bluuebunny/crossref_metadata_2025_split huggingface dataset:
ANTIBACTERIAL PROPERTIES OF NISIN LAYER-BY-LAYER BASED COATING ON TITANIUM K-WIRES IN GALLERIA MELLONELLA IMPLANT-ASSOCIATED INFECTION MODEL
Authors: Mannala Gopala , Labat Beatrice , Ladam Guy , Pascal Thebault , Walter Nike , Szymski Dominik , Riool Martijn , Alt Volker , Rupp Markus 
Published: November 2024

AimOrthopedic implants play a tremendous role in fixing bone damages due to aging as well as fractures. However, these implants tend to get colonized by bacteria on the surface, leading to infections and subsequently prevention of healing and osteointegration. Recently, Roupie et al. showed that a nisin layer-by-layer based coating applied on biomaterials has both osteogenic and antibacterial properties. The Galleria mellonella larva is a well-known insect infection model that has been used to test the virulence of bacterial and fungal strains as well as for the high throughput screening of antimicrobial compounds against infections. Recently, we have developed an insect infection model with G. mellonella larvae to study implant-associated biofilm infections using Kirschner (K)-wires as implant material. Here, we would like to test the antibacterial capacity of nisin layer-by-layer based coatings on K-wires against Staphylococcus aureus in the G. mellonella larva implant infection model.MethodPrior to the implantation procedure, G. mellonella larvae are maintained at room temperature on wheat germ in an incubator. The larvae received bare titanium K-wires (uncoated), or either control-coated or nisin-coated K-wires. After one hour, the larvae were injected with 5×105S. aureus bacteria per larva (i.e., hematogenous implant infection model). Next, the larvae were incubated at 37oC in an incubator and the survival of the larvae was monitored for five days. Moreover, the number of bacteria on the implant surface and in the surrounding tissue was determined after 24h of incubation. Further, scanning electron microscopy (SEM) analyses were performed to study the effect of nisin on biofilm formation.ResultsThe larvae receiving the nisin-coated K-wires showed significantly higher survival rates compared to uncoated titanium K-wires, although not when compared to control-coated K-wires. A more than 1-log reduction in number of bacteria on the implant surface and in the surrounding tissue was observed in larvae receiving the nisin-coated K-wires, when compared to uncoated titanium K-wires SEM analysis showed reduced colonization of the bacteria nisin-coated K-wires compared to the controlsConclusionsIn conclusion, the antimicrobial nisin layer-by-layer based coating applied on titanium surfaces is able to prevent implant-related S. aureus biofilm infection in G. mellonella and is a promising antimicrobial strategy to prevent implant-related infections.
COMBINATION OF SONICATION WITH DITHIOTHREITOL TREATMENT DOES NOT IMPROVE BACTERIAL DISLODGING FROM BIOFILM IN AN IN VITRO MODEL
Authors: De Vecchi Elena , Riccardi Mila , Mastroianni Nicolò 
Published: November 2024

AimDiagnosis of prosthetic joint infection are often complicated by the presence of biofilm, which hampers bacteria dislodging from the implants, thus affecting sensitivity of cultures. In the last 20 years several studies have evidenced the usefulness of implant sonication to improve microbial recovery from biofilm formed on inert substrates. More recently, treatment of prosthetic joints and tissues with Dithiothreitol, a sulphur compound already used in routine diagnostic workflow for fluidification of respiratory samples, has proved to be not inferior to sonication in microbiological diagnosis of prosthetic joint infections.This study aimed to evaluate if the combination of the two treatments could further improve microbial retrieval from biofilm in an in vitro model.MethodThree isolates of Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus lugdunensis, Eschericha coli and Pseudomonas aeruginosa responsible of prosthetic joint infections were used. They were grown onto 3 titanium discs (20 mm diameter) and incubated in 3 sterile plastic containers with 15 mL of Triptyc Soy Broth. After overnight incubation, not adhered cells were removed and fresh broth was added to each sample. After 48 hours incubation, the exausted broth was removed and one sample was used for sonication, one for treatment with 0,1% (v:v) Dithiothreitol and one treated with Dithiothreitol followed by sonication. Treated fluids were plated on Muller Hinton Agar plates for colony count.One-way ANOVA analysis was performed to evidence statistical differences between treatments.ResultsSimilar colony counts were observed for the 3 treatments: 10.1± 0.77 log CFU/mL for Dithiothreitol, 10.0 ± 0.75 for sonication and 10.1 ±0.73 for dithiothreitol + sonication.No statistical differences between the 3 treatments were evidenced by ANOVA analysis.ConclusionsResults seems to confirm that treatment with dithiothreitol is equivalent to sonication in recovering bacteria from biofilm grown on inert surface. Combining dithiotreitol treatment with sonication does not significantly improve bacterial recovery in respect to each treatment alone.
