For an optimal implementation of materials, such as, e.g. medical implants in living environments, a thorough characterization of cell adhesion, kinetics and strength is required, as well as a prerequisite e.g. for bone integration. Here we present a miniaturized (~100 μl) lab-on-a-chip implant hybrid system which allows quantification of cell adhesion under dynamic conditions mimicking those of physiological relevance. Surface acoustic waves are excited and used on optical transparent chips to induce micro acoustic streaming and to create a microfluidic shear spectrum ranging from 0 to ~35 s(-1). We demonstrate its potential for a time-efficient, dynamic screening test of new implant materials using a model of an osseointegration with SAOS-2 cells. The upside-down orientation also allows utilization of the micro reactor on non-transparent materials like titanium and diamond-like-carbon (DLC).

The implant-bone interface is the scene of competition between microorganisms and distinct types of tissue cells. In the past, various strategies have been followed to support bony integration and to prevent bacterial implant-associated infections. In the present study we investigated the biological properties of diamond-like carbon (DLC) surfaces containing silver nanoparticles. DLC is a promising material for the modification of medical implants providing high mechanical and chemical stability and a high degree of biocompatibility. DLC surface modifications with varying silver concentrations were generated on medical-grade titanium discs, using plasma immersion ion implantation-induced densification of silver nanoparticle-containing polyvinylpyrrolidone polymer solutions. Immersion of implants in aqueous liquids resulted in a rapid silver release reducing the growth of surface-bound and planktonic Staphylococcus aureus and Staphylococcus epidermidis. Due to the fast and transient release of silver ions from the modified implants, the surfaces became biocompatible, ensuring growth of mammalian cells. Human endothelial cells retained their cellular differentiation as indicated by the intracellular formation of Weibel-Palade bodies and a high responsiveness towards histamine. Our findings indicate that the integration of silver nanoparticles into DLC prevents bacterial colonization due to a fast initial release of silver ions, facilitating the growth of silver susceptible mammalian cells subsequently.

BackgroundAntibacterial coatings of medical devices have been introduced as a promising approach to reduce the risk of infection. In this context, diamond-like carbon coated polyethylene (DLC-PE) can be enriched with bactericidal ions and gain antimicrobial potency. So far, influence of different deposition methods and ions on antimicrobial effects of DLC-PE is unclear.MethodsWe quantitatively determined the antimicrobial potency of different PE surfaces treated with direct ion implantation (II) or plasma immersion ion implantation (PIII) and doped with silver (Ag-DLC-PE) or copper (Cu-DLC-PE). Bacterial adhesion and planktonic growth of various strains of S. epidermidis were evaluated by quantification of bacterial growth as well as semiquantitatively by determining the grade of biofilm formation by scanning electron microscopy (SEM). Additionally silver release kinetics of PIII-samples were detected.Results(1) A significant (p < 0.05) antimicrobial effect on PE-surface could be found for Ag- and Cu-DLC-PE compared to untreated PE. (2) The antimicrobial effect of Cu was significantly lower compared to Ag (reduction of bacterial growth by 0.8 (Ag) and 0.3 (Cu) logarithmic (log)-levels). (3) PIII as a deposition method was more effective in providing antibacterial potency to PE-surfaces than II alone (reduction of bacterial growth by 2.2 (surface) and 1.1 (surrounding medium) log-levels of PIII compared to 1.2 (surface) and 0.6 (medium) log-levels of II). (4) Biofilm formation was more decreased on PIII-surfaces compared to II-surfaces. (5) A silver-concentration-dependent release was observed on PIII-samples.ConclusionThe results obtained in this study suggest that PIII as a deposition method and Ag-DLC-PE as a surface have high bactericidal effects.

Silver ions (Ag+) have strong bactericidal effects and Ag-coated medical devices proved their effectiveness in reducing infections in revision total joint arthroplasty. We quantitatively determined the antimicrobial potency of different surface treatments on a titanium alloy (Ti), which had been conversed to diamond-like carbon (DLC-Ti) and doped with high (Ag:PVP = 1:2) and low (Ag:PVP = 1:10 and 1:20) concentrations of Ag (Ag-DLC-Ti) with a modified technique of ion implantation. Bacterial adhesion and planktonic growth of clinically relevant bacterial strains (Staphylococcus epidermidis,Staphylococcus aureus, and Pseudomonas aeruginosa) on Ag-DLC-Ti were compared to untreated Ti by quantification of colony forming units on the adherent surface and in the growth medium as well as semiquantitatively by determining the grade of biofilm formation by scanning electron microscopy. (1) A significant (p < 0.05) antimicrobial effect could be found for all Ag-DLC-Ti samples (reduced growth by 5.6–2.5 logarithmic levels). (2) The antimicrobial effect was depending on the tested bacterial strain (most for P. aeruginosa, least for S. aureus). (3) Antimicrobial potency was positively correlated with Ag concentrations. (4) Biofilm formation was decreased by Ag-DLC-Ti surfaces. This study revealed potent antibacterial effects of Ag-DLC-Ti. This may serve as a promising novel approach to close the gap in antimicrobial protection of musculoskeletal implants.

Antibacterial coating of medical devices is a promising approach to reduce the risk of infection but has not yet been achieved on wear surfaces, e.g. polyethylene (PE). We quantitatively determined the antimicrobial potency of different PE surfaces, which had been conversed to diamond-like carbon (DLC-PE) and doped with silver ions (Ag-DLC-PE). Bacterial adhesion and planktonic growth of various strains of S. epidermidis on Ag-DLC-PE were compared to untreated PE by quantification of colony forming units on the adherent surface and in the growth medium as well as semiquantitatively by determining the grade of biofilm formation by scanning electron microscopy. (1) A significant (p < 0.05) antimicrobial effect could be found for Ag-DLC-PE. (2) The antimicrobial effect was positively correlated with the applied fluences of Ag (fivefold reduced bacterial surface growth and fourfold reduced bacterial concentration in the surrounding medium with fluences of 1 × 1017 vs. 1 × 1016 cm−2 under implantation energy of 10 keV). (3) A low depth of Ag penetration using low ion energies (10 or 20 vs. 100 keV) led to evident antimicrobial effects (fourfold reduced bacterial surface growth and twofold reduced bacterial concentration in the surrounding medium with 10 or 20 keV and 1 × 1017 cm−2 vs. no reduction of growth with 100 keV and 1 × 1017 cm−2). (4) Biofilm formation was decreased by Ag-DLC-PE surfaces. The results obtained in this study suggest that PE-surfaces can be equipped with antibacterial effects and may provide a promising platform to finally add antibacterial coatings on wear surfaces of joint prostheses.