Method

Mature methicillin-resistant Staphylococcus aureus biofilms were grown on Ti-6Al-4V coupons by adding 1mL of a 8Log10 ATCC 33591 suspension in TGN (TSB + 1% glucose + 2% NaCl) to 24-wells plates containing the coupons and incubating the plates for 24h at 37°C with a continuous 50rpm agitation. The samples were rinsed and placed in 6 wells plates containing 1ml of the enzymatic cocktail (C.D.D.) solution (tris-buffered (pH 7.0) solution of 400 U/ml of aspecific DNA/RNA endonuclease, 50 U/ml of endo-1,4-b-D-glucanase, and 0.06 U/ml of β-N-acetylhexosaminidase). 9ml of TGN or TGN containing antibiotics RIF/VAN (rifampicin 5µg/mL + vancomycin 8µg/mL) at clinically relevant concentrations found locally in bone or joints, was then added and the samples were incubated in identical conditions for 24h. The samples were then recovered and rinsed. CFU counts were obtained by recovering the bacteria with sonication, serial dilutions, and TSA plating. Biomass was determined via crystal violet staining, followed by dye solubilization in acetic acid, and absorbance measurement using a spectrophotometer.

Material and methods

Medical files of 208 patients (101 knees and 107 hips) who underwent a two-stage revision for late PJI prosthesis infection were retrospectively reviewed. Antibiotic loaded articulated homemade cement spacers were used. Second stage was usually planned on average 4 to 6 weeks after the first stage. Patients were allowed to walk without loading. The success rate was defined as the absence of septic recurrence after a minimum follow-up of two years. Descriptive statistics and uni- and multivariate analysis were conducted.

Material and Methods

We retrospectively reviewed 140 prosthetic joint infection (PJI) treated with a two stage procedure. Patient data has been reviewed to determine which factors would be predictive for failure.

Method

This investigation was set up as a multicenter prospective cohort study. Synovial fluid was obtained by means of joint aspiration or intra-operative tissue biopsies. A presumptive PJI diagnosis was made according to criteria outlined by the Musculoskeletal Infection Society (MSIS), the Infectious Diseases Society of America (IDSA) and the European Bone and Joint Infection Society (EBJIS). The intention to treat by the surgeon was logged. Sensibility and specificity for the ADLF test was plotted for each aforementioned diagnostic algorithm. Spearman correlations between all scores were analyzed. Multiple logistic regression was used to determine the contribution of independent variables to the probability of PJI.

Method

Strains: 1 reference (MSSA: ATCC25923; MRSA: ATCC33591) and 2 clinical MSSA and MRSA isolated from PJI.

Biofilm culture: Coupons were incubated for 24h at 37°C with bacteria (starting inoculum ∼6.6Log₁₀CFU/mL in TGN [TSB + 1% glucose + 2% NaCl]), under shaking at 50rpm.

Treatment: Half of the coupons were irrigated with 50mL physiological serum from 5cm using a Stryker Interpulse; the coupons were then either analysed (ControlT0 and PWT0) or reincubated for 24h in TGN or TGN containing flucloxacillin (MSSA) or vancomycin (MRSA) at MIC or 20mg/L.

Analysis: Coupons were rinsed twice with PBS. Biomass was measured by crystal violet (CV) assay. CFUs were counted after recovering bacteria from coupons using sonication and TSA plating.

Materials and Methods

We reviewed all DAIR procedures for hip PJI performed between 2002–2017 (n=69). Data recorded included all factors correlated with treatment failure. KLIC score and an adapted McPherson score (sum of three criteria where the lower score is three for 3 A and the higher is 9 for 3 C) were analyzed.