Specific discrimination of pathogenic bacteria causing septic arthritis using Raman spectroscopy: In-vitro study.

In this study we performed preliminary experiments using Raman spectroscopy as an evolving technology in biofluid and microbial characterization, to explore its potential for rapid diagnosis of pathogenic bacteria in an in-vitro synovial fluid infection model. Normal human synovial fluids samples were collected from patients undergoing knee surgery and the three most common pathogenic bacteria introduced in-vitro into the samples. The bacterial growth was systematically monitored using a Raman spectroscopy. Multivariate regression analysis of acquired spectra showed bacterial characteristic Raman bands related to bacterial cell membranes and DNA structures to increase continuously as the incubation period was increased. Spectra signature recorded from cultured synovial fluid samples showed a significant loss in synovial quality and protein morphology over time compared to control samples. In this study, Raman spectroscopy shows promise for rapid pathogenic bacteria identification in synovial fluid. Marker peaks distinguished inoculated bacteria, while chemical changes reveal infection dynamics.

Analysis of Bone Microarchitectural Changes and Structural Damage in Sickle Cell Disease-Induced Avascular Necrosis Using Raman Spectroscopy: Is there potential for medical management?

OBJECTIVES: Bone failure due to avascular necrosis (AVN) is a complex pathological phenomenon. Analysis of molecular changes in the bone matrix may help to shed light on the disease process and guide management. This study aimed to explore changes in bone quality and structural damage caused by sickle cell disease (SCD)-induced AVN using Raman spectroscopy. METHODS: A total of 10 necrotic femoral heads were obtained from seven SCD patients who underwent total hip replacements. The femoral heads were cut in half and scanned using Raman spectroscopy in correlation with preoperative magnetic resonance imaging to identify necrotic and healthy control areas. Subsequently, samples were examined to determine changes in bone mineralisation, crystallinity, carbonate content, collagen cross-linking and mineral and collagen fibril orientation. RESULTS: Significant changes were observed in bone mineral content, mineral-to-organic content and collagen fibril orientation in necrotic compared to control areas (P ≤0.050). CONCLUSION: The necrotic samples displayed severe structural damage and loss of mineral and organic contents. Similar Raman signals have been reported in other metabolic bone diseases such as osteoporosis, thereby potentially supporting the use of medical treatment in AVN to promote bone quality.

Investigating the Growth of Pseudomonas aeruginosa and Its Influence on Osteolysis in Human Bone: An In Vitro Study.

BACKGROUND: Isolation of the causal microorganisms in osteomyelitis presents a major challenge for treating clinicians. Several methods have been proposed to rapidly and accurately identify microorganisms. There has been an increasing interest in using Raman spectroscopy in the field of microbial detection and characterisation. This paper explores the use of Raman spectroscopy identification as one of the most difficult-to-isolate microorganisms causing osteomyelitis. METHODS AND RESULTS: Fresh healthy human bone samples were collected from patients undergoing a total knee replacement. These samples were then inoculated with fresh overnight Pseudomonas aeruginosa (PAO) cultures. Bacteria growth and bone ultrastructural changes were monitored over a period of 6 weeks. The experiment demonstrated ultrastructural bony destruction caused by osteolytic PAO secretions. Raman-specific spectral signatures related to the cellular membranes of PAO structures were spotted indicating survival of bacteria on the bone surface. CONCLUSION: This study showed the promising ability of Raman spectroscopy to identify the presence of bacteria on the surface of inoculated bone samples over time. It was able to detect the osteolytic activity of the bacteria as well as ultrastructure specific to PAO virulence. This method may have a role as an aid to existing diagnostic methods for fast and accurate bacterial identification in bone infection. HOW TO CITE THIS ARTICLE: Al Ghaithi A, Al Bimani A, Al Maskari S. Investigating the Growth of Pseudomonas aeruginosa and Its Influence on Osteolysis in Human Bone: An In Vitro Study. Strategies Trauma Limb Reconstr 2021;16(3):127-131.

Good Clinical Success Rates Are Seen 5 Years After Meniscal Repair in Patients Regularly Undertaking Extreme Flexion.

