An effective laser surface treatment method to reduce biofilm coverage of multiple bacterial species associated with medical device infection

Abstract

Device associated infection (DAI) is recognized as a worldwide health challenge in total joint replacement (TJR). Bacteria exhibit very strong antibiotic tolerance when they attach to a device and form a biofilm and thus DAI is difficult to treat. In this study, a one-step, clean (no chemicals or additional materials involved) and effective surface engineering approach via laser surface treatment (LST) to tackle the DAI challenge is reported. Commercially pure (CP) Ti were laser-treated in open air using continuous wave (CW) fibre laser. The laser-treated CP Ti was tested against five bacterial species including Gram positive (Staphylococcus aureus and Staphylococcus epidermidis) and Gram negative (Pseudomonas aeruginosa, Escherichia coli and Proteus mirabilis). Live/Dead staining and image analysis results indicated that LST can significantly reduce biofilm coverage of the five tested bacterial species on the CP Ti surfaces. Overall biofilm coverage as a percentage of the surface reduced after laser treatment averaging from 4.24 % to 17.4 %. This meant that relative to the untreated surface, biofilm coverage was reduced after laser treatment, ranging from 84.9 % to 95.6 % across the five species. Furthermore, cytotoxicity results (using MTT assay) showed that the laser-treated CP Ti is non-toxic across both L929 fibroblast and RAW macrophage cell lines. Surface properties after LST were investigated using WLI and AFM (measuring micro-/nano-surface roughness and topography) as well as ToF-SIMS (measuring surface chemistry and oxide thickness), respectively. The cross-sectional microstructure at the surface was imaged using SEM and analysed by XRD, whilst the surface wettability was measured using sessile drop method. To summarise, the reduction of biofilm coverage can be attributed to the favourable changes in surface roughness and topography together with the increased concentration of oxides at the topmost surface after LST.