Duration of antibiotic treatment and risk of recurrence after surgical management of orthopaedic device infections: a multicenter case-control study.

BACKGROUND: Device-related infections in orthopaedic and trauma surgery are a devastating complication with substantial impact on morbidity and mortality. Systemic suppressive antibiotic treatment is regarded an integral part of any surgical protocol intended to eradicate the infection. The optimal duration of antimicrobial treatment, however, remains unclear. In a multicenter case-control study, we aimed at analyzing the influence of the duration of antibiotic exposure on reinfection rates 1 year after curative surgery. METHODS: This investigation was part of a federally funded multidisciplinary network project aiming at reducing the spread of multi-resistant bacteria in the German Baltic region of Pomerania. We herein used hospital chart data from patients treated for infections of total joint arthroplasties or internal fracture fixation devices at three academic referral institutions. Subjects with recurrence of an implant-related infection within 1 year after the last surgical procedure were defined as case group, and patients without recurrence of an implant-related infection as control group. We placed a distinct focus on infection of open reduction and internal fixation (ORIF) constructs. Uni- and multivariate logistic regression analyses were employed for data modelling. RESULTS: Of 1279 potentially eligible patients, 269 were included in the overall analysis group, and 84 contributed to an extramedullary fracture-fixation-device sample. By multivariate analysis, male sex (odds ratio [OR] 2.06, 95% confidence interval [CI] 1.08 to 3.94, p = 0.029) and facture fixation device infections (OR 2.05, 95% CI 1.05 to 4.02, p = 0.036) remained independent predictors of reinfection. In the subgroup of infected ORIF constructs, univariate point estimates suggested a nearly 60% reduced odds of reinfection with systemic fluoroquinolones (OR 0.42, 95% CI 0.04 to 2.46) or rifampicin treatment (OR 0.41, 95% CI 0.08 to 2.12) for up to 31 days, although the width of confidence intervals prohibited robust statistical and clinical inferences. CONCLUSION: The optimal duration of systemic antibiotic treatment with surgical concepts of curing wound and device-related orthopaedic infections is still unclear. The risk of reinfection in case of infected extramedullary fracture-fxation devices may be reduced with up to 31 days of systemic fluoroquinolones and rifampicin, although scientific proof needs a randomized trial with about 1400 subjects per group. Concerted efforts are needed to determine which antibiotics must be applied for how long after radical surgical sanitation to guarantee sustainable treatment success.

Antibiotics release from cement spacers used for two-stage treatment of implant-associated infections after total joint arthroplasty.

Two-stage revision arthroplasty is the treatment of choice for periprosthetic infection, a serious complication after knee or hip arthroplasty. Our prospective clinical trial aimed to investigate the concentrations of gentamicin and vancomycin in wound exudate and tissue in two-stage revision arthroplasty. Wound exudate and periprosthetic membrane samples were collected from 18 patients (10 hip and eight knee patients), who were due for two-stage treatment after a periprosthetic joint infection. Samples were taken during insertion of antibiotic-impregnated spacers and after their removal. The concentrations of gentamicin and vancomycin in wound exudates and adjacent tissue were analyzed using high-performance liquid chromatography mass spectrometry. Average time period of spacer implantation was 13.6 weeks (9.3-22.6 weeks). The concentration of vancomycin in wound exudate decreased from a median of 43.28 µg/mL (0.28-261.22) after implantation to 0.46 µg/mL (0.13-37.47) after the removal of the spacer. In the adjacent tissue, vancomycin concentration was mainly undetectable prior to spacer implantation (0.003 µg/g [0.003-0.261]) and increased to 0.318 µg/g [0.024-484.16] at the time of spacer removal. This was also observed for gentamicin in the tissue of patients who previously had cement-free implants (0.008 µg/g [0.008-0.087] vs. 0.164 µg/g [0.048-71.75]) while in the tissue of patients with previously cemented prosthesis, baseline concentration was already high (8.451 µg/g [0.152-42.926]). Despite the rapid decrease in antibiotics release from spacer cement observed in vitro, in vivo antibiotics are much longer detectable, especially in the adjacent soft tissue. © 2018 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials Published By Wiley Periodicals, Inc. J Biomed Mater Res B Part B, 2019. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1587-1597, 2019.

Damages with High Consequences: Analysis of Perforations in Surgical Latex Operation Gloves from Orthopedic Surgeries.