INOCULATION OF HOMOGENIZED TISSUE AND BONE BIOPSIES IN BLOOD CULTURE BOTTLES FOR DIAGNOSING ORTHOPAEDIC DEVICE-RELATED INFECTIONS: PRELIMINARY RESULTS FROM AN ONGOING STUDY
Authors: Jacob Ann-Sophie , Onsea Jolien , Bessems Laura , Spoormans Pauline , Vles Georges , Metsemakers Willem-Jan , Depypere Melissa 
Published: November 2024

AimThe primary objective is to evaluate the diagnostic performance of inoculating homogenized tissue and bone biopsies in blood culture bottles (BCB) for patients with (suspected) orthopaedic device-related infections. As secondary objective the time to positivity (TTP) of BCB and Wilkins-Chalgren broth (conventional method) will be evaluated.MethodPatients undergoing revision surgery due to suspected or proven fracture-related infection (FRI) or periprosthetic joint infection (PJI) according to respectively Consensus definition and EBJIS definition are included.1,2 A minimal of three macroscopic infected/inflamed tissue/bone samples are collected in a container with saline and glass beads. 1.5 mL of the homogenized suspension is inoculated in BacT/ALERT FA and FN Plus bottles for 14 days. The remaining suspension is inoculated in Wilkins-Chalgren broth for 10 days and subcultured when cloudy or after 10 days. TTP is defined as the time until definite identification of the pathogen in the Laboratory Information System.ResultsUp to now, 25 patients have been included, 11 (44%) had concordant results in BCB and the CM. In 11 patients cultures showed negative results for both methods. Three patients tested positive with BCB but remained negative with the same pathogen in CM. In the first patient, the CM failed to identify anaerobic bacteria (i.e. Fusobacterium nucleatum). In the second patient, three BCB were positive with Staphylococcus capitis. The third patient showed an infection with Escherichia coli, which was detected in all samples from the BCB, while all cultures obtained with the CM remained negative. A possible explanation for this discrepancy could be that this patient already received antibiotic therapy. BCB contain resins, which are capable of neutralizing antibiotic activity. Another case illustrating superiority of BCB involved an infection with Cutibacterium acnes, which showed positivity in six BCB, while only three were positive using the CM. We observed the shortest TTP with BCB. The median TTP of BCB was 32.0 hours (IQR 29.8) compared to a median TTP of 77.5 hours (IQR 107.6) when culturing with the CM.Contamination was seen in three patients with both methods, in eight patients contamination was only seen with the CM. For the remaining 14 patients no contamination was found.ConclusionsThe results in this ongoing study indicate that the recovery of pathogens and TTP is better using BCB compared to CM. In addition, contamination occurs less frequently with the BCB method. Culturing tissue or bone biopsies in BCB seems a promising and faster detection method.
DEVELOPMENT AND VALIDATION OF A MULTIPLEX PCR FOR THE DETECTION OF THE MOST COMMON PATHOGENS IN PERIPROSTHETIC JOINT INFECTIONS
Authors: Erbeznik Anja , Smrdel Katja Strašek , Kišek Tjasa Cerar , Cvitković-Špik Vesna , Triglav Tina , Vodicar Polona Maver 
Published: November 2024

AimThe aim of this study was to develop an in-house multiplex PCR real-time assay on the LightCycler 480 system (Roche, Basel, Switzerland) with the aim of rapid detection of common pathogens in prosthetic joint infections (PJI), followed by validation on clinical samples (sonication fluid and tissue biopsies) routinely collected for PJI diagnosis.MethodsUsing the PrimerQuest and CLC WorkBench tool, we designed six primer sets with specific fluorescently labelled TaqMan probes for the nuc gene in different Staphylococcus species (S. aureus, S. epidermidis, S. capitis, S. lugdunensis, S. hominis, S. haemolyticus). In addition, primers previously developed by Renz et al. (2022) for C. acnes were integrated into our assay with internal control of isolation, leading to the development of specific mPCR assay with seven included targets. Analytical sensitivity and specificity were evaluated using reference bacterial strains. To determine the assay's limit of detection (LOD), we conducted serial dilutions of eluates containing known concentrations of bacterial DNA copies/µl. The overall LOD in spiked clinical samples, including sample preparation and DNA isolation on MagnaPure24, was measured through 10-fold serial dilutions (from 109 to 10-1 CFU/ml) including additional dilutions of 5000, 500, 50 and 5 CFU/ml.ResultsThe results with LOD in serial dilutions of eluates and spiked clinical samples, together with analytical sensitivity and specificity, are shown in Table 1.ConclusionThe mPCR assay showed excellent analytical sensitivity and specificity, but with considerably lower LOD after sample preparation and further DNA isolation in spiked clinical samples. Although still promising in diagnostics of acute infections, the use of mPCR could be challenging in chronic, low-grade infections with lower microbial burden. Nevertheless, PCR offers significant advantages in terms of speed and can shorten the time to result, especially for C. acnes infections. Additionally, it represents a promising complementary approach in patients with suspected PJI on antibiotic therapy with negative culture results.For any tables or figures, please contact the authors directly.
