Dual release of daptomycin and BMP-2 from a composite of ß-TCP ceramic and ADA gelatin.

BACKGROUND: Antibiotic-containing carrier systems are one option that offers the advantage of releasing active ingredients over a longer period of time. In vitro sustained drug release from a carrier system consisting of microporous ß-TCP ceramic and alginate has been reported in previous works. Alginate dialdehyde (ADA) gelatin gel showed both better mechanical properties when loaded into a ß-TCP ceramic and higher biodegradability than pure alginate. METHODS: Dual release of daptomycin and BMP-2 was measured on days 1, 2, 3, 6, 9, 14, 21, and 28 by HPLC and ELISA. After release, the microbial efficacy of the daptomycin was verified and the biocompatibility of the composite was tested in cell culture. RESULTS: Daptomycin and the model compound FITC protein A (n = 30) were released from the composite over 28 days. A Daptomycin release above the minimum inhibitory concentration (MIC) by day 9 and a burst release of 71.7 ± 5.9% were observed in the loaded ceramics. Low concentrations of BMP-2 were released from the loaded ceramics over 28 days.

Integrative tissue, cellular and molecular responsiveness of an innovative ex vivo model of the Staphylococcus aureus-mediated bone infection.

Osteomyelitis is a pathological condition of the bone, frequently associated with the presence of infectious agents - namely Staphylococcus aureus - that induce inflammation and tissue destruction. Recent advances in the understanding of its pathophysiology and the identification of innovative therapeutic approaches were gathered from experimental in vitro and in vivo systems. However, cell culture models offer limited representativeness of the cellular functionality and the cell-cell and cell-matrix interactions, further failing to mimic the three-dimensional tissue organization; and animal models allow for limited mechanistic assessment given the complex nature of systemic and paracrine regulatory systems and are endorsed with ethical constraints. Accordingly, this study aims at the establishment and assessment of a new ex vivo bone infection model, upon the organotypic culture of embryonic chicken femurs colonized with S. aureus, highlighting the model responsiveness at the molecular, cellular, and tissue levels. Upon infection with distinct bacterial inoculums, data reported an initial exponential bacterial growth, followed by diminished metabolic activity. At the tissue level, evidence of S. aureus-mediated tissue destruction was attained and demonstrated through distinct methodologies, conjoined with decreased osteoblastic/osteogenic and increased osteoclastic/osteoclastogenic functionalities-representative of the osteomyelitis clinical course. Overall, the establishment and characterization of an innovative bone tissue infection model that is simple, reproducible, easily manipulated, cost-effective, and simulates many features of human osteomyelitis, further allowing the maintenance of the bone tissue's three-dimensional morphology and cellular arrangement, was achieved. Model responsiveness was further demonstrated, showcasing the capability to improve the research pipeline in bone tissue infection-related research.

Automatic detection, segmentation, and classification of primary bone tumors and bone infections using an ensemble multi-task deep learning framework on multi-parametric MRIs: a multi-center study.

