ObjectivesOsteoarthritis (OA) patients with a neuropathic pain (NP) component may represent a specific phenotype. This study compares joint damage, pain and functional disability between knee OA patients with a likely NP component, and those without a likely NP component.MethodsBaseline data from the Innovative Medicines Initiative Applied Public-Private Research enabling OsteoArthritis Clinical Headway knee OA cohort study were used. Patients with a painDETECT score ≥19 (with likely NP component, n=24) were matched on a 1:2 ratio to patients with a painDETECT score ≤12 (without likely NP component), and similar knee and general pain (Knee Injury and Osteoarthritis Outcome Score pain and Short Form 36 pain). Pain, physical function and radiographic joint damage of multiple joints were determined and compared between OA patients with and without a likely NP component.ResultsOA patients with painDETECT scores ≥19 had statistically significant less radiographic joint damage (p≤0.04 for Knee Images Digital Analysis parameters and Kellgren and Lawrence grade), but an impaired physical function (p<0.003 for all tests) compared with patients with a painDETECT score ≤12. In addition, more severe pain was found in joints other than the index knee (p≤0.001 for hips and hands), while joint damage throughout the body was not different.ConclusionsOA patients with a likely NP component, as determined with the painDETECT questionnaire, may represent a specific OA phenotype, where local and overall joint damage is not the main cause of pain and disability. Patients with this NP component will likely not benefit from general pain medication and/or disease-modifying OA drug (DMOAD) therapy. Reserved inclusion of these patients in DMOAD trials is advised in the quest for successful OA treatments.Trial registration numberThe study is registered under clinicaltrials.gov nr: NCT03883568.

Objective:To determine the value of serological biomarkers of collagen degradation/ turnover as serum markers of organ involvement in patients with systemic sclerosis (SSc).Methods: Serum samples were obtained from 79 SSc patients and 19 healthy control subjects. Types I to VI collagen turnover, excluding type II collagen, were evaluated using nine serological biomarkers. Organ involvement, such as interstitial lung disease (ILD), pulmonary arterial hypertension (PAH), esophageal motility disorder, lower gastrointestinal lesions, joint contractures and digital ulceration, were evaluated and correlated with serum biomarkers.Results: Among multiple serological biomarkers of collagen turnover, the mean level of C5M was higher in SSc (3.9 ng/mL) than healthy subjects (2.6 ng/mL). In addition, PRO-C6 (12.6 ng/mL vs 5.4 ng/mL) and C6M (26.0 ng/mL vs 16.7 ng/mL) were higher in SSc than the controls. The modified Rodnan skin score correlated with PRO-C3 and PRO-C6. Serum level of C6M was higher in patients with ILD. Furthermore, serum levels of PRO-C3, PRO-C6, and C6M were higher in patients with PAH. The use of high levels of these biomarkers as risk factors of PAH showed that all patients with PAH had high levels of risk factors. Of note, two-third of patients with serum PRO-C3, PRO-C6 and C6M values above the respective cut-off values had PAH. Conclusion:Our study indicated that collagen turnover abnormality, especially type VI collagen, is not only important pathologically in skin sclerosis but also in organ involvement. These biomarkers of collagen turnover are potentially clinically useful as biomarkers of organ involvement in SSc. K E Y W O R D S biomarkers, extracellular matrix, interstitial lung disease, pulmonary arterial hypertension, systemic sclerosis | 533 KUBO et al.

