title: Covalent immobilization of antimicrobial agents on titanium prevents Staphylococcus aureus and Candida albicans colonization and biofilm formation.
abstract: OBJECTIVE
  Biofilm-associated implant infections represent a serious public health problem. Covalent immobilization of antimicrobial agents on titanium (Ti), thereby inhibiting biofilm formation of microbial pathogens, is a solution to this problem.
  
  
  METHODS
  Vancomycin (VAN) and caspofungin (CAS) were covalently bound on Ti substrates using an improved processing technique adapted to large-scale coating of implants. Resistance of the VAN-coated Ti (VAN-Ti) and CAS-coated Ti (CAS-Ti) substrates against in vitro biofilm formation of the bacterium Staphylococcus aureus and the fungal pathogen Candida albicans was determined by plate counting and visualized by confocal laser scanning microscopy. The efficacy of the coated Ti substrates was also tested in vivo using an adapted biomaterial-associated murine infection model in which control-Ti, VAN-Ti or CAS-Ti substrates were implanted subcutaneously and subsequently challenged with the respective pathogens. The osseointegration potential of VAN-Ti and CAS-Ti was examined in vitro using human bone marrow-derived stromal cells, and for VAN-Ti also in a rat osseointegration model.
  
  
  RESULTS
  In vitro biofilm formation of S. aureus and C. albicans on VAN-Ti and CAS-Ti substrates, respectively, was significantly reduced compared with biofilm formation on control-Ti. In vivo, we observed over 99.9% reduction in biofilm formation of S. aureus on VAN-Ti substrates and 89% reduction in biofilm formation of C. albicans on CAS-Ti substrates, compared with control-Ti substrates. The coated substrates supported osseointegration in vitro and in vivo.
  
  
  CONCLUSIONS
  These data demonstrate the clinical potential of covalently bound VAN and CAS on Ti to reduce microbial biofilm formation without jeopardizing osseointegration.
authors.affiliation: Department of Molecular Microbiology, VIB, Kasteelpark Arenberg 31, Box 2438, 3001 Leuven, Belgium Laboratory of Molecular Cell Biology, KU Leuven, Kasteelpark Arenberg 31, Box 2438, 3001 Leuven, Belgium.
authors.forename: Soňa
authors.initials: S
authors.lastname: Kucharíková
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium.
authors.forename: Evelien
authors.initials: E
authors.lastname: Gerits
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium.
authors.forename: Katrijn
authors.initials: K
authors.lastname: De Brucker
authors.affiliation: Department of Materials Engineering (MTM), KU Leuven, Kasteelpark Arenberg 44, Box 2450, 3001 Leuven, Belgium.
authors.forename: Annabel
authors.initials: A
authors.lastname: Braem
authors.affiliation: Center for Animal Genomics, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia.
authors.forename: Katerina
authors.initials: K
authors.lastname: Ceh
authors.affiliation: Center for Animal Genomics, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia.
authors.forename: Gregor
authors.initials: G
authors.lastname: Majdič
authors.affiliation: Center for Animal Genomics, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia.
authors.forename: Tanja
authors.initials: T
authors.lastname: Španič
authors.affiliation: Center for Animal Genomics, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia.
authors.forename: Estera
authors.initials: E
authors.lastname: Pogorevc
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium.
authors.forename: Natalie
authors.initials: N
authors.lastname: Verstraeten
authors.affiliation: Department of Molecular Microbiology, VIB, Kasteelpark Arenberg 31, Box 2438, 3001 Leuven, Belgium Laboratory of Molecular Cell Biology, KU Leuven, Kasteelpark Arenberg 31, Box 2438, 3001 Leuven, Belgium.
authors.forename: Hélène
authors.initials: H
authors.lastname: Tournu
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Delattin
authors.affiliation: Biotech International, Allées de Craponne 305, 13300 Salon-de-Provence, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Impellizzeri
authors.affiliation: Hemoteq AG, Adenauerstrasse 15, 52146 Wuerselen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Erdtmann
authors.affiliation: SP Food and Bioscience, Department of Structure and Material Design, Box 5401, 402 29 Gothenburg, Sweden.
authors.forename: Annika
authors.initials: A
authors.lastname: Krona
authors.affiliation: SP Food and Bioscience, Department of Structure and Material Design, Box 5401, 402 29 Gothenburg, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Lövenklev
authors.affiliation: Educell, d.o.o., Prevale 9, 1236 Trzin, Slovenia.
authors.forename: Miomir
authors.initials: M
authors.lastname: Knezevic
authors.affiliation: Educell, d.o.o., Prevale 9, 1236 Trzin, Slovenia Department of Biochemistry and Molecular and Structural Biology, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Fröhlich
authors.affiliation: Department of Materials Engineering (MTM), KU Leuven, Kasteelpark Arenberg 44, Box 2450, 3001 Leuven, Belgium.
authors.forename: Jef
authors.initials: J
authors.lastname: Vleugels
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium.
authors.forename: Maarten
authors.initials: M
authors.lastname: Fauvart
authors.affiliation: Department of Oral Health Sciences-Biomaterials BIOMAT, KU Leuven, Kapucijnenvoer 33, Box 7001, 3000 Leuven, Belgium FOP-UNICAMP, Department of Prosthodontics and Periodontology, Av. Limeira, 901, 13414-903, Piracicaba-SP, Brazil.
authors.forename: Wander Jose
authors.initials: WJ
authors.lastname: de Silva
authors.affiliation: Department of Oral Health Sciences-Biomaterials BIOMAT, KU Leuven, Kapucijnenvoer 33, Box 7001, 3000 Leuven, Belgium.
authors.forename: Katleen
authors.initials: K
authors.lastname: Vandamme
authors.affiliation: Alhenia AG, Täfernstrasse 39, 5405 Dättwil, Switzerland.
authors.forename: Jordi
authors.initials: J
authors.lastname: Garcia-Forgas
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Ghent, Belgium.
authors.forename: Bruno P A
authors.initials: BP
authors.lastname: Cammue
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Michiels
authors.affiliation: Department of Molecular Microbiology, VIB, Kasteelpark Arenberg 31, Box 2438, 3001 Leuven, Belgium Laboratory of Molecular Cell Biology, KU Leuven, Kasteelpark Arenberg 31, Box 2438, 3001 Leuven, Belgium.
authors.forename: Patrick
authors.initials: P
authors.lastname: Van Dijck
authors.affiliation: Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, Box 2460, 3001 Leuven, Belgium karin.thevissen@biw.kuleuven.be.
authors.forename: Karin
authors.initials: K
authors.lastname: Thevissen
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D002176:Candida albicans / Q000187:drug effects* / Q000502:physiology; D000077336:Caspofungin; D002460:Cell Line; D054714:Echinocandins / Q000494:pharmacology; D005260:Female; D006801:Humans; D055666:Lipopeptides / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D016348:Osseointegration; D019736:Prostheses and Implants / Q000382:microbiology; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000502:physiology; D014025:Titanium / Q000494:pharmacology*; D014640:Vancomycin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D000935:Antifungal Agents; D054714:Echinocandins; D055666:Lipopeptides; D014640:Vancomycin; D014025:Titanium; D000077336:Caspofungin