OBJECTIVES: To develop an ensemble multi-task deep learning (DL) framework for automatic and simultaneous detection, segmentation, and classification of primary bone tumors (PBTs) and bone infections based on multi-parametric MRI from multi-center. METHODS: This retrospective study divided 749 patients with PBTs or bone infections from two hospitals into a training set (N = 557), an internal validation set (N = 139), and an external validation set (N = 53). The ensemble framework was constructed using T1-weighted image (T1WI), T2-weighted image (T2WI), and clinical characteristics for binary (PBTs/bone infections) and three-category (benign/intermediate/malignant PBTs) classification. The detection and segmentation performances were evaluated using Intersection over Union (IoU) and Dice score. The classification performance was evaluated using the receiver operating characteristic (ROC) curve and compared with radiologist interpretations. RESULT: On the external validation set, the single T1WI-based and T2WI-based multi-task models obtained IoUs of 0.71 ± 0.25/0.65 ± 0.30 for detection and Dice scores of 0.75 ± 0.26/0.70 ± 0.33 for segmentation. The framework achieved AUCs of 0.959 (95%CI, 0.955-1.000)/0.900 (95%CI, 0.773-0.100) and accuracies of 90.6% (95%CI, 79.7-95.9%)/78.3% (95%CI, 58.1-90.3%) for the binary/three-category classification. Meanwhile, for the three-category classification, the performance of the framework was superior to that of three junior radiologists (accuracy: 65.2%, 69.6%, and 69.6%, respectively) and comparable to that of two senior radiologists (accuracy: 78.3% and 78.3%). CONCLUSION: The MRI-based ensemble multi-task framework shows promising performance in automatically and simultaneously detecting, segmenting, and classifying PBTs and bone infections, which was preferable to junior radiologists. CLINICAL RELEVANCE STATEMENT: Compared with junior radiologists, the ensemble multi-task deep learning framework effectively improves differential diagnosis for patients with primary bone tumors or bone infections. This finding may help physicians make treatment decisions and enable timely treatment of patients. KEY POINTS: • The ensemble framework fusing multi-parametric MRI and clinical characteristics effectively improves the classification ability of single-modality models. • The ensemble multi-task deep learning framework performed well in detecting, segmenting, and classifying primary bone tumors and bone infections. • The ensemble framework achieves an optimal classification performance superior to junior radiologists' interpretations, assisting the clinical differential diagnosis of primary bone tumors and bone infections.

Naringin exerts antibacterial and anti-inflammatory effects on mice with Staphylococcus aureus-induced osteomyelitis.

Osteomyelitis is an invasive bone infection that can lead to severe pain and even disability, posing a challenge for orthopedic surgery. Naringin can reduce bone-related inflammatory conditions. This study aimed to elucidate the function and mechanism of naringin in a Staphylococcus aureus-induced mouse model of osteomyelitis. Femurs of S. aureus-infected mice were collected after naringin administration and subjected to microcomputed tomography to analyze cortical bone destruction and bone loss. Bacterial growth in femurs was also assessed. Proinflammatory cytokine levels in mouse femurs were measured using enzyme-linked immunosorbent assays. Pathological changes and bone resorption were analyzed using hematoxylin and eosin staining and tartrate-resistant acid phosphatase staining, respectively. Quantitative reverse transcription polymerase chain reaction and western blot analysis were used to quantify the messenger RNA and protein expression of osteogenic differentiation-associated genes in the femurs. The viability of human bone marrow-derived stem cells (hBMSCs) was determined using cell counting kit-8. Alizarin Red S staining and alkaline phosphatase staining were performed to assess the formation of mineralization nodules and bone formation in vitro. Notch signaling-related protein levels in femur tissues and hBMSCs were assessed using western blot analysis. Experimental results revealed that naringin alleviated S. aureus-induced cortical bone destruction and bone loss in mice by increasing the bone volume/total volume ratio. Naringin suppressed S. aureus-induced bacterial growth and inflammation in femurs. Moreover, it alleviated histopathological changes, inhibited bone resorption, and increased the expression of osteogenic markers in osteomyelitic mice. It increased the viability of hBMSCs and promoted their differentiation and bone mineralization in vitro. Furthermore, naringin activated Notch signaling by upregulating the protein levels of Notch1, Jagged1, and Hes1 in the femurs of model mice and S. aureus-stimulated hBMSCs. In conclusion, naringin reduces bacterial growth, inflammation, and bone resorption while upregulating the expression of osteogenic markers in S. aureus-infected mice and hBMSCs by activating Notch signaling.

Detection of Cutibacterium (Propionibacterium) acnes in orthopaedic surgery: serious problem or contamination?

