Aims. To verify whether secretory leucocyte protease inhibitor (SLPI) can promote early tendon-to-bone healing after anterior cruciate ligament (ACL) reconstruction. Methods. In vitro: the mobility of the rat bone mesenchymal stem cells (BMSCs) treated with SLPI was evaluated by scratch assay. Then the expression levels of osteogenic differentiation-related genes were analyzed by real-time quantitative PCR (qPCR) to determine the osteogenic effect of SLPI on BMSCs. In vivo: a rat model of ACL reconstruction was used to verify the effect of SLPI on tendon-to-bone healing. All the animals of the SLPI group and the negative control (NC) group were euthanized for histological evaluation, micro-CT scanning, and biomechanical testing. Results. SLPI improved the migration ability of BMSCs and upregulated the expression of genes related to osteogenic differentiation of BMSCs in vitro. In vivo, the SLPI group had higher histological scores at the tendon-bone interface by histological evaluation. Micro-CT showed more new bone formation and bone ingrowth around the grafted tendon in the SLPI group. Evaluation of the healing strength of the tendon-bone connection showed that the SLPI group had a higher maximum failure force and stiffness. Conclusion. SLPI can effectively promote early tendon-to-bone healing after ACL reconstruction via enhancing the migration and osteogenic differentiation of BMSCs. Cite this article: Bone Joint Res 2022;11(7):503–512

Elevated synovial leukocyte count is a minor criterion derived from the musculoskeletal infection society (MSIS) widely used in clinical practice for diagnosis of prosthetic joint infection. There is evidence to suggest analysis within 1 hour, preferentially within 30 minutes, of aspiration reduces the risk of ex vivo cell lysis occurring during prolonged transport. Multiple site working is more common practice and the availability of a lab on site to perform these tests is not always possible. We aimed to assess whether we could safely perform synovial leukocyte counts within our cold site in the diagnosis of prosthetic joint infection. We reviewed all orthopaedic synovial fluid aspirates within the lower limb arthroplasty unit from April 2021 – April 2022 performed at South Tees NHS Foundation Trust. We assessed time from aspirate to the lab using electronic data resources. This information was compared with the labs ability to perform a synovial leukocyte count to determine the impact of delays on testing. 110 patients (34.5% hips and 63.6% knees) were identified between two sites. Time from aspirate to lab ranged from 0 mins to 26 hrs 34 mins. Mean time to processing was 3hrs 10 mins. 50% of all samples had a synovial leukocyte count performed. 67% of patients had a cell differential performed. There was no difference in the ability to perform a synovial leukocyte count between samples process in < 2hours vs > 6 hours. We conclude that it is safe practice to perform joint aspirates for the work up of periprosthetic joint infections in sites where no laboratory is immediately available as the delay to processing synovial fluid does not alter the ability to perform a synovial leukocyte count. This study will provide evidence to enable the work up of periprosthetic joint infections in cold centres and therefore reduce the delay in diagnosis and proceeding management

Aim. The cut-off values for synovial fluid leukocyte count and neutrophils differential (%PMN) for differentiating aseptic from septic failure in total knee arthroplasties were already defined in the past. Our goal was to determine the cut-off values for synovial fluid leukocyte count and %PMN in failed total hip arthroplasties (THA). Method. Patients undergoing revision THA were prospectively included. In perioperative assessment phase, synovial fluid leukocyte count and %PMN were determined. During the surgery, at least 4 intraoperative samples for microbiological and one for histopathological analysis were obtained. Infection was defined as presence of sinus tract, inflammation in histopathological samples, and ≥2 tissue and/or synovial fluid samples growing the same microorganism. Exclusion criteria were systemic inflammatory diseases, revision surgery performed less than 3 months from index surgery and insufficient tissue sampling. Receiver operating characteristic (ROC) curves were constructed to assess the diagnostic performance and Youden's J statistic was computed to identify optimal cut-off values. Results. During the study period (between June 2006 and June 2011) 227 revision THAs were performed by the senior author. 31 patients were excluded. 196 patients (mean age, 69 years; 68% females) with THA failure were included. Aseptic failure was diagnosed in 150 patients (76,5%) and THA infection was diagnosed in 46 patients (23,5%). Synovial fluid leukocyte counts were significantly higher in the infected group (median, 5.50×10. 6. leukocytes/ml range, 0.05 to 143.9×10. 6. leukocytes/mL) than in the aseptic group (median, 0.23×10. 6. cells/ml; range, 0 to 21.3×10. 6. leukocytes/ml, P<0,0001). The %PMN was also significantly higher in the infected group (median, 83%; range, 6% to 97%) than in the aseptic group (median, 27,5%; range, 0% to 94%, P<0,0001). A synovial fluid leukocyte count of > 1.54×10. 6. leukocytes/ml, had a sensitivity of 63%, specificity of 95%, positive and negative predictive values of 78% and 89%, respectively. A synovial fluid %PMN of > 64%, had a sensitivity of 65%, specificity of 93%, positive and negative predictive values of 73% and 90%, respectively. Conclusion. The synovial fluid leukocyte count of > 1.54×10. 6. leukocytes/ml and %PMN of > 64% are useful and reliable tests for excluding THA infection, having a negative predictive value of around 90%. This tests and calculated cut-off values are highly recommended in the diagnostic process of failed THAs

