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See detailBiomarker for osteoarthritis and/or other ageing-related diseases, and use thereof
Henrotin, Yves ULg; Gharbi, Myriam; DEBERG, Michelle ULg et al

Patent (2014)

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See detailEarly decrease of serum biomarkers of type II collagen degradation (Coll2-1) and joint inflammation (Coll2-1 NO(2) ) by hyaluronic acid intra-articular injections in patients with knee osteoarthritis: a research study part of the Biovisco study.
Henrotin, Yves ULg; Chevalier, X.; DEBERG, Michelle ULg et al

in Journal of orthopaedic research : official publication of the Orthopaedic Research Society (2013), 31(6), 901-7

To measure the evolution of the serum levels of specific Osteoarthritis (OA) biomarker, Coll2-1 and Coll2-1 NO(2) in knee osteoarthritic patients after viscosupplementation (VS). Fifty-one patients with ... [more ▼]

To measure the evolution of the serum levels of specific Osteoarthritis (OA) biomarker, Coll2-1 and Coll2-1 NO(2) in knee osteoarthritic patients after viscosupplementation (VS). Fifty-one patients with unilateral symptomatic knee were recruited for this prospective open label study. They received three intra-articular injections of 2 ml of hyaluronic acid (Hylan GF-20) and were followed for 3 months. Walking pain was evaluated and serum samples were taken at each visit. Coll2-1 and Coll2-1 NO(2) were measured in the serum using specific immunoassays. Variations over time of each parameter and predictive factor of response were studied. Forty-five patients were analyzed. The serum concentrations of Coll2-1 and Coll2-1 NO(2) were significantly higher in KL III/IV patients compared to KL I/II patients at baseline and decreased systematically over time after VS. Its effect was ever more pronounced in patients with KL III/IV. The serum concentration of Coll2-1 was significantly lower at baseline in responders than in non-responders. This study suggests a rapid slowdown of type II collagen degradation and joint inflammation after VS with Hylan G-20 and gives additional information for the validation of accurate biomarkers for OA. The serum level of Coll2-1 appeared to be a predictive factor for response to treatment. [less ▲]

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See detailDetection of specific nitrated markers
REGINSTER, Jean-Yves ULg; DEBERG, Michelle ULg; Henrotin, Yves ULg et al

Patent (2012)

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See detailColl2-1, Coll2-1NO2 and myeloperoxidase serum levels in erosive and non-erosive osteoarthritis of the hands.
Punzi, L.; Ramonda, R.; DEBERG, Michelle ULg et al

in Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society (2012), 20(6), 557-61

OBJECTIVE: Erosive osteoarthritis of the hand (EHOA) is thought to be an aggressive variant of hand osteoarthritis (HOA) characterised by prominent local inflammation and radiographic aspects of bone ... [more ▼]