PURPOSE: Bone and joint infections are an important and increasing problem. Whether intraoperatively detected bacteria should be considered relevant or not is often difficult to assess. This retrospective cohort study analyzes the relevance of C. acnes cultured from deep intraoperative specimens. METHODS: All deep tissue samples collected intraoperatively between 2015 and 2020 from a quartiary care provider were evaluated for detection of C. acnes and its therapeutical consequences. Infection rates were determined according to a standardized definition and protocol and analyzed in dependence of patient's demographic data (age and gender), operative parameters (type of surgery, body region/location of surgery, and impression of the surgeon), and initiated therapy. RESULTS: In 270 cases of more than 8500 samples, C. acnes was detected. In 30%, the detection was considered an infection. The number of samples taken and tested positive for C. acnes correlated significantly with its classification as a cause of infection. If more than one sample of the patient was positive, the detection was significantly more likely to be treated as infection (p < 0.001). In 76% of cases, a consultation to the infectious diseases (ID) department took place regarding the classification of the pathogen detection and the therapy to be carried out. Almost all of the tested isolates demonstrated the wild-type susceptibility for penicillin and clindamycin. CONCLUSION: Intraoperative detection of skin-colonizing bacteria such as C. acnes is not always synonymous with infection. In particular, if other examination results contradict an infection (pathological sample without evidence of an infectious event, detection of malignant cells, etc.), the situation must be considered in a very differentiated manner. Interdisciplinary boards, for example, are suitable for this purpose. Care should be taken to obtain a sufficiently large number of tissue samples for microbiological examination to be able to better classify the result.

Augmentation with a non-vascularized autologous fibular graft for the management of Cierny-Mader type IV chronic femoral osteomyelitis: a salvage procedure.

PURPOSE: The study aimed to evaluate preliminary clinical and radiographic results of patients with Cierny-Mader type IV chronic femoral osteomyelitis and augmented with a non-vascularized fibular autograft as a salvage procedure because of the poorly regenerated new bone after bone transport over an intramedullary nail (BTON). METHODS: Patients diagnosed with CM type IV chronic femoral bone infection and treated with BTON procedure between 2003 and 2020 were retrospectively reviewed. Seven patients were included in the study whose distraction gap was poorly regenerated and then augmented with a non-vascularized fibular autograft. A three-stage treatment was administered. First, the infection was eradicated. Second, BTON was performed. Third, the poorly regenerated distraction gap was augmented with a fibular autograft before removing the external fixator (EF). Clinical and radiological results were evaluated based on the criteria described by Paley-Maar and Li classification. RESULTS: The mean patient age was 52 years. The mean treatment time was 24.8 months, with a mean femoral lengthening of 12.6 cm. The mean EF and bone healing indexes were 0.57 months/cm and 0.8 months/cm, respectively. The mean length of the fibular graft was 13 cm. The bone healing of new bones was achieved in all patients with good quality after grafting. Functional scores were excellent in four patients. No patients experienced any sequelae. CONCLUSIONS: Non-vascularized fibular autograft augmentation may be an effective salvage procedure for poorly regenerated new bone after BTON to manage chronic femoral bone infection.

Etiology and antimicrobial resistance patterns in chronic osteomyelitis of the tibia: an 11-year clinical experience.

PURPOSE: To analyze changes in tendency of etiology and of antimicrobial resistance patterns to most common local and systemic antibiotics in chronic osteomyelitis of the tibia (COM-T) in a Level I trauma center over an 11-year period. METHODS: A retrospective review including all patients with COM-T who were surgically treated from January 2009 to December 2019. Patients were divided into two period groups: 2009-2014 and 2015-2019. Microbiologic etiology was analyzed. Bacterial resistance patterns evaluation was based on the Magiorakos et al. classification, including proportions of multidrug-resistant organisms (MDROs, acquired non-susceptibility to at least one agent in three or more antimicrobial categories), extensively drug-resistant (XDR) and pan drug-resistant (PDR) organisms encountered. RESULTS: A total of 173 episodes of COM-T were identified. Monomicrobial infections represented 47.4% of all cases, while 28.3% had polymicrobial infections. Negative deep-bone cultures were identified in 24.3% of the patients. The most commonly isolated microorganisms were coagulase-negative Staphylococci (24.5%) and S. aureus (20.5%). No differences were found when comparing Gram-positive infections between periods (58.3% for 2009-2014 vs. 46.7% for 2015-2019; p = 0.10). Findings were similar for Gram-negative infections (37% vs. 33.7%; p = 0.62), although more polymicrobial infections were detected (24.7% vs. 33.3%, respectively; p = 0.359). MDROs were involved in 15% of the cases, with an upward trend when comparing both periods (12.8% vs. 23.6%; p = 0.07). The most-used combination of local antibiotics-glycopeptide (vancomycin) plus aminoglycoside (gentamicin or tobramycin)-was met with low rates of resistance in the most frequently isolated microorganisms. CONCLUSION: According to the results of the present study, rates of Gram-positive and Gram-negative infections remained consistent during the two study periods, but with an upward trend in MDRO and polymicrobial infections detected. The local combination of a glycopeptide plus an aminoglycoside was effective in treating the most frequently isolated microorganisms.

