Abhilash Jain

Production of cytokines, vascular endothelial growth factor, matrix metalloproteinases, and tissue inhibitor of metalloproteinases 1 by tenosynovium demonstrates its potential for tendon destruction in rheumatoid arthritis.

OBJECTIVE To investigate the role of proinflammatory cytokines, vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMPs), and tissue inhibitor of metalloproteinases 1 (TIMP-1) in the destruction of tendons by tenosynovium in rheumatoid arthritis (RA). METHODS Synovial specimens were obtained from encapsulating tenosynovium (n = 17), invasive tenosynovium (n = 13), and wrist joints (n = 17) in 18 RA patients undergoing wrist extensor tenosynovectomy. Synovial membrane cells were dissociated from connective tissue by enzyme digestion and cultured in vitro for 48 hours, and harvested supernatants were assayed for the cytokines tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6), VEGF, MMPs 1, 2, 3, and 13, and TIMP-1 by enzyme-linked immunosorbent assay. Gelatin zymography was performed to demonstrate enzyme activity. Statistical analysis was performed using Students paired 2-tailed t-tests for parametric data and the Wilcoxon signed rank test for nonparametric data. RESULTS MMP-1 and MMP-13 levels were approximately 2.5-fold higher in invasive tenosynovium compared with encapsulating tenosynovium. Levels of MMP-2 were approximately 1.5-fold higher in invasive tenosynovium compared with both encapsulating tenosynovium and wrist joint synovium. MMP-13 (P = 0.009) and IL-6 (P = 0.03) levels were significantly lower in encapsulating tenosynovium compared with wrist joint synovium. Levels of VEGF, TIMP-1, TNFalpha, and MMP-3 were similar in all synovial sample groups. Zymography demonstrated enzyme activity in all synovium samples from all 9 patients assessed. CONCLUSION Tenosynovium produces proinflammatory cytokines and proteolytic enzymes that are important in the tissue degradation seen in RA. Increased production of the enzymes MMP-1, MMP-2, and MMP-13 by invasive tenosynovium suggests a possible explanation for the worse prognosis and increased rupture rate associated with invasive tenosynovitis in RA. Production of VEGF by tenosynovium suggests that angiogenesis may have a role in tenosynovial proliferation and invasion of tendons.

Membrane type 1 matrix metalloproteinase is a crucial promoter of synovial invasion in human rheumatoid arthritis

OBJECTIVE A hallmark of rheumatoid arthritis (RA) is invasion of the synovial pannus into cartilage, and this process requires degradation of the collagen matrix. The aim of this study was to explore the role of one of the collagen-degrading matrix metalloproteinases (MMPs), membrane type 1 MMP (MT1-MMP), in synovial pannus invasiveness. METHODS The expression and localization of MT1-MMP in human RA pannus were investigated by Western blot analysis of primary synovial cells and immunohistochemical analysis of RA joint specimens. The functional role of MT1-MMP was analyzed by 3-dimensional (3-D) collagen invasion assays and a cartilage invasion assay in the presence or absence of tissue inhibitor of metalloproteinases 1 (TIMP-1), TIMP-2, or GM6001. The effect of adenoviral expression of a dominant-negative MT1-MMP construct lacking a catalytic domain was also examined. RESULTS MT1-MMP was highly expressed at the pannus-cartilage junction in RA joints. Freshly isolated rheumatoid synovial tissue and isolated RA synovial fibroblasts invaded into a 3-D collagen matrix in an MT1-MMP-dependent manner. Invasion was blocked by TIMP-2 and GM6001 but not by TIMP-1. Invasion was also inhibited by the overexpression of a dominant-negative MT1-MMP, which inhibits collagenolytic activity and proMMP-2 activation by MT1-MMP on the cell surface. Synovial fibroblasts also invaded into cartilage in an MT1-MMP-dependent manner. This process was further enhanced by removing aggrecan from the cartilage matrix. CONCLUSION MT1-MMP serves as an essential collagen-degrading proteinase during pannus invasion in human RA. Specific inhibition of MT1-MMP-dependent invasion may represent a novel therapeutic strategy for RA.

