ChatGPT May Offer an Adequate Substitute for Informed Consent to Patients Prior to Total Knee Arthroplasty-Yet Caution Is Needed.

Prior to undergoing total knee arthroplasty (TKA), surgeons are often confronted with patients with numerous questions regarding the procedure and the recovery process. Due to limited staff resources and mounting individual workload, increased efficiency, e.g., using artificial intelligence (AI), is of increasing interest. We comprehensively evaluated ChatGPT's orthopedic responses using the DISCERN instrument. Three independent orthopedic surgeons rated the responses across various criteria. We found consistently high scores, predominantly exceeding a score of three out of five in almost all categories, indicative of the quality and accuracy of the information provided. Notably, the AI demonstrated proficiency in conveying precise and reliable information on orthopedic topics. However, a notable observation pertains to the generation of non-existing references for certain claims. This study underscores the significance of critically evaluating references provided by ChatGPT and emphasizes the necessity of cross-referencing information from established sources. Overall, the findings contribute valuable insights into the performance of ChatGPT in delivering accurate orthopedic information for patients in clinical use while shedding light on areas warranting further refinement. Future iterations of natural language processing systems may be able to replace, in part or in entirety, the preoperative interactions, thereby optimizing the efficiency, accessibility, and standardization of patient communication.

Candida spondylodiscitis: a systematic review and meta-analysis of seventy two studies.

OBJECTIVES: Knowledge of Candida spondylodiscitis is limited to case reports and smaller case series. Controversy remains on the most effective diagnostical and therapeutical steps once Candida is suspected. This systematic review summarized all cases of Candida spondylodiscitis reported to date concerning baseline demographics, symptoms, treatment, and prognostic factors. METHODS: A PRISMA-based search of PubMed, Web of Science, Embase, Scopus, and OVID Medline was performed from database inception to November 30, 2022. Reported cases of Candida spondylodiscitis were included regardless of Candida strain or spinal levels involved. Based on these criteria, 656 studies were analyzed and 72 included for analysis. Kaplan-Meier curves, Fisher's exact, and Wilcoxon's rank sum tests were performed. RESULTS: In total, 89 patients (67% males) treated for Candida spondylodiscitis were included. Median age was 61 years, 23% were immunocompromised, and 15% IV drug users. Median length of antifungal treatment was six months, and fluconazole (68%) most commonly used. Thirteen percent underwent debridement, 34% discectomy with and 21% without additional instrumentation. Median follow-up was 12 months. The two year survivorship free of death was 80%. The two year survivorship free of revision was 94%. Younger age (p = 0.042) and longer length of antifungal treatment (p = 0.061) were predictive of survival. CONCLUSION: Most patients affected by Candida spondylodiscitis were males in their sixties, with one in four being immunocompromised. While one in five patients died within two years of diagnosis, younger age and prolonged antifungal treatment might play a protective role.

Periprosthetic Joint Infections of the Knee Lastingly Impact the Bone Homeostasis.

After periprosthetic joint infection (PJI)-dependent revision surgery, a significantly elevated number of patients suffer from prosthesis failure due to aseptic loosening and require additional revision surgery despite clearance of the initial infection. The mechanisms underlying this pathology are not well understood, as it has been assumed that the bone stock recovers after revision surgery. Despite clinical evidence suggesting decreased osteogenic potential in PJI, understanding of the underlying biology remains limited. In this study, we investigated the impact of PJI on bone homeostasis in a two-stage exchange approach at explantation and reimplantation. Sixty-four human tibial and femoral specimens (20 control, 20 PJI septic explantation, and 24 PJI prosthesis reimplantation samples) were analyzed for their bone microstructure, cellular composition, and expression of relevant genetic markers. Samples were analyzed using X-ray microtomography, Alcian blue and tartrate-resistant acid phosphatase staining, and RT-qPCR. In patients with PJI, bone volume (BV/TV; 0.173 ± 0.026; p < 0.001), trabecular thickness (164.262 ± 18.841 µm; p < 0.001), and bone mineral density (0.824 ± 0.017 g/cm2 ; p = 0.049) were reduced; trabecular separation (1833.939 ± 178.501 µm; p = 0.005) was increased. While prevalence of osteoclasts was elevated (N.Oc/BS: 0.663 ± 0.102, p < 0.001), osteoblast cell numbers were lower at explantation (N.Ob/BS: 0.149 ± 0.021; p = 0.047). Mean expression of bone homeostasis markers osteocalcin, osteopontin, Runx2, TSG-6, and FGF-2 was significantly reduced at prosthesis explantation. Despite partial recovery, all analyzed parameters were still significantly impacted at reimplantation. In contrast, mean expression of osteoclastogenesis-stimulating cytokine IL-17a was significantly increased at both explantation and reimplantation. In this study, we found a strong and lasting impact of PJI on the bone homeostasis on a molecular, cellular, and microstructural level. These changes may be responsible for the increased risk of prosthesis failure due to aseptic loosening. Our data suggest there is significant potential in modulating bone homeostasis to improve prosthesis fixation and long-term clinical outcome in affected patients. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Combining radiographic and CT measurements to rival MRI for the diagnosis of acute isolated syndesmotic injury.