Objectives. Cortical and cancellous bone healing processes appear to be histologically different. They also respond differently to anti-inflammatory agents. We investigated whether the leucocyte composition on days 3 and 5 after cortical and cancellous injuries to bone was different, and compared changes over time using day 3 as the baseline. Methods. Ten-week-old male C56/Bl6J mice were randomized to either cancellous injury in the proximal tibia or cortical injury in the femoral diaphysis. Regenerating tissues were analyzed with flow cytometry at days 3 and 5, using panels with 15 antibodies for common macrophage and lymphocyte markers. The cellular response from day 3 to 5 was compared in order to identify differences in how cancellous and cortical bone healing develop. Results. Between day 3 and 5, the granulocytes increased in the cancellous model, whereas the lymphocytes (T cells, B cells, NK cells) and monocytes (CD11b+, F4/80+, CD206+, CD14+) increased in the cortical model. Conclusion. These results suggest an acute type of inflammation in cancellous bone healing, and a more chronic inflammation in cortical healing. This might explain, in part, why cancellous healing is faster and more resistant to anti-inflammatory drugs than are diaphyseal fractures. Cite this article: L. Tätting, O. Sandberg, M. Bernhardsson, J. Ernerudh, P. Aspenberg. Different composition of leucocytes in cortical and cancellous bone healing in a mouse model. Bone Joint Res 2018;7:620–628. DOI: 10.1302/2046-3758.712.BJR-2017-0366.R2

Aims. Current guidelines consider analyses of joint aspirates, including leucocyte cell count (LC) and polymorphonuclear percentage (PMN%) as a diagnostic mainstay of periprosthetic joint infection (PJI). It is unclear if these parameters are subject to a certain degree of variability over time. Therefore, the aim of this study was to evaluate the variation of LC and PMN% in patients with aseptic revision total knee arthroplasty (TKA). Methods. We conducted a prospective, double-centre study of 40 patients with 40 knee joints. Patients underwent joint aspiration at two different time points with a maximum period of 120 days in between these interventions and without any events such as other joint aspirations or surgeries. The main indications for TKA revision surgery were aseptic implant loosening (n = 24) and joint instability (n = 11). Results. Overall, 80 synovial fluid samples of 40 patients were analyzed. The average time period between the joint aspirations was 50 days (SD 32). There was a significantly higher percentage change in LC when compared to PMN% (44.1% (SD 28.6%) vs 27.3% (SD 23.7%); p = 0.003). When applying standard definition criteria, LC counts were found to skip back and forth between the two time points with exceeding the thresholds in up to 20% of cases, which was significantly more compared to PMN% for the European Bone and Joint Infection Society (EBJIS) criteria (p = 0.001), as well as for Musculoskeletal Infection Society (MSIS) (p = 0.029). Conclusion. LC and PMN% are subject to considerable variation. According to its higher interindividual variance, LC evaluation might contribute to false-positive or false-negative results in PJI assessment. Single LC testing prior to TKA revision surgery seems to be insufficient to exclude PJI. On the basis of the obtained results, PMN% analyses overrule LC measurements with regard to a conclusive diagnostic algorithm. Cite this article: Bone Jt Open 2021;2(8):566–572