Impact of VIP and cAMP on the regulation of TNF-α and IL-10 production: implications for rheumatoid arthritis

Vasoactive intestinal peptide (VIP) is an anti-inflammatory immunomodulatory neuropeptide with therapeutic potential demonstrated for collagen-induced arthritis. The aim of this study was to characterise its potential anti-arthritic effect on human monocytes, macrophages, T cells, and rheumatoid arthritis synovial membrane cells. Monocytes, macrophages, and T cells derived from human peripheral blood were treated with VIP and compared with other cAMP-elevating drugs for a range of activating stimuli. Cytokine production was assessed for cell cultures and, in addition, the ability of VIPs to activate cAMP response element binding protein. VIP partially suppressed monocyte- and macrophage-derived tumour necrosis factor α (TNF-α) with no effect on IL-10, whereas VIP fails to regulate IL-10 and TNF-α production by T lymphocytes. No such modulation of cytokine profile was observed for rheumatoid arthritis synovial membrane cells. Elevation of intracellular cAMP, on the other hand, potently suppressed macrophage TNF-α production and modulated T-cell response by inhibiting TNF-α and IFN-γ. VIPs lack of effect on IL-10 and its slight effect on TNF-α results from cAMP being rapidly degraded as the phosphodiesterase IV inhibitor, rolipram, rescues cAMP-dependent activation of cAMP response element binding protein. Interestingly, macrophages stimulated with phorbol 12-myristate 13-acetate/ionomycin displayed an augmented IL-10 response upon addition of dibutyryl cAMP, with corresponding downregulation in TNF-α, suggesting a complex interaction between protein kinase C and protein kinase A in cytokine regulation. In conclusion, VIP may represent an efficaceous anti-arthritic treatment modulating macrophage and T-cell cytokine profiles when used alongside a phosphodiesterase inhibitor.

Systematic review: Anastomotic microvascular device

BACKGROUND Anastomotic microvascular device has gained popularity in reconstructive microsurgery over the last two decades. A systematic literature search has been carried out in the use of the venous coupler device in microsurgery to assess its impact on patency rate in microsurgery. METHOD Using key words related to the topic, a literature search of major databases was carried out. Selection was undertaken on two level screening. From the literature search 53 potential articles were identified. Of these 13 met both levels of screening for the final critical analysis. RESULTS A total of 2976 venous anastomoses with coupler device were carried out in the 13 studies with a combined thrombosis in 46 veins reported, giving an average 98.5% patency and a variation in thrombosis rate ranging from 0 to 3%. CONCLUSION The venous coupler device serves as a powerful instrument in microsurgery by achieving high patency rate. More importantly the data suggests less variation in thrombosis rate compared to hand-sewn venous anastomosis.

Functional outcome following extensor synovectomy and excision of the distal ulna in patients with rheumatoid arthritis

We prospectively measured hand and wrist function in rheumatoid patients undergoing excision of the distal ulna. Range of motion, visual analogue pain scores and grip strength were measured in 22 wrists, and the Jebsen hand function test was administered to seven patients, preoperatively and at 3 and 12 months. At 1 year there were improvements in forearm pronation (P = 0.04), supination (P = 0.03) and wrist extension (P = 0.02), but a reduction in flexion (P = 0.009). Active radial deviation was reduced and ulnar deviation increased. There was a significant improvement in grip strength (P = 0.05) and reduction in wrist pain (P = < 0.0001). At 1 year the Jebsen hand function test showed improvements in simulated feeding, stacking checkers, and lifting large empty cans. Excision of the distal ulna in rheumatoid patients results in an improvement in some aspects of hand function.

Treatment of rheumatoid tenosynovitis with cytokine inhibitors

Hand function depends on tendon integrity, but in rheumatoid arthritis tenosynovitis can result in tendon adhesions and rupture. Cytokine inhibitors have proved effective in rheumatoid joint disease; however, their effect on the tenosynovium is not well understood. We investigated the ability of inhibitors of tumour necrosis factor alpha and interleukin 1 to reduce production of collagenolytic matrix metalloproteinases 1 and 13 in tenosynovial tissue obtained from patients with rheumatoid arthritis. Our data show that cytokine blockade can reduce collagenase concentrations in tenosynovial tissue, suggesting cytokine inhibitors could be effective in reduction of tendon damage.

Outcomes of anterolateral thigh free flap thinning using liposuction following lower limb trauma

BACKGROUND Whilst soft tissue closure is the priority to prevent infection in open fractures of the lower limb, some patients find that bulky flaps interfere with function and dislike the appearance. We report the outcomes of delayed free anterolateral thigh flap thinning with liposuction. MATERIAL AND METHODS 38 patients treated between 2006 and 2009 were offered flap contouring. 23 chose flap thinning and 15 did not. We measured outcomes using the SF-36v2 questionnaire and cosmetic outcome scores pre and postoperatively at a mean follow up of 12 weeks (range 10-16 weeks). RESULTS SF-36v2 physical health (PH) scores improved from a mean of 67 preoperatively to 80 postoperatively (p = 0.01) in the thinned group, while mental health (MH) scores remained unchanged (74-72). The mean SF-36v2 scores for the non-thinned group were 77 (PH) and 86 (MH). Following liposuction the median cosmetic outcome scores out of 5 improved from 1 (not at all satisfied) to 4 (very satisfied) postoperatively (p = 0.0005), which was also higher than the non-thinned group (3) [moderately satisfied], p = 0.004). There was no difference in sex, age, BMI and region on the leg of free flap reconstruction between the non-thinned and thinned groups. CONCLUSIONS Delayed contouring of free ALT flaps used for lower limb reconstruction results in improvements in physical health measures and cosmetic outcomes. Patients not requesting thinning are generally satisfied with their reconstruction.