OBJECTIVE: Erosive osteoarthritis of the hand (EHOA) is thought to be an aggressive variant of hand osteoarthritis (HOA) characterised by prominent local inflammation and radiographic aspects of bone erosions in interphalangeal (IP) joints. However, rare studies have until now investigated the value of biomarkers in these patients. Thus, we determined Coll2-1, a marker of type II collagen denaturation, its nitrated form (Coll2-1NO2) and myeloperoxidase (MPO) levels in serum of patients with EHOA vs non-EHOA and subsequently evaluated their relationships with disease indices of severity and activity. METHODS: Coll2-1, Coll2-1NO2 and MPO were measured using specific immunoassays in 82 patients, 57 with EHOA, all females, median age 59 (41-74 yrs) and 20 with non-EHOA, all females, median age 55 (43-73 yrs), fulfilling the American College of Rheumatology (ACR) criteria for hand OA. EHOA was characterized by the presence of at least one central bone erosion on radiograph in the IP joints. Patients were also evaluated for disease duration, number of affected (swollen and painful or tender) joints, radiographic score (RS) by Kallman scale and high sensitivity C-reactive protein (hsCRP). RESULTS: Serum levels of MPO were higher in EHOA (230.0 +/- 152.1 ng/ml) than in non-EHOA (160.2 +/- 111.5 ng/ml, P=0.037). Coll2-1NO2 levels trended towards an elevation in EHOA compared non-EHOA (0.40 +/- 0.86 vs 0.22 +/- 0.14 nmol/l, P=0.06), while Coll2-1 levels were not different. Correlations were found for disease duration and both MPO (R(2)=0.48, P=0.001) and Coll2-1NO2 (R(2)=0.73, P=0.01) after the splitting of the population in subgroups according to a cut off value above the 50th percentile. A correlation was found between hsCRP and MPO (R(2)=0.57, P=0.01). CONCLUSIONS: This study clearly demonstrates an elevation of some serum biomarkers in EHOA, in comparison with non-EHOA. In particular, MPO, hsCRP and the ratio Coll2-1NO2/Coll2-1 discriminated the two subsets of hand osteoarthritis (HOA), and a trend was also observed for Coll2-1NO2. These data suggest that these biomarkers could be helpful for the diagnosis of EHOA. [less ▲]

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See detailUsefulness of specific OA biomarkers, Coll2-1 and Coll2-1NO(2), in the anterior cruciate ligament OA canine model.
Henrotin, Yves ULg; Martel-Pelletier, Johanne; Msika, Philippe et al

in Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society (2012), 20(7), 787-90

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See detailFibulin 3 peptides Fib3-1 and Fib3-2 are potential biomarkers of osteoarthritis.
Henrotin, Yves ULg; Gharbi, Myriam; Mazzucchelli, Gabriel ULg et al

in Arthritis and Rheumatism (2012), 64(7), 2260-7

OBJECTIVE: This study was undertaken to identify new biomarkers of osteoarthritis (OA) by proteomics analysis and to develop specific immunoassays to detect and quantify them. METHODS: Proteomics analysis ... [more ▼]

OBJECTIVE: This study was undertaken to identify new biomarkers of osteoarthritis (OA) by proteomics analysis and to develop specific immunoassays to detect and quantify them. METHODS: Proteomics analysis was performed in urine samples from 10 women (mean+/-SD age 76.0+/-5.0 years) undergoing knee replacement surgery due to severe OA and 5 healthy women (mean+/-SD age 25.6+/-2.6 years). Protein content was analyzed by 2-dimensional differential gel electrophoresis. Protein spots that exhibited an OA:control abundance ratio of >/=1.5 were identified by mass spectrometry. Specific enzyme-linked immunosorbent assays were developed and validated in serum obtained from 236 healthy subjects ages 20-64 years and from 76 patients with severe radiologic knee OA (mean+/-SD age 68.8+/-11.9 years). Immunohistochemical analysis was performed on articular cartilage from tibial plateaus. RESULTS: Thirteen proteins within spots that were significantly modified between groups were identified. Two peptides of fibulin 3, named Fib3-1 and Fib3-2, were of particular interest. Two antisera directed against these peptides were used to develop immunoassays. Compared with age-matched healthy subjects, median levels of serum Fib3-1 and Fib3-2 were elevated in OA patients (54.6 pM versus 85.1 pM [P<0.0001] and 144.4 pM versus 191.4 pM [P<0.0001], respectively). Using area under the receiver operating characteristic curve analysis, we demonstrated that Fib3-1 and Fib3-2 levels discriminate between OA and normal populations. Immunostaining revealed the presence of Fib3-1 and Fib3-2 in chondrocytes and in the extracellular matrix of the superficial layer of the fibrillated cartilage. CONCLUSION: Our findings indicate that Fib3-1 and Fib3-2 are potential biochemical markers for the diagnosis of OA. [less ▲]

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See detailDetection of specific nitrated markers
Reginster, Jean-Yves ULg; DEBERG, Michelle ULg; Henrotin, Yves ULg et al

Patent (2011)