No single test has demonstrated absolute accuracy in the diagnosis of periprosthetic joint infection (PJI). Leukocyte esterase (LE) is a synovial marker that has proven utility in the diagnosis of PJI. The purpose of this prospective study was to (1) identify the optimal cutoff for the use of LE in the diagnosis of PJI and (2) determine whether performance of the LE strip test varied by infecting organism. This prospective study enrolled 1,015 patients undergoing hip or knee revision arthroplasty at a single institution from 2009 to September 2021. PJI was defined using a modified version of 2018 International Consensus Meeting (ICM) criteria that excluded LE when calculating the ICM score. Receiver operating characteristic curves were used to assess the utility of the LE strip test in the diagnosis of PJI. 973 patients were included in the analyses. 246 (25.4%) were classified as ICM-positive and 727 (74.6%) were classified as ICM-negative. An LE cutoff of “1+” (AUC 0.819, sensitivity 73.2%, specificity 90.6%) had superior accuracy to an LE cutoff of “2+” (AUC 0.713, sensitivity 43.9%, specificity 98.8%) in the overall diagnosis of PJI (p<0.001). When stratifying by organism type, an LE cutoff of “1+” had the best diagnostic utility for PJI caused by methicillin resistant Staphylococcus aureus (AUC 0.888, sensitivity 87.0%, n=23) followed by Streptococcus spp. (AUC 0.882, sensitivity 85.7%, n=28), coagulase negative Staphylococci (AUC 0.836, sensitivity 76.6%, n=47), methicillin sensitive Staphylococcus aureus (AUC 0.806, sensitivity 70.6%, n=34), culture negative (AUC 0.793, sensitivity 67.9%, n=56), and gram negative rods (AUC 0.763, sensitivity 61.9%, n=21). To our knowledge, this is the largest study evaluating the utility of the LE strip test in the diagnosis of PJI. Based on our findings, it appears that a “1+” cutoff has higher diagnostic utility than a cutoff of “2+”

Aim. The aim of the study is to evaluate the specificity and sensibility of leukocyte esterase for the diagnosis of periposthetic joint infection (PJI). Method. Between October 2016 and April 2017 we enrolled 65 patients underwent to hip and knee revision arthroplasty due to uncertain joint infection. Synovial fluid was obtained from 64 joints that underwent revision arthroplasty. Each patient was evaluated in the preoperative time with CRP, ESR and leukoscan, in the intraoperative time with frozen section and leukocyte esterase strip and post-operative with sonication fluid culture, periprosthetic tissues cultures and histological examination. Results of all of these exams were compared to assess the specificity, the sensibility, the positive and negative predicting values of leukocyte esterase for the diagnosis of PJI. Results. The leukocyte esterase test with a threshold of +/++ had a sensitivity of 80.2%, a specificity of 82.8%, a positive predictive value of 63.8%, and negative predictive value of 92.1%. Using the threshold of ++ as a positive leukocyte esterase result, the specificity reached 97.8%, the positive predictive value 90.8%, and the negative predictive value 89.0%. Conclusions. These results demonstrate that leukocyte esterase is a quite accurate, effective marker of periprosthetic joint infection and that it is a valuable tool that can be used in conjunction with the the other tests for diagnosis of PJI

Leucocytes represent a very important host defence against a number of invading pathogens and neoplasia. However, the activity of phagocytic leucocytes has been heavily implicated in the development of ischaemia-reperfusion injury, and as an aetiological factor in the pathology of other clinically important inflammatory conditions. Ischaemia-reperfusion injury occurs in diseases such as stroke and ischaemic heart disease (IHD), and during surgical procedures such as orthopaedic surgery. Investigations presented here employed a model of tourniquet-induced forearm ischaemia-reperfusion injury to investigate the effect on leucocyte adhesion and trapping (n=20). Neutrophil and monocyte leucocyte subpopulations were isolated by density gradient centrifugation techniques. Neutrophil and monocyte cell surface expression of the adhesion molecule CD11b was measured by labelling with fluorescent anti-CD11b monoclonal antibody via flow cytometry. Plasma concentrations of the soluble intercellular adhesion molecule-1 (sICAM-1) and soluble L-selectin (sL-selectin) adhesion molecules were measured using commercially available ELISA kits. Leucocyte trapping was investigated by measuring the concentration of leukocytes in venous blood leaving the arm. During ischaemia-reperfusion there was an increase in CD11b expression on neutrophils (p=0.040) and monocytes (p=0.049), a decrease in sL-selectin (p=0.387) and sICAM-1 (p=0.089) concentrations, and a decrease in peripheral blood leucocyte concentration (p=0.019). Evidence of increased leucocyte adhesion and trapping during ischaemia-reperfusion injury was supported by an increase in CD11b cell surface expression of neutrophils and monocytes. CD11b is expressed on phagocytic leucocytes and binds to ICAM-1 expressed on the surface of vascular endothelium. This increased expression of CD11b on leucocytes may therefore play a central role as the mechanism by which leucocyte trapping in the microcirculation occurs. The measured decrease in plasma concentration of sICAM-1 and sL-selectin suggests that these adhesion molecules retain their functional activity, and may bind to their corresponding cell surface ligands. It is therefore reasonable to believe that ICAM-1 expressed on the endothelium and L-selectin expressed on leucocytes is also binding to their corresponding cell surface ligands. A decrease in the number of leucocytes in the peripheral circulation may be due to increased trapping of leucocytes in the microcirculation. When leucocytes become trapped their concentration in blood leaving the microcirculation decreases, resulting in the measured decrease in leucocyte concentration. In conclusion, this study confirms the important role of leucocytes during ischaemia-reperfusion injury, which could allow for the possibility of future research that may provide therapeutic intervention for inflammatory conditions

