epidermal-growth-factor and Osteoarthritis

Osteoarthritis (OA) is defined as a degenerative joint disease that can affect all tissues of the joint, including the articular cartilage, subchondral bone, ligaments capsule, and synovial membrane. The conventional nonoperative treatments are ineffective for cartilage repair and induce only symptomatic relief. Platelet-rich plasma (PRP) is a platelet concentrate derived from autologous whole blood with a high concentration of platelets, which can exert anti-inflammatory and regenerative effects by releasing multiple growth factors and cytokines. Recent studies have shown that PRP exhibits clinical benefits in patients with OA. However, high operational and equipment requirements greatly limit the application of PRP to OA treatment. Past studies have indicated that high-concentration PRP growth factors and cytokines may be applied as a commercial replacement for PRP. We reviewed the relevant articles to summarize the feasibility and mechanisms of PRP-based growth factors in OA. The available evidence suggests that transforming growth factor-α and β, platelet-derived growth factors, epidermal growth factor, insulin-like growth factor-1, and connective tissue growth factors might benefit OA, while vascular endothelial growth factor, tumor necrosis factor-α, angiopoietin-1, and stromal cell derived factor-1α might induce negative effects on OA. The effects of fibroblast growth factor, hepatocyte growth factor, platelet factor 4, and keratinocyte growth factor on OA remain uncertain. Thus, it can be concluded that not all cytokines released by PRP are beneficial, although the therapeutic action of PRP has a valuable potential to improve.

Topics: Chemokine CXCL12; Epidermal Growth Factor; Humans; Osteoarthritis; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Synovial fibroblasts are critical for maintaining homeostasis in major autoimmune diseases involving joint inflammation, including osteoarthritis and rheumatoid arthritis. However, little is known about the interactions among different cell subtypes and the specific sets of signaling pathways and activities that they trigger.. Using social network analysis, pattern recognition, and manifold learning approaches, we identified patterns of single-cell communication in OA (osteoarthritis) and RA (rheumatoid arthritis).. Increased understanding of the interaction networks between synovial fibroblast subtypes, particularly the shared and unique cellular communication features between osteoarthritis and rheumatoid arthritis and their hub cells, should help inform the design of therapeutic agents for inflammatory joint disease.

Topics: Arthritis, Rheumatoid; Cell Communication; Collagen Type VI; Communication; Epidermal Growth Factor; Fibroblasts; Humans; Laminin; Osteoarthritis; Synovial Membrane

Chondrocytes in mice developing osteoarthritis (OA) exhibit an aberrant response to the secreted cytokine transforming growth factor (TGF)-β, consisting in a potentiation of intracellular signaling downstream of the transmembrane type I receptor kinase activin receptor-like kinase (ALK)1 against canonical TGF-β receptor ALK5-mediated signaling. Unfortunately, the underlying mechanisms remain elusive. In order to identify novel druggable targets for OA, we aimed to investigate novel molecules regulating the ALK1/ALK5 balance in OA chondrocytes. We performed gene expression analysis of TGF-β signaling modulators in joints from three different mouse models of OA and found an upregulated expression of the TGF-β co-receptor Cripto (Tdgf1), which was validated in murine and human cartilage OA samples at the protein level. In vitro and ex vivo, elevated expression of Cripto favors the hypertrophic differentiation of chondrocytes, eventually contributing to tissue calcification. Furthermore, we found that Cripto participates in a TGF-β-ALK1-Cripto receptor complex in the plasma membrane, thereby inducing catabolic SMAD1/5 signaling in chondrocytes. In conclusion, we demonstrate that Cripto is expressed in OA and plays a functional role promoting chondrocyte hypertrophy, thereby becoming a novel potential therapeutic target in OA, for which there is no efficient cure or validated biomarker. © 2021 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.