BACKGROUND: Acute isolated syndesmotic injuries (AISIs) have a high potential to be misdiagnosed or underdiagnosed at initial presentation to the hospital. Although magnetic resonance imaging (MRI) is the gold standard in noninvasive diagnostics, it is not always available immediately and is much more expensive than other imaging modalities. This study identifies improvements in conventional radiography and computed tomography (CT) to diagnose AISI and aims to reduce the number of MRI scans needed to verify the diagnosis. METHODS: A retrospective case match control study was conducted by searching our trauma database between 2008 and 2022. A study group of patients with AISI (n = 64) and a control group of patients without AISI (n = 76) were formed to generate an equal number of images from both groups (62 radiographs and 22 CT scans). A total of 16 parameters that quantify the distal tibiofibular relation in injured and uninjured ankles were analyzed. For statistical analysis, a two-sided t-test was applied to calculate significant differences (p < 0.05). In a further step, a receiver operating characteristic curve (ROC) was used to determine cut-off values for the most significant parameters. RESULTS: The most significant measurement (p < 0.001) on axial CT scans was the syndesmotic area (SA). The ROC curve revealed an area under the curve (AUC) of 0.94 (95% CI 0.86-1.0) and a cut-off value of 71.68 mm2 that shows a sensitivity and specificity of 95.5% and 81.8%, respectively. CONCLUSION: This study suggests that radiographic imaging could represent an equally accurate alternative to MRI. These methods might generate the correct diagnosis faster due to their availability and inexpensiveness. By applying our new cut-off values in a clinical setting, the number of underdiagnosed and untreated unstable syndesmotic injuries could be reduced. LEVEL OF EVIDENCE: III, retrospective comparative study.

Exertional Rhabdomyolysis in Athletes: Systematic Review and Current Perspectives.

OBJECTIVE: Exertional rhabdomyolysis results from a breakdown of skeletal muscle cells after intense exercise in otherwise healthy patients, causing increased levels of creatine kinase (CK) or myoglobin, as well as urine dipstick positive for blood, and may result in kidney insufficiency. The aim of this study was to outline the current perspectives of exertional rhabdomyolysis in athletes and subsequent treatment based on the current literature. DATA SOURCES: We searched the MEDLINE/PubMed and Google databases for ([exercise] OR [exertional]) AND rhabdomyolysis following the PRISMA guidelines. All abstracts were reviewed by 2 independent examiners. Inclusion criteria consisted of original articles presenting studies on exertional rhabdomyolysis or exercise-induced rhabdomyolysis with 7 or more cases. All case reports, case series, or editorials were excluded. MAIN RESULTS: A total of 1541-abstracts were screened, leaving 25 studies for final inclusion and analysing 772patients. Especially, young male patients were affected at a mean age of 28.7 years (range 15.8-46.6 years). Most of the athletes performed running, including marathons in 54.3% of cases (n = 419/772), followed by weightlifting in 14.8% (n = 114/772). At the time of presentation, the mean creatine kinase was 31 481 IU/L (range 164-106,488 IU/L). Seventeen studies reported the highest level of CK, which was 38 552 IU/L (range 450-88,496 IU/L). For treatment, hydration was the most common method of choice reported by 8 studies. CONCLUSIONS: Exertional rhabdomyolysis seems to be underestimated, and it is essential to screen patients who present with muscle soreness/cramps and/or dark urine after heavy endurance events to avoid any further complications. LEVEL OF EVIDENCE: II; systematic review.