Aims. The purpose of this study was to validate our hypothesis that centrifugation may eliminate false-positive leucocyte esterase (LE) strip test results caused by autoimmune diseases in the diagnosis of knee infection. Methods. Between January 2016 and May 2019, 83 cases, including 33 cases of septic arthritis and 50 cases of aseptic arthritis, were enrolled in this study. To further validate our hypothesis, another 34 cases of inflammatory arthritis from the Department of Rheumatology of our institution were also included. After aspiration, one drop of synovial fluid was applied to LE strips before and after centrifugation. The results were recorded after approximately three minutes according to the different colour grades on the colour chart. The differences of LE results between each cohort were analyzed. Results. Before centrifugation, 46% (23/50) of the LE strip tests in the aseptic arthritis group were false-positives. Most of the false-positive results were due to inflammatory arthritis; after centrifugation, 78.3% (18/23) of the tests yielded negative results. Similar results were observed in cases from the Department of Rheumatology. The sensitivity of the centrifuged LE strip test was 0.818 (0.639 to 0.924), which is still an acceptable level compared with the uncentrifuged results, which yielded a sensitivity of 0.909 (0.745 to 0.976). However, the specificity was increased from 0.540 (0.395 to 0.679) to 0.900 (0.774 to 0.963) after centrifugation. Conclusion. Although inflammatory arthritis can yield a false-positive LE strip test result in the diagnosis of knee infection, centrifugation may eliminate these false-positive results. Cite this article: Bone Joint Res. 2020;9(5):236–241

Leukocyte esterase (LE) has shown to be an accurate marker of prosthetic joint infections and has been proposed as an alternative to frozen section (FS) for intra-operative diagnosis. In this study, intra-operative determination of LE was compared with FS for the diagnosis of periprosthetic hip infections. One hundred and nineteen patients undergoing hip revision surgery due to prosthetic joint failure from June 2015 to December 2017 were considered. Joint fluids were collected before the arthrotomy for determination of LE which was performed by using enzymatic colorimetric strips. Four to six samples were stained with hematoxylin eosin for FS and considered suggestive for infection when at least 5 polymorphonuclear leukocytes in 5 fields at high power fields were found. Sensitivity and specificity of LE were 100% and 93.8 %, respectively. The positive predictive value was 79.3 %, while the negative predictive value was 100%. Time from collection to response was 20.1 ± 4.4 minutes. Sensitivity and specificity of FS were 83.3 % and 92.1 %, respectively. The positive and negative predictive values were 84.6 % and 97.1%. Time from sample collection response was 27.2 ± 6.9 minutes. LE showed a higher sensitivity and a slightly lower specificity and the same diagnostic accuracy of intraoperative FS. The faster turnaround time (about 20 minutes from receiving of sample by the laboratory), its ease of use and the low costs make this test a valuable alternative to frozen sections and is going to replace FS in our clinical practice

