Laurence Pesesse

Regulation of subchondral bone osteoblast metabolism by cyclic compression

OBJECTIVE Recent data have shown that abnormal subchondral bone remodeling plays an important role in osteoarthritis (OA) onset and progression, and it was suggested that abnormal mechanical pressure applied to the articulation was responsible for these metabolic changes. This study was undertaken to evaluate the effects of cyclic compression on osteoblasts from OA subchondral bone. METHODS Osteoblasts were isolated from sclerotic and nonsclerotic areas of human OA subchondral bone. After 28 days, the osteoblasts were surrounded by an abundant extracellular matrix and formed a resistant membrane, which was submitted to cyclic compression (1 MPa at 1 Hz) for 4 hours. Gene expression was evaluated by reverse transcription-polymerase chain reaction. Protein production in culture supernatants was quantified by enzyme-linked immunosorbent assay or visualized by immunohistochemistry. RESULTS Compression increased the expression of genes coding for interleukin-6 (IL-6), cyclooxygenase 2, RANKL, fibroblast growth factor 2, IL-8, matrix metalloproteinase 3 (MMP-3), MMP-9, and MMP-13 but reduced the expression of osteoprotegerin in osteoblasts in both sclerotic and nonsclerotic areas. Colα1(I) and MMP-2 were not significantly affected by mechanical stimuli. Nonsclerotic osteoblasts were significantly more sensitive to compression than sclerotic ones, but after compression, differences in messenger RNA levels between nonsclerotic and sclerotic osteoblasts were largely reduced or even abolished. Under basal conditions, sclerotic osteoblasts expressed similar levels of α5, αv, β1, and β3 integrins and CD44 as nonsclerotic osteoblasts but 30% less connexin 43, an important mechanoreceptor. CONCLUSION Genes involved in subchondral bone sclerosis are mechanosensitive. After compression, nonsclerotic and sclerotic osteoblasts expressed a similar phenotype, suggesting that compression could be responsible for the phenotype changes in OA subchondral osteoblasts.

Subchondral bone and osteoarthritis: biological and cellular aspects

The subchondral bone is involved in the pathophysiology of osteoarthritis (OA), both by biochemical and mechanical pathways. Overloaded OA subchondral bone osteoblasts express a pro-angiogenic and pro-inflammatory phenotype which contributes to explain the structural changes (sclerosis and bone marrow lesion) visible in OA subchondral bone. Further, microfractures and conjonctivo-vascular structures constitute exchange routes between bone and the overlying cartilage for mediators produced by osteoblasts. This narrative review describes these physiopathological mechanisms and identifies possible therapeutic targets for pharmacological modalities.

Osteochondral plate angiogenesis: A new treatment target in osteoarthritis.

Healthy adult joint cartilage contains neither blood vessels nor nerves. Osteoarthritic cartilage, in contrast, may be invaded by blood vessels from the subchondral bone. The mechanisms underlying cartilage angiogenesis in osteoarthritis are unclear but may involve hypertrophic chondrocyte differentiation. Active research is under way to identify the factors involved in cartilage angiogenesis. Here, we discuss the pathophysiological mechanisms of osteoarthritic cartilage angiogenesis based on evidence from a systematic literature review of articles retrieved via PubMed and ISI Web of Knowledge. Our conclusions suggest new research perspectives and treatment options.

Consequences of chondrocyte hypertrophy on osteoarthritic cartilage: potential effect on angiogenesis

OBJECTIVE The aim of this study was to investigate the link between the hypertrophic phenotype of chondrocytes and angiogenesis in osteoarthritis (OA) and more particularly to demonstrate that OA hypertrophic chondrocytes potentially express a phenotype promoting angiogenesis through the expression of factors controlling endothelial cells migration, invasion and adhesion. METHOD Human OA chondrocytes were cultivated in alginate beads in medium supplemented with 10% fetal bovine serum (FBS) to induce chondrocyte hypertrophy. The hypertrophic phenotype was characterized throughout 28 days of culture by measuring the expression of specific genes and by a microscopic observation of cellular morphology. The effect of media conditioned by OA hypertrophic chondrocyte on endothelial cells migration, invasion and adhesion was evaluated in functional assays. Moreover, hypertrophic OA chondrocytes were tested for the expression of angiogenic factors by real-time RT-PCR. RESULTS Specific markers of hypertrophy and observation of cellular morphology attested of the hypertrophic phenotype of chondrocytes in our culture model. Functional angiogenesis assays showed that factors produced by hypertrophic chondrocytes stimulated migration, invasion and adhesion of endothelial cells. Among the evaluated angiogenic factors, bone sialoprotein (BSP) was the most highly upregulated in hypertrophic chondrocytes. The inhibition of endothelial cell adhesion by a GRGDS peptide confirmed the implication of RGD domain proteins, like BSP, in hypertrophic chondrocyte-induced adhesion of endothelial cells. CONCLUSION Hypertrophic differentiation of chondrocyte may promote angiogenesis. Our findings established the relation of BSP with OA chondrocyte hypertrophy and suggested that this factor could constitute a potential target to control cartilage neovascularisation in OA.

