lactobacillus paracasei, Streptococcus anginosus and Streptococcus gordonii isolated from root canals with persistent infections. Biofilms formed for 24h were exposed for 5 min to alkali (pH 12), chlorhexidine digluconate (2.5%), EDTA (50mM) and sodium hypochlorite (1%). The biofilms were then characterized using fluorescent markers targeting cell membrane integrity (LIVE/DEAD) and metabolic activity (CTC and FDA). In addition, the biofilm architecture and the extent to which coating of the substrate surface with collagen influenced the resistance pattern to the chemicals were also analyzed. NaOCl (1%) affected membrane integrity of all organisms and removed most biofilm cells. Exposure to EDTA (50mM) affected the membrane integrity in all organisms, but failed to remove more than a few cells in biofilms of E. faecalis, L. paracasei and S. anginosus. Chlorhexidine (2.5%) had a mild effect on the membrane integrity of E. faecalis and removed only 50% of its biofilm cells The effects were substratum-dependent and most organisms displayed increased resistance to the antimicrobials on collagen-coated surfaces. The biofilm system developed here was sensitive and differences in cell membrane integrity and removal of biofilm cells after exposure to antimicrobials commonly used in endodontics was discernable. Chávez de Paz LE, Bergenholtz G, Svensäter G. The effects of antimicrobials on endodontic biofilm bacteria.J Endod 2010;36:70-7.

Figure 4.7. Three-dimensional reconstructions from confocal micrographs showing the effect of chlorhexidine and alkali on biofilms of Enterococcus faecalis and S. gordonii uing LIVE/DEAD staining. The green cells represent cells with intact membranes, wheras the red cells are damaged or dead. Images were constructed using the software bioImage_L.



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4.8 A novel method to detect gene expressions in biofilms

Microbial biofilms constitute a major medical problem when formed on human tissues due to their high resistance to an unlimited number of environmental stress including antibiotic and biocide treatments. This decreased susceptibility of biofilm-forming bacteria to host defenses and antibiotic treatments is related to a heterogeneous distribution of sub-populations of micro-organisms with altered patterns of gene expression. This biological heterogeneity in biofilm populations need to be studied by techniques which do not disrupt the natural distribution of biofilm cells. We have developed a fluorescense in situ hybridization (FISH) method to detect gene expression in situ at early stages of biofilm formation. In this protocol we use multiple probes targeting mRNA sequences in mono and dual species cultures. As observed in Figure 4.8, probes targeting the gene expression of serine protease challisin in Streptococcus

gordonii (yellow signals) were simultaneously visualized with 16S rRNA probes that target Streptococcus gordonii (red cells) and Lactobacillus salivarius (blue cells). By using this combined protocol we could successfully monitored in situ gene expression in microbial biofilms. Luis Chávez de Paz, Julia Davies, Gunnel Svensäter (To be published)

Figure 4.8. Probes targeting the gene expression of serine protease challisin in Streptococcus gordonii (yellow signals) were simultaneously visualized with 16S rRNA probes that target Streptococcus gordonii (red cells) and Lactobacillus salivarius (blue cells).



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4.9 Effect of fluoride and Lactobacillus rhamnosus on plaque acid tolerance

Fluoride has long been used in preventive strategies against dental caries due to its hydroxyapatite incorporation that lower the solubility of the enamel and dentine. From in

vitro-studies it is known that fluoride also affects the physiology of oral bacteria making them acid sensitive but data on the effect of acid tolerance in plaque in vivo is limited. The aim of the present investigation was to study the effect of fluoride and probiotic strain Lactobacillus rhamnosus LB21 on plaque acid tolerance and lactic acid production in vivo. Subjects participating in this study were instructed to drink milk containing fluoride or/and L. rhamnosus on a daily basis for 15 months. Plaque samples were taken at baseline and after 15 months. Plaque acid tolerance was measured using LIVE/DEAD® BacLightTM Viability Stain after exposure to pH 3.5 for 2 hours. The results showed that dental plaque from subjects that had ingested milk containing fluoride or fluoride in combination with L. rhamnosus exhibited a significant decrease in acid tolerance after 15 months compared baseline (see Figure 4.9). Subjects that had been drinking milk without fluoride or with lactobacilli only did not show any decrease in acid tolerance. Lactic acid production from glucose was also lower in the groups that had been exposed to fluoride. Figure 4.9. Examples on plaque acid tolerance in two different subjects visualized using LIVE/DEAD® BacLight

TM Viability stain after the exposure to pH 3.5 fro two hours.

Conclusively, this pilot study shows that fluoride affects plaque acid tolerance and lactic acid production in vivo. L. rhamnosus on the other hand does not seem to affect plaque acid tolerance or lactic acid production in either direction. Welin-Neilands J, G. Svensäter G To be presented at the IADR congress in Barcelona, 2010

DECREASED PLAQUE ACID TOLERANCE

INCREASED PLAQUE ACID TOLERANCE

BASELINE AFTER 15 MONTHS



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4.10 Mucin-bacterial interactions – degradation of MUC5B by dental biofilms

Members of the indigenous oral microbiota must interact to degrade and utilize complex salivary components in order to survive and grow in the oral cavity. We have shown for the first time, that a naturally occurring substrate, MUC5B, is proteolytically digested preferentially by cooperation of several members of the dental plaque microbiota. Interspecies cooperation appears to overcome the high level of molecular complexity in MUC5B and makes the digestion products available as a nutrient source. Endopeptidase activities in conjunction with full or partial deglycosylation, may be the first step in the degradation of salivary proteins by the oral biofilm. This first step in degradation would increase the availability and number of N- and C-termini to serve as substrates for aminopeptidases, which liberate amino acids and short peptides to support bacterial growth. Mono-cultures was not able to degrade the protein backbone of the MUC5B mucin (a), but a consortia of four species efficiently degraded the protein (b). When incubated with pooled dental plaque, MUC5B was degraded less efficiently than with the four-species consortium (c). This illustrates the diversified functions of the polymicrobial species in dental plaque and that a balance is established between the multispecies biofilm and host glycoproteins, that is beneficial to oral health. Wickström, C., Herzberg, M. C., Beighton, D. & Svensäter, G. Proteolytic degradation of human salivary MUC5B by dental biofilms (2009) Microbiology 155, 2866-2872.

Figure 4.10. SDS/PAGE – Western blot showing degradation of the MUC5B polypeptide backbone. MUC5B was incubated with (a) Streptococcus gordonii (b) a four-species consortium containing Streptococcus gordonii, Steptococcus biovar 2, Streptococcus cristatus and Actinomyces naeslundii and (c) supragingival dental plaque.