rFSAV promotes Staphylococcus aureus-infected bone defect healing via IL-13- mediated M2 macrophage polarization.

Staphylococcus aureus (S. aureus) contamination commonly occurs in orthopedic internal fixation operations, leading to a delayed healing of the defected bone tissue. However, antibiotic treatments are ineffective in dealing with S. aureus bone infections due to the rise in multiple antimicrobial resistances. Here, we reported the protective effects of a recombinant five-antigen S. aureus vaccine (rFSAV) in an S. aureus infected bone defect model. In this study, we found the number of M2 macrophages markedly increased in the defect site and played a critical role in the healing of defected bone mediated by rFSAV. Mechanistically, rFSAV mediated increased level of IL-13 in bone defect site predominant M2 macrophage polarization. In summary, our study reveals a key role of M2 macrophage polarization in the bone regeneration process in S. aureus infection induced bone defect, which provide a promising application of rFSAV for the treatment of bone infection for orthopedic applications.

Diagnostic performance comparison of conventional radiography to magnetic resonance imaging for suspected osteomyelitis of the extremities: a multi-reader study.

OBJECTIVE: To determine whether MRI provides improved diagnostic accuracy compared to radiography for the diagnosis of extremity osteomyelitis (OM) with multi-reader analysis. METHODS: In this cross-sectional study, three musculoskeletal fellowship-trained expert radiologists evaluated cases of suspected OM in two rounds-first using radiographs (XR), then with conventional MRI. Radiologic features consistent with OM were recorded. Each reader recorded individual findings on both modalities and rendered a binary diagnosis along with certainty of final diagnosis on a confidence scale of 1-5. This was compared with the pathology-proven diagnosis of OM to determine diagnostic performance. Intraclass correlation (ICC) and Conger's Kappa were used for statistics. RESULTS: XR and MRIs of 213 pathology proven cases (51.5 years ± 14.0 years, mean ± St.Dev.) were included in this study, with 79 tested positive for OM and 98 were positive for a soft tissue abscess, with 78 patients being negative for both. In total, 139 were males and 74 females with bones of interest in the upper and lower extremities in 29 and 184 cases, respectively. MRI showed significantly higher sensitivity and negative predictive value than XR (p < 0.001 for both metrics). Conger's Kappa for OM diagnosis were 0.62 and 0.74 on XR and MRI, respectively. Reader confidence improved slightly from 4.54 to 4.57 when MRI was used. CONCLUSIONS: MRI is a diagnostically more effective imaging modality than XR for finding extremity osteomyelitis with better inter-reader reliability. CLINICAL RELEVANCE STATEMENT: This study validates the diagnosis of OM with MRI over XR but adds novelty because it is the largest study of its kind with a clear reference standard to guide clinician decision making. KEY POINTS: • Radiography is the first-line imaging modality for musculoskeletal pathology but MRI can add value for infections. • MRI shows greater sensitivity for the diagnosis of osteomyelitis of the extremities than radiography. • This improved diagnostic accuracy makes MRI a better imaging modality for patients with suspected osteomyelitis.

Incremental value of diffusion weighted imaging over conventional MRI for the diagnosis of osteomyelitis of extremities.

