header advert
Results 21 - 40 of 61
Results per page:
Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 66 - 66
1 Dec 2018
Karbysheva S Di Luca M Butini ME Trampuz A
Full Access

Aim

To compare the performance of sonication and chemical methods (EDTA and DTT) for biofilm removal from artificial surface.

Method

In vitro a mature biofilms of Staphylococcus epidermidis (ATCC 35984) and P. aeruginosa ATCC®53278) were grown on porous glass beads for 3 days in inoculated brain heart infusion broth (BHI). After biofilm formation, beads were exposed to 0.9% NaCl (control), sonication (40 kHz, 1 min, 0.2 W/cm2), EDTA (25 mM/15 min) and DTT (1 g/L/15 min). Quantitative and qualitative biofilm analysis were performed with viable counts (CFU/ml) and microcalorimetry using time to detection (TTD), defined as the time from insertion of the ampoule into the calorimeter until the exponentially rising of heat flow signal exceeded 100 μW, which is inversely proportional to the amount of remaining bacterial biofilm on the beads. All experiments were performed in triplicate.


Aim

Aim of this study was to evaluate the ability of Sb-1 to enhance the antibiotic activity (tested in combination) degrading the biofilm matrix (impairing the freely diffusion of antimicrobials) and specifically targeting “persister” cells (biofilm sub-population tolerant to most antibiotics and responsible for the infection recalcitrance) of methicillin-resistant Staphylococcus aureus.

Method

MRSA ATCC 43300 24h-old biofilm was treated for 18h with Sb-1 titers (from 104 to 106 pfu/ml). Biofilm matrix was evaluated by confocal laser scanning microscopy after staining with wheat germ agglutinin conjugate with Alexafluor488 (WGA488) to label exopolysaccharide matrix and Syto 85 to label bacterial cells. Persister status was induced using two different protocols: i) by exposing stationary phase S. aureus to 400 µg/ml carbonyl cyanide m-chlorophenylhydrazone (CCCP) in PBS for 3h at 37°C and ii) by treatment of 24h old biofilm with 512 µg/ml ciprofloxacin for further 24h at 37°C. Then, induced persister cells and non-induced controls (106 CFU/ml) were treated with 104 PFU/ml and 107 PFU/ml Sb-1 for 3h, followed by CFU counting. Alternatively, bacteria were washed and incubated in fresh BHI medium for the resumption of normal growth and the bacterial growth assessed after further 24 hours.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 29 - 29
1 Dec 2018
Margaryan D Renz N Kendlbacher P Vajkoczy P Trampuz A
Full Access

Aim

Spinal implant-associated infections (SIAI) require combined surgical and antimicrobial treatment and prolonged hospital stay. We evaluated the clinical, laboratory, microbiological and radiological characteristics and treatment approaches in patients with SIAI.

Method

Consecutive adult patients with SIAI treated between 2015 and 2017 were prosepctively included. SIAI was defined by: (i) significant microbial growth from intraoperative tissue or sonication fluid, (ii) intraoperative purulence, secondary wound dehiscence or implant on view, (iii) radiographic evidence of infection and fever (>38°C) without other recognized cause, increasing back pain or neurologic impairment, (iv) peri-implant tissue inflammation in histopathology.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 58 - 58
1 Dec 2018
Sigmund IK Önder N Winkler T Perka C Trampuz A Renz N
Full Access

Aim

Two stage revision is the most commonly used surgical treatment strategy for periprosthetic hip infections (PHI). The aim of our study was to assess the intra- and postoperative complications during and after two stage revision using resection arthroplasty between ex- and reimplantation.

Method

In this retrospective cohort study, all patients treated with a two stage revision using resection arthroplasty for PHI were included from 2008 to 2014. During the first stage, the prosthesis was removed resulting in a resection arthroplasty without the use a PMMA spacer. During second stage, (cemented or uncemented) reimplantation of the hip prosthesis was performed. The cohort was stratified into two groups according to the length of prosthesis-free interval (≤10 weeks and >10 weeks). Data on complications during explantation, prosthesis-free interval, reimplantation, and after reimplantation was collected. The overall complication rate between both groups was compared using the chi-squared test. The revision-free and infection-free survival was estimated using Kaplan-Meier survival analysis.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 65 - 65
1 Dec 2018
Tkhilaishvili T Di Luca M Trampuz A
Full Access

Aim

Staphylococcus aureus and Pseudomonas aeruginosa are ubiquitous pathogens often found together in polymicrobial, biofilm-associated infections. The mixed-species biofilm are significantly more resistant to antimicrobial treatment and are associated with failures. Bacteriophages present a promising alternative to treat biofilm-related infections due to their rapid bactericidal activity on multi-drug resistant bacteria. In this study, we assess the simultaneous or sequential application of phages and ciprofloxacin on the mixed-species biofilm in vitro.

