The efficacy of saline irrigation for the treatment of periprosthetic infection (PJI) is limited in the presence of infected implants. This study evaluated the efficacy of vancomycin/tobramycin-doped polyvinyl alcohol (PVA)/ceramic composites (PVA-VAN/TOB-P) after saline irrigation in a mouse pouch infection model. 3D printed porous titanium (Ti) cylinders (400, 700 and 100 µm in pore size) were implanted into mice pouches, then inoculated with S. aureus at the amounts of 1X10. 3. CFU and 1X10. 6. CFU per pouch, respectively. Mice were randomized into 4 groups (n=6 for each group): (1) no bacteria; (2) bacteria without saline wash; 3) saline wash only, and (4) saline wash+PVA-VAN/TOB-P. After seven days, pouches were washed out alone or with additional injection of 0.2 ml of PVA-VAN/TOB-P. Mice were sacrificed 14 days after pouch wash. Bacteria cultures of collected Ti cylinders and washout fluid and histology of pouch tissues were performed. The low-grade infection (1X10. 3. CFU) was more significant in 400 µm Ti cylinders than that in Ti cylinders with larger pore sizes (700 and 1000 µm (p<0.05). A similar pattern of high-grade infection (1X10. 6. CFU) was observed (p<0.05). For the end wash, the bacteria burden (0.49±0.02) in saline wash group was completely eradicated by the addition of PVA-VAN/TOB-P (0.005±0.001, p<0.05). We noticed that 400 µm Ti cylinders have the highest risk of implant infection. Our data supported that the effect of saline irrigation was very limited in the presence of contaminated porous Ti cylinders. PVA-VAN/TOB-P was biodegradable, biocompatible, and was effective in eradicating bacteria retention after saline irrigation in a mouse model of low grade and high-grade infection. We believe that PVA-VAN/TOB-P represents an alternative to reduce the risk of PJI by providing a sustained local delivery of antibiotics

Summary Statement. CXCR4 gene and protein expression is regulated in a dose and time-dependent manner by metallic wear debris but not polyethylene wear debris in vitro and in vivo. Introduction. Progressive osteolysis leading to aseptic loosening among metal-on-metal (MoM) total hip arthroplasties (THA's), and adverse reactions to metallic debris (ARMD) are increasing causes for concern among existing patients who have been implanted with MoM hip replacements. Close surveillance of these patients is necessary and difficulties lie in early detection as well as differentiating low-grade infection from ARMD in the early stages. Several inflammatory markers have been investigated in this context, but to date, none is specific with regards to the offending material. In earlier studies, it has been shown that osteoblastic phenotypes and differentiation are regulated by different types of wear particles. Methods. In vitro experiments were performed using MG63 and SaOs-2 osteoblast-like cells co-cultured with increasing concentrations of metallic (Co-35Ni-20Cr-10Mo and Co-28Cr-6Mo) and polyethylene (UHMWPE-GUR1020) particles simulating periprosthetic wear debris. Real-time Polymerase Chain Reaction (RT-PCR) and Western Blotting were used to quantify gene and protein expression of CXCR4. The expression of TNF-a and the effects of AMD3100 on both CXCR4 and TNF-a expression among these cells was also investigated. Immunohistochemical techniques were used to investigate the in-vivo expression of CXCR4 in retrieval tissues obtained from 2 cohorts of failed metal-on-metal and ceramic-on-polyethylene THA's. Results. In-vitro RT-PCR and experiments demonstrated a dose-dependent increase in CXCR4 mRNA (7.5 fold for MG63 and 4.0 fold for SaOs-2 cells) among cells co-cultured with metal alloy particles. Western blotting also showed a time-dependent increase in protein expression of CXCR4. No regulatory effects on CXCR4 gene expression were seen among cells co-cultured with UHMWPE particles. The attempted blockade of CXCR4 by it's known competitive receptor agonist AMD3100 (bicyclam) led to a significant inhibition of metal particle induced TNF-a mRNA expression. In-vivo immunohistochemical data from the 2 cohorts of patients with failed THA's showed CXCR4 positivity among 83% of patients with metal-on-metal hip replacements but none among ceramic-on-polyethylene hip replacements. Discussion/Conclusion. CXCR4, the chemokine receptor for the chemokine SDF-1 (stromal cell derived factor-1), has been shown to play a pivotal role in bone metastasis, inflammatory and autoimmune conditions but has not been investigated in the context of periprosthetic osteolysis in failed joint replacements. Our in-vivo and in-vitro findings collectively suggest that the CXCR4 chemokine is specifically upregulated in a dose and time-dependent manner in the presence of metallic (cobalt-chrome) wear debris but not by polyethylene wear debris. The CXCR4 chemokine receptor may be a selective and specific biomarker for progressive osteolysis seen in failed MoM hip replacements and this phenomenon could potentially have a translational effect on the practice of orthopaedic surgery. Further research is needed to evaluate the interactions of CXCR4 with osteoclast activation and signalling pathways

Summary Statement. In vivo microCT allows monitoring of subtle bone structure changes around infected implants in a rat model. Introduction. The principal causes of orthopedic implant revisions are periprosthetic bone loss and infections. Immediately after implantation, a dynamic process of bone formation and resorption takes place around an orthopedic implant, influencing its mechanical fixation. Despite its importance, the effect of bacteria on the temporal pattern of periprosthetic remodeling is still unknown. The aim of this study was to evaluate the morphological changes of bone adjacent to an implant in the presence and absence of infection using micro computed tomography (microCT). Materials and methods. Twenty-four three-month-old female Wistar rats were used in this study. Twelve rats received a single control screw (sterile) in the proximal part of the right tibia while the other twelve received an infected screw (1×10. 4. CFU Staphylococcus aureus). The self-tapping cancellous bone screws, custom made of PEEK and coated with 30µm of titanium, were 2mm in outer diameter and 5mm in length. Bone changes around the screws were assessed using in vivo microCT with a nominal isotropic resolution of 12mm (at 70 kV, 300 ms integration time, 1000 projections) at days 0, 3, 6, 9, 14, 20 and 27. Each measurement took approximately 30 min while the animal was anesthetised via isoflurane inhalation. After reconstruction, these data were registered in space. The screw was segmented and dilated to define a region surrounding the coating. Bone-implant contact (BIC) was defined as the bone volume fraction (BV/TV) within this region. The changes in bone structure were computed from the differences between two consecutive time points. After sacrifice, in each group six tibiae were prepared for histology and six were used for mechanical pullout of the screw from the tibia, then quantitative microbiological analysis was carried-out after homogenization of the bone sample and sonication of the screw. Results. In the control group, no animal showed an infection, while all animals in the infected group developed an infection. In the uninfected group, BIC increased from 35±5% to 55±10% between day 0 and day 27 (p<0.05); at day 27 pullout stiffness was 220±48 N/mm and the maximal force 120±16 N. The microstructural changes were most prominent between day 0 and day 14. In the infected group, BIC dramatically dropped to zero within 14 days and the animals were sacrificed. Histology revealed that in the infected group there was marked osteolysis, purulent inflammation and a fibrous capsule around the screws. The pullout stiffness and maximal force were not significant (respectively 39±54 N/mm and 12±16 N). While 1×10. 4. CFU were introduced at day 0, at day 27, microbiological analysis revealed 1×10. 6. CFU on the screws and 5×10. 5. CFU in the neighboring bone. Conclusion. High-resolution in vivo microCT shows in the current model a rapid progression of osteolysis. This new approach allows a better understanding of the changes in bone structure around S. aureus infected implants. It may be particularly useful in detecting low-grade infections, such as S. epidermidis infections in the same model