PURPOSE: To report the functional and radiologic outcomes of meniscal repair healing in a cohort of patients with a high demand for loaded extreme flexion angles after undergoing meniscal repair. METHODS: We performed a retrospective clinical and radiologic evaluation of patients who perform extreme knee flexion activities on a daily basis at a minimum follow-up of 2 years after meniscal repair. International Knee Documentation Committee, Lysholm, and Tegner scores were obtained, and clinical examinations and radiologic (magnetic resonance imaging and radiography) evaluations were performed. RESULTS: Of 47 eligible patients, 39 patients (40 knees) were available for review with an average follow-up time of 5 years (range, 2-9 years). The average age was 26.7 years (range, 19-39 years); 38 patients were men. The average time from injury to surgery was 20.9 months (range, 3 days to 120 months). Associated anterior cruciate ligament injury was present in 31 knees, but only 20 underwent simultaneous anterior cruciate ligament surgery. The mean International Knee Documentation Committee score was 88.9 (range, 53-99). The mean Lysholm score was 90.9 (range, 48-100). The mean Tegner activity level dropped from 6.18 before injury to 5.51 at the time of evaluation. According to the Barrett criteria for clinical outcomes, complete healing was observed in 29 of 40 knees (72.5%). There was a statistically significant correlation between the functional outcomes and the clinical outcomes (P = .008). On magnetic resonance imaging, 22 of 38 knees (57.9%) showed completely healed menisci. CONCLUSIONS: The results of this study suggest that loaded deep knee flexion may be safe after a period of restricted rehabilitation, and clinical and radiologic tissue healing is independent of the overall functional outcome. LEVEL OF EVIDENCE: Level IV, case series with subgroup analysis.

Thirty-Day Mortality in COVID-19 Positive Patients With Hip Fractures: A Case-Series and Literature Review.

INTRODUCTION: The novel coronavirus has spread rapidly around the world with particularly high mortality in the elderly. Care and nursing homes have become the sites of greatest concentration of cases. We intended to review 30-day mortality of COVID-19 patients with hip fractures. This is relevant given the disease impact in this age group. METHODS: Medline and Embase databases were searched for relevant studies linked to mortality and morbidity in COVID-19 patients who have undergone non-elective hip surgeries using the keywords "COVID-19"OR "SARS-cov-2"OR "Coronavirus Infections"; AND "Surgery"OR "Hip"OR "Fracture"OR "Orthopedics." We included all patients with hip fractures, but excluded pathological fractures and other non-traumatic hip pathologies. Four-hundred and eighty-one articles were identified for screening, in addition to an unpublished case-series of 67 patients that have 3 cases turned positive for COVID 19, yielding a total of 50patients for the final review. RESULTS: The study included 4 articles published until May 9th 2020 and a case-series: 26(52%) patients were females; the median age was 86years; hypertension(53.6%), diabetes mellitus type II(28.6%), and coronary artery heart disease(25%) were the most common comorbidities; 34(68%) patients had intertrochanteric hip fracture and 16(32%) patients had femoral neck fractures; 22(59.5%) patients underwent cephalomedullary nail fixation, 12(32.4%) patients had hemiarthroplasty, and for 3(8.1%) patients, the type of surgery was not documented; 20(40%) patients died(12patients died before the surgery and 8died after surgery at a median time of 3days), 29(76.3%) patients had an unremarkable course throughout hospitalization and were discharged including 1(2.6%) patient managed non-operatively, and 1(2.6%) patient was admitted to the ICU after the surgery but eventually discharged. CONCLUSION: COVID-19 infected elderly patients have a higher 30-days mortality rate compared to non-COVID-19 infected cases. Further studies are warranted to look at the morbidity and mortality rates in COVID-19 positive patients with hip fractures and to investigate how these outcomes can be improved. LEVEL OF EVIDENCE: Level IV.

Retrograde femoral nails for emergency stabilization in multiply injured patients with haemodynamic instability.

INTRODUCTION: The purpose of this study is to retrospectively evaluate the immediate effect of retrograde intramedullary femoral nail (RIMFN) fixation technique on patient's hemodynamic status as documented by vital signs (blood pressure and pulse) intraoperatively in all patients with femoral shaft fractures with multiple injuries and hemodynamic instability who were treated with RIMFN at our institution on emergency basis as part of damage control orthopaedics. PATIENTS AND METHODS: A retrospective review of intra operative vital signs obtained from patient records was completed at a Level 1 trauma center in a university hospital.In all, 11 multiply injured patients with (14) femur fractures with hemodynamic instability were identified. Of those, 3 had bilateral femur fractures. Closed reduction and retrograde femoral nailing without proximal locking was performed to achieve immediate skeletal and haemodynamic stability. Pulse rate and BP measurements were noted for all patients starting from the time patient would enter the operating room till the patient was shifted back to the recovery ward. RESULTS: The average cohort age was 28 years (20-36 years). The average Injury Severity Score was 28 (16-50). Statistically significant improvement in pulse rate and blood pressure was noted following femoral fracture fixation with intramedullary nail. No cases of infection or symptomatic fat or pulmonary embolism were encountered. One patient required exchange nailing for non-union and one femur underwent later lengthening. CONCLUSIONS: Retrograde Intramedullary femoral nail can be an effective alternative to external fixator as damage control device and is associated with immediate improvement in vital signs (pulse and blood pressure) intra operatively.