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4.11 Antibody binding kinetics in ELISA

The purpose of an ELISA (Enzyme Linked Immunosorbent Assay) is usually to detect an antigen or antibody in a biological sample. In the detection of a specific antibody a surface pre-coated with the antigen is exposed to the sample. The binding is then detected by a second antibody that is covalently linked to an enzyme. The final step in the assay is to determine the enzymatic activity, which is a measure of the amount of the specific antibody in the sample. In this respect an ELISA can be regarded as a black box, where only the end result is obtained. Therefore, the objective of the study was to elucidate the possibilities to follow the binding kinetics with ellipsometry. This is an optical technique that does not require labeling of the molecules involved, which can be used to follow the mass and thickness of thin films on solid substrates in situ. The binding study was performed on commercial surfaces used for ELISA and the experiment followed the standard procedure that is used in this assay. In the figure below the thickness of the protein layer is presented during the complete ELISA procedure. The addition of the specific antibody did not result in an increase in the layer thickness. However, after the addition of the secondary antibody a clear increase was obtained proving the existence the specific antibody on the surface. Information on the binding kinetics may be useful to improve existing assays for specific antibodies with respect to incubation times and concentrations. Also, it can provide information on the role of other components such as blocking proteins and detergents that are used in these assays.

0

5

10

15

20

25

30

35

40

0 50 100 150 200 250

Th

ickn

ess

(n

m)

Time (min)

Specific antibody Secondary conjugated antibody

Antigen

Figure 4.11. Thickness of the protein layer versus time for an ELISA used to detect a specific antibody. Washing steps (not indicated) preceded the additions of the specific and secondary antibodies.



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5 PhD Theses Supervised by Center Members 1. Olof Svensson, Malmö University (Supervisors Thomas Arnebrant and Krister

Thuresson (Camurus AB)): “Interactions of mucins with biopolymers and drug delivery particles” (2003-2008)

2. Jessica Neilands, Malmö University (Supervisors: Gunnel Svensäter and Thomas Arnebrant ) “Acid tolerance of streptococcus mutans biofilms” (2003-2007)

3. Christer Spegél, Malmö University (Supervisors: Tautgirdas Ruzgas and Rafael Taboryski (Sophion Bioscience A/S)) “Electrochemical monitoring of living cells” (2003-2007)

4. Erik Alpkvist, Malmö University (Supervisors: Anders Heyden and Niels Overgaard) “Mathematical Modeling of Biofilms: Theory, Numerics and Applications” (2002-2006)

5. Ida Svendsen, Malmö University (Supervisors Thomas Arnebrant and Liselott Lindh):"In vitro and in vivo studies of salivary films at solid/liquid interfaces" (2003-2009)

6. Karolina Haberska, Lund University (Supervisor Tautgirdas Ruzgas): " Protein and electrolyte layer-by layer films: Assembly and electron transfer” (2005-2009)

7. Linda Andersson, Malmö University (Supervisor Håkan Eriksson): “Endocytosis by human dendritic cells” (2003-2009)

8. Peter Hellman, Malmö University (Supervisor Håkan Eriksson): “Human dendritic cells- A study of early events during pathogen recognition and antigen endocytosis” (2003-2009)

9. Jildiz Hamit, 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)

10. Maria Pihl, Malmö University (Supervisors Gunnel Svensäter, Bertil Kinnby, Thomas Arnebrant) “Biofilms on Peritoneal Dialysis Catheters” (2005-)

11. Ulf Hejman, Malmö University (Supervisor Per Ståhle, co-supervisor Christina Bjerkén): "Biologivcally induced stress corrosion" (2005-)

12. Alma Masic, Lunds University (Supervisor Anders Heyden, co-supervisor Niels Chr. Overgaard) "Mathematical modeling of biofilms" (2007-)

13. 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-)

14. Adnan Safdar, Lunds University (Supervisor Liu-Ying Wei, co-supervisor Per Ståhle) "Biocompatibility of ion beam melted materials" (2008-)

15. Anton Fagerström, Malmö University (Supervisors Johan Engblom, Vitaly Kocherbitov and Karin Bergström (AkzoNobel)): “Bioavailability of active ingredients used in agriculture” (2009-)

16. Yana Znamenskaya, Malmö University (Supervisors Vitaly Kocherbitov and Johan Engblom): “Hydration of mucous gel” (2009-)



20

17. 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-)

18. Marjan Dorkhan, Malmö University (Supervisors Julia Davies, Gunnel Svensäter, Ann Wennerberg): “Activities of microbial biofilms on bioactive implant surfaces” (2009-)

19. Kostas Bougas, Malmö University (Supervisors Ann Wennerberg, Pentti Tengvall GU), Victoria Franke Stenport (GU)): “Protein coat and bone incorporation” (2009-)

20. Lory Melin, Malmö University (Supervisors Ann Wennerberg, Martin Andersson (Promimic AB): “On the importance of nanometer structures for implant incorporation in bone tissue” (2009-)

21. Magnus Falk, Malmö University (Supervisors Tautgirdas Ruzgas, Sergey Shleev): “Three-dimensional nanobiostructure-based self-contained devices for biomedical application” (2009-)

22. Rickard Hägglund, LTH (Supervisors P Ståhle, P Isaksson): “Damage of paper materials” (2009-)

23. Jon Lind, LTH (Supervisors A Massih, C Bjerkén): “Methods for evaluation of evaluation of the hydride embrittlement of Ni-based super alloys” (2009-)

24. Tuerdi Maymaytilli, LTH (Supervisors C Bjerkén, P Ståhle): Influence of plastic deformation on the formation and growth of embritteling metal hydride's" (2009-)

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. AstraZeneca R&D Lund* 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. Sinclair Pharma AB* 29. Stora Enso AB 30. Studsvik Nuclear AB 31. TetraPak AB 32. Volvo Aero AB 33. YKI AB



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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. Professor Gunnar Dahlén, Microbiology/Endodontics University of Gothenburgh 5. Prof. Lo Gorton, Biochemistry, Lund University 6. Prof. Christer Hansson, Dermatology, Lund University 7. Prof. Göran Lundborg, Hand Surgery, Malmö, Lund University 8. Prof. Martin Malmsten, Pharmacy, Uppsala University 9. Prof. Jan Nilsson, CRC UMAS, Lund University 10. Prof. Tommy Nylander, Physical Chemistry 1, Lund University 11. Prof. Adrian Rennie, Physics, Uppsala University 12. Prof. Mark Rutland, The Royal Institute of Technology (KTH) and Surface Chemistry Institute