CAN MOLECULAR TECHNIQUES BECOME THE NEW “GOLD STANDARD” IN DIAGNOSTICS OF PERIPROSTHETIC JOINT INFECTIONS?
Authors: Erbeznik Anja , Šturm Andraž Celar , Smrdel Katja Strašek , Triglav Tina , Cvitković-Špik Vesna , Kišek Tjasa Cerar , Kocjancic Boštjan , Pompe Borut , Dolinar Drago , Mavcic Blaz , Mercun Aljaz , Kolar Matic , Avsec Klemen , Papst Lea , Vodicar Polona Maver 
Published: November 2024

AimWe prospectively evaluated four different microbiological tools for diagnostics of prosthetic joint infections (PJI), and assessed their impact on the categorization of infection according to EBJIS guidelines. We compared culture, in-house real-time mPCR for S. aureus, S. lugdunensis, S. hominis, S. epidermidis, S. capitis, S. haemolyticus, C. acnes (mPCR), broad-spectrum PCR (Molzym) with 16S rRNA V3-V4 amplicon Sanger sequencing (16S PCR), and 16S rRNA V3-V4 amplicon next-generation sequencing (16S NGS) on MiSeq (Ilumina).MethodsA total of 341 samples (sonication fluid, tissue biopsy, synovial fluid) were collected from 32 patients with suspected PJI who underwent 56 revision surgeries at the Orthopaedic Centre University Hospital Ljubljana, between 2022 and 2024. Samples were processed using standard protocols for routine culture, followed by DNA isolation using the MagnaPure24 (Roche). All samples were tested with mPCR, and an additional ≥4 samples from each revision (244 in total) were subjected to further metagenomic analysis. Culture results were considered positive if the same microorganism was detected in ≥2 samples, ≥50 CFU/ml were present in the sonication fluid, or ≥1 sample was positive for a more virulent microorganism or if the patient had received antibiotic treatment.ResultsEach tool demonstrated high sensitivity for correct EBJIS categorization (100% culture and 16S NGS, 96.88% mPCR and 16S PCR). The highest specificity was observed with mPCR and 16S PCR (87.5%), while culture (79.17%) and NGS (37.5%) showed lower specificity. In 27% (15/56) of revisions, all microbiological tests were negative, although infection was confirmed with histology in one case, and four cases were classified as infection-likely based on clinical signs. In 20% (11/56) of cases, all microbiological tests were positive; in three cases a combination of other EBJIS criteria (without microbiology) categorized the episodes as infection-likely and one as infection-unlikely, emphasizing the importance of microbiological tests in diagnostic criteria. In 43% (24/56) of revisions categorized as infection-unlikely using a combination of other EBJIS criteria, five had positive culture, and three had positive mPCR and 16S PCR. Fifteen (62%) had positive 16S NGS, 12 due to a low number of reads, which may indicate low-grade infection or possible contamination.ConclusionTo date, no test can be established as the ultimate gold standard. The lack of interpretation criteria can result in low specificity of some methods, as the threshold is difficult to determine. A multidisciplinary approach with combination of microbiological tools is still considered the most efficient.