Leucocytes are white blood cells that help the body fight against bacteria, viruses and tumour cells. However, the activity of leucocytes has been implicated in other clinically important inflammatory conditions such as ischaemic heart disease, stroke, and during cardio-aortic and orthopaedic surgery. The main objectives of this study was to optimise methods for the isolation of leucocyte subpopulations (neutrophils and monocytes), and to assess in vitro the effects of PMA and fMLP on markers of leucocyte adhesion (CD11b, CD62L) and activation (intracellular hydrogen peroxide) (n=10). Leucocyte subpopulations were labelled by incubation with fluorescein isothiocya-nate (FITC) conjugated anti-human CD11b and CD62L antibodies. The cell surface expression of these labelled adhesion molecules were measured by flow cytometry. Intracellular production of hydrogen peroxide by neutrophils and monocytes was measured by flow cytometry, using the fluorochrome dichloroflurorescin diacetate (DCFH-DA). These were visualised by Immunofluorescence microscopy. During this study, methods of isolating leucocyte subpopulations from whole blood were optimised. This ensured that these cells were isolated with consistently high yields, purity and with no changes in cellular function. Following incubation with PMA and fMLP, neutrophils and monocytes displayed an increase in CD11b cell surface expression; a decrease in CD62L cell surface expression; and increased leucocyte activation. Leucocyte activation was represented by the intracellular production of hydrogen peroxide. In conclusion this study confirms that both PMA and fMLP have an intrinsic effect on markers of leucocyte function. These findings are in agreement with previous studies performed

Background. Periprostetic joint infections (PJI) are often difficult to diagnose, to treat and often leave the patient with severe impaired function. The presence of low virulent bacteria is frequently discovered in apparent aseptic revisions of shoulder arthroplasties and pose a challenge to diagnose preoperatively. Dual Isotope In111 Leucocyte/ Tc99 Bone Marrow SPECT CT scan (L/BMS) is considered the radionuclide gold standard in preoperative diagnosing PJI with reported high specificity and sensitivity in hip and knee arthroplasties. Unfortunately, it is labour-intensive and expensive to perform and documentation using L/BMS on shoulder arthroplasties lack. Aim. To investigate if L/BMS succeeds in detecting shoulder PJI compared to tissue cultures obtained perioperatively. Method. All patients referred to a highly-specialised shoulder department with a painful or stiff shoulder-arthroplasty were included in the cohort. To diagnose infection as a possible cause of arthroplasty failure a L/BMS was planned for all patients. If the arthroplasty was revised, 5 tissue biopsies were obtained from the most infection-suspicious site during revision. Biopsies were cultured in broth and on plates for 14 days due to the high frequency of low virulent infection in shoulder revisions. Infection was defined as growth of the same bacteria in 3 or more of 5 the biopsies. Results. During the observation period 71 patients were referred. Revision surgery was performed in 62% of the patients (44/71) of which 29 also had been examined by L/BMS. A microbiological diagnose was available for all. The most predominant organism isolated was P. Acnes. Two patients both had a positive L/BMS and positive cultures. Negative L/BMS and negative cultures were found in 20 patients. The remaining 7 patients had negative L/BMS, but positive cultures. The two patients with a positive L/BMS both showed overt clinical signs of infection. L/BMS show a sensitivity 0.22 95%CI(0–0.49) and specificity 1.00 95%CI(1.00–1.00) in detecting shoulder PJI. The Positive Predictive Value is 1.00 95%CI(1.00–1.00) and Negative Predictive Value 0.74 95%CI(0.57–0.90). No patients infected with P. Acnes resulted in a positive scintigraphy nor had they preoperative or perioperative signs of infection. Conclusion. Only patients with severe infectious symptoms of shoulder PJI resulted in positive L/BMS. Hence, the scan added nothing to the preoperative clinical diagnose. In111 Leucocyte/ Tc99 Bone Marrow SPECT CT scan cannot be recommended as a standard screening procedure when evaluating failed shoulder arthroplasties for possible infection