(YKI), Stockholm 13. Prof. Olle Söderman, Physical Chemistry 1, Lund University 14. Prof. Per Uvdal, MAX lab, Lund University 15. Prof. Martin Andersson, Dept Applied Chemistry, Chalmers university of Technology. 16. Prof. Pentti Tengvall, Dept Biomaterials, Sahlgrenska Academy, Göteborg University 17. Prof. Tomas Albrektsson, Dept Biomaterials, Sahlgrenska Academy, Göteborg University 18. Prof Marie Wahlgren, Food Technology, LTH, Lund University 19. Prof. Ingegerd Johansson, Cariology, Umeå University 20. Assoc. Prof. Birgitta Rosén, Hand Surgery, Malmö, Lund University 21. Assoc. Prof. Lars Norlén, Cell- and Molecular biology, Karolinska Institute 22. Assoc. Prof. Viveka Alfredsson, Physical Chemistry 1, Lund University 23. Assoc. Prof. Ola Bergendorff, Dermatology, Lund University 24. Assoc. Prof. Emma Sparr, Physical Chemistry 1, Lund University 25. Assoc.Prof. Peter Siesjö, Department of Clinical Sciences, BMC, Lund University 26. Assoc. Prof.Eva Blomberg, The Royal Institute of Technology (KTH) and Surface Chemistry

Institute (YKI), Stockholm 27. Associate Professor Victoria Franke-Stenport, Dept Prosthodontics, Sahlgrenska Academy,

Göteborg University 28. Dr Yngve Cerenius, MAX lab, Lund University 29. Dr Adam Feiler, The Royal Institute of Technology (KTH) and Surface Chemistry Institute

(YKI), Stockholm 30. Dr Anders Björkman, Hand Surgery, Malmö, Lund University 31. Dr Isabel Goncalves, CRC UMAS, Lund University 32. Dr Lars G Petersson, Specialist Clinic for Oral Health Care, Hallands Läns Landsting, Halmstad 33. Dr. Justas Barauskas, Institute of Biochemistry, Vilnius, Lithuania 34. Dr Robert Corkery, The Royal Institute of Technology (KTH) and Surface Chemistry Institute

(YKI), Stockholm 35. Dr Anna Westerlund, Odontology, Gothenburg University 36. Dr. Ivan Maximov, Solid State Physics, Lund University 37. Dr. Johan Drott, Lund University Innovation, Lund University 38. Dr Valera Veryazov, Theoretical Chemistry, Lund University



22

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, Singapore National University, Singapore 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. S. Surresh, MIT, Cambridge, Materials Science, USA 10. Prof. Bent Sörensen, Denmark Technical University, Riso Labs, Denmark 11. Prof. R. K. Thomas, Oxford University, Dept of Physical Chemistry, and ISIS at RAL (Neutron

reflection facility), Oxford 12. Prof. Regine Willumeit, GKSS Research Centre, Geesthacht, Germany 13. Prof. Mark Hertzberg, University of Minnesota, Department of Microbiology, Minneapolis, USA 14. Prof. A. Needleman, Brown university, Providence, USA 15. Prof. Jukka Meurman, Helsinki University Central Hospital, Dept of Oral and Maxillofacial

Diseases, Helsinki, Finland 16. Prof. Christopher Exley, Keele University, UK 17. Prof. Takashi 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. Assoc. Prof. Dennis Cvitkovitch, University of Toronto, Dept of Microbiology, Toronto, Canada 23. Assoc. Prof. Jeannine Brady, University of Florida, Dept of Oral Biology, Gainesville, USA 24. Assoc. Prof. Marie Ranson, School of Biological Sciences, Scientific Director – Cancer, Illawarra

Health and Medical Research Institute, University of Wollongong, Wollongong, Australia 25. Assoc. Prof. Kamal Mustafa, Bergen University, Bergen, Norway. 26. Assoc. Prof. Duncan Sutherland, Aarhus University, Denmark 27. Dr Michael Ortize, Caltech, USA 28. Dr Andrey Jivkov, Manchester University, UMIST, UK 29. Dr. Isaac Klapper, Montana State University , Center of Biofilms Research, Montana, USA 30. Dr Juozas Kulys, Inst. of Biochemistry, Vilnius, Lithuania 31. Dr. Anne Meyer, University of Buffalo, Industry/University Cooperative Research Center for

Biosurfaces, Buffalo, USA 32. Dr Srikumar Banerjee, Indian Atomic Research Centre, India 33. Dr. G. Fragneto, ILL (Neutron reflection facility), Grenoble 34. Dr Adam Heller, University of Texas at Austin, TX, USA 35. Dr Kenneth Holmberg, Tekniska högskolan i Helsinki, Finland 36. Dr Rafael Taborisky, Denmark Technical University, Riso Labs, Denmark 37. Dr Alexander Yaropolov, Inst. of Biochemistry, Moscow 38. Dr. Sergei Lobov, School of Biological Sciences, University of Wollongong, Wollongong,

Australia 39. Dr. Miguel Alcalde, Consejo Superior de Investigaciones Científicas, Applied Biocatalysis group,

Spain 40. Dr. Antonio L. De Lacey, , Consejo Superior de Investigaciones Científicas, Bioelectrocatalysis

group, Spain 41. Dr. Donal Leech, National University of Ireland, Galway, Ireland



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7 List of Publications of the Center from 2009 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 2005-2008 can be found at http://www.mah.se/biofilms.

7.1 Journal articles and invited review papers in journals

1. Chávez de Paz LE, Bergenholtz G, Svensäter G. The effects of antimicrobials on endodontic biofilm bacteria.J Endod 36 (2010) 70-7.

2. Chavez de Paz LE. Image analysis software based on color segmentation for characterization of viability and physiological activity of biofilms. Appl Environ Microbiol 75 (2009) 1734-9.

3. Davies J, Svensäter G, Herzberg MC. Identification of novel surface proteins from Streptococcus gordonii involved in adhesion to oral surfaces. Microbiology 155 (2009) 1977–88.

4. Wickström C, Herzberg MC, Beighton D, Svensäter G. Proteolytic degradation of human salivary MUC5B by dental biofilms. Microbiology 155 (2009) 2866-72.

5. Wickström C, Hamilton I, Svensäter G. Differential metabolic activity by dental plaque bacteria in association with two preparations of MUC5B mucins in solution and in biofilms. Microbiology 155 (2009) 53-60.