The role of myeloid derived suppressor cells in musculoskeletal disorders.

The immune system is closely linked to bone homeostasis and plays a pivotal role in several pathological and inflammatory conditions. Through various pathways it modulates various bone cells and subsequently sustains the physiological bone metabolism. Myeloid-derived suppressor cells (MDSCs) are a group of heterogeneous immature myeloid-derived cells that can exert an immunosuppressive function through a direct cell-to-cell contact, secretion of anti-inflammatory cytokines or specific exosomes. These cells mediate the innate immune response to chronic stress on the skeletal system. In chronic inflammation, MDSCs act as an inner offset to rebalance overactivation of the immune system. Moreover, they have been found to be involved in processes responsible for bone remodeling in different musculoskeletal disorders, autoimmune diseases, infection, and cancer. These cells can not only cause bone erosion by differentiating into osteoclasts, but also alleviate the immune reaction, subsequently leading to long-lastingly impacted bone remodeling. In this review, we discuss the impact of MDSCs on the bone metabolism under several pathological conditions, the involved modulatory pathways as well as potential therapeutic targets in MDSCs to improve bone health.

Inflammation of Bone in Patients with Periprosthetic Joint Infections of the Knee.

Despite the general success of total knee arthroplasty (TKA), addressing periprosthetic joint infection (PJI) and the resulting long-term complications is a growing medical need given the aging population and the increasing demand for arthroplasty. A larger proportion of patients face revision surgery because of the long-term complication of aseptic loosening despite clearance of the infection. The pathomechanisms leading to prosthetic loosening are not understood as it has been widely assumed that the bone stock recovers after explantation revision surgery. While clinical observations suggest a reduced osteogenic potential in patients with PJI, knowledge regarding the relevant biology is sparse. In the present study, we investigated the inflammatory impact of PJI on the bone and bone marrow in the vicinity of the joint. Additionally, we evaluated changes in the local inflammatory environment in a 2-stage exchange at both explantation and reimplantation. Methods: In this study, we analyzed 75 human bone and bone-marrow specimens (obtained from 65 patients undergoing revision arthroplasty with cement for the treatment of PJI) for markers of inflammation. Samples were analyzed using hematoxylin and eosin overview staining, fluorescent immunohistochemical staining, flow cytometry, and polymerase chain reaction (PCR). Results: Leukocyte prevalence was significantly elevated at explantation (femur, +218.9%; tibia, +134.2%). While leukocyte prevalence decreased at reimplantation (femur, -49.5%; tibia, -34.2%), the number of cells remained significantly higher compared with the control group (femur, +61.2%; tibia, +54.2%). Expression of inflammatory markers interleukin (IL)-1α (femur, +2,748.7%; tibia, +1,605.9%), IL-6 (femur, +2,062.5%; tibia, +2,385.7%), IL-10 (femur, +913.7%; tibia, +897.5%), IL-12 (femur, +386.1%; tibia, +52.5%), IL-18 (femur, +805.3%; tibia, +547.7%), and tumor necrosis factor (TNF)-α (femur, +296.9%; tibia, +220.9%) was significantly elevated at prosthesis explantation in both femoral and tibial specimens. Expression remained significantly elevated at reimplantation for all inflammatory markers except IL-12 compared with the control group. Conversely, there were only limited inflammatory changes in the bone marrow environment. Conclusions: The present study demonstrated a strong and lasting upregulation of the proinflammatory environment in the joint-surrounding osseous scaffold in patients with PJI. Our data suggest that modulating the inflammatory environment has substantial potential to improve the clinical outcome in affected patients.

Mechanical failure of total hip arthroplasties and associated risk factors.