Aim. Treatment of complicated wound healing after total joint arthroplasty is controversial. What exactly constitutes prolonged wound drainage is matter of debate and recommendations to manage it vary considerably. Nonoperative measures are often recommended. If drainage persists, surgery may be indicated. To further intricate decision-making, differentiating superficial from deep surgical site infection is also controversial and inherently complex. Specific cutoffs for synovial fluid leukocyte count and blood C-reactive protein (CRP) in the acute stage have been suggested as a way to superficial infection requiring superficial wound washout from deep infection requiring a formal debridement, antibiotics and implant retention (DAIR) procedure. The goal of this study is to analyze clinical and laboratory findings of an institutional protocol of “aggressively” proceeding with formal DAIR in all patients with complicated wound healing. Method. Our indications for DAIR in suspected acute postoperative periprosthetic joint infection (PJI) are: 1)prolonged wound drainage and CRP upward trend after day-3; 2)persistent wound drainage by day-10 regardless of CRP; 3)wound healing disturbance (e.g. “superficial” infection, “superficial” skin necrosis) anytime in early postoperative weeks. We retrospectively evaluated patients undergoing DAIR in the first 60 postoperative days between 2014–2018. Patients without multiple deep tissue cultures obtained intraoperative were excluded. Deep infection was defined by at least two positive deep tissue cultures or one positive deep culture and positive leukocyte count (>10,000 cells/mL or >90% PMN). Results. A total of 44 DAIR procedures were included. Deep infection was confirmed in 79.5%(35/44) of cases. Mean CRP in infected cases was 93mg/L with 63%(19/30) of them below the 100 mg/L threshold. Unfortunately, only a small proportion of cases (10/44) had synovial fluid leukocyte counts available. Mean leukocyte count was 15,558 cells/mL and mean proportion of PMN was 65.3%. Of these ten, six confirmed deep infections were below the proposed >10,000 cells/mL or >90% PMN cutoff. Conclusions. Early diagnosis of acute postoperative PJI is often hampered by its very subtle presentation. This study confirms that more often than not, deep infection is present when facing complicated wound healing after total joint arthroplasty, supporting our institutional “aggressive” protocol. We have been surprised by the number of confirmed acute PJI with low blood CRP levels and low synovial leukocyte counts. We hypothesize that the proposed acute PJI specific thresholds may lead to misinterpret a significant proportion of cases as superficial infections thus compromising timely intervention. The findings of this study lack confirmation in larger cohorts

Introduction: Labelled leukocyte scintigraphy has been shown to be a sensitive and specific technique for the detection of pedal osteomyelitis in patients with diabetes mellitus. There has however been little data relating the efficacy of the technique to outcomes. Aim: To examine the prognostic value of sequential 99m Tc labelled leukocyte scans at diagnosis and after 3–4 weeks of appropriate antibiotic therapy. Method: Twenty-three patients with proven pedal osteomyelitis or persistent uptake on the sequence of scans were studied. Results: Five additional episodes of osteomyelitis developed in the group over the period of the study. Eleven patients demonstrated persistent uptake in the sequential scans. Nine progressed to amputation. The remaining two patients were biopsy negative for infection, did not have cutaneous ulceration and were thought to have rapidly progressive arthropathy. Conclusion: Sequential leukocyte scintigraphy accurately predicted the need for amputation and circumvented ineffective prolonged antibiotic therapy

Culture examination is still considered the gold standard for diagnosis of bone and joint infections, including prosthetic ones, even if in up to 20–30% of cases, particularly prosthetic joint infections, it fails to yield microbial growth. To overcome this limitation, determination of markers of inflammation and or infection directly in joint fluid has been proposed. Aim of this study was to evaluate the applicability of measurement of lecukocyte esterase (LE), C-reactive protein (CRP) and glucose in synovial fluid for diagnosis of bone and joint infections. Synovial fluids from 80 patients were aseptically collected and sent to laboratory for microbiological cultures. After centrifugation at 3000 rpm for 10 minutes, pellet was used for cultures, while the surnatant was used for determination of LE, CRP and glucose. LE and glucose were evaluated by means of enzymatic colorimetric strips developed for urinanalysis. One drop of synovial fluid was placed on the LE and on the glucose pads and the results were read after about 120 seconds. A LE test graded + or ++, and a glucose test equal to trace or negative were considered suggestive for infection. CRP was measured by an automated turbidimetric method. On the basis of clinical findings, microbiological, haematological and histological analyses patients were retrospectively divided into 2 groups. Group 1 comprised 19 infected patients (12 males, 7 females age: 70.6 ± 10.3 yrs, range: 47 – 88 yrs) while Group 2 included 61 aseptic patients (32 males and 29 females, age: 61.5 ± 16.3 yrs, range: 15 – 84). Sensitivity of the three tests was 89.5%. 84% and 73,7% for LE, CRP and glucose, respectively. Specificity was 98.4%, 88.5% and 70% for LE, CRP and glucose, respectively. Positive and negative predictive values were 94.4% and 96.8% for LE, 69.6% and 94.6% for CRP and 77.8% and 89.6% for glucose test. When LE was combined with CRP, sensitivity increased to 94.7%, while no differences were observed for LE combined with glucose. Leukocyte esterase has proven to be a rapid, simple and inexpensive test to rule in or out bone and joint infections. Combination of its measurement with that of CRP increased sensitivity. In conclusion, the combination of leukocyte esterase and CRP may represent a simple and useful tool for diagnosis of bone and joint infections