Method

Ciprofloxacin was tested alone and in combination with Pyo-bacteriophage cocktail against P.aeurginosa ATCC 27853 and MRSA ATCC 43300 mixed-species biofilm. In order to evaluate the effect of combined treatment on biofilm-embedded cells, mature biofilms were grown on porous glass beads with MRSA (106 CFU/ml) and P.aeruginosa (103 CFU/ml) and incubated for 24h at 37° C in LB broth. The beads were then washed and placed in fresh LB in the presence of sub-eradicating titers/concentrations of phages and ciprofloxacin (corresponding to 1/4, 1/8, 1/16, 1/32, 1/64, 1/128 × MBECbiofilm), respectively, simultaneous or in order (pretreated with phages for 3-6-12-24 hours) at 37°C. In all cases, heat flow produced by the viable cells still embedded in the biofilm was measured for 48 hours by isothermal microcalorimetry


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 20 - 20
1 Dec 2018
Ojeda-Thies C Li C Renz N Trampuz A
Full Access

Aim

Radiologic signs such as radiolucent lines around the implant, hardware fracture or displacement and periosteal reaction have been considered suggestive of implant-associated infection. The goal of this study is to assess the correlation of these signs with confirmed internal fixation-associated infection evaluated in a prospective cohort.

Method

We evaluated the radiologic appearance of preoperative standard x-ray images in 421 surgeries performed in 380 patients with internal fixation device in place (56.8% male, mean age 53 ± 17 years). This prospective study was performed in a large single center for musculoskeletal surgery from 2013–2017. Infection was suspected preoperatively in only 23.8% of the surgeries. The most common indications for surgeries in which infection was not suspected were nonunion (84 cases) and symptomatic hardware (57 cases). All removed implants were sent to sonication for biofilm removal and detection. In addition, several peri-implant tissue samples were collected. Radiographs were analyzed in a blinded fashion for signs of radiolucent lines around the implant before removal, hardware fracture or displacement, and soft periosteal reactions suggestive of infection. Diagnosis was established according to the IDSA criteria for PJI. Contingency tables were constructed to determine sensitivity and specificity, and to perform Chi-square tests to compare the presence of infection with radiological signs of infection.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 10 - 10
1 Dec 2018
Renz N Trebse R Akgün D Perka C Trampuz A
Full Access

Aim

Treatment of enterococcal periprosthetic joint infections (PJI) is challenging due to heterogeneous pathogenesis, non-standardized management strategies and lack of biofilm-active antibiotics. Previous studies report treatment success from 50–76%. We evaluated the characteristics and outcome of enterococcal PJI, in particular the influence of antimicrobial treatment regimens.

Method

Consecutive patients with enterococcal PJI treated at two specialized orthopaedic institutions were retrospectively included from 2010 to 2017. PJI was defined by the proposed European Bone and Joint Infection Society (EBJIS) criteria. Adequate antimicrobial treatment was considered when the antibiotic was appropiate for the treatment of enterococcal bone infections (activity, dose, oral bioavailability, bone penetration). The treatment success (defined as no relapse of enteroccal infection) and clinical success(i.e. infection-free status) was evaluated and compared using Fishers exact test.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 35 - 35
1 Dec 2018
Karbysheva S Schütz M Trampuz A
Full Access

Aim

To assess the analytical performance and to establish the cut-off of synovial fluid D-lactate concentration for the diagnosis of prosthetic joint infection (PJI) and septic arthritis (SA) using commercial kits provided by two manufacturers (A and B).