Brucella Periprosthetic Joint Infection Involving Bilateral Knees with Negative Synovial Fluid Alpha-Defensin.

Periprosthetic joint infection (PJI) due to Brucella is uncommon despite relatively high endemicity of human brucellosis and its osteoarticular predilection. We report a case of a 57-year-old woman with bacteraemic brucellosis complicated by Brucella periprosthetic infection of both knee joints occurring a decade after bilateral knee arthroplasty and associated with a negative synovial fluid alpha-defensin test. The patient was successfully treated with anti-Brucella therapy alone and without surgical revision, resulting in clinical and microbiological cure. We propose that Brucella should be considered as a possible cause of prosthetic joint infection in the appropriate clinical and epidemiological settings. A negative synovial fluid alpha-defensin (Synovasure AD test) should not be used as a rule-out test for Brucella PJI. Brucella PJI without radiological loosening may be treated conservatively and solely with antimicrobial therapy.

Folate/Vitamin B12 Supplementation Combats Oxidative Stress-Associated Carcinogenesis in a Rat Model of Colon Cancer.

Folate and vitamin B12 deficiency is associated with depletion of the major intracellular antioxidant glutathione, and oxidative stress is emerging as an etiological mechanism for colon cancer. Azoxymethane (AOM), a potent carcinogen, induces colon cancer in rats by causing pathophysiological changes and oxidative stress. We investigated the synergistic effect of folate and vitamin B12 supplementation against AOM-induced carcinogenesis and oxidative stress in rat colon. Adult male rats were distributed into four groups: 1) Basal diet only; 2) AOM injection (15 mg/kg once per week in weeks 5 and 6); 3) Folate and vitamin B12 supplemented diet; 4) Folate and B12 diet with AOM injection. After 16 weeks, rats were sacrificed, colon tissue dissected, indicators of oxidative stress were measured, and immunohistochemical and ultrastructural changes were evaluated. AOM-injected rats showed oxidative stress, evident by glutathione depletion, oxidation of cellular proteins, and DNA oxidative damage. AOM increased mucosal levels of antiapoptotic and proapoptotic proteins Bcl2 and Bax and caused ultrastructure changes in colonic cell organelles. Folate and vitamin B12 supplementation decreased the level of oxidative stress and ameliorated the cytotoxic effects of AOM. In this in vivo experimental model of colon cancer, folate and vitamin B12 supplementation combats carcinogen-induced oxidative stress.

Broccoli (Brassica oleracea) Reduces Oxidative Damage to Pancreatic Tissue and Combats Hyperglycaemia in Diabetic Rats.

Oxidative stress plays a pivotal role in the development of diabetes and hyperglycaemia. The protective effects of natural extracts against diabetes are mainly dependent on their antioxidant and hypoglycaemic properties. Broccoli (Brassica oleracea) exerts beneficial health effects in several diseases including diabetes; however, the mechanism has not been elucidated yet. The present study was carried out to evaluate the potential hypoglycaemic and antioxidant properties of aqueous broccoli extracts (BEs) in diabetic rats. Streptozotocin (STZ) drug was used as a diabetogenic agent in a single intraperitoneal injection dose of 50 mg/kg body weight. The blood glucose level for each rat was measured twice a week. After 8 weeks, all animals were fasted overnight and sacrificed; pancreatic tissues were homogenized and used for measuring oxidative DNA damage, biochemical assessment of glutathione (GSH), and total antioxidant capacity (TAC) as well as histopathological examination for pancreatic tissues was examined. Diabetic rats showed significantly higher levels of DNA damage, GSH depletion, and impaired TAC levels in comparison to non-diabetics (P<0.05). The treatment of diabetic rats with BE significantly reduced DNA damage and conserved GSH and TAC values (P<0.01). BE attenuated pancreatic histopathological changes in diabetic rats. The results of this study indicated that BE reduced the STZ mediated hyperglycaemia and the STZ-induced oxidative injury to pancreas tissue. The used in vivo model confirmed the efficacy of BE as an anti-diabetic herbal medicine and provided insights into the capacity of BE to be used for phytoremediation purposes for human type 2 diabetes.