INTRODUCTION: To prevent surgical site infections (SSIs) during operation, the use of sterile surgical latex gloves is common. The aim of this study was to examine the damage of the gloves in surgeries with different mechanical stress and the influence on the kind of damages. Gloves were collected during primary arthroplasty, revision arthroplasty (hip and knee), and arthroscopy (shoulder, hip, and knee). MATERIALS AND METHODS: Surgical latex operation gloves were collected from surgeons after the operation and were tested with watertightness test (ISO EN 455-1:2000). RESULTS: A total of 1460 surgical gloves were retrieved from 305 elective operations. On average, 15.9% of the gloves showed postoperative lesions, with the highest incidence occurring in revision arthroplasty with 25%. In primary and revision arthroplasty, the index finger of the dominant hand was most frequently affected (62.7% and 58.6%); in contrast, gloves from arthroscopies had most lesions on thumb and middle finger (42.9% each). Tear and perforation size differed from ≤1 mm to >5 mm, and primary and revision arthroplasty showed bigger damages. CONCLUSIONS: Surgical gloves have a high malfunction, which increases with growing mechanical stress. A high rate of perforation occurred mostly in revision arthroplasty. Breaching the integrity of the gloves, especially by high mechanical loads, could lead to an increased rate of infection.

Copper as an alternative antimicrobial coating for implants - An in vitro study.

AIM: To investigate osteoconductive and antimicrobial properties of a titanium-copper-nitride (TiCuN) film and an additional BONIT® coating on titanium substrates. METHODS: For micro-structuring, the surface of titanium test samples was modified by titanium plasma spray (TPS). On the TPS-coated samples, the TiCuN layer was deposited by physical vapor deposition. The BONIT® layer was coated electrochemically. The concentration of copper ions released from TiCuN films was measured by atomic absorption spectrometry. MG-63 osteoblasts on TiCuN and BONIT® were analyzed for cell adhesion, viability and spreading. In parallel, Staphylococcus epidermidis (S. epidermidis) were cultivated on the samples and planktonic and biofilm-bound bacteria were quantified by counting of the colony-forming units. RESULTS: Field emission scanning electron microscopy (FESEM) revealed rough surfaces for TPS and TiCuN and a special crystalline surface structure on TiCuN + BONIT®. TiCuN released high amounts of copper quickly within 24 h. These release dynamics were accompanied by complete growth inhibition of bacteria and after 2 d, no planktonic or adherent S. epidermidis were found on these samples. On the other hand viability of MG-63 cells was impaired during direct cultivation on the samples within 24 h. However, high cell colonization could be found after a 24 h pre-incubation step in cell culture medium simulating the in vivo dynamics closer. On pre-incubated TiCuN, the osteoblasts span the ridges and demonstrate a flattened, well-spread phenotype. The additional BONIT®coating reduced the copper release of the TiCuN layer significantly and showed a positive effect on the initial cell adhesion. CONCLUSION: The TiCuNcoating inhibits the formation of bacterial biofilms on orthopedic implants by influencing the "race for the surface" to the advantage of osteoblasts.

Thin magnesium layer confirmed as an antibacterial and biocompatible implant coating in a co­culture model.

Implant-associated infections commonly result from biofilm­forming bacteria and present severe complications in total joint arthroplasty. Therefore, there is a requirement for the development of biocompatible implant surfaces that prevent bacterial biofilm formation. The present study coated titanium samples with a thin, rapidly corroding layer of magnesium, which were subsequently investigated with respect to their antibacterial and cytotoxic surface properties using a Staphylococcus epidermidis (S. epidermidis) and human osteoblast (hOB) co­culture model. Primary hOBs and S. epidermidis were co­cultured on cylindrical titanium samples (Ti6Al4V) coated with pure magnesium via magnetron sputtering (5 µm thickness) for 7 days. Uncoated titanium test samples served as controls. Vital hOBs were identified by trypan blue staining at days 2 and 7. Planktonic S. epidermidis were quantified by counting the number of colony forming units (CFU). The quantification of biofilm­bound S. epidermidis on the surfaces of test samples was performed by ultrasonic treatment and CFU counting at days 2 and 7. The number of planktonic and biofilm­bound S. epidermidis on the magnesium­coated samples decreased by four orders of magnitude when compared with the titanium control following 7 days of co­culture. The number of vital hOBs on the magnesium­coated samples was observed to increase (40,000 cells/ml) when compared with the controls (20,000 cells/ml). The results of the present study indicate that rapidly corroding magnesium­coated titanium may be a viable coating material that possesses antibacterial and biocompatible properties. A co­culture test is more rigorous than a monoculture study, as it accounts for confounding effects and assesses additional interactions that are more representative of in vivo situations. These results provide a foundation for the future testing of this type of surface in animals.

Fast corroding, thin magnesium coating displays antibacterial effects and low cytotoxicity.