Topics: Animals; Chondrocytes; Epidermal Growth Factor; GPI-Linked Proteins; Humans; Hypertrophy; Intercellular Signaling Peptides and Proteins; Membrane Glycoproteins; Mice; Neoplasm Proteins; Osteoarthritis; Signal Transduction; Smad Proteins; Transforming Growth Factor beta

This study has aimed to study different culture systems that might stimulate an increase in cell proliferation of normal and osteoarthritis chondrocytes from articular cartilage in rat model.. Three culture systems using chondrocytes embedded in alginate beads were tested: chondrocytes cultured in Dulbecco's modified Eagle's medium (DMEM) as control, a co-culture system consisting of a monolayer of de-differentiated chondrocytes as a source of mitotic factors, and an enriched medium containing culture medium obtained from a monolayer of chondrocytes and DMEM. Normal and osteoarthritis chondrocytes were stained with 5-carboxyfluorescein diacetate succinimidyl ester and were cultured in each of the three systems. After 5 days of culture cell, proliferation was detected by flow cytometry. Chondrocyte phenotype was confirmed by collagen type II and MMP-3 expression. To determine possible molecules released into the medium by the cultured chondrocyte monolayer and which would probably be involved in cell proliferation, a study of mRNA and expression of transforming growth factor-beta1 (TGF-beta1), fibroblastic growth factor-2 (FGF-2), epidermal growth factor (EGF), platelet derived growth factor-A (PDGF-A) and insulin-like growth factor-1 (IGF-1) proteins was conducted.. Chondrocytes in the co-culture system or in enriched medium showed an increase in proliferation; only when osteoarthritis chondrocytes were cultured in enriched medium would they display a statistically significant increase in their proliferation rate and in their viability. When chondrocytes from the monolayer were analysed, differential mRNA expression of TGF-beta1 and IGF-1 was found during all passages, which suggests that these two growth factors might be involved in chondrocyte proliferation.

Topics: Alginates; Animals; Cartilage, Articular; Cell Culture Techniques; Cell Dedifferentiation; Cell Proliferation; Chondrocytes; Coculture Techniques; Collagen Type II; Culture Media, Conditioned; Epidermal Growth Factor; Fibroblast Growth Factor 2; Gene Expression; Glucuronic Acid; Hexuronic Acids; Insulin-Like Growth Factor I; Male; Matrix Metalloproteinase 3; Mitosis; Osteoarthritis; Platelet-Derived Growth Factor; Rats; Rats, Wistar; Transforming Growth Factor beta1

To identify regulators of the cartilaginous phenotype, on the basis of their differential expression in human conventional chondrogenic tumors compared with articular cartilage.. Differential proteomics analysis revealed matrilin-3 (MATN3) as a candidate regulator of the cartilaginous phenotype. Its capacity to modulate gene expression was investigated in human HCS-2/8 chondrosarcoma cells and transfected chondrocytes, using cell culture fractionation, reverse transcription-polymerase chain reaction, and Western blot analyses.. Increased expression of the cartilage-specific matrix protein MATN3 was specifically observed in enchondromas and conventional chondrosarcomas. A substantial fraction of MATN3 was found in cytoplasmic structures of tumor cells, as demonstrated by immunohistochemistry. Analyses of intracellular MATN3 revealed that it corresponded to an imperfectly maturated MATN3 polypeptide, both in HCS-2/8 human chondrosarcoma cells and in transfected human chondrocytes. Moderately increased expression of MATN3 resulted in its intracellular retention. Antibody-mediated blockade of soluble, extracellular MATN3 in HCS-2/8 cell cultures resulted in increased expression of MATN3 and the chondrogenic transcription factor SOX9. Conversely, increased ectopic expression of MATN3 resulted in decreased expression of MATN3 and SOX9 in primary chondrocytes, while a mutant MATN3 lacking its first epidermal growth factor (EGF)-like domain failed to down-regulate SOX9.. Aberrant expression and processing of MATN3 are hallmarks of conventional cartilaginous neoplasms. A particular step in the maturation of MATN3 limits its processing through the secretion machinery, resulting in its intracellular accumulation upon increased expression. Soluble, secreted MATN3, however, down-regulates SOX9 at the messenger RNA and protein levels. The first EGF-like domain of MATN3 is a critical determinant of its regulatory activity toward SOX9. These activities of MATN3 suggest that its increased expression in osteoarthritis might contribute to the degeneration of articular cartilage.