OBJECTIVE: To determine the incremental value of diffusion weighted imaging (DWI) over conventional MR imaging in diagnosing extremity osteomyelitis (OM). MATERIALS AND METHODS: In this cross-sectional study, three experienced musculoskeletal radiologists evaluated clinically suspected cases of extremity OM in two rounds-first on conventional MR imaging, and then conventional MR imaging combined with DWI 4-6 weeks later. The readers recorded a result of the presence or absence of OM and their diagnostic confidence on a 1-5 scale. Mean and minimum apparent diffusion coefficient (ADC) were measured. Pathology diagnosis served as the reference standard. Statistical analysis utilized intraclass correlation (ICC) and Conger's kappa. RESULTS: A total of 213 scans of suspected OM were reviewed by three musculoskeletal radiologists with no significant changes in sensitivity (0.97, 0.97), specificity (0.97, 0.94), positive predictive value (0.91, 0.87), or negative predictive value (0.98, 0.98) between conventional MR imaging and MR imaging combined with DWI, respectively. Reader confidence did not significantly change with the addition of DWI (4.55 and 4.70, respectively). A high inter-reader agreement was observed for the diagnosis of OM, soft tissue abscess, and intraosseous abscess in both rounds. A higher mean (1.46+/-0.43 × 10-3 mm2/s > 0.64+/-0.47 × 10-3mm2/s) and minimum (1.18+/-0.45 × 10-3mm2/s > 0.37+/-0.44 × 10-3mm2/s) ADC value was associated with OM (p-value < 0.0001) with odds ratios of 1.34 and 1.31, respectively, for mean and minimum ADC of the involved bone. CONCLUSION: DWI-derived ADC increase is associated with OM. The use of DWI slightly increases reader confidence in the diagnosis of OM; however, no significant incremental value over conventional MR imaging is seen for the final diagnosis of OM.

Localization of osteomyelitis lesions for operative eradication of chronic osteomyelitis of the lower extremities by bone SPECT/CT: a feasibility study.

PURPOSE: This study was performed to investigate the feasibility of bone single-photon emission tomography/computed tomography (SPECT/CT) for pre-operative planning of chronic osteomyelitis (COM) of the lower extremities by localization of osteomyelitis lesions. METHODS: From January 2016 to January 2020, we surgically treated ten adult patients with Cierny-Mader type III COM in the tibia or femur for a mean duration of 24.4 months (range 7.0-70.0 months). We conducted pre-operative planning by bone SPECT/CT and localization of osteomyelitis lesions. The treatment consisted of intra-operative eradication of the infective focus and antibiotic administration. The clinical and radiological outcomes were retrospectively analyzed after a minimum of one year of follow-up. RESULTS: The patients were surgically treated by thorough debridement, dead space management, and appropriate antibiotics without bone transport or an external fixator. The location of the hot uptake region on bone SPECT/CT coincided with that of the osteomyelitis lesion, which was confirmed intra-operatively in all patients. At an average of 16.5 ± 4.3 months (range, 13.0-25.0 months), clinical eradication of osteomyelitis was achieved in nine of the ten patients. One patient required amputation due to recurrence of osteomyelitis. A successful clinical outcome was achieved in eight patients; one suffered persistent ankle pain due to a destructive change in the ankle joint despite eradication of the infection. CONCLUSION: Bone SPECT/CT is a feasible method for the localization and eradication of osteomyelitis lesions in COM of the lower extremities and has favourable clinical outcomes. It can also be applied in cases of distorted bony structures caused by previous trauma or surgery, or in the presence of implants.

Enhancing Bone Infection Diagnosis with Raman Handheld Spectroscopy: Pathogen Discrimination and Diagnostic Potential.

Osteomyelitis is a bone disease caused by bacteria that can damage bone. Raman handheld spectroscopy has emerged as a promising diagnostic tool for detecting bone infection and can be used intraoperatively during surgical procedures. This study involved 120 bone samples from 40 patients, with 80 samples infected with either Staphylococcus aureus or Staphylococcus epidermidis. Raman handheld spectroscopy demonstrated successful differentiation between healthy and infected bone samples and between the two types of bacterial pathogens. Raman handheld spectroscopy appears to be a promising diagnostic tool in bone infection and holds the potential to overcome many of the shortcomings of traditional diagnostic procedures. Further research, however, is required to confirm its diagnostic capabilities and consider other factors, such as the limit of pathogen detection and optimal calibration standards.