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See detailFibulin-3 fragments (FIB3-1 and FIB3-2) are potential new biomarkers for the diagnosis of osteoarthritis
Gharbi, M; Dubuc, JE; Deberg, Michelle ULg et al

in Annals of the Rheumatic Diseases (2011), 70(Suppl 3), 354

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See detailColl2-1, Coll2-1NO2 and myeloperoxidase concentrations in the synovial fluid of equine tarsocrural joints affected with osteochondrosis.
Verwilghen, Denis ULg; Martens, Ann; Busschers, Evita et al

in Veterinary Research Communications (2011), 35(7), 401-8

The measurement of biomarkers that reflect cartilage breakdown is a powerful tool for investigating joint damage caused by disease or injury. Particularly in cases of osteochondrosis, synovial ... [more ▼]

The measurement of biomarkers that reflect cartilage breakdown is a powerful tool for investigating joint damage caused by disease or injury. Particularly in cases of osteochondrosis, synovial concentrations of these biomarkers may reveal the presence of osteoarthritic changes. Coll2-1, Coll2-1 NO2 and myeloperoxidase have recently been introduced in equine osteoarticular research but comparison between the concentrations of these markers in OCD affected and healthy joints has not been made. Therefore, this study aimed at reporting the synovial concentrations of these biomarkers in joints affected with osteochondral fragments in the tarsocrural joint compared to unaffected joints. Myeloperoxidase and Coll2-1NO2 revealed to have similar levels between affected joints and controls. However, in contrast to previous studies using C2C the present study demonstrated that synovial levels of Coll2-1 were significantly elevated in tarsocrural joints affected with osteochondrosis. Thus, Coll2-1 may be an earlier marker of cartilage degeneration than other cartilage degradation markers that have been previously used in equine medicine. [less ▲]

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See detailApplication for proteomic techniques in studying osteoarthritis: a review
Gharbi, Myriam; Deberg, Michelle ULg; Henrotin, Yves ULg

in Frontiers in Physiology (2011), 2(90), 1-11

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See detailMitochondrial DNA haplogroups and serum levels of proteolytic enzymes in patients with osteoarthritis.
Rego-Perez, I.; Fernandez-Moreno, M.; Deberg, Michelle ULg et al

in Annals of the Rheumatic Diseases (2011), 70(4), 646-52

OBJECTIVE: To analyse the influence of mitochondrial DNA haplogroups, as well as the radiographic grade, on serum levels of proteolytic enzymes in patients with osteoarthritis (OA). METHODS: Serum levels ... [more ▼]

OBJECTIVE: To analyse the influence of mitochondrial DNA haplogroups, as well as the radiographic grade, on serum levels of proteolytic enzymes in patients with osteoarthritis (OA). METHODS: Serum levels of metalloproteinase-1 (MMP-1), MMP-3, MMP-13, myeloperoxidase and cathepsin K were analysed in 73 patients with OA and 77 healthy controls carrying the haplogroups J, U and H, by ELISA. Knee and hip radiographs were classified according to Kellgren and Lawrence (K/L) scoring from grade 0 to grade IV. Non-parametric and multiple regression analyses were performed to test the effects of clinical variables, including gender, age, smoking status, diagnosis, haplogroups and radiological K/L grade on serum levels of these enzymes. RESULTS: A significant influence of the haplogroups on the serum levels of MMP-3 and MMP-13 was detected (p=0.027 and p=0.035, respectively). Patients with OA with haplogroup H showed higher serum levels of MMP-3 than healthy controls. Serum levels of MMP-13 were significantly higher in patients with OA (p<0.001), and carriers of the haplogroup J showed lower levels than H carriers. Besides, levels of MMP-13 were proportionally higher in radiological groups B (K/L grade II and III) and C (K/L grade IV) than in group A (K/L grade 0 and I) (p=0.005). CONCLUSIONS: This study shows that haplogroups have a significant influence on serum levels of MMP-3 and MMP-13. The influence of the haplogroups on serum levels of MMP-3 is clearly dependent on the diagnosis, whereas the influence of the haplogroups on serum levels of MMP-13 is independent of diagnosis. [less ▲]