6. Holmberg K, Ronkainen H, Laukkanen A, Wallin K, Hogmark S, Jacobson S, Wiklund S, Wiklund U, Souza RM, Ståhle P. Residual stresses in TiN, DLC and MoS2 coated surfaces with regard to their tribological fracture behaviour Journal of Wear (2009) 267

7. Singh RN, Ståhle P, Chakravartty JK, Shmakov AA. Threshold stress intensity factor for delayed hydride cracking in Zr-2.5%Nb pressure tube alloy. Materials Science and Engineering a-Structural Materials Properties Microstructure and Processing, 523 (2009) 112-117

8. Massih AR, Diffusion-controlled phase growth on dislocations, Philosophical Magazine, 89:33 (2009) 3075-3086

9. Massih AR, Jernkvist LO, Stress orientation of second-phase in alloys: Hydrides in zirconium alloys, Computational Materials Science, 46:4 (2009) 1091-1097

10. Massih AR, Jernkvist LO, Transformation kinetics of alloys under non-isothermal conditions, Modelling and Simulation in Materials Science and Engineering, 17:5 (2009)

11. Massih AR, Transformation kinetics of zirconium alloys under non-isothermal conditions, Journal of Nuclear Materials, 384:3 (2009) 330-335

12. Hellman P, Andersson L, Eriksson H. Ligand surface density is important for efficient capture of immunoglobulin and phosphatidylcholine coated particles by human peripheral dendritic cells. Cellular Immunology, 258 (2009) 123-130

13. Gauffin F, Diffner E, Gustafsson B, Nordgren E, Gjörloff Wingren A, Sander B, Liao Persson J Gustafsson B. Expression of PTEN and SHP1, investigated from tissue micro arrays in pediatric acute lymphoblastic leukaemia. Pediatric Hematology and Oncology, 26 (2009) 48-56

14. Göransson A, Arvidsson A, Currie F, Franke-Stenport V, Kjellin P, Mustafa K, Sul T, Wennerberg A. An in vitro comparison of possibly bioactive titanium implant surfaces. J Biomed Mater Res A 15:88(4) (2009)1037-1047

15. Eliasson A, Blomqvist F, Wennerberg A, Johansson A. A Retrospective Analysis of Early and Delayed Loading of Full-Arch Mandibular Prostheses Using Three Different Implant Systems: Clinical Results with Up to 5 Years of Loading. Clin Implant Rel Res 11:2 (2009) 134-148

16. Wennerberg A, Albrektsson T. Effects of titanium surface topography on bone integration. Review Clin Oral Impl Research 20(suppl 4) (2009) 172-184

17. Wennerberg A, Fröjd V, Olsson M, Nannmark U, Emanuelsson L, Johansson P, Josefsson Y, Kangasniemi I, Peltola T, Tirri Teemu, Pänkäläinen T, Thomsen P. Nanoporous TiO2 thin film on titanium oral implants for enhanced human soft tissue contact. A light and electron microscopy study. Clin Impl Dent Rel Res (2009)



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18. Albrektsson, T, Brunski J, Wennerberg A. A requiem for the periodontal ligament revisited. Invited commentary Int J Prostodontics 22 (2009) 120-122

19. Edblad T, Hoffman M, Hakeberg M, Örtengren U, Milleding P, Wennerberg A. Micro-topography of dental enamel and root cementum. Swed Dent J. 33 (2009) 41-48

20. Wennerberg A, Albrektsson T. Structural influence from Calcium Phosphate coatings and its possible effect on enhanced bone integration. Review Acta Odont Scand 31 (2009) 1-8

21. Dunér P, To F, Alm R, Gonçalves I, Fredrikson GN, Hedblad B, Berglund G, Nilsson J, Bengtsson E. Immune responses against fibronectin modified by lipoprotein oxidation and their association with cardiovascular disease. Journal of Internal Medicine, 265 (2009) 593-603

22. Wigren M, Bengtsson D, Dunér P, Olofsson K, Björkbacka H, Bengtsson E, Fredrikson GN, Nilsson J. Atheroprotective effects of Alum are associated with capture of oxidized LDL antigens and activation of regulatory T cells. Circulation Research, 104:12 (2009) 62-70

23. Goncalves I, Cherfan P, Söderberg I, Fredrikson GN and Jonasson L. Effects of simvastatin on circulating levels of autoantibodies to oxidized LDL antigens – relationships with immune stimulation markers. Autoimmunity, 42(3) (2009) 203-208

24. Fredrikson GN, Anand DV, Alm R, Shah PK, Lahiri A and Nilsson J. Associations between autoantibodies against apo B-100 peptides and vascular complications in patients with type 2 diabetes. Diabetologia, Jul;52:7 (2009) 1426-33

25. Goncalves I, Nitulescu M, Ares MPS, Fredrikson GN, Jansson B, Li Z-C and Nilsson J. Identification of the target for therapeutic recombinant anti- apo B-100 peptide antibodies in human atherosclerotic plaques. Atherosclerosis, Jul;205:1 (2009) 96-100

26. Nilsson-Öhman J, Fredrikson GN, Gustavsson C, Nilsson LM, Bengtsson E, Smith M-L, Agardh C-D, Agardh E, Jovinge S, Gomez MF, and Nilsson J. Tumor necrosis factor-� does not mediate diabetes-induced vascular inflammation in mice. Arteriosclerosis Thrombosis and Vascular Biology, Oct;29:10 (2009) 1465-70

27. Santos O, Arnebrant T. Silica supported phospholipid layers doped with GM1. A comparison between different methods, Journal of Colloid and Interface Science, 329 (2009), 213-221

28. Gorbacheva M, Morozova O, Shumakovich G, Streltsov A, Shleev S, Yaropolov A. Enzymatic oxidation of manganese ions catalysed by laccase. Bioorganic Chemistry, 37:1 (2009) 35-49

29. Haberska K, Vaz-Domínguez C, De Lacey AL, Dagys M, Reimann CT, Shleev S. Direct electron transfer reactions between human ceruloplasmin and electrodes. Bioelectrochemistry, 76:1-2 (2009) 34-41

30. Coman V, Vaz-Domínguez C, 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, (2009) DOI: 10.1002/fuce200900121.

31. 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) 1001-1007

32. Stoica L, Ruzgas T, Gorton L. Electrochemical evidence of self-substrate inhibition as functions regulation for cellobiose dehydrogenase from Phanerochaete chrysosporium. BIOELECTROCHEMISTRY, 76:1-2 (2009) 42-52

33. Haberska K, Ruzgas T. Polymer multilayer film formation studied by in situ ellipsometry and electrochemistry, BIOELECTROCHEMISTRY, 76:1-2 (2009) 153-161

34. Heiskanen A, Spegel C, Kostesha N, Lindahl S, Ruzgas T, Emneus J. Mediator-assisted simultaneous probing of cytosolic and mitochondrial redox activity in living cells. ANALYTICAL BIOCHEMISTRY, 384:1 (2009) 11-19

35. Svendsen IE, Lindh L. The composition of enamel salivary films is different from the ones formed on dental materials. Biofouling 25 (2009) 255 – 261

36. Lindh L, Svendsen IE., The composition of the salivary pellicle is substrate specific. Swed. Dent. J., 33 (2009) 218

37. Bjerkén C, Melin S. Growth of a short fatigue crack - A long term simulation using a dislocation technique. International Journal of Solids and Structures, 46 (2009) 1196–1204



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38. Tassidis H, Brokken L, Jirström K, Ehrnström R, Ponté F, Ulmert D, Bjartell A, Härkönen P, Gjörloff Wingren A. Immunohistochemical detection of tyrosine phophatase SHP-1 predicts outcome after radical prostatectomy for localized prostate cancer. The International Journal of Cancer, in press