NOVEL MOLECULAR APPROACH IS USEFUL IN CULTURE-NEGATIVE PERIPROSTHETIC HIP AND KNEE JOINT INFECTIONS
Authors: Gardete-Hartmann Susana , Sebastian Sujeesh , Berdalli Sofia , Simon Sebastian , Hofstaetter Jochen 
Published: November 2024

AimUnexpected negative-cultures (UNC) are a common diagnostic problem in periprosthetic joint infection (PJI) of the hip and knee when using culture-based methods. A novel molecular approach (MC)1 based on the identification of the vast majority of bacterial species in a single assay using species-specific bacterial interspacing region length polymorphisms and phylum-specific 16S rDNA sequence polymorphisms has demonstrated clinical utility in PJI diagnostics (1). In addition, MC provides an estimate of the leukocyte concentration in the specimen analysed. The aim of this retrospective, blinded study was to evaluate the performance of MC in identifying the microbiological content and determining the leukocyte count in synovial fluid (SF) collected from hip and knee revision arthroplasty cases with UNC. It was also assessed whether antibiotic treatment would have been changed if the result from MC had been known.MethodA total of 89 SF samples from 70 patients (43 female; 27 male) who underwent revision arthroplasty (14 hip; 75 knee) were included. Using European and Bone Joint Infection Society (EBJIS) criteria, 82 cases were classified as infected (77 UNC and 5 septic culture-positive controls), five as non-infected (aseptic culture-negative controls), and two as likely infected, but infected by clinical observation. MC was performed and evaluated together with SF parameters. Antibiotic treatment, clinical outcome, patient demographics and surgical details were analysed.ResultsOverall, 29.1% (23/79) of UNC had a positive yield by MC, of which 2/23 (8.7%) had two microorganisms detected simultaneously. Of the 25 microorganisms identified by MC, 12/25 (48%) were clinically relevant after re-evaluation of the patients’ microbiological history. The microorganisms detected were 5/25 (20%) Streptococcus pneumoniae/mitis, 4/25 (16%) Staphylococcus epidermidis, 3/25 (12%) Cutibacterium acnes, 3/25 (12%) Streptococcus agalactiae, 2/25 (8%) Streptococcus bovis, 2/25 (8%) Staphylococcus aureus, and 2/25 (8%) Haemophilus parainfluenzae. The prevalence of Enterococcus faecalis, Bacteroides fragillis, Staphylococcus lugdunensis, Corynebacterium striatum among all MC results was 1/25 (4%) each species. In total, 13/23 (56%) cases were associated with patients receiving antibiotic therapy at the time of SF collection. The yield for leukocyte counts provided the molecular technique was consistently much higher in the UNC and clearly septic groups than in the clearly aseptic group. Overall, 20/61 (32.8%) patients with UNC could have been managed differently and more accurately after MC assessment.ConclusionsMC shows clinical value in the diagnosis and management of PJI with UNC. The included leukocyte count shows promising results.AcknowledgmentsThis work was partially funded by Inbiome.
IS ONE DOSE NON-INFERIOR TO FOUR DOSES OF SYSTEMIC ANTIBIOTIC PROPHYLAXIS AGAINST PERIPROSTHETIC JOINT INFECTION IN PRIMARY ARTHROPLASTY?