INTRODUCTION: Mechanical failure of total hip arthroplasties is a rare but devastating complication. With increasing numbers in primary arthroplasty implantation, revision surgeries are indicated more often. Therefore, understanding the mechanism and the location of failure is essential in determining proper treatment. Aim of this study was to identify mechanical failures of all total hip arthroplasties performed in a major academic center as well as the associated risk factors such as BMI and sports. METHODS: A retrospective trial was conducted using our prospective arthroplasty database. Database was searched for all patients presenting with mechanical failures of total hip arthroplasty (THA) to the emergency department between 2011 and 2019. All medical charts and radiographs as well as surgical reports were analyzed to identify demographics, implant choice in addition to location of failure and subsequent treatment. RESULTS: In total, 13 patients suffering from mechanical total hip implant failure were found. The femoral neck (conus) was broken in four patients, the stem in five cases, one broken inlay, two cup failures and one conus dislocation. The mean BMI was 31.42 ± 5.29 kg/m2 including five patients who have obesity class II. In all cases, revision surgeries were indicated. No structural causes or underlying risk factors such as repeated physical load (i.e. in sports) were identified. CONCLUSION: Implant failure does not seem to correlate with participation in sports or BMI. Catastrophic failure of implants is a technical challenge requiring special extraction instruments that can be difficult even for experienced surgeons. It should be noted that functional outcome is often worse for this group of patients after surgery than comparing against those revised for loosening. LEVEL OF EVIDENCE: III, Retrospective Trial.

Serum and Synovial Markers in Patients with Rheumatoid Arthritis and Periprosthetic Joint Infection.

Current diagnostic standards for PJI rely on inflammatory markers that are typically elevated in autoimmune diseases, thus making the diagnosis of PJI in patients with rheumatoid arthritis and joint replacement particularly complicated. There is a paucity of data on differentiating PJI from rheumatoid arthritis in patients with previous arthroplasty. In this study, we retrospectively analyzed the cases of 17 patients with rheumatoid arthritis and 121 patients without rheumatoid disease who underwent surgical intervention due to microbiology-positive PJI of the hip or knee joint. We assessed clinical patient characteristics, laboratory parameters, and prosthesis survival rates in patients with and without rheumatoid arthritis and acute or chronic PJI. ROC analysis was conducted for the analyzed parameters. In patients with chronic PJI, peripheral blood CRP (p = 0.05, AUC = 0.71), synovial WBC count (p = 0.02, AUC = 0.78), synovial monocyte cell count (p = 0.04, AUC = 0.75), and synovial PMN cell count (p = 0.02, AUC = 0.80) were significantly elevated in patients with rheumatoid arthritis showing acceptable to excellent discrimination. All analyzed parameters showed no significant differences and poor discrimination for patients with acute PJI. Median prosthesis survival time was significantly shorter in patients with rheumatoid arthritis (p = 0.05). In conclusion, routinely used laboratory markers have limited utility in distinguishing acute PJI in rheumatoid patients. In cases with suspected chronic PJI but low levels of serum CRP and synovial cell markers, physicians should consider the possibility of activated autoimmune arthritis.

Altered molecular pathways and prognostic markers in active systemic juvenile idiopathic arthritis: integrated bioinformatic analysis.

Systemic juvenile idiopathic arthritis (SJIA) is a severe childhood-onset inflammatory disease characterized by arthritis accompanied by systemic auto-inflammation and extra-articular symptoms. While recent advances have unraveled a range of risk factors, the pathomechanisms involved in SJIA and potential prognostic markers for treatment success remain partly unknown. In this study, we included 70 active SJIA and 55 healthy control patients from the National Center for Biotechnology Information to analyze for differentially expressed genes (DEGs) using R. Functional enrichment analysis, protein-protein interaction (PPI), and gene module construction were performed for DEGs and hub gene set. We additionally examined immune system cell composition with CIBERSORT and predicted prognostic markers and potential treatment drugs for SJIA. In total, 94 upregulated and 24 downregulated DEGs were identified. Two specific modules of interest and eight hub genes (ARG1, DEFA4, HP, MMP8, MMP9, MPO, OLFM4, PGLYRP1) were screened out. Functional enrichment analysis suggested that complex neutrophil-related functions play a decisive role in the disease pathogenesis. CIBERSORT indicated neutrophils, M0 macrophages, CD8+ T cells, and naïve B cells to be relevant drivers of disease progression. Additionally, we identified TPM2 and GZMB as potential prognostic markers for treatment response to canakinumab. Moreover, sulindac sulfide, (-)-catechin, and phenanthridinone were identified as promising treatment agents. This study provides a new insight into molecular and cellular pathogenesis of active SJIA and highlights potential targets for further research.