Topics: Adolescent; Adult; Aged; Blotting, Western; Bone Neoplasms; Cartilage, Articular; Cell Differentiation; Cells, Cultured; Child; Child, Preschool; Chondrocytes; Chondroma; Chondrosarcoma; Down-Regulation; Epidermal Growth Factor; Extracellular Matrix Proteins; Female; Flow Cytometry; Humans; Immunohistochemistry; Male; Matrilin Proteins; Middle Aged; Osteoarthritis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; SOX9 Transcription Factor

Angiogenesis is a process stimulated in inflamed synovium of patients with osteoarthritis (OA), and contributes to the progression of the disease. Synovial fibroblasts secrete angiogenic factors, such as vascular endothelial growth factor (VEGF), an up-regulator of angiogenesis, and this ability is increased by interleukin (IL)-1beta. The purpose of this study was to verify whether IL-17 contributes and/or synergizes with IL-1beta and tumor necrosis factor (TNF)-alpha in vessel development in articular tissues by stimulating the secretion of proangiogenic factors by synovial fibroblasts.. We stimulated in vitro synovial fibroblasts isolated from OA, rheumatoid arthritis (RA) and fractured patients (FP) with IL-17 and IL-1beta and from OA patients with IL-17, IL-1beta and TNF-alpha. In the supernatants from the cultures, we assayed the amount of VEGF by immunoassay and other angiogenic factors (keratinocyte growth factor, KGF; hepatocyte growth factor, HGF; heparin-binding endothelial growth factor, HB-EGF; angiopoietin-2, Ang-2; platelet-derived growth factor B, PDGF-BB; thrombopoietin, TPO) by chemiluminescence; semiquantitative RT-PCR was used to state mRNA expression of nonreleased angiogenic factors (Ang-2 and PDGF-BB) and tissue inhibitors of metalloproteinase (TIMP)-1.. IL-17, TNF-alpha and IL-1beta increased VEGF secretion by synovial fibroblasts from OA patients. IL-17 and IL-1beta also increased VEGF secretion in RA and FP. Besides, IL-17 increased KGF and HGF secretions in OA, RA and FP; in OA and RA, IL-17 also increased the HB-EGF secretion and the expression of TIMP-1 as protein and mRNA. In OA patients IL-17 had an additive effect on TNF-alpha-stimulated VEGF secretion.. These results suggest that IL-17 is an in vitro stimulator of angiogenic factor release, both by its own action and by cooperating with TNF-alpha.

Topics: Adult; Aged; Aged, 80 and over; Angiogenesis Inducing Agents; Arthritis, Rheumatoid; Epidermal Growth Factor; Fibroblast Growth Factor 7; Fibroblasts; Fractures, Bone; Heparin-binding EGF-like Growth Factor; Hepatocyte Growth Factor; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-1; Interleukin-17; Middle Aged; Osteoarthritis; RNA, Messenger; Synovial Membrane; Tissue Inhibitor of Metalloproteinase-1; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

