Evaluation of antimicrobial effects of novel implant materials by testing the prevention of biofilm formation using a simple small scale medium-throughput growth inhibition assay

Staphylococcal colonization of implants is a serious complication of orthopaedic surgery. Anti-infectious modification of implant surfaces may serve to prevent bacterial colonization. The authors set out to develop an in vitro test system for the analysis of prevention of biofilm formation by Staphylococcus epidermidis and Staphylococcus aureus on implant materials. Biofilm growth was monitored over 10 days on titanium disks in order to develop appropriate test parameters. Bacterial cell counts following ultrasonic treatment of the colonized samples were compared with scanning electron microscope images of the specimens. Copper ion containing surfaces (ie copper [Cu] and inter-metallic Ti-Cu films) were used for growth inhibition assays: Copper ion releasing specimens led to reduced bacterial numbers in biofilms and decreased bacterial persistence in the model used. The assay used represents an inexpensive and quick in vitro screen for the antibacterial effects of novel implant surface materials.

[1]  P. Giusti,et al.  Combined drug release from biodegradable bilayer coating for endovascular stents. , 2010, Journal of biomedical materials research. Part B, Applied biomaterials.

[2]  N. Südkamp,et al.  Antibiotics in trauma and orthopedic surgery -- a primer of evidence-based recommendations. , 2006, Injury.

[3]  W. Zimmerli Implantatinfektionen. Was muss der Internist wissen , 2005 .

[4]  C. Yan,et al.  Antibiotic prophylaxis after total joint replacements. , 2009, Hong Kong medical journal = Xianggang yi xue za zhi.

[5]  M. Assal,et al.  Low incidence of haematogenous seeding to total hip and knee prostheses in patients with remote infections. , 2009, The Journal of infection.

[6]  G. Rogers,et al.  Enhancing the utility of existing antibiotics by targeting bacterial behaviour? , 2012, British journal of pharmacology.

[7]  H. Tsuchiya,et al.  Prevention of pin tract infection with titanium-copper alloys. , 2009, Journal of biomedical materials research. Part B, Applied biomaterials.

[8]  J. Zenilman,et al.  The estimated magnitude and direct hospital costs of prosthetic joint infections in the United States, 1997 to 2004. , 2010, The Journal of arthroplasty.

[9]  D. Allison,et al.  Resistance of bacterial biofilms to antibiotics: a growth-rate related effect? , 1988, The Journal of antimicrobial chemotherapy.

[10]  H. Ceri,et al.  The Calgary Biofilm Device: New Technology for Rapid Determination of Antibiotic Susceptibilities of Bacterial Biofilms , 1999, Journal of Clinical Microbiology.

[11]  F. Garcia-alvarez,et al.  Evaluation of four experimental osteomyelitis infection models by using precolonized implants and bacterial suspensions , 2002, Acta orthopaedica Scandinavica.

[12]  P. Choong,et al.  Risk factors associated with acute hip prosthetic joint infections and outcome of treatment with a rifampinbased regimen , 2007, Acta orthopaedica.

[13]  E. Schwarz,et al.  Quantitative mouse model of implant‐associated osteomyelitis and the kinetics of microbial growth, osteolysis, and humoral immunity , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[14]  J. Costerton,et al.  Bacterial biofilms: a common cause of persistent infections. , 1999, Science.

[15]  R. Landmann,et al.  Weak effect of metal type and ica genes on staphylococcal infection of titanium and stainless steel implants. , 2008, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[16]  H. Ceri,et al.  Microtiter susceptibility testing of microbes growing on peg lids: a miniaturized biofilm model for high-throughput screening , 2010, Nature Protocols.

[17]  R. Bader,et al.  Analysis of the Release Characteristics of Cu-Treated Antimicrobial Implant Surfaces Using Atomic Absorption Spectrometry , 2011, Bioinorganic chemistry and applications.

[18]  K. Yanagihara,et al.  Quantitative analysis of Staphylococcus epidermidis biofilm on the surface of biomaterial , 2009, Journal of orthopaedic science : official journal of the Japanese Orthopaedic Association.

[19]  G. Christensen,et al.  Methods for studying microbial colonization of plastics. , 1995, Methods in enzymology.

[20]  J. Costerton,et al.  Bacterial adherence to biomaterials and tissue. The significance of its role in clinical sepsis. , 1985, The Journal of bone and joint surgery. American volume.

[21]  J. Costerton,et al.  Antibiotic resistance of bacteria in biofilms , 2001, The Lancet.

[22]  M. B. Coventry Treatment of infections occurring in total hip surgery. , 1975, The Orthopedic clinics of North America.

[23]  B. Ratner,et al.  Design of infection-resistant antibiotic-releasing polymers. II. Controlled release of antibiotics through a plasma-deposited thin film barrier. , 1999, Journal of controlled release : official journal of the Controlled Release Society.

[24]  W. Mittelmeier,et al.  A novel antibacterial titania coating: Metal ion toxicity and in vitro surface colonization , 2005, Journal of materials science. Materials in medicine.

[25]  J. Morrissey,et al.  Copper Stress Induces a Global Stress Response in Staphylococcus aureus and Represses sae and agr Expression and Biofilm Formation , 2009, Applied and Environmental Microbiology.

[26]  S. Molin,et al.  The clinical impact of bacterial biofilms , 2011, International Journal of Oral Science.

[27]  Xuanyong Liu,et al.  Biological and antibacterial properties of plasma sprayed wollastonite/silver coatings. , 2009, Journal of biomedical materials research. Part B, Applied biomaterials.

[28]  Rainer Bader,et al.  Deposition of thin titanium–copper films with antimicrobial effect by advanced magnetron sputtering methods , 2011 .

[29]  M. Rupp,et al.  Coagulase-negative staphylococci: pathogens associated with medical progress. , 1994, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[30]  R. Mason,et al.  Hydroxyl radical formation from cuprous ion and hydrogen peroxide: a spin-trapping study. , 1995, Archives of biochemistry and biophysics.

[31]  P Stoodley,et al.  Survival strategies of infectious biofilms. , 2005, Trends in microbiology.

[32]  A. Gristina,et al.  In vitro and in vivo comparative colonization of Staphylococcus aureus and Staphylococcus epidermidis on orthopaedic implant materials. , 1989, Biomaterials.

[33]  E. Wickstrom,et al.  Vancomycin covalently bonded to titanium beads kills Staphylococcus aureus. , 2005, Chemistry & biology.

[34]  P. Fey,et al.  Genetic and phenotypic analysis of biofilm phenotypic variation in multiple Staphylococcus epidermidis isolates. , 2004, Journal of medical microbiology.

[35]  R. Grimer,et al.  The incidence of deep prosthetic infections in a specialist orthopaedic hospital: a 15-year prospective survey. , 2006, The Journal of bone and joint surgery. British volume.

[36]  Robin Patel,et al.  Clinical practice. Infection associated with prosthetic joints. , 2009, The New England journal of medicine.

[37]  W. Mittelmeier,et al.  Antibacterial poly(D,L-lactic acid) coating of medical implants using a biodegradable drug delivery technology. , 2003, The Journal of antimicrobial chemotherapy.

[38]  J. Pratten,et al.  Use of biofilm model systems to study antimicrobial susceptibility. , 2010, Methods in molecular biology.

[39]  P. Choong,et al.  Diagnosis and management of prosthetic joint infection , 2012, Current opinion in infectious diseases.

[40]  E. Witsø,et al.  Sonication is superior to scraping for retrieval of bacteria in biofilm on titanium and steel surfaces in vitro , 2009, Acta orthopaedica.