Method

We prospectively included patients with native or prosthetic joints undergoing synovial fluid aspiration as routine diagnostic procedure. Among 224 patients with prosthetic joints, 137 patients had aseptic loosening (AL) and 87 were diagnosed with PJI. Among 71 patients with native joints, 39 were diagnosed with osteoarthrosis (OA) and 32 with SA.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 60 - 60
1 Dec 2018
Ojeda-Thies C Li C Renz N Trampuz A
Full Access

Aim

Unexpected positive infections are distinct entity in prosthetic revision surgery. The prevalence and characteristics of unexpected positive cultures in internal fixation are however less established. The aim of this study was to describe the prevalence and characteristics of unexpected diagnosis of infection in a prospective cohort of revision surgeries following internal fixation.

Method

We reviewed the microbiological results following 356 surgeries that included partial or complete removal of internal fixation, performed in 328 patients (54% male, mean age 53 ± 17 years), in which infection was not initially suspected. This prospective study was performed in a large single center for musculoskeletal surgery from 2013–2017. The implants most commonly removed were plate and/or screws (281 cases, 78,9%), followed by intramedullary nails (64 cases, 18,0%). The main indications for surgery were nonunion (89 cases, 25%) and symptomatic hardware (70 cases, 19,7%). All removed implants were sonicated, and tissue cultures were obtained depending on the surgeon's criteria. Diagnosis of infection was established by the presence of 2 or more positive tissue cultures (1 with a highly virulent microorganism), or ≥ 50 colony-forming units found in the sonication fluid.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_1 | Pages 20 - 20
1 Jan 2018
Renz N Mudrovcic S Trampuz A Perka C
Full Access

Cutibacterium species (formerly Propionibacterium species) are increasingly recognized as causative pathogens of low-grade periprosthetic joint infections (PJI). The clinical manifestation of infections caused by this low virulent microorganism is nonspecific and the interpretation remains challenging. In this prospective cohort study from 01/2012 to 07/2017 we analyzed the clinical and diagnostic characteristics of microbiologically proven hip PJI caused by Cutibacterium species. PJI was defined by growth of Cutibacteria in ≥2 periprosthetic tissue samples or in sonication fluid of the removed implant (>50 CFU/ml) at revision surgery. If Cutibacteria grew only in synovial fluid at least one other positive microbiological specimen or non-microbiological criterion was required.

We included 26 patients suffering from Cutibacterium hip PJI, among them 19 were males (73%). The majority of PJI (24, 92%) presented delayed (3–24 months) or late (>24 months) after implantation. Sinus tract was present in 4 patients (16%) and radiological implant loosening in 16 patients (62%). Among non-microbiological diagnostic tests, increased synovial fluid leukocyte count showed the highest sensitivity (82%), followed by tissue histology (71%) and serum C-reactive protein (58%). After 7 days of incubation Cutibacterium grew in synovial fluid, periprosthetic tissue and sonication fluid culture in 20%, 42% and 32%, respectively, and in 43%, 76% and 83%, respectively, after 14 days of incubation.

We conclude that Cutibacterium PJI was diagnosed late in the disease course and presented with subtle clinical signs. Prolonged culture incubation and implant sonication improved the poor performance of conventional microbiological tests. Due to lack of reliable diagnostic tests, Cutibacterium remains difficult to detect making the diagnosis challenging.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 8 - 8
1 Dec 2017
Tkhilaishvili T Di Luca M Trampuz A Gaudias J
Full Access

Aim

The increase of antimicrobial resistance reduces treatment options for implant-associated infections caused by methicillin-resistant Staphylococcus aureus (MRSA). Bacteriophages present a promising alternative to treat biofilm-related infections due to their rapid bactericidal activity and activity on multi-drug resistant bacteria. In this study, we investigated the synergistic activity of lytic bacteriophage Sb-1 with different antibiotics against MRSA biofilm, using a real-time highly sensitive assay measuring growth-related heat production (microcalorimetry).

Methods

Rifampin, fosfomycin, vancomycin and daptomycin were tested alone and in combination with S. aureus specific phage, Sb-1, against MRSA (Staphylococcus aureus*). MRSA biofilm was formed on porous glass beads (Φ 4 mm, pore size 60 µm) and incubated for 24 h at 37° C in BHI. After 3 times washing biofilms were exposed first to different titers of bacteriophages, ranging from 102 to104 plaque-forming unite (pfu)/ml and after 24h treated again with subinhibitory concentration of antibiotics (corresponding to 1/4, 1/8, 1/16, 1/32 × MHICbiofilm). After 24h antibiotic treatment, the presence of biofilm on glass beads was evaluated by isothermal microcalorimetry for 48h. Heat flow (µW) and total heat (J) were measured.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 100 - 100
1 Dec 2017
Karbysheva S Yermak K Grigoricheva L Trampuz A
Full Access

The diagnosis of prosthetic-joint infection (PJI) is challenging, as bacteria adhere on implant and form biofilm. Therefore, current diagnostic methods, such as preoperative culture of joint aspirate have limited sensitivity with false-negative results.