EMR2 and CD97 are closely related members of the epidermal growth factor (EGF)-TM7 family of adhesion class 7-span transmembrane (TM7) receptors. Chondroitin sulfates (CS) have recently been identified as ligands for EMR2 and CD97. CS have been implicated in the pathogenesis of rheumatoid arthritis (RA). We undertook this study to determine the expression of EMR2 and the distribution of EMR2 and CD97 ligands within RA synovial tissue (ST).. ST samples were obtained by arthroscopy from 19 patients with RA, 13 patients with inflammatory osteoarthritis (OA), and 13 patients with reactive arthritis (ReA). Immunohistochemistry was performed with a monoclonal antibody against EMR2, and stained STs were analyzed by digital image analysis. Coexpression of EMR2 with cell lineage- and activation-specific markers was determined by double immunofluorescence microscopy. To evaluate the expression of EMR2 and CD97 ligands in RA synovium, binding assays were performed using EMR2- and CD97-specific multivalent fluorescent probes.. EMR2 expression in the synovial sublining was found to be significantly higher in RA patients compared with OA and ReA control patients. Most EMR2+ cells were macrophages and dendritic cells expressing costimulatory molecules and tumor necrosis factor alpha. Dermatan sulfate was shown to be the ligand of the largest isoforms of EMR2 and CD97 in rheumatoid synovium. In addition, the smaller isoforms of CD97, but not those of EMR2, bound CD55 on fibroblast-like synoviocytes.. The EGF-TM7 receptors EMR2 and CD97 are abundantly expressed on myeloid cells in ST of RA patients where their cognate ligands dermatan sulfate and CD55 are detected. These results suggest that these interactions may facilitate the retention of activated macrophages in the synovium.

Topics: Aged; Antigens, CD; Arthritis, Rheumatoid; CD55 Antigens; Dermatan Sulfate; Epidermal Growth Factor; Female; Fluorescent Dyes; Humans; Immunohistochemistry; Ligands; Male; Membrane Glycoproteins; Microscopy, Interference; Middle Aged; Osteoarthritis; Prohibitins; Receptors, G-Protein-Coupled; Synovial Membrane

Since many cytokines have been identified in chronically inflamed human synovium, it is possible that particular cytokines or combinations of cytokines play dominant roles in driving or inhibiting metabolic processes important to inflammation. To assess these possibilities, we compared selected effects of individual cytokines and their binary, ternary, and higher combinations in human synovial cell cultures.. Cytokines studied known to occur in human synovial tissue included: interleukin 1beta (IL-1beta), IL-6, tumor necrosis factor-alpha, granulocyte macrophage colony stimulating factor, interferon-gamma, acidic fibroblast growth factor (aFGF), basic FGF (bFGF), platelet derived growth factor, transforming growth factor-beta1, connecting tissue activating peptide-III, and epidermal growth factor. The growth related effects of these agents singly and in combinations were assessed by measuring newly synthesized [3H]DNA and [14C]GAG (glycosaminoglycan) in human synovial cell cultures. Cytokine induced synthesis of prostaglandin E2 (PGE2) was measured by ELISA.. Most cytokine combinations resulted in additive/synergistic anabolic effects, except when IL-1beta was present; IL-1beta was markedly antagonistic to the mitogenic effects of other cytokines tested. Combinations of platelet derived cytokines were the most potent stimulators of DNA synthesis, while combinations of synovial derived cytokines were more active in stimulating GAG synthesis. Synovial cells exposed simultaneously to both platelet and synovial derived cytokines produced large quantities of [14C]GAG and showed a modest increase in [3H]DNA synthesis. IL-1beta, alone or in combinations, was dominant with respect to stimulation of PGE2 synthesis. Acetylsalicylic acid substantially interfered with all the effects of cytokine combinations measured.. Quantitative alterations in synovial cell synthesis of GAG and DNA varied greatly depending on the ambient mixture of cytokines. Virtually all combinations of cytokines tested gave rise to large increases in synovial cell synthesis of GAG. Four platelet derived cytokines, a "physiologic combination," appeared to be dominant agents in stimulating DNA synthesis. This effect was profoundly reduced by the antagonistic effect of IL-1beta, mediated in part by PGE2. The patterns of cytokine combination induced metabolic effects suggest that the "cytokine network" has a significant measure of redundancy with respect to control of synovial cell metabolism.