Bacterial colonisation and biofilm formation are characteristics of implant-associated infections. In search of candidates for improved prosthetic materials, fast corroding Mg-based coatings on titanium surfaces were examined for their cytotoxic and antimicrobial properties. Human osteoblasts and Staphylococcus epidermidis were each cultured on cylindrical Ti samples coated with a thin layer of Mg/Mg45Zn5Ca, applied via magnetron sputtering. Uncoated titanium samples served as controls. S. epidermidis was quantified by counting colony forming units. The biofilm-bound fraction was isolated via ultrasonic treatment, and the planktonic fraction via centrifugation. Biofilm-bound S. epidermidis was significantly decreased by approximately four to five orders of magnitude in both Mg- and Mg45Zn5Ca-coated samples after seven days compared to the control. The osteoblast viability was within the tolerance threshold of 70% stated in DIN EN ISO 10993-5:2009-10 for Mg (~80%) but not for Mg45Zn5Ca (~25%). Accordingly, Mg-coated titanium was identified as a promising candidate for an implant material with antibacterial properties and low cytotoxicity levels. The approach of exploiting fast corrosion contrasts with existing methods, which have generally focused on reducing corrosion.

Co-Culture of S. epidermidis and Human Osteoblasts on Implant Surfaces: An Advanced In Vitro Model for Implant-Associated Infections.

OBJECTIVES: Total joint arthroplasty is one of the most frequent and effective surgeries today. However, despite improved surgical techniques, a significant number of implant-associated infections still occur. Suitable in vitro models are needed to test potential approaches to prevent infection. In the present study, we aimed to establish an in vitro co-culture setup of human primary osteoblasts and S. epidermidis to model the onset of implant-associated infections, and to analyze antimicrobial implant surfaces and coatings. MATERIALS AND METHODS: For initial surface adhesion, human primary osteoblasts (hOB) were grown for 24 hours on test sample discs made of polystyrene, titanium alloy Ti6Al4V, bone cement PALACOS R®, and PALACOS R® loaded with antibiotics. Co-cultures were performed as a single-species infection on the osteoblasts with S. epidermidis (multiplicity of infection of 0.04), and were incubated for 2 and 7 days under aerobic conditions. Planktonic S. epidermidis was quantified by centrifugation and determination of colony-forming units (CFU). The quantification of biofilm-bound S. epidermidis on the test samples was performed by sonication and CFU counting. Quantification of adherent and vital primary osteoblasts on the test samples was performed by trypan-blue staining and counting. Scanning electron microscopy was used for evaluation of topography and composition of the species on the sample surfaces. RESULTS: After 2 days, we observed approximately 104 CFU/ml biofilm-bound S. epidermidis (103 CFU/ml initial population) on the antibiotics-loaded bone cement samples in the presence of hOB, while no bacteria were detected without hOB. No biofilm-bound bacteria were detectable after 7 days in either case. Similar levels of planktonic bacteria were observed on day 2 with and without hOB. After 7 days, about 105 CFU/ml planktonic bacteria were present, but only in the absence of hOB. Further, no bacteria were observed within the biofilm, while the number of hOB was decreased to 10% of its initial value compared to 150% in the mono-culture of hOB. CONCLUSION: We developed a co-culture setup that serves as a more comprehensive in vitro model for the onset of implant-associated infections and provides a test method for antimicrobial implant materials and coatings. We demonstrate that observations can be made that are unavailable from mono-culture experiments.

Prospective data collection and analysis of perforations and tears of latex surgical gloves during primary endoprosthetic surgeries.

Introduction: Surgical gloves are used to prevent contamination of the patient and the hospital staff with pathogens. The aim of this study was to examine the actual effectiveness of gloves by examining the damage (perforations, tears) to latex gloves during surgery in the case of primary hip and knee prosthesis implantation. Materials and methods: Latex surgical gloves used by surgeons for primary hip and knee replacement surgeries were collected directly after the surgery and tested using the watertightness test according to ISO EN 455-1:2000. Results: 540 gloves were collected from 104 surgeries. In 32.7% of surgeries at least one glove was damaged. Of all the gloves collected, 10.9% were damaged, mainly on the index finger. The size of the perforations ranged from ≤1 mm to over 5 mm. The surgeon's glove size was the only factor that significantly influenced the occurrence of glove damage. Surgeon training level, procedure duration, and the use of bone cement had no significant influence. Conclusions: Our results highlight the high failure rate of surgical gloves. This has acute implications for glove production, surgical practice, and hygiene guidelines. Further studies are needed to detect the surgical steps, surface structures, and instruments that pose an increased risk for glove damage.

A Novel In Vitro System for Comparative Analyses of Bone Cells and Bacteria under Electrical Stimulation.