Purpose: The diagnosis of chronic bone and joint infections, particularly in patients with implants, can be a difficult task. Among the clinical and laboratory tests proposed for the diagnosis of infection, 99mTc HMPOA labelled leukocyte scintigraphy is one of the least invasive examinations available. We evaluated its efficacy in terms of reliability. Material and methods: Ninety patients with suspected bone and joint infections were included in this study: 53% men and 47% women. Mean age was 56.6 years and 80% had osteosynthesis implants. Mean duration of clinical signs before scintigraphy was 6.5 months. The suspected site was the hip in 49%, the knee in 28% and another in 23%. Physical examination (local aspect, temperature) and laboratory tests (differential count, platelets, CRP, ESR) as well as standard radiographs were performed in addition to labelled scintigraphy. These patients were operated and bone samples were taken for bacteriology studies to confirm or infirm the presence of infection. In this series, 73% of the patients were found to have a real infection (73% staphylococcal, 17% multiple germs, 20% other). Results: The following variables were included in the multivariate analysis: fever, standard radiographs, polynuclear neutrophil count, CRP, ESR, leukocyte-labelled scintigraphy. Sensitivity (Se), specificity (Sp), and odds ratio (OR) were determined. The multivariate analysis showed: fever (Se=0.48; Sp=0.59; OR=1.3); abnormal radiograph (Se=0.71; Sp=0.62; OR=4; p=0.02); polynuclear neutrophil count (OR=1; p=0.19); CRP (OR=1.02; p=0.06); ESR (OR=1.03; p=0.04); leukocyte-labelled scintigraphy (Se=0.71; Sp=0.82; OR=11.6; p< 0.001). Discussion and conclusion: These findings demonstrate the efficacy of 99mTc HMPOA-labelled leukocyte scintigraphy in terms of reliability for the diagnosis of chronic bone infection compared with other clinical (fever), laboratory (ESR, CRP), and radiographic indicators

The aim of the study to analyze the circulating white blood cells including the intensity expression of surface receptors and cytoplasmic molecules in patients underwent total hip replacement, with either aseptic or septic loosening of hip prostheses in order to identify cell-surface and cytoplasmic markers that could be indicative of early loosening. Flow cytometry was performed in whole peripheral blood samples of 20 patients with loosening (10 septic and 10 aseptic). Ten healthy individuals served a control group. The CD62L, CD18, CD11a, CD11b and CD11c expressions were evaluated. The mean fluorescence intensity (MFI) of CD 18 was decreased on all leukocytes subsets compared to control group. For patients with aseptic loosening we demonstrated an increase of MFI for CD11b in granulocytes and for CD11c in monocytes and granulocytes compared to control group. In patients with septic loosening an increase of MFI for CD 11c was observed in monocytes compared to control group. The comparison between aseptic and septic loosening showed a statistically significant lower CD18 MFI value in granulocytes for aseptic loosening. A trend towards lower MFI values of CD 62L in lymphocytes and granulocytes were observed in aseptic but not in septic loosening patients compared to control group. The present study is the first study in published literature to demonstrate cell surface and cytoplasmic markers in peripheral blood indicative of loosening of THAs by means of flow cytometry