Topics: Arthritis, Rheumatoid; Aspirin; Cells, Cultured; Connective Tissue; Culture Media, Conditioned; Cytokines; Dinoprostone; DNA; Drug Synergism; Epidermal Growth Factor; Fibroblast Growth Factors; Glycosaminoglycans; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hydrocortisone; Inflammation; Insulin-Like Growth Factor I; Interferon-gamma; Interleukin-6; Osteoarthritis; Peptides; Platelet-Derived Growth Factor; Recombinant Proteins; Synovial Membrane; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To compare, by immunohistochemistry, the cellular and cytokine profile in rheumatoid arthritis (RA) and osteoarthritis (OA) synovial membranes (SMs). Synovium was obtained at knee arthroplasty from 10 patients with RA and 10 with OA.. Synovial membranes were stained with a panel of monoclonal antibodies (MAb) to assess cytokine expression (IL-1 alpha, IL-1 beta, IL-6, GM-CSF, TNF-alpha and EGF) and the intensity of the mononuclear cellular infiltrate (MNC).. Significantly greater percentages of IL-1 alpha, IL-1 beta, IL-6, TNF-alpha, GM-CSF and EGF cells were detected in all areas of the rheumatoid SMs when compared with osteoarthritic SMs. Five RA but only one OA SM demonstrated focal lymphoid aggregates. Lining layer thickening was noted in RA SMs only. The intensity of the MNC and number of blood vessels were greater in the RA group.. The results suggest that the differences in cytokine production by RA and OA SMs are quantitative but that the greater thickness of the synovial lining layer and higher vascularity may be specific to RA.

Topics: Adult; Aged; Arthritis, Rheumatoid; Cytokines; Epidermal Growth Factor; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunoenzyme Techniques; Interleukins; Male; Middle Aged; Osteoarthritis; Synovial Membrane; Tumor Necrosis Factor-alpha

The immuno-reactive human epidermal growth factor (h-EGF) level, measured by a sensitive enzyme immunoassay, was significantly higher in synovial fluids or synovial tissue extracts from 89 patients with rheumatoid arthritis (RA) than in those from 53 patients with osteoarthritis. RA synovial immuno-reactive h-EGF was predominantly of a low molecular weight form (LMW h-EGF) on gel filtration chromatography. Furthermore, in the RA group, the synovial immuno-reactive h-EGF correlated positively with C-reactive protein, an acute-phase reactant, in the blood examination and with synovial immunoglobulin M. These results suggest that synovial h-EGF is specifically produced by RA synovium from the initial stage of arthritis, and that the measurement of synovial h-EGF serves as an early indicator to appraise the RA activity. A pathogenesis of RA including growth factors and growth inhibitory factors are discussed.

Topics: Adult; Aged; Arthritis, Rheumatoid; Epidermal Growth Factor; Female; Humans; Immunoenzyme Techniques; Immunoglobulins; Male; Middle Aged; Osteoarthritis; Synovial Fluid; Synovial Membrane

Conditioned media obtained from fibroblasts cultured from rheumatoid and certain other inflammatory synovia were observed to stimulate [3H]thymidine incorporation in an indicator murine fibroblast line. Synovial fibroblasts derived from the joints of patients with osteoarthritis did not display this property. This effect persisted in culture for many weeks and occurred in the absence of co-stimulatory immune cells. Antibody neutralization studies implicated a role for basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-beta), granulocyte/macrophage colony-stimulating factor (GM-CSF), and interleukin 1 beta (IL-1 beta) in the increased proliferative activity of synovial fibroblast-conditioned media. Synovial cell synthesis of bFGF, TGF beta 1, GM-CSF, IL-1 beta, and IL-6 was confirmed by 35S-methionine labeling and immunoprecipitation. The constitutive production of inflammatory and mitogenic cytokines by synovial fibroblasts may represent the result of long-term, phenotypic changes that occurred in vivo. Persistent cytokine production by synovial fibroblasts may play an important role in the continued recruitment and activation of inflammatory cells in chronic arthritis and in the formation of rheumatoid pannus.