Candida periprosthetic joint infections - risk factors and outcome between albicans and non-albicans strains.

BACKGROUND: Despite its scarcity, fungal periprosthetic joint infection (PJI) is of great clinical relevance as diagnosis and treatment are highly challenging. Previous analyses focused on the treatment rather than the role of the causative fungal agent on clinical outcome. This is the largest study of its kind to evaluate Candida strain-dependent differences in patients with fungal PJI. METHODS: We retrospectively analyzed 29 patients who underwent surgical intervention due to Candida hip or knee PJI in our department from 2010 to 2018. PJI was defined according to IDSA, recurrent PJI according to modified Delphi consensus criteria. Statistical analysis was performed using t-test, chi-square test with Yates correction, and log rank test. RESULTS: Besides age and affected joint, no significant differences were found between Candida albicans and non-albicans PJI patients (75.83 versus 64.11 years, p = 0.012; 12 hip versus two knee cases, p = 0.013). Most patients received two- (27.59%) or three-stage exchange surgery (41.38%). There was a statistical trend towards an increase in surgery needed in non-albicans Candida PJI (2.92 versus 2.12; p = 0.103). After initial Candida PJI treatment, functional prosthesis implantation was achieved in 72.41% of all patients. At last follow-up, infection-free survival was at 26.79% in Candida albicans versus 72.00% in non-albicans PJI (p = 0.046). CONCLUSIONS: In this study, we found infection-free survival rates to be significantly decreased in patients with albicans compared to non-albicans Candida PJI. While age and affected joint might play a confounding role, we speculate the causative pathogen to play a decisive role in disease progression.

Public Interest in Knee Pain and Knee Replacement during the SARS-CoV-2 Pandemic in Western Europe.

Due to the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) pandemic, a large number of elective knee replacement procedures had to be postponed in both early and late 2020 in most western countries including Germany and the UK. It is unknown how public interest and demand for total knee arthroplasties was affected. Public interest in knee pain, knee osteoarthritis and knee arthroplasty in Germany and the UK was investigated using Google Trend Analysis. In addition, we monitored for changes in patient composition in our outpatient department. As of early March in Germany and of late March in the UK, until the lockdown measures, a 50 to 60% decrease in relative search frequency was observed in all categories investigated compared to the beginning of the year. While public interest for knee pain rapidly recovered, decreased interest for knee osteoarthritis and replacement lasted until the easing of measures. Shortly prior to and during the first lockdown mean search frequency for knee replacement was significantly decreased from 39.7% and 36.6 to 26.9% in Germany and from 47.7% and 50.9 to 23.7% in the UK (Germany: p = 0.022 prior to lockdown, p < 0.001 during lockdown; UK: p < 0.0001 prior to and during lockdown). In contrast, mean search frequencies did not differ significantly from each other for any of the investigated time frames during the second half of 2020 in both countries. Similarly, during the first lockdown, the proportion of patients presenting themselves to receive primary knee arthroplasty compared to patients that had already undergone knee replacement declined markedly from 64.7% to 46.9%. In contrast, patient composition changed only marginally during the lockdown measures in late 2020 in both Germany and the UK. We observed a high level of public interest in knee arthroplasty despite the ongoing pandemic. The absence of a lasting decline in interest in primary knee arthroplasty suggests that sufficient symptom reduction cannot be achieved without surgical care for a substantial number of patients.

High Rates of Aseptic Loosening After Revision Total Knee Arthroplasty for Periprosthetic Joint Infection.