Background In patients with inflammatory bowel disease (IBD), up to 30% do not achieve response, 66% do not achieve remission, and 40% experience loss of response to anti-TNFα treatment. In Crohn’s disease (CD), fecal calprotectin is the most widely used fecal biomarker for diagnosis and monitoring of disease activity. Fecal sampling and processing are tedious compared to serum sampling, and biomarker quantification in serum is more efficient, convenient, and also more acceptable for patients. Therefore, there is a need for a robust and reliable serum calprotectin biomarker that performs better than conventional serum biomarkers. The aim of this study was to investigate the performance of a novel serum calprotectin assay, which quantifies a specific human neutrophil elastase (HNE) generated calprotectin (S100a9) fragment (CPa9-HNE), as a measure of true neutrophil granulocyte activity, for monitoring treatment responses in patients with CD. Methods We included 30 CD patients with clinically active (Harvey-Bradshaw Index (HBI)≥5) luminal inflammation scheduled for anti-TNF treatment (adalimumab or infliximab). Serum samples where obtained at baseline (n=30), week 1 (n=29), week 8 (n=28), week 26 (n=15), and week 56 (n=7). CPa9-HNE (Calprotectin, S100a9, specific fragment generated by human neutrophil elastase), CRP and fecal calprotectin were quantified. Remitters (n=11) were defined as patients in clinical remission (HBI&lt;5) at week 8 with no flares at week 26 or week 56. Non-remitters (n=19) were defined by having clinical active disease (HBI≥5) at week 8 or having a flare at week 26 or week 56. Results Patients with severe disease activity (HBI&gt;16) had significantly elevated serum levels of CPa9-HNE compared with patients in remission (p&lt;0.05), with mild (p&lt;0.05) or moderate (p&lt;0.01) disease activity. CPa9-HNE serum levels were significantly suppressed in remitters compared with non-remitters at week 8 (p&lt;0.001), week 26 (p&lt;0.01) and week 56 (p&lt;0.05) (Figure 1). Fecal calprotectin and CRP were significantly suppressed at week 8, week 26 and week 56 compared with baseline levels for remitters and non-remitters, but there was no significant difference between remitters and non-remitters (Figure 1). Non-remitters were more likely to have high levels of CPa9-HNE at baseline (tertile 1,2 vs tertile 3; OR: 6.8 (CI:1.22-36.5), p=0.027) and week 8 (tertile 1,2 vs tertile 3; OR: 24 (CI:2.56-279), p=0.0014). Conclusion CPa9-HNE is a novel non-invasive biomarker for monitoring treatment response to anti-TNFα for CD patients and may be used as a surrogate marker for assessing disease activity and to predict outcome of anti-TNFα treatment.

Background The MANKIN score is the most widely used histology evaluation on osteoarthritis (OA). It has been modified in number of studies by different research groups since it was first published 40 years ago. The MANKIN score investigate the degradation level of the articular cartilage by assessing four parameters including: cartilage structure, cellularity, proteoglycan depletion and tidemark integrity. However, it is well-established that the interaction between cartilage and subchondral bone, such as the new vessel invasion from subchondral bone into calcified cartilage and the accompanied nerve formation, play essential role in the onset and progression of OA. Objectives As to better assess the pathologic changes in joint during OA, we developed a new modified MANKIN scoring system with special attention to the subchondral bone. Methods Besides 4 categories of evaluation on cartilage, we also performed type X collagen immunohistochemical staining, tartrate-resistant acidic phosphatase (TRAP) staining, cartilage and subchondral bone thickness measurement, the number of vessels counting (CD34), Vascular Endothelial Growth Factor Receptor (VEGFR) immunostaining and nerve growth factor receptor (NGFR, P75staining) immunostaining in 2 areas (lateral femur condyle and lateral tibia plateau) of 11 OA patients undergoing knee replacement surgery at Frederikshavn hospital. Each new evaluation was analyzed the correlation with MANKIN score, of which with moderate correlation would be included into the new scoring table. Furthermore, statistical analysis was performed for inter-/intra- observers variability according to the criteria of new modified scoring system. Results After carful assessment of the different novel parameters, following were found to be associated with cartilage quality: 1) number of vessels (moderate correlation with MANKIN score, Spearman's r=0.6), 2) number of positive cells expressing NGFR (moderate correlation with MANKIN score, Spearman's r=0.4), and 3) the thickness of cartilage/subchondral bone (moderate correlation with MANKIN score, Spearman's r=0.7). A final scoring scheme was put together (table 1) and tested in 11 patients. The inter-observer variability among two observers for the new score system showed reliability (Person's r=0.80). Repeating scoring by one of two observers showed good reproducibility (Person's r=0.73). Conclusions The new scoring system was constructed with valuable information to indicate the changes in subchondral bone and the severity of OA, which might be used as a rapid and reliable histology evaluation on both human OA and experiment induced OA animal models. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.4224

