<i>In vitro</i> response of osteoarthritic chondrocytes and fibroblast‐like synoviocytes to a 500–730 kDa hyaluronan amide derivative

Brun, Paola; Zavan, Barbara; Vindigni, Vincenzo; Schiavinato, Antonella; Pozzuoli, Assunta; Iacobellis, Claudio; Abatangelo, Giovanni

doi:10.1002/jbm.b.32771

Cited by 41 publications

(37 citation statements)

References 47 publications

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“…rhPRG4 prevents IκBα degradation in a CD44-dependent manner; this is consistent with its ability to inhibit IL-1-induced NFκB nuclear translocation. This observation extends the efficacy of rhPRG4 as a biologic agent that inhibits synoviocyte proliferation in RA and OA, and establishes a role for CD44 in modulating OA synoviocyte proliferation [38]. The role of PRG4 in modulating OA synoviocyte proliferation is further highlighted by the ability of PRG4 in SF to reduce OA synoviocyte proliferation.…”

Section: Discussionsupporting

confidence: 57%

The autocrine role of proteoglycan-4 (PRG4) in modulating osteoarthritic synoviocyte proliferation and expression of matrix degrading enzymes

et al. 2017

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BackgroundLubricin/proteoglycan 4 (PRG4) is a mucinous glycoprotein secreted by synovial fibroblasts and superficial zone chondrocytes. Recently, we showed that recombinant human PRG4 (rhPRG4) is a putative ligand for CD44 receptor. rhPRG4-CD44 interaction inhibits cytokine-induced rheumatoid arthritis synoviocyte proliferation. The objective of this study is to decipher the autocrine function of PRG4 in regulating osteoarthritic synoviocyte proliferation and expression of catabolic and pro-inflammatory mediators under basal and interleukin-1 beta (IL-1β)-stimulated conditions.MethodsCytosolic and nuclear levels of nuclear factor kappa B (NFκB) p50 and p65 subunits in Prg4 +/+ and Prg4 -/- synoviocytes were studied using western blot. Nuclear translocation of p50 and p65 proteins in osteoarthritis (OA) fibroblast-like synoviocytes (FLS) in response to IL-1β stimulation in the absence or presence of rhPRG4 was studied using DNA binding assays. OA synoviocyte (5000 cells per well) proliferation following IL-1β (20 ng/ml) treatment in the absence or presence of rhPRG4 (50–200 μg/ml) over 48 hours was determined using a colorimetric assay. Gene expression of matrix metalloproteinases (MMPs), tissue inhibitor of metallproteinases-1 (TIMP-1), TIMP-2, IL-1β, IL-6, IL-8, TNF-α, cycloxygenae-2 (COX2) and PRG4 in unstimulated and IL-1β (1 ng/ml)-stimulated OA synoviocytes, in the presence or absence of rhPRG4 (100 and 200 μg/ml), was studied following incubation for 24 hours.Results Prg4 -/- synoviocytes contained higher nuclear p50 and p65 levels compared to Prg4 +/+ synoviocytes (p < 0.05). rhPRG4 (100 μg/ml) reduced p50 and p65 nuclear levels in Prg4 +/+ and Prg4 -/- synoviocytes (p < 0.001). Similarly, rhPRG4 (200 μg/ml) inhibited NFκB translocation and cell proliferation in OA synoviocytes in a CD44-dependent manner (p < 0.001) via inhibition of IκBα phosphorylation. IL-1β reduced PRG4 expression in OA synoviocytes and rhPRG4 (100 μg/ml) treatment reversed this effect (p < 0.001). rhPRG4 (200 μg/ml) reduced basal gene expression of MMP-1, MMP-3, MMP-13, IL-6, IL-8, and PRG4 in OA synoviocytes, while increasing TIMP-2 and cycloxygenase-2 (COX2) expression (p < 0.001). rhPRG4 (200 μg/ml) reduced IL-1β induction of MMP-1, MMP-3, MMP-9, MMP-13, IL-6, IL-8, and COX2 expression in a CD44-dependent manner (p < 0.001).ConclusionPRG4 plays an important anti-inflammatory role in regulating OA synoviocyte proliferation and reduces basal and IL-1β-stimulated expression of catabolic mediators. Exogenous rhPRG4 autoregulates native PRG4 expression in OA synoviocytes.

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Section: Discussionsupporting

confidence: 57%

The autocrine role of proteoglycan-4 (PRG4) in modulating osteoarthritic synoviocyte proliferation and expression of matrix degrading enzymes

et al. 2017

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“…Cells remained viable throughout all experiments, with preliminary trials revealing no change in cell numbers with any treatment (data not shown) as previously published [22]. Some HSF detached from the culture well surface in the absence of FBS and this phenomenon was exacerbated in the presence of HA.…”

Section: Resultssupporting

confidence: 79%

“…The interaction of unmodified HA with CD44 is important in suppression of IL-1β-stimulated effects in cultured chondrocytes [15,38,42,53] and synovial fibroblasts [49]. CD44 is also involved in the stimulation of OA chondrocyte proliferation by the amide derivative of HA [22]. We were able to demonstrate amelioration of a few of the effects of the derivatized HA on gene expression using a CD44 blocking antibody in confluent cultures of HAC and HSF supporting a role for this cell-surface receptor.…”