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See detailDe nouveaux outils d’évaluation de l’arthrose
Henrotin, Yves ULg; Deberg, Michelle ULg

in Ortho-Rhumato (2010), 8

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See detailMitochondrial DNA haplogroups modulate the serum levels of biomarkers in patients with osteoarthritis.
Rego-Perez, I.; Fernandez-Moreno, M.; Deberg, Michelle ULg et al

in Annals of the Rheumatic Diseases (2010), 69(5), 910-7

OBJECTIVE: To analyse the influence of mitochondrial DNA (mtDNA) haplogroups on serum levels of molecular biomarkers in patients with osteoarthritis (OA). METHODS: Serum levels of molecular biomarkers of ... [more ▼]

OBJECTIVE: To analyse the influence of mitochondrial DNA (mtDNA) haplogroups on serum levels of molecular biomarkers in patients with osteoarthritis (OA). METHODS: Serum levels of molecular biomarkers of cartilage metabolism (collagen type II markers: C-terminal neoepitope generated by the collagenase-mediated cleavage of collagen type II triple helix (C2C), collagen type II (Coll2-1, and its nitrated form, Coll2-1NO(2)), procollagen type II (CPII)), synovial metabolism (hyaluronic acid (HA)) and cartilage and synovial turnover (cartilage glycoprotein 39 (YKL-40)) were analysed in 73 patients with OA and 77 healthy controls using ELISAs. All participants had been previously genotyped for the mtDNA haplogroups J, U and H. Non-parametric and multivariate analysis were performed to test the effects of the clinical variables, including gender, age, smoking status, diagnosis, mtDNA haplogroups and radiological Kellgren and Lawrence (K/L) grade on the serum levels of the molecular markers. RESULTS: Non-parametric analysis found increased serum levels of HA in patients with OA, while the values for C2C and the C2C/CPII ratio were significantly higher in the healthy controls. A multiple regression analysis showed a relationship between the mtDNA haplogroups and serum levels of the typical collagen type II markers. Carriers of the mtDNA haplogroup H had higher levels while carriers of the mtDNA haplogroup J showed lower levels. Statistically significant interactions between mtDNA haplogroups and diagnosis and between mtDNA haplogroups and radiological K/L grade in the serum levels of molecular markers were also found. CONCLUSION: A new role for mtDNA haplogroups emerges from this work. The results suggest that the mtDNA haplogroups interact significantly with the serum levels of OA-related molecular markers, suggesting the possibility of their use as a complementary assay with these molecular markers. [less ▲]

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See detailCollagen fibril disruption occurs early in primary guinea pig knee osteoarthritis.
Huebner, J. L.; Williams, J. M.; Deberg, Michelle ULg et al

in Osteoarthritis and Cartilage (2010), 18(3), 397-405

OBJECTIVE: A major barrier inhibiting the discovery of structural modifying agents for osteoarthritis (OA) is an incomplete understanding of early disease events. Herein, we investigated the time course ... [more ▼]