Insights into S. aureus-Induced Bone Deformation in a Mouse Model of Chronic Osteomyelitis Using Fluorescence and Raman Imaging.

Osteomyelitis is an infection of the bone that is often difficult to treat and causes a significant healthcare burden. Staphylococcus aureus is the most common pathogen causing osteomyelitis. Osteomyelitis mouse models have been established to gain further insights into the pathogenesis and host response. Here, we use an established S. aureus hematogenous osteomyelitis mouse model to investigate morphological tissue changes and bacterial localization in chronic osteomyelitis with a focus on the pelvis. X-ray imaging was performed to follow the disease progression. Six weeks post infection, when osteomyelitis had manifested itself with a macroscopically visible bone deformation in the pelvis, we used two orthogonal methods, namely fluorescence imaging and label-free Raman spectroscopy, to characterise tissue changes on a microscopic scale and to localise bacteria in different tissue regions. Hematoxylin and eosin as well as Gram staining were performed as a reference method. We could detect all signs of a chronically florid tissue infection with osseous and soft tissue changes as well as with different inflammatory infiltrate patterns. Large lesions dominated in the investigated tissue samples. Bacteria were found to form abscesses and were distributed in high numbers in the lesion, where they could occasionally also be detected intracellularly. In addition, bacteria were found in lower numbers in surrounding muscle tissue and even in lower numbers in trabecular bone tissue. The Raman spectroscopic imaging revealed a metabolic state of the bacteria with reduced activity in agreement with small cell variants found in other studies. In conclusion, we present novel optical methods to characterise bone infections, including inflammatory host tissue reactions and bacterial adaptation.

Spacer exchange in persistent periprosthetic joint infection: microbiological evaluation and survivorship analysis.

PURPOSE: The purposes of this study were to determine demographics and characteristics of patients who underwent spacer exchange for persistent infection in the setting of two-stage arthroplasty for periprosthetic joint infection, to describe the microbiology of pathogens involved, to analyze survivorship free from infection in these patients. METHODS: The institutional prospectively collected database was reviewed to enroll patients with minimum 2 years follow-up. Patients who underwent two-stage procedure for septic arthritis were excluded, as were patients who had spacer fracture or dislocation. RESULTS: A total of 34 patients (41 procedures) were included. Mean age was 65.0 ± 12.8 years. Mean follow-up was 53.4 ± 24.8 months. Mean number of previous procedures was 3.6 ± 1.2. A total of 27 (79.4%) patients underwent final reimplantation. The most frequently isolated pathogen in spacer exchange was Staphylococcus epidermidis (10 cases, 28.6%). Polymicrobial cultures were obtained from 9 (25.71%) patients, 10 (28.6%) presented culture-negative infections. A total of 11 (32.4%) resistant pathogens were isolated, and 16 (47.0%) difficult to treat pathogens were detected. Eradication rate was 78.8%. Overall survivorship of implants after final reimplantation was 72.8% at 51.8 months. CONCLUSION: Surgeons should be aware that subjects necessitating spacer exchange often present multiple comorbidities, previous staged revision failures, soft-tissue impairment and difficult to treat infection. In these patients, spacer exchange provides good clinical results and infection eradication, preventing arthrodesis or amputation.

A major trauma centre experience with gentamicin-coated tibial intramedullary nails (ETN PROtect™) in acute primary open fracture fixation and complex revision surgery.