Electrical stimulation is a promising approach to enhance bone regeneration while having potential to inhibit bacterial growth. To investigate effects of alternating electric field stimulation on both human osteoblasts and bacteria, a novel in vitro system was designed. Electric field distribution was simulated numerically and proved by experimental validation. Cells were stimulated on Ti6Al4V electrodes and in short distance to electrodes. Bacterial growth was enumerated in supernatant and on the electrode surface and biofilm formation was quantified. Electrical stimulation modulated gene expression of osteoblastic differentiation markers in a voltage-dependent manner, resulting in significantly enhanced osteocalcin mRNA synthesis rate on electrodes after stimulation with 1.4VRMS. While collagen type I synthesis increased when stimulated with 0.2VRMS, it decreased after stimulation with 1.4VRMS. Only slight and infrequent influence on bacterial growth was observed following stimulations with 0.2VRMS and 1.4VRMS after 48 and 72 h, respectively. In summary this novel test system is applicable for extended in vitro studies concerning definition of appropriate stimulation parameters for bone cell growth and differentiation, bacterial growth suppression, and investigation of general effects of electrical stimulation.

Augmented recovery of microorganisms from swabs by homogenization: a novel standardizable high-throughput approach.

A new approach introducing a quantitative and standardizable step into sample processing was evaluated by homogenizing in vitro inoculated swab tips with Precellys 24 high-throughput homogenizer. Recovery of microorganisms from homogenized swab tips was significantly higher as compared to conventional processing methods. Thus, swab homogenization is a promising approach introducing a new quality in microbial analysis.

Examination of cross contamination risks between hospitals by external medical staff via cross-sectional intercept survey of hand hygiene.

INTRODUCTION: Work in hospitals is supported by contributions of life sciences industry representatives (IR) in various ways of fields. Close contact between them, caretakers and patients is unavoidable, even in situations where hygiene is critical. The present study investigates whether IR display comparable levels of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) contamination after being exposed to a shared environment for a minimum of 4 hours. MATERIAL AND METHODS: An anonymous survey to sample a group of healthcare professionals for traces of fingertip contamination was performed. We used dip slides (S. aureus and MRSA) to evaluate 311 healthcare professionals at the medical exhibition MEDICA. After applying exclusion criteria 298 participants remained valid, they consisted of 208 industry representatives, 49 nurses and 41 physicians. RESULTS: IR where engaged in hospitals, operating rooms and outpatient clinics (82%, 41.8%, 51.9% respectively). 65.9% of IR (vs. 48.8% physicians and 40.8% nurses) carried a microbiological burden ≥10(4) CFU (colony forming units). Neither S. aureus (≥10(4) CFU) in IR (40.9%) did show statistical differences in contamination patterns in comparison to physicians (43.9%, p=0.346) and nurses (36.7%, p=0.878) nor did MRSA (physicians p=0.579, nurses p=0.908). We were unable to differentiate transient from pre-existing permanent colonization. CONCLUSION: Exposure to the same environment may result in similar hand contamination patterns of IR when compared caregivers. This supports the concern that industry representatives can cause cross infection between hospitals and hygiene sensitive areas like operation room, intensive care unit and central sterilization units particularly. Further study is required to clarify whether pre-existing bacterial colonization is an influencing factor and how industry is taking care of this to create a safe working environment for their employees, the customers and ultimately the patients.

Autoimmune pancreatitis in MRL/Mp mice is a T cell-mediated disease responsive to cyclosporine A and rapamycin treatment.

BACKGROUND: Autoimmune pancreatitis (AIP) in humans invariably responds to steroid treatment, but little is known about the underlying pathogenesis and the benefits of alternative treatments. OBJECTIVE: To study the pathogenesis, and the efficacy of alternative immunosuppressant agents in the MRL/Mp mouse model of AIP. DESIGN: MRL/Mp mice were pretreated for 4 weeks with polyinosinic:polycytidylic acid to induce AIP. Pancreatic sections of mice genetically deleted for CTLA-4 were analysed. Blockage of CTLA-4 was achieved by intraperitoneal antibody treatment with 2 µg/g anti-mouse-CD152. Subsequent therapeutic studies were performed for a period of 4 weeks using cyclosporine A (40 µg/g), rapamycin (1 µg/g) or azathioprine (15 µg/g). RESULTS: Blockage of CTLA-4 in MRL/Mp mice suppressed regulatory T cell (Treg) function and raised the effector T cell (Teff) response with subsequent histomorphological organ destruction, indicating that AIP is a T cell-driven disease. Using an established histopathological score, we found that dexamethasone, cyclosporine A and rapamycin, but less so azathioprine, reduced pancreatic damage. However, the beneficial effects of cyclosporine A and rapamycin were achieved via different mechanisms: cyclosporine A inhibited Teff activation and proliferation whereas rapamycin led to selective expansion of Tregs which subsequently suppressed the Teff response. CONCLUSIONS: The calcineurin inhibitor cyclosporine A and the mammalian target of rapamycin (mTOR) inhibitor, rapamycin, improve the course of AIP in MRL/Mp mice via different mechanisms. These findings further support the concept of autoreactive T cells as key players in the pathogenesis of AIP and suggest that cyclosporine A and rapamycin should be considered for treatment of AIP in humans.