Topics: Animals; Arthritis, Rheumatoid; Cell Division; Cell Line; Cells, Cultured; Cytokines; DNA Replication; Epidermal Growth Factor; Fibroblast Growth Factor 2; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-1; Interleukin-6; Mice; Osteoarthritis; Synovial Membrane; Transforming Growth Factor beta

When stimulated with increasing amounts of interleukin 1 beta (IL 1 beta) rheumatoid arthritis (RA), as compared with osteoarthritis (OA), synovial cells grown in RPMI plus fetal bovine serum (FBS), released significantly more prostaglandin E2 (PGE2) (p less than 0.05; paired t test, two-tailed). PGE2 release by IL 1 beta-stimulated RA synovial cells grown for 14 days in serum-free RPMI was significantly less than that released by the same cells grown in medium plus 10% FBS (p less than 0.03; two-tailed). Since these data suggest that growth factors present in FBS may augment the effects of IL 1 beta, experiments were conducted to study the influence of four polypeptide growth factors--transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF), on IL 1 beta-induced release of PGE2 by cultured RA synovial cells. Both EGF and bFGF significantly enhanced IL 1 beta-induced release of PGE2 (p less than 0.05; paired t test, one-tailed), while PDGF was synergistic with IL 1 beta, significantly increasing release of PGE2 by these cultured cells (p less than 0.02; two-tailed). No such effect was seen when TGF-beta was added to the culture medium. Taken together, these data lend support to the concept that within the synovial micro-environment small quantities of individual growth factors may potentiate the effects of IL 1 beta to amplify intra-articular inflammation.

Topics: Arthritis, Rheumatoid; Cells, Cultured; Dinoprostone; Epidermal Growth Factor; Fibroblast Growth Factor 2; Growth Substances; Humans; In Vitro Techniques; Interleukin-1; Osteoarthritis; Platelet-Derived Growth Factor; Synovial Membrane; Transforming Growth Factor beta

Immunohistochemical study showed selective localisation of human epidermal growth factor (hEGF) to the synovial lining layer. Although the synovial lining layer of the rheumatoid, osteoarthritic, and traumatic joints was hEGF positive, hEGF staining was especially dense at the rheumatoid synovial lining layer; the staining increasing linearly according to the degree of stratification of the lining layer (r = 1). Human epidermal growth factor was ultrastructurally localised to cytoplasm, especially to rough endoplasmic reticulum, of the synovial lining fibroblast-like (type B) cell. Only the cell surface of macrophage-like (type A) cells was hEGF positive. When different histological variables were compared with each other a positive correlation was found between hEGF staining of the synovial lining layer and the degree of neovascularisation of rheumatoid synovium (r = 0.72). Although some lymphocytes were weakly hEGF positive, neovascularisation did not correlate with the extent of lymphocyte infiltration or of hEGF staining of lymphocytes. Lymphocyte infiltration or hEGF staining of lymphocytes did not correlate with hEGF staining of the synovial lining layer, whereas the lymphocyte infiltration correlated positively with the extent of perivascular accumulation of lymphocytes (r = 0.89). These findings suggest that (a) hEGF is synthesised by and secreted through endoplasmic reticulum and Golgi apparatus from the synovial lining type B cell; (b) hEGF is at least partially responsible for the pathogenesis of stratification of the rheumatoid synovial lining layer, and perhaps of neovascularisation of the rheumatoid synovium, whereas it is not responsible for lymphocyte accumulation to the rheumatoid synovium.

Topics: Arthritis, Rheumatoid; Epidermal Growth Factor; Humans; Immunohistochemistry; Lymphocytes; Neovascularization, Pathologic; Osteoarthritis; Synovial Membrane