Targeting rheumatoid tenosynovial angiogenesis with cytokine inhibitors.

Proliferation and invasion of the tenosynovial lining of tendons in patients with rheumatoid arthritis can result in tendon damage and rupture, leading to decreased hand function. Angiogenesis is an important process in rheumatoid joint disease; however, the role of angiogenesis in tendon disease is unknown. Our aim was to determine whether rheumatoid tenosynovial lining could produce angiogenic proteins, and if inhibition of tumor necrosis factor-α and interleukin-1 could decrease vascular endothelial growth factor production. Samples of encapsulating and invasive tenosynovial lining taken from the same hand and wrist synovial lining were harvested from 58 patients with rheumatoid arthritis having wrist surgery. Ex vivo samples were studied to quantify vascularity, angiogenic protein production under normoxic and hypoxic conditions, and the effect of inhibiting tumor necrosis factor-α and interleukin-1 on vascular endothelial growth factor production. Rheumatoid tenosynovial lining was more vascular than rheumatoid joint synovial lining and produced high levels of angiogenic factors such as vascular endothelial growth factor, interleukin-1β, fibroblast growth factor-2, and angiopoietin-2. Hypoxia induced an increase in production of vascular endothelial growth factor by ex vivo tenosynovial lining cells. Inhibition of the cytokines interleukin-1 and tumor necrosis factor-α effectively reduced vascular endothelial growth factor production by tenosynovial samples.Level of Evidence: Therapeutic study. Level II (Prospective comparative study). See the Guidelines for Authors for a complete description of levels of evidence.

The surgical management of injectional anthrax

Anthrax is caused by the spore forming, gram-positive aerobic organism Bacillus anthracis. Three clinical forms of anthrax are recognised; respiratory, gastrointestinal and cutaneous, with the latter being the most common. An isolated fatal case of injectional anthrax as a result of heroin use has been previously reported. Recently, there has been an outbreak of injectional anthrax in Europe (www.hps.scot.nhs.uk/anthrax) and we describe our experience of the surgical management of two cases.

The iBRA (implant breast reconstruction evaluation) study: protocol for a prospective multi-centre cohort study to inform the feasibility, design and conduct of a pragmatic randomised clinical trial comparing new techniques of implant-based breast reconstruction

BackgroundImplant-based breast reconstruction (IBBR) is the most commonly performed reconstructive procedure in the UK. The introduction of techniques to augment the subpectoral pocket has revolutionised the procedure, but there is a lack of high-quality outcome data to describe the safety or effectiveness of these techniques. Randomised controlled trials (RCTs) are the best way of comparing treatments, but surgical RCTs are challenging. The iBRA (implant breast reconstruction evaluation) study aims to determine the feasibility, design and conduct of a pragmatic RCT to examine the effectiveness of approaches to IBBR.Methods/designThe iBRA study is a trainee-led research collaborative project with four phases:Phase 1 – a national practice questionnaire (NPQ) to survey current practicePhase 2 – a multi-centre prospective cohort study of patients undergoing IBBR to evaluate the clinical and patient-reported outcomesPhase 3– an IBBR-RCT acceptability survey and qualitative work to explore patients’ and surgeons’ views of proposed trial designs and candidate outcomes.Phase 4 – phases 1 to 3 will inform the design and conduct of the future RCTAll centres offering IBBR will be encouraged to participate by the breast and plastic surgical professional associations (Association of Breast Surgery and British Association of Plastic Reconstructive and Aesthetic Surgeons).Data collected will inform the feasibility of undertaking an RCT by defining current practice and exploring issues surrounding recruitment, selection of comparator arms, choice of primary outcome, sample size, selection criteria, trial conduct, methods of data collection and feasibility of using the trainee collaborative model to recruit patients and collect data.DiscussionThe preliminary work undertaken within the iBRA study will determine the feasibility, design and conduct of a definitive RCT in IBBR. It will work with the trainee collaborative to build capacity by creating an infrastructure of research-active breast and plastic surgeons which will facilitate future high-quality research that will ultimately improve outcomes for all women seeking reconstructive surgery.Trial registrationISRCTN37664281