Authors: Lutro Olav , Tjørhom Marianne Bollestad , Fenstad Anne Marie , Leta Tesfaye H. , Hallan Geir , Bruun Trond , Furnes Ove , Gjertsen Jan-Erik , Dale Håvard 
Published: November 2024

AimThe current recommendation in Norway is to use four doses of a first-generation cephalosporin (cefazolin or cephalotin) as systemic antibiotic prophylaxis (SAP) the day of surgery in primary joint arthroplasty. Due to shortage of supply, scientific development, changed courses of treatment and improved antibiotic stewardship, this recommendation has been disputed. We therefore wanted to assess if one dose of SAP was non-inferior to four doses in preventing periprosthetic joint infection (PJI) in primary joint arthroplasty.MethodWe included patients with primary hip- and knee arthroplasties from the Norwegian Arthroplasty Register and the Norwegian Hip Fracture Register for the period 2005-2023. We included the most used SAPs (cephalotin, cefazolin, cefuroxime, cloxacillin and clindamycin), administered as the only SAP in 1-4 doses, starting preoperatively. Risk of revision (Hazard rate ratio; HRR) for PJI was estimated by Cox regression analyses with adjustment for sex, age, ASA class, duration of surgery, reason for- and type of arthroplasty, and year of primary arthroplasty. The outcome was 1-year reoperation or revision for PJI. Non-inferiority margins were calculated for 1, 2 and 3 doses versus reference of 4 doses of SAP at the day of surgery, against a predetermined limit of 15% increased risk of PJI.ResultsIn total 274,188 primary arthroplasties (total hip 133,985, hemi hip 51,442, and total knee 88,761) were included. Of these primary arthroplasties, 2,996 (1.1%) had subsequent revisions for PJI during the first postoperative year. One dose of SAP was given in 9,603 arthroplasties, two doses in 10,068, three doses in 18,351, and four doses in 236,166 arthroplasties. With the recommended four doses as reference, the HRR (95% CI) for 1-year revision for infection was 0.9 (0.7-1.1) for one dose, 1.0 (0.8-1.2) for two doses, and 0.9 (0.8-1.1) for three doses. The corresponding adjusted 1-year revision incidences for PJI was 0.9 (0.7-1.1), 1.0 (0.8-1.2), 0.9 (0. 8-1.1) and 1.0 (1.0-1.1) for one, two, three and four doses respectively, and less than four doses was found to be non-inferior.ConclusionsOne preoperative dose of SAP in primary joint arthroplasty surgery seems to be non-inferior to the current recommendation of four doses of a first-generation cephalosporin as PJI-prophylaxis. This finding may simplify the course of treatment for arthroplasty patients, save costs, and improve antibiotic stewardship.
POSTOPERATIVE ANTIBIOTIC TREATMENT DOES NOT LOWER RE-REVISION RATE IN PRESUMED ASEPTIC HIP AND KNEE REVISION ARTHROPLASTIES WITH UNEXPECTED POSITIVE INTRAOPERATIVE CULTURES: A PROPENSITY SCORE-MATCHED COHORT STUDY
Authors: Simon Sebastian , Wouthuyzen-Bakker Marjan , Mitterer Jennyfer A , Gardete-Hartmann Susana , Frank Bernhard J.H. , Hofstaetter Jochen 
Published: November 2024

AimIt still remains unclear whether postoperative antibiotic treatment is advantageous in presumed aseptic revision-arthroplasties of the hip (rTHA) and knee (rTKA) with unexpected-positive-intraoperative-cultures (UPIC). The aim of this study was to evaluate if there is a difference in the septic and/or aseptic re-revision rate in patients with or without postoperative antibiotics.MethodIn this retrospective propensity-score (PS) matched cohort-study we compared the re-revision rate and the microbiological spectrum in rTHA and rTKA treated with (AB-Group; n=70) and without (non-AB-Group; n=70) antibiotic treatment in patients with UPIC. Baseline covariates for PS-matching were type of revision, sex, Body-Mass-Index, age, Surgical-Site-Infection-Score, American-Society-of-Anesthesiologists-Classification, serum C-reactive-protein.All patients received routine antibiotic prophylaxis, but empiric AB treatment was started only in patients in the AB-Group. Post-operative treatment was decided on an individual basis according to the preference of the surgeon and the infectious disease specialist for a minimum duration of two weeks. In total, 90 rTHA (45 AB-Group, 45 in non-AB-Group) patients with UPICs and 50 rTKA (25 AB-Group, 25 in non-AB-Group) were included in the study. There was no significant variation in patient demographics.ResultsAfter a median follow-up of 4.1 (IQR: 2.9-5.5) years after rTHA and rTKA, there was no higher re-revision rate (p=0.813) between the AB-group 10/70 (14.3%), and the non-AB-group 11/70 (15.7%). In the AB group, 4.3% (3/70) of patients underwent revision due to septic complications compared to 5.7% (4/70) in the non-AB group (survival log-rank: p=0.691).In total, 30/70 (42.9%) of patients in the AB-group and 23/70 (32.9%) of patients in the non-AB group were diagnosed as having an “infection likely” according to the PJI diagnostic criteria of EBJIS (p=0.223). All UPICs comprised low virulent microorganisms and were considered as a contaminant. In total, 68/70 (97.1%) of the patients in the AB-group received a dual antibiotic treatment for a mean duration of 41 (IQR: 23.5-56.5) days.ConclusionPostoperative antibiotic treatment did not result in a decreased re-revision rate compared to non-antibiotic treatment in patients with UPIC in presumed aseptic rTHA and rTKA. UPICs with pathogens are likely to be a containment and therefore the classification of “infection likely” according to the EBJIS definition can be safely ignored.