With increasing life expectancy, the demand for knee replacement is continuously rising. Despite the use of antibiotic prophylaxis and improved aseptic surgical techniques, periprosthetic joint infection (PJI) still occurs in 1% to 5% of patients after primary arthroplasty. An open question is the influence of PJI and resulting surgical procedures on the occurrence of long-term complications such as aseptic loosening. Patients needing multiple revision surgeries are especially at risk for decreases in bone mass and damage to the medullary cavity. Thus, we theorized that prior surgeries on the affected knee increase the risk of aseptic loosening in patients with PJI. METHODS: We retrospectively analyzed the cases of 100 patients who underwent total knee replacement exchange surgery as a result of PJI. In addition to clinical, paraclinical, and radiographic examination, we assessed comorbidities and the number of prior surgeries. RESULTS: Prosthetic survival was drastically decreased after PJI-related revision arthroplasty: during the first 7.3 years after reimplantation, 22% and 16% of all patients had aseptic loosening and recurrent PJI, respectively. The prevalence of aseptic loosening was 27.8% for female and 15.2% for male patients. A significant association between increasing patients' American Society of Anesthesiologists (ASA) classification and prosthetic failure rates was found, as was a strong correlation between number of prior surgeries and aseptic loosening. CONCLUSIONS: In this study, we found notable rates of aseptic loosening and recurrent PJI following PJI-related revision arthroplasty. The difference in the rate of aseptic loosening among male and female patients supports theories of the role of bone metabolism in the development of aseptic loosening. The economic and clinical burdens of prosthetic failure make it paramount to gain a better understanding of bone metabolism in PJI. Additional research should address the need to optimize treatment strategies to increase prosthetic survival. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Antibiotic treatment of postoperative spinal implant infections.

Postoperative spinal implant infection (PSII) is a serious complication after spinal surgery. It is associated with increased morbidity and mortality for affected patients as well as significant costs for the healthcare system. Due to the formation of biofilm on foreign material, both diagnosis and treatment of PSII can pose a considerable challenge. Modern treatment protocols allow efficient eradication and good clinical outcomes in the majority of patients. In this article, we review the current antibiotic treatment concepts for PSII including the correct choice of antibiotics and their combination. In cases of late-onset PSII where the implants can be removed, two weeks of intravenous (IV) antibiotics followed by 4 weeks of oral antibiotics seem appropriate. If the implant needs to be retained, a 2-week IV antibiotic treatment should be followed by 10 weeks of oral antibiotic therapy with biofilm activity or, in case of problematic pathogens, a long-term suppression therapy. Initial empiric antibiotic therapy should cover staphylococci, streptococci, enterococci and Gram-negative bacilli as the most common pathogens. Antibiotic adjustments according to the type of pathogen and its antimicrobial susceptibility are essential for successful eradication of infection.

Structural heteropolysaccharides as air-tight sealants of the human pleura.

Pulmonary "air leaks," typically the result of pleural injury caused by lung surgery or chest trauma, result in the accumulation of air in the pleural space (pneumothorax). Air leaks are a major source of morbidity and prolonged hospitalization after pulmonary surgery. Previous work has demonstrated structural heteropolysaccharide (pectin) binding to the mouse pleural glycocalyx. The similar lectin-binding characteristics and ultrastructural features of the human and mouse pleural glycocalyx suggested the potential application of these polymers in humans. To investigate the utility of pectin-based polymers, we developed a simulacrum using freshly obtained human pleura. Pressure-decay leak testing was performed with an inflation maneuver that involved a 3 s ramp to a 3 s plateau pressure; the inflation was completely abrogated after needle perforation of the pleura. Using nonbiologic materials, pressure-decay leak testing demonstrated an exponential decay with a plateau phase in materials with a Young's modulus less than 5. In human pleural testing, the simulacrum was used to test the sealant function of four mixtures of pectin-based polymers. A 50% high-methoxyl pectin and 50% carboxymethylcellulose mixture demonstrated no sealant failures at transpleural pressures of 60 cmH2 O. In contrast, pectin mixtures containing 50% low-methoxyl pectin, 50% amidated low-methoxyl pectins, or 100% carboxymethylcellulose demonstrated frequent sealant failures at transpleural pressures of 40-50 cmH2 O (p < 0.001). Inhibition of sealant adhesion with enzyme treatment, dessication and 4°C cooling suggested an adhesion mechanism dependent upon polysaccharide interpenetration. We conclude that pectin-based heteropolysaccharides are a promising air-tight sealant of human pleural injuries. © 2018 Wiley Periodicals, Inc. J. Biomed. Mater. Res. Part B, 2018. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 799-806, 2019.