39. Björklund S, Engblom J, Thuresson K and Sparr E, A water gradient can be used to regulate drug transport across skin, J Control Release, in press

40. Exley C, Siesjö P and Eriksson H. The Immunobiology of Aluminium Adjuvants: How do they really work? Trends in Immunology, in press

41. 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, in press

42. Gonçalves I, Stollenwerk M, Lindholm MW, Dias N, Pedro L, Fernandes e Fernandes J, Moses J, Fredrikson GN, Nilsson J, Ares MPS. AP-1 increased in carotid plaques associated with cerebrovascular symptoms and cholesterol ester content. Cardiovascular Pathology, in press

43. Nilsson LM, Nilsson-Öhman J, Sigvardsson M, González Bosc LV, Smith M-L, Agardh E, Fredrikson GN, Agardh C-D, Nilsson J, Wamhoff BR, Hultgårdh-Nilsson A, and Gomez MF. Hyperglycemia induces osteopontin expression via NFATc3 in vascular smooth muscle. Arteriosclerosis Thrombosis and Vascular Biology, in press

44. 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, in press

45. Zhao M, Wigren M, Dunér P, Bengtsson D, Olofsson K, Björkbacka H, Nilsson J, Fredrikson GN. Fc�RIIB inhibits atherosclerosis development in LDLR deficient mice. Journal of Immunology, in press

46. Kocherbitov V, Ulvenlund S, Briggner L-E, Kober M, Arnebrant T. Hydration of a natural polyelectrolyte Xanthan Gum: comparison with non-ionic carbohydrates, submitted

47. 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, submitted

48. Pihl M, Davies JR, Chávez de Paz LE, Svensäter G. Differential effects of Pseudomonas aeruginosa on biofilm formation by fresh isolates and laboratory strains of Staphylococcus epidermidis. submitted

49. Chávez de Paz LE, Davies J, Svensäter G. Fluorescence in situ hybridization for intracellular mRNA detection in adherent bacteria. Appl Environ Microbiol, submitted

50. Pihl M, Davies J, Chavez de Paz LE, Svensäter G. FEMS Microbiol Immunol, submitted 51. A. Masic, J. Bengtsson, N.C. Overgaard, M. Christensson, A. Heyden Measuring and modeling the

oxygen profile in a nitrifying Moving Bed Biofilm Reactor. Mathematical Biosciences, submitted

Proceeding papers:

52. Singh RN, Chakravartty JK, Ståhle P. Interpretation of hydrogen induced sub-critical crack growth in Zirconium alloys. 3rd International Congress on Computational Mechanics and Simulation (ICCMS09). IIT-Bombay, Mumbai-400 076, INDIA (2009)

53. Singh AK, Hussain MM, Singh RN, Chakravartty JK, Shah BK, Singh RP, Ståhle P. Stress field of blister forming in a metallic fuel and its interaction with clad. Characterization and Quality Control of Nuclear Fuels -CQCNF. Hyderabad, India (2009)

54. Safdar A, He HZ, Wei L-Y, Snis A, Zolotarevskii A, Chávez de Paz LE. Effect of EBM processing parameters on surface roughness and microstructure of Ti-6Al-4V. Proc. 22nd European Conference for Biomaterials, Lausanne, Switzerland, 7-11 Sep. (2009) No. 442

55. Sadar, A, Wei L-Y. Microstructures of electron beam melted (EBM) biomaterial Ti-6Al-4V. Proc. The Materials Research Society (MRS) 2008 annual fall meeting, Mater. Res. Soc. Symp. Proc. Vol. 1132 (2009) Z09-06.

56. Bjerkén C. Branching of a dissolution driven stress corrosion crack (2009), Conf Proc of Conf. Crack Paths, Vicenza, Italy (2009)



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57. Bjerkén C, Rimoli J, Ortiz M. Stress corrosion crack growth beneath a stiff coating – influence of chemical potential and interface toughness. International Conference on Fracture, Ottawa, Canada (2009)

7.2 Books and book chapters

1. Svensäter G, Chávez de Paz LE, Theilade E. The microbiology of the necrotic pulp in Textbook of Endodontology 2nd edition. Eds G Bergenholtz, Preben Horsted-Bindslev, Claes Reit. Blackwell Publishing Ltd (2009) 95-112.

2. Jernkvist LO, Massih AR. Nuclear Fuel Behaviour under Reactivity Initiated Accident Conditions, The OECD Nuclear Energy Agency Organisation for Economic Co-operation and Development, France (2009)

7.3 Popular articles

3. Svensäter G, Johansson I. Munnens mikroflora på gott och ont. Nordisk nutrition nr 3 (2009) 15-18 (In Swedish)

4. Petersson LG, Magnusson K, Hakestam U, Gustafsson L, Neilands J, Svensäter G. Prevention av rotkaries med hjälp av fluor och probiotika. Tandläkartidningen 3 (2009) 64-5. (In Swedish)

5. Gjörloff Wingren A. Digtal holografi – innovativ teknik för cellstudier. Laboratoriet 5 (2009) 6-8 (In Swedish)

8 Visitors and Seminars at the Center 1. Prof Renata Bilewicz, Warsaw University, 2009-12-09--11. 2. Profs Alexander Toikka and Irina Zvereva, Saint-Petersburg State University, 2009-06-17--25 3. Prof RN Singh, Indian Institute of Technology, Mumbai, Bahbah Atomic Reserach Centre 4. Prof J K Chakravartty, Bahbah Atomic Reserach Centre, 5. Prof L. Banks Sills, Tel Aviv University 6. Prof. Kåre Larsson, Camurus Lipid Research Foundation, Mentor at PhD-student day 2009-10-06 7. Prof Martin Malmsten, Pharmacy, Uppsala University, Seminar: “Protein interfacial behaviour in

microfabricated analysis systems and microarrays” 2009-10-06 8. Prof Håkan Olsson, Oncology, Lund University, Seminar: “DNA-arrays and their applications”

2009-10-07 9. Prof Christer Wingren, Imunotechnology. LTH, Seminar: “Protein arrays and their applications”

2009-10-07 10. Prof Johan Nilsson, Electrical Measurements, LTH, Seminar: “Acoustic manipulation and

arraying in microfluidics” 2009-10-07 11. Prof. Gunnar Bergenholtz, University of Gothenburg, 2009-02-23 & 2009-10-13--15 12. Prof. David Beighton, Guy’s Kings and St Thomas’ Dental Institute, London, Experimental work

with confocal microscopy, Seminars: “The role of the complex oral microflora in the caries process” & “The complex oral microflora of high-risk individuals and groups and its role in the caries process” 2009-02-25, 2009-04-16—17 & 2009-12-08—09