PURPOSE: Fracture-related infections (FRI) following intramedullary nailing for tibial shaft fractures remain challenging to treat with associated high patient morbidity and health care costs. Recently, antibiotic-coated nails have been introduced as a strategy to reduce implant related infection rates in high-risk patients. We present the largest single-centre case series on ETN PROtect® outcomes reporting on fracture union, infection rates and treatment complications. METHODS: Fifty-six adult patients underwent surgery with ETN PROtect® between 01/09/17 and 31/12/20. Indications consisted of acute open fractures and complex revision cases (previous FRI, non-union surgery and re-fracture) with a mean of three prior surgical interventions. We report on patient demographics, union rates and deep infection. Minimum follow-up was one year. RESULTS: One (1.8%) patient developed a deep surgical infection and associated non-union requiring further surgery. In addition, we identified three cases (5.4%) of aseptic non-union following facture treatment with ETN PROtect®. Of the five patients who underwent staged complex revision surgery for established FRI with ETN PROtect®, all had treatment failure with ongoing symptoms of deep infection requiring implant removal and further treatment. CONCLUSION: Use of the ETN PROtect® nail in high-risk patients (open fractures and those initially treated with external fixation) and in those patients with aseptic non-unions, demonstrates promising outcomes in the prevention of implant-related infection. In our limited series we have failed to observe any benefit over uncoated nails, when used in treating cases of previously established FRI/osteomyelitis and would therefore advise caution in their use, especially in view of the high cost.

Relevance of using both aerobic and anaerobic enrichment vials for optimizing rapid diagnosis of osteoarticular infections.

The performance of a pair of blood culture vials (BACTEC® Plus Aerobic/F, and Anaerobic Lytic/F) were analyzed in 496 osteoarticular specimens (246 synovial fluids and 250 crushed bone samples), obtained in patients during routine diagnostic procedure at the Teaching Hospital of Rennes (France). The positive detection times were recorded for a 14 day-incubation period, and compared between both vials and with agar cultures. For samples from infected patients, the positive detection time was significantly shortened when vials were used compared to agar plates (p < 0.001). Median positive detection time was later with the Anaerobic Lytic/F vials (15.0 h) compared to the Plus Aerobic/F (13.0 h). Positivity rate was similar for Anaerobic Lytic/F vials (80.4%) and Plus Aerobic/F vials (83.2%) (p = 0.25). Some microorganisms were only identified from aerobic vials (15.5%) or from anaerobic vials (12.7%). The use of both atmosphere conditions for optimal positive detection time is therefore critical.

AmpC hyperproduction in a Cedecea davisae implant-associated bone infection during treatment: a case report and therapeutic implications.

BACKGROUND: Data on antimicrobial resistance mechanisms are scanty for Cedecea spp., with very variable antibiotic resistance patterns documented. Here we report the first in vivo resistance evolution of a C. davisae clinical isolate in a patient with a complex hand trauma and provide insight in the resistance mechanism, leading to therapeutic implications for this pathogen. CASE PRESENTATION: Cedecea davisae was isolated from a patient with hand trauma during a first surgical debridement. Six days after primary surgical treatment and under antimicrobial treatment with amoxicillin-clavulanic acid and later cefepime, follow up cultures yielded C. davisae which demonstrated a resistance development. The susceptible parental isolate and its resistant derivative were characterized by whole genome sequencing, ampC, ompC and ompF by RT- PCR. The resistant derivative demonstrated an A224G SNP in ampD, the transcriptional regulator of ampC, leading to a His75Arg change in the corresponding AmpD protein. AmpC transcription of the resistant derivative was 362-times higher than the susceptible isolate. Transcription levels of ompF and ompC were 8.5-fold and 1.3-fold lower, respectively, in the resistant derivative. Downregulation of OmpF putatively resulted from a mutation in the presumed promoter region upstream of the dusB-Fis operon, a proposed regulator for ompF. CONCLUSIONS: This case demonstrates the in vivo resistance development of C. davisae within 7 days similar to that of the members of the Enterobacter cloacae complex. Our findings add valuable information for future therapeutic management of these opportunistic pathogens as they warrant the same empirical treatment as AmpC producers.

Implantable drug delivery systems for the treatment of osteomyelitis.