Background:Many systemic sclerosis (SSc) patients develop lung fibrosis, which contribute significantly to increased mortality1. Activated and proliferating fibroblasts are responsible for the excessive extracellular matrix (ECM) formation and stiffening of the connective tissue leading to skin and lung fibrosis. There is currently no effective treatment for the fibrosis in SSc and there is therefore a medical need for further understanding the pathogenesis of fibrosis. Fibrosis is associated with different growth factors, including tumor growth factor beta 1 (TGF-β1) and platelet derived growth factor-ab (PDGF-ab)2.Objectives:We investigated how stimulation with TGF-β1 and PDGF-ab affected the migration capacity and the ECM production using translational biomarkers of type I, III and VI collagens in healthy human dermal and lung fibroblasts.Methods:The fibroblasts were grown in DMEM media containing 0.4% fetal calf serum, ficoll (to produce a crowded environment) and ascorbic acid for up to 12 days. The cells were stimulated with TGF-β1 [0.04-1 nM] or PDGF-ab [3 nM] at treatment initiation and changed twice a week. Non-stimulated fibroblasts (w/o) were used as control. A wound was induced by scratching the cells at day 1 after treatment initiation and the migration was followed for 2 days. Type I, III and VI collagen formation (PRO-C1, PRO-C3 and PRO-C6, respectively) were evaluated by validated ELISAs (Nordic Bioscience) in supernatant from day 0, 4, 8 and 12. Statistical analysis included 2-way ANOVA and Dunnett’s test.Results:The PDGF-ab stimulated dermal fibroblasts migrated significantly more than the non-stimulated (p<0.0001) and TGF-β1 stimulated (p<0.001) dermal fibroblasts 48 hours after the scratch (migration app. 70%, 30% and 30% respectively). There was no difference between the migration of the non-stimulated, TGF-β1 and PDGF-ab stimulated lung fibroblasts after 48 hours, as all migrated to approximately 70%.TGF-β1 stimulation led to a significant increase in type I collagen formation (PRO-C1) in both dermal and lung fibroblasts from day 4 and onwards compared to w/o (p<0.0001). TGF-β1 also lead to a significant increase in type III collagen formation (PRO-C3) from day 8 in lung fibroblasts compared to w/o (p<0.0001). PDGF-ab stimulation led to a significant increase in type III collagen formation in dermal fibroblasts from day 8 compared to w/o (p<0.0001). PDGF-ab stimulation led to a significant increase in type VI collagen formation (PRO-C6) in both dermal and lung fibroblasts from day 4 and onwards compared to w/o (p<0.0001).Conclusion:PDGF-ab increased the migration activity of the dermal fibroblasts, where the lung fibroblasts had a general high migration activity. The dermal and lung fibroblasts showcase the same ECM production within both type I and type VI collagen formation. The two fibroblasts types did however react opposite each other regarding the type III collagen formation: the dermal fibroblasts responded to PDGF-ab stimulation, where the lung fibroblasts responded to the TGF-β1 stimulation. The clear differences in the ECM production between the dermal and lung fibroblasts can be important in the search for an effective treatment for fibrosis in SSc and related lung fibrosis.References:[1]McNearney, T. A. et al. Pulmonary involvement in systemic sclerosis: Associations with genetic, serologic, sociodemographic, and behavioral factors. Arthritis Care Res.57, 318–326 (2007).[2]Wynn, T. A. Common and unique mechanisms regulate fibrosis in various fibroproliferative diseases. J. Clin. Invest.117, 524–529 (2007).Disclosure of Interests:Sofie Falkenløve Madsen Employee of: Nordic Bioscience and University of Copenhagen, Anne Sofie Siebuhr Employee of: Nordic Bioscience A/S, Henrik Jessen: None declared, Jannie Marie Bülow Sand Employee of: Nordic Bioscience A/S, Morten Karsdal Shareholder of: Nordic Bioscience A/S, Employee of: Nordic Bioscience A/S, Anne-Christine Bay-Jensen Shareholder of: Nordic Bioscience A/S, Employee of: Nordic Bioscience A/S