Section: Discussionmentioning

confidence: 99%

“…In HSF from 1 patient and HAC from 2 patients, CD44 antibody (R&D systems BBA10; 5 or 10 μg/mL) or IgG isotype control antibody at the same concentration (R&D Systems MAB003) were added to the cultures 1 or 3 hours before the addition of the hexadecylamide derivative of HA (HYADD4®-G; HYMOVIS® at 0.5 mg/ml), ± IL-1β (2 ng/mL) one hour later, as previously described [22]. These cultures (n = 6 replicates for each treatment in each patient) were terminated at 24 hours for gene expression analysis.…”

Section: Methodsmentioning

confidence: 99%

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A hexadecylamide derivative of hyaluronan (HYMOVIS®) has superior beneficial effects on human osteoarthritic chondrocytes and synoviocytes than unmodified hyaluronan

et al. 2013

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BackgroundIntra-articular hyaluronan (HA) injection provides symptomatic benefit in the treatment of osteoarthritis (OA). Previously we found superior beneficial effects in a large animal OA model of a hexadecylamide derivative compared with unmodified HA of the same initial molecular weight. The current study sought to define possible molecular mechanisms whereby this enhanced relief of symptoms was occurring.MethodsChondrocytes and synovial fibroblasts were isolated from tissues of patients undergoing arthroplasty for knee OA. Monolayer cultures of cells were treated with 0, 0.5, 1.0 or 1.5 mg/mL of unmodified HA (500–730 kDa) or a hexadecylamide derivative of HA of the same initial molecular weight (HYADD4®-G; HYMOVIS®) simultaneously or 1 hour before incubation with interleukin (IL)-1beta (2 ng/mL). Cultures were terminated 15 or 30 minutes later (chondrocytes and synovial fibroblasts, respectively) for quantitation of phosphorylated-(p)-JNK, p-NFkappaB, p-p38, or at 24 hours for quantitation of gene expression (MMP1 &13, ADAMTS4 &5, TIMP1 &3, CD44, COL1A1 &2A1, ACAN, PTGS2, IL6, TNF) and matrix metalloproteinase (MMP)-13 activity.ResultsThe hexadecylamide derivative of HA had significantly better amelioration of IL-1beta-induced gene expression of key matrix degrading enzymes (MMP1, MMP13, ADAMTS5), and inflammatory mediators (IL6, PTGS2) by human OA chondrocytes and synovial fibroblasts. Pre-incubation of cells with the derivatized HA for 1 hour prior to IL-1beta exposure significantly augmented the inhibition of MMP1, MMP13, ADAMTS4 and IL6 expression by chondrocytes. The reduction in MMP13 mRNA by the amide derivative of HA was mirrored in reduced MMP-13 protein and enzyme activity in IL-1beta-stimulated chondrocytes. This was associated in part with a greater inhibition of phosphorylation of the cell signalling molecules JNK, p38 and NF-kappaB.ConclusionsThe present studies have demonstrated several potential key mechanisms whereby the intra-articular injection of a hexadecylamide derivative of HA may be acting in joints with OA.

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“…Smith et al demonstrated that this HA derivative reduced the expression in OA of key proteolytic enzymes implicated in cartilage degradation such as matrix metalloproteinase (MMP)1, MMP13, disintegrin and metalloproteinases with thrombospondin motifs (ADAMTS)4, ADAMTS5, as well as the production of inflammatory mediators such as IL‐6 . An upregulation of anti‐inflammatory cytokine expression such as IL‐10 has also been shown …”

Section: Introductionmentioning

confidence: 99%

IL‐1ß and IL‐8 are scavenged by the hexadecylamide derivative of hyaluronic acid: A new mechanism

Oliviero

Scanu

Ramonda

et al. 2015

J Biomedical Materials Res

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This study aimed to investigate, using an in vitro model, the mechanisms involved in the effects linked to a novel hexadecylamide derivative of hyaluronic acid (HA), HYADD®4 (HS), on some inflammatory aspects related to the osteoarthritis process. The human leukemic monocytic cell line THP-1 was stimulated with calcium pyrophosphate (CPP) crystals or lipopolysaccaride (LPS) and cultured in the presence of HS or two unmodified HAs (500-730 kDa and >1500 kDa, respectively). The effects of the three HA derivatives were compared by examining the inhibition of IL-1ß and IL-8 release, the phagocytic capacity of THP-1, and HA's physical interference with the cytokines and their biological activity. Adding HS simultaneously with the stimuli led to a marked (nearly 100%) decrease in cytokine release and biological activity with respect to the two unmodified HAs. The effect was not altered when a CD44 function-blocking monoclonal antibody was used. Incubation of the three derivatives with IL-1ß and IL-8 led to a reduced bioavailability of the cytokines in the medium in the presence of HS but not of unmodified HA. This study examines a novel mechanism inhibiting cytokine bioactivity. The HA hexadecylamide derivative was found to suppress, in vitro, the inflammatory response induced by CPP crystals and LPS by reducing the bioavailability of the two cytokines that were analyzed.

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In vitro response of osteoarthritic chondrocytes and fibroblast‐like synoviocytes to a 500–730 kDa hyaluronan amide derivative

Cited by 41 publications

References 47 publications

The autocrine role of proteoglycan-4 (PRG4) in modulating osteoarthritic synoviocyte proliferation and expression of matrix degrading enzymes

The autocrine role of proteoglycan-4 (PRG4) in modulating osteoarthritic synoviocyte proliferation and expression of matrix degrading enzymes

A hexadecylamide derivative of hyaluronan (HYMOVIS®) has superior beneficial effects on human osteoarthritic chondrocytes and synoviocytes than unmodified hyaluronan

IL‐1ß and IL‐8 are scavenged by the hexadecylamide derivative of hyaluronic acid: A new mechanism

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