Targeting the synovial angiogenesis as a novel treatment approach to osteoarthritis

Synovitis is a key feature in osteoarthritis and is associated with symptom severity. Synovial membrane inflammation is secondary to cartilage degradation which occurs in the early stage and is located adjacent to cartilage damage. This inflammation is characterized by the invasion and activation of macrophages and lymphocytes, the release in the joint cavity of large amounts of pro-inflammatory and procatabolic mediators, and by a local increase of synovial membrane vascularity. This latter process plays an important role in the chronicity of the inflammatory reaction by facilitating the invasion of the synovium by immune cells. Therefore, synovial membrane angiogenesis represents a key target for the treatment of osteoarthritis. This paper is a narrative review of the literature referenced in PubMed during the past 5 years. It addresses in particular three questions. What are the mechanisms involved in synovium blood vessels invasion? Are current medications effective in controlling blood vessels formation and invasion? What are the perspectives of research in this area?

Subchondral bone in osteoarthritis physiopathology: state-of-the art and perspectives.

Osteoarthritis (OA) is the most common form of arthritic disease, and it is a major cause of disability and impaired quality of life in the elderly. A hallmark of the disease is progressive degeneration of articular cartilage and subsequent joint space narrowing. However, OA is a complex disease not limited to cartilage degeneration, but involving also synovial membrane and subchondral bone, thereby presenting alternatives approaches for treatment. In this paper, we propose a short review of the recent advances in the understanding of the role played by subchondral bone in OA.

Bone sialoprotein as a potential key factor implicated in the pathophysiology of osteoarthritis

OBJECTIVE We previously identified an association between bone sialoprotein (BSP) and osteoarthritic (OA) chondrocyte hypertrophy but the precise role of BSP in ostearthritis (OA) has not been extensively studied. This study aimed to confirm the association between BSP and OA chondrocyte hypertrophy, to define its effect on molecules produced by chondrocytes and to analyse its association with cartilage degradation and vascular density at the osteochondral junction. METHOD Human OA chondrocytes were cultivated in order to increase hypertrophic differentiation. The effect of parathyroid hormone-related peptide (PTHrP), interleukin (IL)-1β or tumour necrosis factor (TNF)-α on BSP was analysed by real-time reverse transcription polymerase chain reaction (RT-PCR) and western blot. The effects of BSP on OA chondrocytes production of inflammatory response mediators (IL-6, nitric oxide), major matrix molecule (aggrecan), matrix metalloprotease-3 and angiogenic factors (vascular endothelial growth factor, basic fibroblast growth factor, IL-8, and thrombospondin-1) were investigated. BSP was detected by immunohistochemistry and was associated with cartilage lesions severity and vascular density. RESULTS PTHrP significantly decreased BSP, confirming its association with chondrocyte hypertrophy. In presence of IL-1β, BSP stimulated IL-8 synthesis, a pro-angiogenic cytokine but decreased the production of TSP-1, an angiogenesis inhibitor. The presence of BSP-immunoreactive chondrocytes in cartilage was associated with the severity of histological cartilage lesions and with vascular density at the osteochondral junction. CONCLUSION This study supports the implication of BSP in the pathology of OA and suggests that it could be a key mediator of the hypertrophic chondrocytes-induced angiogenesis. To control chondrocyte hypertrophic differentiation is promising in the treatment of OA.

Chapiter 10: Identification of mechanosensitive genes in chondrocytes and osteoblasts and their role in OA pathogenesis

Cytokines can be marked out as a new separate regulatory system which, along with nervous and endocrine systems, helps to maintain homeostasis, and these three systems are tightly interconnected and interdependent. Cytokines are the most universal regulatory system, because they can possess both distant biological action after secretion by cell-producer (locally and systemically), and action by intercellular contact as membrane-bound biologically active form. Cytokines include interferons, interleukins, growth and colony-stimulating factors, chemokines, mediators from tumor necrosis factor group, transforming growth factors and some other molecules. Cytokines are not primary mediators of pathology. However cytokines can participate in immunopathologic processes formation and function as diagnostic markers in some diseases. On the other hand the promising perspective of cytokine clinical usage for treatment of widespread diseases, including infectious and cancer, always was the driving force for cytokine study.

Identification of Mechanosensitive Genes in Chondrocytes and Osteoblasts and Their Role in OA Pathogenesis

Osteoarthritis is a pathological condition resulting of the deregulation of the homeostatic mechanisms of the whole joint, triggered by mechanical and biochemical factors. Mechanical factors play a key role in this process by directly breaking the matrix scaffold or by altering the production of matrix components and anabolic/catabolic factors. However, recent studies have shown that only some, but not all genes and regulatory pathways are sensitive to mechanical stimuli. It is also emphasized that cell responses to mechanical stimuli are dependent of their physical characteristics including the magnitude and the frequency. This chapter proposes a review of the responses of chondrocytes and osteoblasts to mechanical stimuli described in the literature recently.

215 STUDY OF THE ASSOCIATION BETWEEN BONE SIALOPROTEIN, HYPERTROPHIC DIFFERENTIATION OF CHONDROCYTES AND CARTILAGE LESIONS IN OSTEOARTHRITIC CARTILAGE

P U R P O S E Chondrocyte hypertrophy is commonly observed in OA cartilage, associated with matrix mineralization and vascularization. In previous work, we suggested a role played by blood supply in the hypertrophic differentiation of osteoathritic chondrocytes. This study aims to investigate the production of Bone SialoProtein (BSP), an angiogenesis enhancer, during hypertrophic differentiation of OA chondrocytes and its expression in cartilage according to the severity of osteoarthritic lesions.