13. Prof. Jeannine Brady, University of Florida, 2009-06-28--31 14. Prof. Ian Hamilton, University of Manitoba, 2009-10-03--11 15. Prof. Dave Thornton, University of Manchester, 2009-10-06--07 16. Prof. Jukka Meurman, Helsinki University Central Hospital, Helsinki, 2009-10-22 17. Prof. Mark Herzberg, University of Minneapolis, Seminar: “Characterization of hydrogen

peroxide-induced DNA release by streptococci” 2009-12-05--09 18. Prof. Mogens Kilian, University of Aarhus, Seminar: “The etiology of periodontal disease

revisited by population genetic analysis” 2009-12-07—08



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19. Prof. Måns Rosén, Director of the Swedish Council on Technology Assessment in Health Care, Seminar: “When can RCTs and observational intervention studies mislead us and what can we do about it?” 2009-12-09

20. Prof. Ingegerd Johansson, Umeå University, 2009-12-07--09 21. Prof. Leslie Banks Sills, Tel Aviv University, Israel, Seminar: “Interfacial fracture of a

piezoelectric material” 22. Prof. Y K Charkravartty, Mumbai Insitute of Technology, Bombay, India, Seminar: “Fracture

Behaviour of Dilute Zirconium Alloys : Role of Hydrogen and Trace Element Impurities 23. Prof. R N Singh, Mumbai Insitute of Technology, Bombay, India, Seminar: “Accommodation of

hydrides, its stress-reorientation behavior and threshold stress intensity factor for delayed hydride cracking in Zr-alloys”

24. Prof. A. Needleman, Brown University, USA, Seminar: “Cohesive zone modeling - A review” 25. Prof. Ravi Chandar, University of Texas at Austin, Seminar: “Diffuse dynamic stress waves” 26. Prof. Beatrix Alsanius, SLU, Alnarp, Seminar:”Design av biofilm för att öka verkningsgraden av

sjukdomsmotverkande (antagonistiska) organismer alternativt av biologiska bekämpningsmedel i hortikulturella produktionssystem?” 2009-04-23

27. Prof Jan-Ingvar Jönsson, Linköping University, Seminar: “Mechanisms of transformation by oncogenic FLT3 receptor tyrosine kinase signaling in leukemia and its pharmaceutical intervention”

28. Associate Professor Kamal Mustafa, Bergen University, Seminar: “Cell culture studies and stem cells for improved biomaterials”

29. Assoc. Prof. Marie Ranson, University of Wollongong, Australia, 2009-09-21--10-29 30. Assoc. Prof. Johannes Persson, Lund University, Seminar: “Epistemic Risk: The significance of

knowing what one does not know” 2009-12-07 31. Dr Lars G Petersson, Specialist Clinic for Oral Health Care, Hallands Läns Landsting, Halmstad,

2009-11-18 32. Dr Sergei Lobov, University of Wollongong, Australia, 2009-04-14--05-09 33. Dr. Anne Meyer, Research Center for Biointerfaces, Buffalo, 2009-06-12 34. Dr Peter Kleinsmit, Roche Diagnostics Scandinavia AB, Seminar: “NimbleGen DNA microarrays

of the human genome” 2009-10-07 35. Dr Marie Skepö, Medeon AB, Mentor at PhD-student day 2009-10-06 36. Dr Justas Barauskas, Institute of Biochemistry, Vilnius, 2009-02-25--03-01, 2009-05-14--25 37. Dr Peter Nilsson, Div. Physical Chemistry, Lund University, 2009-07-01--28. 38. Dr Salla Marttila and Kertstin Brissmar, SLU, Alnarp, 2009-11-23 39. Ulf Brogren, Promimic AB, Seminar: “Nanostructured implant surfaces” 2009-10-07 40. Marius Dagys, PhD student at Institute of Biochemistry, Vilnius, 2009-09-13--15 and 2009-12-14-

-19. 41. Maksym Golub, PhD student at GKSS Research Center Geesthacht, Germany, 2009-09-20--30. 42. Sadia Niazi, PhD student at Guy’s Kings and St Thomas’ Dental Institute, London, 2009-02-25 43. Maria Toikka, PhD student at Saint-Petersburg State University, 2009-09-23--30 44. Gediminas Niaura, Institute of Biochemistry, Vilnius, 2009-12-09--11 45. Richard Mair, University of Toronto, Canada, Seminar: “Microarrays in biofilms” 2009-01-11--18

Internal seminars at the Center 46. Andrea Möllenkvist "Identification and characterization of the Moraxella catarrhalis IgD-binding

protein MID" 2009-02-18 47. Tautgirdas Ruzgas and Sergey Shleev, ”3D nanobiostructure-based self-contained devices for

biomedical applications (TR, SS) 2009-03-16 48. Gabriella Sinkiewitz “Functional properties of the probiotic Lactobacillus reuteri” 2009-03-18 49. Olof Svenson “Antibody binding kinetics” 2009-04-01 50. Tautgirdas Ruzgas ”Biofuel cells” 2009-04-15 51. Anna Ketelsen, "From intestinal T cell homing to lipid based immune assays" 2009-04-29 52. Ida Svedsen “Interfacial behaviour of salivary proteins” 2009-05-13



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53. Thomas Arnebrant and Ann Wennerberg ”Atomic Force Micrioscopy and applications” 2009-06-09

54. Annika Jögi, post-doc at CMP, UMAS “Hypoxia-impaired acinar morphogenesis in mammary epithelial cells” 2009-10-14

55. Javier Sotres, “Force microscopy and spectroscopy of single biological molecules” 2009-10-28 56. Karolina Haberska “Protein and polyelectrolyte layer-by-layer films: Assembly and electron

transfer” 2009-11-25 57. Anton Fagerström, “Bioavailability of Active Ingredients used in Agriculture”, 2009-12-08

9 Workshops and Conferences Organized by the Center During 2010 the Center has launched the 5th Annual workshop, this year with focus on Microarrays. 67 participants listened to invited speakers from both industry and academia, as well as talks given by the researchers at Mah. The workshop was preceeded by a Ph-D-student day with Prof. Kåre Larsson (Camururs Lipid Research Foundation) and Dr Marie Skepö (Medeon AB) as mentors. The Center also launched kick-off meetings within the two new EU-projects; “Three-dimensional nanobiostructure-based self-contained devices for biomedical application” (EU FP7, Dr Sergey Shleev, coordinator), and “VALOR – Valorisation of knowledge intensive ideas in the South Baltic area” (EU South Baltic programme, Assoc. Prof. Johan Engblom, local coordinator). The potential benefit to individual researchers from the VALOR project was communicated within Biofilms at a subsequent event. Center members have also been responsible for arranging a workshop within the National Research School of Odontology; “Oral Biofilm and Risk”, and an external course on “Cell Culture” at IBL (Institutet för Biomedicinsk Laboratorievetenskap), and two practical training courses in confocal microscopy. Together with Medicon Valley Alliance and Trial Form Support AB we arranged a meeting to present a new MSc-program in Clinical Research Administration, developed during 2008.