Acute effects of exogenous hormone administration on postprandial acylation stimulating protein levels in ovariectomized rats after a fat load.

Background. ASP, a potent lipogenic factor, was linked to female fat metabolism in association studies. Aim. To investigate acute effects of sex hormone treatment on postprandial ASP levels in vivo. Methods. 24 female rats were randomly divided into 4 groups including controls. The rats were ovariectomized and injected with progesterone, estrogen, or testosterone. An hour later, olive oil was administered orally. Plasma ASP and triglycerides were measured at several postprandial time points. Area under the curve (TG-AUC) represented TG clearance. Results. Only the progesterone treated group had a significant postprandial ASP increase at two hours compared to basal levels (439.8 ± 62.4 versus 253.4 ± 59.03 µg/mL, P = 0.04). Interestingly, increased ASP levels coordinated negatively with corresponding TG levels and TG-AUC postprandially, mostly evident in the opposite effects in the progesterone and testosterone treated groups. ASP levels increased 3-fold in the progesterone versus testosterone treated groups, whereas TG-AUC was significantly lower. Conclusion. These findings suggest that progesterone enhances ASP production and TG clearance simultaneously, supporting the notion of a stimulatory role for progesterone on ASP-mediated TG clearance. This is the first functional study demonstrating a cause-effect relationship between hormone treatment and ASP levels in vivo and may contribute to understanding the mechanism of progesterone function as a female lipogenic hormone.

Cardiovascular and lung inflammatory effects induced by systemically administered diesel exhaust particles in rats.

Pollution by particulates has consistently been associated with increased cardiorespiratory morbidity and mortality. It has been suggested that ultrafine particles, of which diesel exhaust particles (DEP) are significant contributors, are able to translocate from the airways into the bloodstream in vivo. In the present study, we assessed the effect of systemic administration of DEP on cardiovascular and respiratory parameters. DEP were administered into the tail vein of rats, and heart rate, blood pressure, blood platelet activation, and lung inflammation were studied 24 h later. Doses of 0.02, 0.1, or 0.5 mg DEP/kg (8, 42, or 212 microg DEP/rat) induced a significant decrease of heart rate and blood pressure compared with saline-treated rats. Although the number of platelets was not affected, all the doses of DEP caused a shortening of the bleeding time. Similarly, in addition to triggering lung edema, the bronchoalveolar lavage analysis revealed the presence of neutrophil influx in DEP-treated rats in a dose-dependent manner. We conclude that the presence of DEP in the systemic circulation leads not only to cardiovascular and haemostatic changes but it also triggers pulmonary inflammation.

Arthroscopic capsular shrinkage of the shoulder for the treatment of patients with multidirectional instability: Minimum 2-year follow-up.

PURPOSE: The study goal was to evaluate the arthroscopic treatment of patients with multidirectional instability of the shoulder (MDI). MDI is a disabling condition as a result of pain and restriction of activity. The treatment of MDI is primarily nonsurgical (supervised physiotherapy). TYPE OF STUDY: Prospective cohort study. METHODS: Arthroscopic capsular shrinkage, using a bipolar radiofrequency system, was performed in 32 patients (33 shoulders). The mean age was 27 years (range, 15 to 49). The Constant score was used for assessment preoperatively and at 6-month intervals. In 8 patients, labral lesions were revealed at arthroscopy and were addressed using arthroscopic methods. RESULTS: The follow-up was from 24 to 33 months (26 mean). Patients without labral pathology (group A) and patients with labral lesions (group B) are discussed separately. Group A showed an increase in the mean Constant score at 6 months, but the scores reached a plateau after 6 months. Statistical analysis using 1-way analysis of variance (ANOVA) showed that the mean difference between the preoperative Constant score and the score at latest follow-up was statistically significant (P <.0001; 95% confidence interval, 13.4 to 31.6; standard error, 2.2). The rate of failure or complication was 16%. Three patients experienced recurrence of instability and another developed adhesive capsulitis. Overall, patient satisfaction was 83%. Group B showed an improvement in the Constant scores, and all 8 patients were satisfied. No clinical recurrence of the instability was seen in this group. However, 1 patient developed adhesive capsulitis. This group is too small to draw any statistically valid conclusions. CONCLUSIONS: The early results are encouraging. Postoperative physiotherapy is of paramount importance. Continued long-term follow-up of this relatively new technique is required to determine the failure rate increase that is likely to occur at longer time intervals.