Summary. This is the first ever study to report the successful elimination of malignant cells from salvaged blood obtained during metastatic spine tumour surgery using a leucocyte depletion filter. Introduction. Catastrophic bleeding is a significant problem in metastatic spine tumour surgery (MSTS). However, intaoperative cell salvage (IOCS) has traditionally been contraindicated in tumour surgery because of the theoretical concern of promoting tumour dissemination by re-infusing tumour cells into the circulation. Although IOCS has been extensively investigated in patients undergoing surgery for gynaecological, lung, urological, gastrointestinal, and hepatobiliary cancers, to date, there is no prior report of the use of IOCS in MSTS. We conducted a prospective observational study to evaluate whether LDF can eliminate tumour cells from blood salvaged during MSTS. Patients & Methods. After Institutional Review Board (IRB) approval, 21 consecutive patients with metastatic spinal tumours from a known epithelial primary (defined as originating from breast, prostate, thyroid, renal, colorectal, lung, nasopharyngeal) who were scheduled for MSTS were recruited with informed consent. During surgery, a IOCS device (Dideco, Sorin Group, Italy) was used to collect shed blood from the operative field. Salvaged blood was then passed through a leucocyte depletion filter (RS1VAE, Pall Corporation, UK). 15-ml specimens of blood were taken from each of three consecutive stages: (i) operative field prior to cell saver processing (Stage A); (ii) transfusion bag post-cell saver processing (Stage B); (iii) filtered blood after passage through LDF (Stage C). Cell blocks were prepared by the pathology department using a standardised laboratory protocol. From each cell block, 1 haematoxylin and eosin (H&E) slide, and 3 slides each labelled with one of the following monoclonal mouse cytokeratin antibodies AE1/3, MNF 116 and CAM 5.2 were prepared. The cytokeratin antibodies are highly sensitive and specific markers to identify tumour cells of epithelial origin. These slides were read by one of two consultant pathologists who were provided full access to information on operative notes, but were blinded to the actual stages from which the slides were derived. Results. One case was excluded when the final diagnosis was revised to infection instead of metastatic spine tumour. Of the remaining cases, 7/21 tested positive for tumour cells in Stage A, 2 positive in Stage B. No specimen tested positive for tumour cells in Stage C. In 5 cases, posterior instrumentation without tumour manipulation was performed. Discussion/Conclusion. In this first-ever study of cell saver use in spine tumour surgery, we prove that leucocyte-depletion filters (LDF) can effectively eliminate tumour cells from blood salvaged during MSTS. It is now possible to conduct a clinical trial to evaluate IOCS-LDF use in MSTS. Our results are consistent with published results of similar studies performed on IOCS and LDF use outside the field of orthopaedic surgery. Spinal metastases originate from a myriad of primary cancers across various organ systems. If LDF can remove tumour cells from blood salvaged during surgery for spinal metastasis of different histological origin, then the finding can likely be extrapolated to several other fields of surgery where IOCS and LDF have not yet been attempted such as: neurosurgery, otolaryngology and general musculoskeletal oncology. Our results form a proof-of-concept for a paradigm shift in thinking regarding autotransfusion during spine tumour surgery

Injection before total knee arthroplasty(TKA) is the one of the postoprative risk factors after TKA and Infection after TKA can result in disastrous consequences. When the duration between injection and TKA is longer than 6 months, the risk is no longer elevated. Evaluation of synovial WBC number in frozen section slide is needed to check the presence of infection in revision total knee arthroplasty. Currently many patients have a history of multiple intraarticular injection before the primary TKA. Purpose of this study is to evaluate the synovial WBC findings in primary TKA and compare between injection group and no injection group. Materials and Methods. The synovial specimen(suprapatella pouch and posterior capsule) of 68 primary total knee arthroplasty were evaluated by the pathologist and reported the number of the WBC in frozen section /5 separate high power fields(HPF) (500x).. Injection group were 37 cases and non -injection group were 31 cases. Preoperative CRP and ESR were recorded and followe-up duration was more than 2 years. Joint fluid was sent to be cultured and analysed. Results. WBC count in frozen section shoed was average 4 WBCs/HPF (range < 0∼ 25) in both specimen and the suprapatella specimen was 3 WBCs/HPW (range 0∼25) and posterior capsule specimen was 1 WBCs/HPF(range 0∼14). The WBC count of injection group was 8 (range, 0∼25) and that of no injection group was 1.2 cells (range 0∼12) (p<0.05). The WBC counts in joint fluid was average 240 cells/ml (range. 1∼300) in non injection group and 643 cells/ml(range, 50∼1000) (p<0.05). The duration from the intraarticular injection to index surgery was 9 months(range, 6 weeks∼ 7 momths). The number of injection and duration bwtween injection and operationto has no significant correlation with the WBC counts. Eight percentage of specimen showed more than 10 WBCs in injection group and these patients have been not infected after more than 24 moths after TKA. Conclusion. The WBC count of the synovium in priamry TKA with injection history for degenerative osteoarthritis is variable and we could not recommend the routine frozen section analysis in primary TKA who have a history of intraarticualr injection..

Aim

The purpose of this work is to study whether there is or not, in the case of an aseptic arthroplasty exchange, a relationship between positive cultures and an early periprosthetic joint infection.