EGFL7 enhances surface expression of integrin α5ß1 to promote angiogenesis in malignant brain tumors.

Glioblastoma (GBM) is a typically lethal type of brain tumor with a median survival of 15 months postdiagnosis. This negative prognosis prompted the exploration of alternative treatment options. In particular, the reliance of GBM on angiogenesis triggered the development of anti-VEGF (vascular endothelial growth factor) blocking antibodies such as bevacizumab. Although its application in human GBM only increased progression-free periods but did not improve overall survival, physicians and researchers still utilize this treatment option due to the lack of adequate alternatives. In an attempt to improve the efficacy of anti-VEGF treatment, we explored the role of the egfl7 gene in malignant glioma. We found that the encoded extracellular matrix protein epidermal growth factor-like protein 7 (EGFL7) was secreted by glioma blood vessels but not glioma cells themselves, while no major role could be assigned to the parasitic miRNAs miR-126/126*. EGFL7 expression promoted glioma growth in experimental glioma models in vivo and stimulated tumor vascularization. Mechanistically, this was mediated by an upregulation of integrin α5ß1 on the cellular surface of endothelial cells, which enhanced fibronectin-induced angiogenic sprouting. Glioma blood vessels that formed in vivo were more mature as determined by pericyte and smooth muscle cell coverage. Furthermore, these vessels were less leaky as measured by magnetic resonance imaging of extravasating contrast agent. EGFL7-inhibition using a specific blocking antibody reduced the vascularization of experimental gliomas and increased the life span of treated animals, in particular in combination with anti-VEGF and the chemotherapeutic agent temozolomide. Data allow for the conclusion that this combinatorial regimen may serve as a novel treatment option for GBM.

Pressure-decay testing of pleural air leaks in intact murine lungs: evidence for peripheral airway regulation.

The critical care management of pleural air leaks can be challenging in all patients, but particularly in patients on mechanical ventilation. To investigate the effect of central airway pressure and pleural pressure on pulmonary air leaks, we studied orotracheally intubated mice with pleural injuries. We used clinically relevant variables - namely, airway pressure and pleural pressure - to investigate flow through peripheral air leaks. The model studied the pleural injuries using a pressure-decay maneuver. The pressure-decay maneuver involved a 3 sec ramp to 30 cmH2 0 followed by a 3 sec breath hold. After pleural injury, the pressure-decay maneuver demonstrated a distinctive airway pressure time history. Peak inflation was followed by a rapid decrease to a lower plateau phase. The decay phase of the inflation maneuver was influenced by the injury area. The rate of pressure decline with multiple injuries (28 ± 8 cmH2 0/sec) was significantly greater than a single injury (12 ± 3 cmH2 O/sec) (P < 0.05). In contrast, the plateau phase pressure was independent of injury surface area, but dependent upon transpulmonary pressure. The mean plateau transpulmonary pressure was 18 ± 0.7 cm H2 O. Finally, analysis of the inflation ramp demonstrated that nearly all volume loss occurred at the end of inflation (P < 0.001). We conclude that the air flow through peripheral lung injuries was greatest at increased lung volumes and limited by peripheral airway closure. In addition to suggesting an intrinsic mechanism for limiting flow through peripheral air leaks, these findings suggest the utility of positive end-expiratory pressure and negative pleural pressure to maintain lung volumes in patients with pleural injuries.

Free-Floating Mesothelial Cells in Pleural Fluid After Lung Surgery.