10 Contributions to and Participation in Conferences and Workshops

10.1 Oral presentations 2009

1. Jernkvist LO, Massih AR and In de Betou J. A review of experiments and computer analyses on RIAs. OECD Nuclear Energy Agency Workshop on Nuclear Fuel Behaviour under Reacivity Initiated Accidents, Sept. 9-11, 2009, Paris, France.

2. Jernkvist LO, Massih AR. A comparison between in-pile and out-of-pile radial DHC in Zircaloy-2 cladding. OECD/NEA Studsvik Cladding Integrity Program workshop on Hydrogen Induced Failures, 2009 Nov. 17-18, Nyköping, Sweden.

3. Massih AR and Jernkvist LO. Stress reorientation of hydrides in zirconium alloys. OECD/NEA Studsvik Cladding Integrity, Program workshop on Hydrogen Induced Failures, 2009 Nov. 17-18, Nyköping, Sweden.

4. Ståhle P. Broberg Memorial symposium, Hydride formation in elastic plastic materials. Trolleholm, 25-26 May, 2009



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5. Safdar A, He HZ, Wei L-Y, Snis A, Zolotarevskii A, Chávez de Paz LE. Effect of EBM processing parameters on surface roughness and microstructure of Ti-6Al-4V. 7-11 Sep. 2009, Lausanne, Switzerland

6. Gjörloff Wingren A. A novel label-free technique of digital holography to study cellular parameters such as growth, viability and antibody interactions of eukaryotic cells. Nordisk konferens i Biomedicinsk laboratorievetenskap 2009.

7. Wennerberg A. Invited lecture: Biomaterials surface topography. Biomaterials in Medicine 2nd Scientific Meeting, 29-30 April, 2009. DTU, Kgs Lyngby, Denmark

8. Wennerberg A. Invited lecture: Do nanometer surface modifications matter? ICP 13th Biennial Meeting Cape Town, South Africa September 10-13, 2009,

9. Wennerberg A. Invited lecture: Implant surfaces. 33rd Annual Congress. European Prosthodontic Association. October 1-3, 2009 Innsbruck. Austria,

10. Wennerberg A. Invited lecture: Vad är god klinisk dokumentation. Svenska Tandläkarstämman Stockholm 091113.

11. Engblom J. Invited lecture: Effects on drug bioavailability from topical formulations and skin barrier response. at The natural protection – from skin to evidence (SCANCOS) Malmö, Sweden, 2009-11-05—06

12. Björklund S. The effect of water on skin permeation of the model drug metronidazole, Skin and Formulation 3rd symposium and Skin Forum 10th Annual Meeting, Versailles, 9-10 March 2009

13. Björklund S. Skin permeability can be regulated by the gradient in chemical potential of water. OMM mini-symposium on diffusive transport, Lund, 3 June 2009

14. Ruzgas T. Invited lecture: Biofuel cells. Conference in memory of Bo Håkansson: Innovative electrochemistry, KTH, Stockholm 2009-01-29

15. Lindh L. The composition of the salivary pellicle is substrate specific. Odontologisk Riksstämma, Stockholm, 2009,

16. Chavez de Paz L. In situ hybridization of mRNA and rRNA in microbial biofilms. Biofilms 5th Annual workshop: Microarrays, Malmö 2009-10-06--07

17. Shleev S. Potentially implantable miniature biofuel cells. Biofilms 5th Annual workshop: Microarrays, Malmö 2009-10-06—07

18. El-Schish Z, Mölder A, Sebesta M, Gisselsson L, Härkönen P, Wingren C, and Gjörloff Wingren A. Developing innovative clinical tools for cancer cell analysis: a label-free technique of digital holography to study cellular parameters such as growth, viability and antibody interactions. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

19. Safdar A, He HZ, Wei L-Y, Snis A, Chàvez de Paz LE. Effects of EBM Processing Parameters on Surface and Microstructure of Biomaterial Ti-6Al-4V. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

20. Davies JR, Chàvez de Paz LE and Svensäter G. Surface and cell wall proteins from Streptococcus gordonii. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

21. Svensson O, and Arnebrant T. Controlling the Adsorption of Serum Albumin on Silica. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

10.2 Posters 2009

1. Pihl M, Chávez de Paz L,E Svensäter G. Mutual effects of P. aeruginosa and S. epidermidis in dual-species biofilms. Eurobiofilms 2009, First European Congress on Microbial Biofilms, Rome, 2-5 September 2009.

2. Chávez de Paz LE, Svensäter G, Wickström C. Surface-associated human salivary MUC5B induces expression of endopeptidase in Lactobacillus fermentum biofilm cells. Eurobiofilms 2009, 1st European Congress on Microbial Biofilms, Rome, 2-5 September 2009.

3. Davies JR, Chávez de Paz LE, Svensäter G. Surface and cell wall proteins from Streptococcus gordonii. Eurobiofilms 2009, 1st European Congress on Microbial Biofilms, Rome, 2-5 September 2009.



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4. Chávez de Paz LE, Svensäter G. BioImage_L: Image Analysis Software for Analysis of Physiological Activities and Gene Expression in Microbial Biofilms. 1st European Congress on Microbial Biofilms, Rome, 2-5 September 2009.

5. Gjörloff Wingren A et al. A novel label-free technique of digital holography to study cellular parameters such as growth, viability and antibody interactions of eukaryotic cells Nordisk konferens i Biomedicinsk laboratorievetenskap 2009

6. Gjörloff Wingren A et al. Ectopic expression of ZAP70 leads to decreased B-cell receptor internalization after IgM stimulation in Diffuse Large B-cell lymphoma. Nordisk konferens i Biomedicinsk laboratorievetenskap 2009

7. Björklund S, Engblom J, Sparr E, and Thuresson K. The effect of water on the skin permeation of a model drug. Gordon Research conference on ”Skin barrier”, Waterville, Boston, USA 2009

8. Znamenskaya Y, Engblom J, Sotres J, Arnebrant T, Kocherbitov V, Effect of Hydration on Structural Properties of Mucuous Gel, Dynamics, Steady State and Arrest, 9th Annual Surface and Colloid Symposium, 18-20 November 2009, Lund, Sweden

9. Björklund S, Engblom J, Sparr E, and Thuresson K. The effect of water on the skin permeation of a model drug, , Dynamics, Steady State and Arrest, 9th Annual Surface and Colloid Symposium, 18-20 November 2009, Lund, Sweden

10. Sjödin T, Chávez de Paz, Troedsson U, Svensäter G. The effect of delmopinol on acid adaptation by dental plaque. International Association of Dental Research General Session, Miami, USA, 2009.

11. Sjödin T. Determination of inhibitory concentrations of delmopinol, triclosan, chlorhexidine and cetylpyridine on planktonic and biofilm bacteria. Eurobiofilms 2009; First European Congress on Microbial Biofilms. Rome, September 2-5, 2009.