Osteomyelitis is an infection of the bone tissue and bone marrow which is becoming increasingly difficult to treat due to the infection causing pathogens associated. Staphylococcus aureus is one of the main bacteria that causes this infection, which has a broad spectrum of antibiotic resistance making it extremely difficult to treat. Conventional metal implants used in orthopedic applications often have the drawback of implant induced osteomyelitis as well as the requirement of a second surgery to remove the implant once it is no longer required. Recently, attention has been focused on the design and fabrication of biodegradable implants for the treatment of bone infection. The main benefit of biodegradable implants over polymethylmethacrylate (PMMA) based non-degradable systems is that they do not require a second surgery for removal and so making degradable implants safer and easier to use. The main purpose of a biodegradable implant is to provide the necessary support and conductivity to allow the bone to regenerate whilst themselves degrading at a rate that is compatible with the rate of formation of new bone. They must be highly biocompatible to ensure there is no inflammation or irritation within the surrounding tissue. During this review, the latest research into antibiotic loaded biodegradable implants will be explored. Their benefits and drawbacks will be compared with those non-degradable PMMA beads, which is the stable material used within antibiotic loaded implants. Biodegradable implants most frequently used are based on biodegradable natural and synthetic polymers. Implants can take the form of many different structures; the most commonly fabricated structure is a scaffold. Other structures that will be explored within this review are hydrogels, nanoparticles and surface coatings, all with their own benefits/drawbacks.

The Role of a Virtual Noncalcium Dual-Energy CT Application in the Detection of Bone Marrow Edema in Peripheral Osteomyelitis.

Purpose: To determine the sensitivity and specificity of dual-energy CT (DECT) virtual noncalcium images (VNCa) with bone and soft tissue reconstructions in the diagnosis of osteomyelitis. Materials & Methods: Between December 1, 2014 to December 1, 2020, 91 patients who had 99 DECT performed for a clinical indication of osteomyelitis with corresponding MRI, triphasic bone scan and/or white blood cell scintigraphy with CT/SPECT performed either 2 weeks before or 1 month after the DECT were retrospectively identified. The presence or absence of osteomyelitis was established using a second imaging test, bone biopsy or surgery. Two radiologists interpreted VNCa images alone and with bone and soft tissue reconstructions for osteomyelitis. Fleiss k statistics was used to assess inter-level agreement. Results: Osteomyelitis was present in 26 cases (26.2%), of which 4 cases (4%) had co-existing septic arthritis. DECT was performed at the following sites: ankle/foot (n = 59), calf (n = 12), knee (n = 3), thigh (n = 7), hip (n = 9), pelvis (n = 6), wrist/hand (n = 1), and shoulder (n = 2). Sensitivity with VNCa images alone was 53.8% and 73.1% and specificity was 84.9% and 71.2%. Sensitivity with VNCa images and bone and soft tissue reconstructions was 80.8% and 80.8% and specificity was 80.8% and 72.6%. Interobserver agreement was 76.7% (76 of 99 cases), for VNCa images alone (k = .487), and 66.7% (66 of 99 patients) for bone and soft tissue reconstructions with VNCa images together (k = .390). Conclusion: When VNCa images were combined with bone and soft tissue reconstructions, there is improved sensitivity in the diagnosis of osteomyelitis.

Surgical treatment outcome after serial debridement of infected nonunion-A retrospective cohort study.

PURPOSE: Reported outcome after multiple staged surgical treatment of infected nonunion is scarce. We, therefore, asked: (1) What is the clinical outcome in infected nonunion patients after multiple staged revision surgery? (2) Are different pathogens evidenced after surgical treatment in patients who have undergone more or less surgeries? METHODS: All enrolled patients were surgically treated for long bone-infected nonunion between January 2010 and March 2018. Besides patients´ demographics outcome in terms of bony consolidation and major complications defined as death during inward treatment, amputation and recurrence of infection during follow-up of at least 12 months were assessed. Microbiological findings were assessed and compared between two groups with less than five versus five or more surgical revisions. RESULTS: Bone consolidation was achieved in 86% of the patients while complications such as femoral or transtibial amputation, recurrence of infection or even death during inpatient treatment could be evidenced in six patients (14%). In patients who underwent multiple-stage surgery for five or more times, germ changes and repeated germ detection was more common than in patients with less surgeries. CONCLUSIONS: Surgical treatment of infected nonunions poses a high burden on the patients with major complications occurring in about 14% of the patients using a multiple staged treatment concept. Future prospective studies comparing outcomes after limited with multiple staged revision surgeries are necessary.