Aim

To evaluate the performance of measurement synovial fluid (SF) D-lactate (as a pathogen-specific marker) for the diagnosis of PJI and estimate of treatment success.

Method

224 patients undergoing removal knee or hip prosthesis were included in the study between January 2015 and March 2017. 173 patients of this group had aseptic loosening of prosthesis and 87 were diagnosed with PJI. Prior to surgery, synovial fluid routine culture, D-lactate test, leukocyte count and neutrophils (%) were performed for each patient. In order to evaluate a treatment success, the measurement of SF D-lactate before second two-stage exchange procedure (after treatment) was implemented in 30 patients. Diagnosis of PJI was established according to modified Zimmerli criteria.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 36 - 36
1 Dec 2017
Trampuz A Klatt A Luca MD
Full Access

Aim

Phage therapy has attracted attention as a promising alternative treatment option for biofilm infections.

To establish a successful phage therapy, a comprehensive stock of different phages covering a broad bacterial spectrum is crucial. We screened human and environmental sources for presence of lytic phages against selected bacteria.

Methods

Saliva collected from 10 volunteers and 500 ml of sewage water were screened for the presence of lytic phages active against 20 clinical strains of Staphylococcus aureus and 10 of Escherichia coli, both isolated from patients with prosthetic joint infection. Laboratory strains of methicillin-resistant S. aureus (MRSA)*1 and E. coli*2 were also tested. Screening was performed plaque-assay to detect phages for different strains. Isolated plaques were collected and phages were enriched to determine their activity against their bacterial host strains. The activity of bacteriophages against adherent E. coli and MRSA was evaluated by crystal violet, staining bacterial biofilms grown on glass beads.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 53 - 53
1 Dec 2017
Akgün D Rakow A Perka C Trampuz A Renz N
Full Access

Background

Periprosthetic joint infections (PJI), caused by pathogens, for which no biofilm-active antibiotics are available, are often referred to as difficult-to-treat (DTT). It is unclear whether DTT PJI has worse outcome due to unavailability of biofilm-active antibiotics. We evaluated the outcome of DTT and non-DTT PJI managed according to a standardized treatment regimen.

Methods

Patients with hip and knee PJI from 2013 to 2015 were prospectively included and followed-up for ≥2 years. DTT PJI was defined as growth of microorganism(s) resistant to biofilm-active antibiotics. The Kaplan-Meier survival analysis was used to compare the probability of infection-free survival between DTT and non-DTT PJI.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 76 - 76
1 Dec 2017
Renz N Mudrovcic S Perka C Trampuz A
Full Access

Aim

To assess the clinical characteristics, diagnostic tests and treatment strategies in orthopedic implant-associated infections (OIAI) caused by Cutibacterium spp.

Method

We retrospectively included consecutive patients with OIAI caused by Cutibacterium spp. treated at our institution from January 2012 to January 2017. OIAI was diagnosed when: (i) macroscopic purulence, sinus tract or exposed implant was present; (ii) acute inflammation in peri-implant-tissue was documented; (iii) Cutibacterium spp. grew in joint aspirate, ≥2 intraoperative peri-implant tissue samples or in sonication fluid of the removed implant (>50 CFU/ml).


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 84 - 84
1 Dec 2017
Rakow A Perka C Akgün D Schütz M Trampuz A Renz N
Full Access

Aim

The incidence of hematogenous periprosthetic joint infections (hPJI) is unknown and the cases probably largely underreported. Unrecognized and untreated primary infectious foci may cause continuous bacteremia, further spread of microorganisms and thus treatment failure or relapse of infection. This study aimed at improving knowledge about primary foci and microbiological characteristics of this entity to establish preventive measures and improve diagnostic and therapeutic strategies to counteract hPJI.