OBJECTIVES: The mesothelium, the surface layer of the heart, lung, bowel, liver, and tunica vaginalis, is a complex tissue implicated in organ-specific diseases and regenerative biology; however, the mechanism of mesothelial repair after surgical injury is unknown. Previous observations indicated seeding of denuded mesothelium by free-floating mesothelial cells may contribute to mesothelial healing. In this study, we investigated the prevalence of mesothelial cells in pleural fluid during the 7 days following pulmonary surgery. STUDY DESIGN: Flow cytometry was employed to study pleural fluid of 45 patients after lung resection or transplantation. We used histologically validated mesothelial markers (CD71 and WT1) to estimate the prevalence of mesothelial cells. RESULTS: The viability of pleural fluid cells approached 100%. Leukocytes and mesothelial cells were identified in the pleural fluid within the first week after surgery. The leukocyte concentration was relatively stable at all time points. In contrast, mesothelial cells, identified by CD71 and WT1 peaked on POD3. The broad expression of CD71 molecule in postoperative pleural fluid suggests that many of the free-floating non-leukocyte cells were activated or proliferative mesothelial cells. CONCLUSION: We demonstrated that pleural fluid post lung surgery is a source of mesothelial cells; most of these cells appear to be viable and, as shown by CD71 staining, activated mesothelial cells. The observed peak of mesothelial cells on POD3 is consistent with a potential reparative role of free-floating mesothelial cells after pulmonary surgery.

Structural Heteropolysaccharide Adhesion to the Glycocalyx of Visceral Mesothelium.

Bioadhesives are biopolymers with potential applications in wound healing, drug delivery, and tissue engineering. Pectin, a plant-based heteropolysaccharide, has recently demonstrated potential as a mucoadhesive in the gut. Since mucoadhesion is a process likely involving the interpenetration of the pectin polymer with mucin chains, we hypothesized that pectin may also be effective at targeting the glycocalyx of the visceral mesothelium. To explore the potential role of pectin as a mesothelial bioadhesive, we studied the interaction of various pectin formulations with the mesothelium of the lung, liver, bowel, and heart. Tensile strength, peel strength, and shear resistance of the bioadhesive-mesothelial interaction were measured by load/displacement measurements. In both high-methoxyl pectins (HMP) and low-methoxyl pectins, bioadhesion was greatest with an equal weight % formulation with carboxymethylcellulose (CMC). The tensile strength of the high-methoxyl pectin was consistently greater than low-methoxyl or amidated low-methoxyl formulations (p < 0.05). Consistent with a mechanism of polymer-glycocalyx interpenetration, the HMP adhesion to tissue mesothelium was reversed with hydration and limited by enzyme treatment (hyaluronidase, pronase, and neuraminidase). Peel and shear forces applied to the lung/pectin adhesion resulted in a near-interface structural failure and the efficient isolation of intact en face pleural mesothelium. These data indicate that HMP, in an equal weight % mixture with CMC, is a promising mesothelial bioadhesive for use in experimental and therapeutic applications.

Functional Mechanics of a Pectin-Based Pleural Sealant after Lung Injury.

Pleural injury and associated air leaks are a major influence on patient morbidity and healthcare costs after lung surgery. Pectin, a plant-derived heteropolysaccharide, has recently demonstrated potential as an adhesive binding to the glycocalyx of visceral mesothelium. Since bioadhesion is a process likely involving the interpenetration of the pectin-based polymer with the glycocalyx, we predicted that the pectin-based polymer may also be an effective sealant for pleural injury. To explore the potential role of an equal (weight%) mixture of high-methoxyl pectin and carboxymethylcellulose as a pleural sealant, we compared the yield strength of the pectin-based polymer to commonly available surgical products. The pectin-based polymer demonstrated significantly greater adhesion to the lung pleura than the comparison products (p < 0.001). In a 25 g needle-induced lung injury model, pleural injury resulted in an air leak and a loss of airway pressures. After application of the pectin-based polymer, there was a restoration of airway pressure and no measurable air leak. Despite the application of large sheets (50 mm2) of the pectin-based polymer, multifrequency lung impedance studies demonstrated no significant increase in tissue damping (G) or hysteresivity (η)(p > 0.05). In 7-day survival experiments, the application of the pectin-based polymer after pleural injury was associated with no observable toxicity, 100% survival (N = 5), and restored lung function. We conclude that this pectin-based polymer is a strong and nontoxic bioadhesive with the potential for clinical application in the treatment of pleural injuries.