OBJECTIVE: A major barrier inhibiting the discovery of structural modifying agents for osteoarthritis (OA) is an incomplete understanding of early disease events. Herein, we investigated the time course of collagen II cleavage and fibril disruption in the well-validated Hartley guinea pig model of spontaneous OA of the knee. METHODS: Knee joints of 46 male Hartley guinea pigs were analyzed at 3 weeks, 2, 4, 7, 10, 12, and 18 months of age for histological severity of OA, cartilage collagen fibril disruption by semi-quantitative polarized light microscopy, and expression of type II collagen degradation biomarkers, 9A4 and Coll2-1, by immunohistochemistry. In addition, serum biomarkers specific for collagen II degradation, CTX-II, C2C, and Coll2-1 were quantified. RESULTS: Collagen fibril disruption and expression of the collagenase-generated cleavage neoepitope, 9A4, were observed as early as 2 months of age, despite the appearance of histological OA at 4 months of age. Only serum Coll2-1 increased coincident with the early disruption of the collagen fibril between 3 weeks and 7 months, in contrast to serum C2C, which did not change significantly or correlate with histological severity. Inversely, CTX-II declined dramatically from 3 weeks to 4 months and remaining low thereafter, coincident with growth plate turnover. CONCLUSIONS: Collagenase cleavage and disruption of the type II collagen network are early OA disease events in this model, preceding histological evidence of proteoglycan loss. The markedly different serum profiles of collagen II-related biomarkers during the early stages of disease development suggest compartmental segregation and temporal regulation of collagen degrading enzymes. [less ▲]

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See detailIncrease in type II collagen turnover after iron depletion in patients with hereditary haemochromatosis.
Richette, Pascal; Eymard, Claire; Deberg, Michelle ULg et al

in Rheumatology (2010), 49(4), 760-6

OBJECTIVE: To determine the effects of iron depletion on serum levels of joint biomarkers and on joint symptoms in patients with hereditary haemochromatosis (HH). METHODS: Levels of biomarkers were ... [more ▼]

OBJECTIVE: To determine the effects of iron depletion on serum levels of joint biomarkers and on joint symptoms in patients with hereditary haemochromatosis (HH). METHODS: Levels of biomarkers were measured in 18 patients with HH at the time of diagnosis and after iron depletion. The markers were type II collagen degradation (Coll2-1) and its nitrated form (Coll2-1NO(2)), type II procollagen synthesis (CPII), MPO, COMP and HA. For each patient, demographic data were collected and the global joint pain (visual analogue scale) was assessed before and after iron depletion by phlebotomy. RESULTS: A total of 18 patients [10 males; mean (s.d.) age 48 (11) years] were homozygous for the C282Y mutation. No patient had liver dysfunction. Ferritin level before iron removal was 627.5 (range 133-3276) microg/l, and duration of the iron depletion phase was 295 (70-670) days. Serum levels of both Coll2-1 and CPII were significantly increased from diagnosis after iron depletion: 80.1 (55.6-113.5) vs 96.0 (48.8-136.3) nM (P = 0.004) and 731.4 (374.2-1012.3) vs 812.8 (535.8-1165.6) ng/ml (P = 0.03), respectively. Levels of other biomarkers were not modified by iron depletion. Ferritin level, which at baseline was correlated with body iron store (r = 0.63; P = 0.008), was significantly correlated with HA level measured before iron depletion (r = 0.60; P = 0.01). Global joint pain was not correlated with ferritin concentration and did not significantly decrease after iron depletion: 43 (19-73) vs 36 (16-67) mm (P = 0.07). CONCLUSIONS: In patients with HH, cartilage homoeostasis is modified by iron excess and an increase in type II collagen turnover occurs after excess iron removal. [less ▲]

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See detailFollow-up of COLL2-1, COLL2-1NO2 and myeloperoxydase in dogs after transection of the cruciate ligament of the knee
Kesteloot, Frédéric ULg; Pelletier, Jean-Pierre; Martel-Pelletier, Johanne et al

Poster (2009, May 23)

Purpose: To determine the profile of Coll2-1, Coll2-1NO2 and myeloperoxydase (MPO) serum concentrations in experimental knee OA induced in the dog by transection of the anterior cruciate ligament. Methods ... [more ▼]