Method

We retrospectively analysed all consecutive patients with hPJI, who were treated at our institution from January 2010 until December 2016. Diagnosis of PJI was established if 1 of the following criteria applied:(i) macroscopic purulence, (ii) presence of sinus tract, (iii) positive cytology of joint aspirate (>2000 leukocytes/μl or >70% granulocytes), (iv) significant microbial growth in synovial fluid, periprosthetic tissue or sonication culture of retrieved prosthesis components, (v) positive histopathology. PJI was classified as hematogenous if the following criteria were fulfilled additionally: (1) onset of symptoms more than 1 month after arthroplasty AND (2) i) isolation of the same organism in blood cultures OR ii) evidence of a distant infectious focus consistent with the pathogen.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 18 - 18
1 Dec 2017
Sigmund IK Renz N Yermak K Trampuz A
Full Access

Aim

Alpha-defensin is a new synovial fluid biomarker for the diagnosis of periprosthetic joint infections (PJI). We compared the performance of two different alpha-defensin assays: quantitative ELISA test and qualitative lateral flow test.

Method

In this prospective cohort study, consecutive patients with a painful prosthesis of the lower limb were eligible for inclusion. In addition to standard diagnostics of PJI, alpha-defensin was determined in the aspirated synovial fluid between October 2016 and April 2017. PJI was defined according to the modified Zimmerli criteria, the Musculoskeletal Infection Society (MSIS) criteria and the Infectious Disease Society of America (IDSA) criteria. A positive quantitative alpha-defensin test was defined at a cut-off value of 5.2 mg/L. The sensitivity, specificity, accuracy and area under the curve of each test were determined and the AUCs were compared among each other.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 19 - 19
1 Dec 2017
Renz N Yermak K Perka C Trampuz A
Full Access

Aim

The aim of the study was to assess the accuracy of the alpha defensin lateral flow test for diagnosis of periprosthetic joint infection (PJI) using an optimized diagnostic algorithm and three classification systems. In addition, we compared the performance with synovial fluid leukocyte count, the most sensitive preoperative test.

Method

In this prospective multicenter study we included all consecutive patients with painful prosthetic hip and knee joints undergoing diagnostic joint aspiration. Alpha defensin lateral flow test was used according to manufacturer instructions. The following diagnostic criteria were used to confirm infection: Musculoskeletal Infection Society (MSIS), Infectious Diseases Society of America (IDSA) and Swiss orthopedics and Swiss Society of Infectious Diseases (SOSSID). In the latter, PJI was confirmed when at least one of following criteria applied: macroscopic purulence, sinus tract, positive cytology of joint aspirate (>2000 leukocytes/μl or >70% granulocytes), histological proof of acute inflammation in periprosthetic tissue, positive culture (from aspirate, tissue or sonication fluid). Infection was classified as chronic, if symptom duration was more than 3 weeks or if infection manifested after more than 1 month after surgery. The sensitivity and specificity of the alpha defensin lateral flow test and leukocyte count in synovial fluid were calculated and compared using McNemar Chi-square test.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 11 - 11
1 Dec 2017
Kocjancic B Jeverica S Trampuz A Simnic L Avsec K Dolinar D
Full Access

Aim

The aim of our study was to evaluate culture-negative prosthetic joint infections in patients who were pre-operatively evaluated as aseptic failure.

Method

For the purpose of the study we included patients planed for revision surgery for presumed aseptic failure. Intraoperatively acquired samples of periprosthetic tissue and explanted prosthesis were microbiologicaly evaluated using standard microbiologic methods and sonication. If prosthetic joint infection was discovered, additional therapy was introduced.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 26 - 26
1 Dec 2017
Vacha E Deppe H Wantia N Trampuz A
Full Access

Aim

The risk of haematogenic periprosthetic joint infection (PJI) after dental procedures is discussed controversially. To our knowledge, no study has evaluated infections according to the origin of infection based on the natural habitat of the bacteria. We investigated the frequency of positive monomicrobial cultures involving bacteria from oral cavity in patients with suspected PJI compared to bone and joint infections without joint prosthesis.

Method

In this retrospective study we included all patients with suspected PJI or bone and joint infection without endoprosthesis, hospitalized at our orthopaedic clinic from January 2009 through March 2014. Excluded were patients with superficial surgical site infections or missing data. Demographic, clinical and microbiological data were collected using a standardized case report form. Groups were compared regarding infections caused by oral bacteria. χ2 test or Fisher's exact test was employed for categorical variables and t-test for continuous variables.