12. Davies JR, Càvez de Paz L and Svensäter G. Surface and cell wall proteins from Streptococcus gordonii. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

13. Fridberg M, Berglund M, Berglund M, Söderberg O, Enblad G, Jirström K, Härkönen P, Gjörloff Wingren A. Ectopic expression of ZAP70 leads to decreased B-cell receptor internalization after IgM stimulation in Diffuse Large B-cell lymphoma. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

14. El-Schish Z, Mölder A, Sebesta M, Gisselsson L, Härkönen P, Wingren C and Gjörloff Wingren A. Developing innovative clinical tools for cancer cell analysis: a label-free technique of digital holography to study cellular parameters such as growth, viability and antibody interactions. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

15. Pihl M, Chaves de Paz L, Svensäter G. Mutual effects of P. aeruginosa and S. epidermidis in dual-species biofilms. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

16. Safdar A, He HZ, Wei L-Y, Snis A, Chaves de Paz L. Effects of EBM Processing Parameters on Surface and Microstructure of Biomaterial Ti-6Al-4V. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

17. Svensson O and Arnebrant T, Controlling the Adsorption of Serum Albumin on Silica, Biofilms 5th Annual workshop, Malmö 2009-10-06--07

18. Znamenskaya Y, Engblom J, Kocherbitov V. Effect of Hydration on Structural Properties of Mucuous Gel. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

19. Kocherbitov V and Arnebrant T. Hydration of lysozyme: enthalpy – entropy compensation and the Langmuir sorption isotherm. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

20. Masic A, Bengtsson J, Overgaard NC, Christensson M, Heyden A. Determination of the oxygen profile in a nitrifying Moving Bed Biofilm Reactor by microelectrodes and a mathematical model. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

21. Sjödin T. Determination of inhibitory concentrations of delmopinol, triclosan, chlorhexidine and cetylpyridine on planktonic and biofilm bacteria. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

22. Björklund S, Engblom J, Sparr E, and Thuresson K. The effect of water on the skin permeation of a model drug. Biofilms 5th Annual workshop, Malmö 2009-10-06--07



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23. Pettersson A, Kocherbitov V, Westbye P, Bergström K, Engblom J. Bioavailability of active ingredients used in agriculture. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

24. Chavez de Paz LE and Svensäter G. bioImage_L: Image Analysis Software for Analysis of Physiological Activities and Gene Expression in Microbial Biofilms. Biofilms 5th Annual workshop, Malmö 2009-10-06--07

25. Chavez de Paz L, Svensäter G and Wickström C. Surface-associated human salivary MUC5B induces expression of endopeptidase in Lactobacillus fermentum biofilm cell. Biofilms 5th Annual workshop, Malmö 2009-10-06—07

11 BSc and MSc education

11.1 BSc-level

1. Biomedical laboratory science (HS) 2. TELMah-Biomedical Technology (CTS & HS) 3. Dentistry program (OD)

11.2 MSc-level

1. Biomedical Methods and Technology (HS) 2. Materials science (CTS & HS) 3. Dentistry program (OD)

12 Other Activities by Center Members Thomas Arnebrant and Tautgirdas Ruzgas have been opponents at dissertations for doctors degree in Sweden (KTH) and Denmark (Copenhagen University). Gunilla Nordin Fredrikson and Julia Davies have been opponents at half-time controls (LU, Mah). Center members have been board members at dissertations at 13 occasions (LuTH, UU, LiU, KI, LU), and Julia Davies and Gunnel Svensäter have been external examiners at Leeds Dental Institute, UK. Gunilla Nordin Fredrikson, Per Ståhle, Christina Bjerkén and Thomas Arnebrant have been external reviewers for promotions to professor (3), associate professor (1) and employment as university lecturer (2). Center members have been referees for international journals for more than 60 papers. The list of journals include; Caries Research, Journal of Medical Microbiology, Microbiology, Archives Oral Biology, Oral diseases, Journal of Clinical Periodontology, European Journal of Oral Science, Journal of Endodontology, Acta Biomaterialia Analytical Chemistry, Arteriosclerosis, Thrombosis and Vascular Biology (ATVB), Atherosclerosis, Biofouling, Biosensors and Bioelectronics, Biotechnology Progress,



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Bioelectrochemistry, Caries research, Clinical Immunology, Colloid and Polymer Science, Colloids and Surfaces B: Biointerfaces, Colloids and Surfaces A, Electroanalysis, Electrochimica Acta, Electrochemical Communications, European Journal of Oral Sciences, European Journal of Histochemistry, Future Cardiology, International Journal of Biomedical Science (IJBS), Journal of the American Chemical Society, Journal of Biomedical Materials Research: Part A, 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, International Journal of Solids and Structures, Journal of Applied Mechanics, Journal of the Mathematics and Physics of Solids, International Journal of Fracture, Annals of Nuclear Energy, and Journal of Nuclear Materials. Tautgirdas Ruzgas is on the editorial board for Nonlinear analysis: Modelling and control, and Per Ståhle was guest editor of International Journal of Fracture. Per Ståhle has been Chairman of the National committee of Mechanics, Royal Academy of Science. Gunnel Svensäter was coordinator of the Swedish National Graduate School in Odontological Science, and member of the scientific advisory board at the Swedish Council on Technology Assessment in Health Care (SBU). Svensäter has been evaluator at the Swedish Dental Association (SDA) and was member of the Research committee at Faculty of Odontology (OD, Mah). Tautgirdas Ruzgas was member of the docent board at Malmö University. Johan Engblom was member of the Quality assurance group at Faculty of Health and Society and member of the Faculty Board at School of Technology (CTS, Mah). Gunilla Nordin Fredrikson was convener of the Evaluation committee of DPLU (Diabetes Program Lund University).

Cover story

Dr Ida Svendsen (right) got the award for the best PhD-thesis at Malmö University 2009. Her thesis “In vitro and in vivo studies of salivary films at solid/liquid interfaces” is a good example of the potential of interfaculty collaboration within Biofilms-Research Center for Biointerfaces. Here with her supervisors Assoc. Professor Liselott Lindh (OD) and Professor Thomas Arnebrant (HS).

Center Mission Statement

Biofilms – Research Center for Biointerfaces is a translational research programme covering six research groups within three faculties/schools (HS, OD & CTS) 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 Centre for Biointerfaces Malmö University SE-205 06 MALMÖ, Sweden Center director: Assoc. Prof. Johan Engblom (JE) Tel: +46-(0)706-08 75 25 (JE); +46-(0)40-66 57 486 (Adm. Coordinator Eva Nilsson) e-mail: [email protected] www.mah.se/biofilms Visiting adress: University Hospital MAS (Entrance 49) MALMÖ

biofilms â€“research center for biointerfaces

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