Purpose: To determine the profile of Coll2-1, Coll2-1NO2 and myeloperoxydase (MPO) serum concentrations in experimental knee OA induced in the dog by transection of the anterior cruciate ligament. Methods: Surgical transection of the ACL of the right knee was performed on 16 adult crossbred dogs. The dogs were sacrificed 8 weeks after the surgical procedure. Coll2-1, Coll2-1NO2 and MPO were measured by specific immunoassays in 16 dogs at baseline and every 2 weeks during the 8 weeks. The results were expressed as median (range). Results: Immunostainings with D3 and D37, the antiserum recognizing Coll2-1 and Coll2-1NO2, respectively, labelled extracellular matrix in the superficial layer of fibrillated cartilage. After the transection of the ACL, the concentration of 3 biomarkers increased significantly (Friedman test: p<0.001). The concentrations of Coll2-1 and MPO were significantly increased at week 2 compared to baseline [Coll2-1 baseline: 281.57 (131.02-384.67) nM vs Coll2-1 week 2: 345.52 (181.15-589.25) nM (p<0.01) and MPO baseline: 5.16 (<0.4-14.7) ng/ml vs MPO week 2: 14.54 (3.28-31.50) ng/ml (p<0.001)] and remained stable until week 8 [Coll2-1 week 8:318.89 (117.95-492.28) nM and MPO week 8: 11.55 (2.87-42.94) ng/ml]. The Coll2-1NO2 concentration increased significantly at weeks 6 and 8 compared to baseline [Coll2-1NO2 baseline: 0.54 (0.29-1.48) nM vs Coll2-1NO2 week 6: 0.64 (0.40-1.9) nM (p<0.001) vs week 8: 0.61 (0.37-1.79) nM]. Conclusions: These findings suggest that Coll2-1 is a relevant marker for the detection of early structural changes in OA dogs. Interestingly, MPO and Coll2-1NO2 are increased in OA dogs indicating that an oxidative stress occurs in this OA model. [less ▲]

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See detailRelationship between biochemical markers and radiographic scores in the evaluation of the osteoarticular status of Warmblood stallions
Verwilghen, Denis ULg; Busoni, Valeria ULg; Gangl, Monika et al

in Research in Veterinary Science (2009), 87(2), 319-28

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could reveal to be an interesting alternative to this ... [more ▼]

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could reveal to be an interesting alternative to this procedure which is time consuming and sometimes technically difficult. The aim of this study was to investigate the relationship between the radiological status of the horses and the levels of biochemical markers of cartilage degradation and synovial inflammation. A specific radiological scoring and classification system was therefore developed and applied on 63 stallions presented for studbook admission. Additionally, groups of horses were established according to the occurrence of osteochondrosis, degenerative joint disease and distal interphalangeal joint effusion. Insulin growth factor-I, myeloperoxidases, Coll2-1 and Coll2-1NO(2) were used as blood markers. The combination of the blood parameters did not seem to correlate with the used scoring system. Coll2-1NO(2) levels however tended to increase with poorer radiological class and this could therefore potentially be a useful predictor of the osteoarticular status in the horse. Coll2-1 levels were significantly higher in the degenerative joint disease group. A high percentage of horses with distal interphalangeal joint effusion was present in this study and was associated with decreased IGF-I and increased Coll2-1 levels. [less ▲]

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See detailBiochemical biomarkers of oxidative collagen damage.
Henrotin, Yves ULg; Deberg, Michelle ULg; Mathy, Marianne ULg et al

in Advances in Clinical Chemistry (2009), 49

Collagens are major constituents of connective tissues in the animal kingdom. During aging and inflammatory-related diseases, the collagen network undergoes oxidation that leads to structural and ... [more ▼]

Collagens are major constituents of connective tissues in the animal kingdom. During aging and inflammatory-related diseases, the collagen network undergoes oxidation that leads to structural and biochemical alterations within the collagen molecule. Collagen oxidation appears to be a key determinant of aging and a critical physiopathologic mechanism of numerous diseases. Further, the detection of oxidized-collagen peptides seems to be a promising approach for the diagnosis and the prognosis of inflammatory diseases. This chapter reviews the structural and biochemical changes to collagen induced by reactive oxygen and nitrogen species and discusses recent data on the use of collagen-derived biomarkers for measuring oxidative damage. [less ▲]

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