Tailor-made inflammation: how neutrophil serine proteases modulate the inflammatory response

Neutrophil granulocytes are important mediators of innate immunity, but also participate in the pathogenesis of (auto)inflammatory diseases. Neutrophils express a specific set of proteolytic enzymes, the neutrophil serine proteases (NSPs), which are stored in cytoplasmic granules and can be secreted into the extra- and pericellular space upon cellular activation. These NSPs, namely cathepsin G (CG), neutrophil elastase (NE), and proteinase 3 (PR3), have early been implicated in bacterial defense. However, NSPs also regulate the inflammatory response by specifically altering the function of cytokines and chemokines. For instance, PR3 and NE both inactivate the anti-inflammatory mediator progranulin, which may play a role in chronic inflammation. Here, we provide a concise update on NSPs as modulators of inflammation and discuss the biological and pathological significance of this novel function of NSPs. Mounting evidence support an important proinflammatory function for PR3, which may have been underestimated in the past

Potential synergies between matrix proteins and soluble factors on resorption and proteinase activities of rabbit bone cells

Human growth hormone (GH) has recently been found to stimulate osteoclastic resorption, cysteineproteinase and metalloproteinase activities (MMP-2 and MMP-9) in vitro via insulin-like growth factor-1 (IGF-1) produced by stromal cells. The present study investigated the effects of two extracellular matrix components (vitronectin and type-1 collagen) on hGHand hIGF-1-stimulated osteoclastic resorption and proteinase activities in a rabbit bone cell model. After 4 days of rabbit bone cell culture on dentin slices with vitronectin coating, hGH and hIGF-1 stimulated bone resorption and hIGF-1 upmodulated cysteine-proteinase activities. MMP-2 expression (but not resorption, cathepsin or MMP-9 activities) was upmodulated by hGH and hIGF-1 on dentin slices coated with type 1 collagen as compared to those without coating. Then, vitronectin was synergistic with hIGF-1 in the regulation of cysteine-proteinase production whereas collagen showed synergy with hGH and hIGF-1 in the regulation of MMP-2 production. Anti-αvβ3 totally abolished the effects of hGH and hIGF-1 on metalloproteinase release, but had no influence on cathepsin release. The results suggest that cysteine-proteinase modulation is not mediated by αvβ3 integrin (strongly expressed on osteoclastic surface) whereas the resorption process and metalloproteinase modulation are clearly'mediated by this integrin. Our finding about the collagen coating also suggests that hGH- and hIGF-1-stimulated MMP-2 activity are mediated, along with αvβ3 integrin, by another adhesion molecule

IL-6, RANKL, TNF-alpha/IL-1: interrelations in bone resorption pathophysiology.

All osteogenic cells (osteoclasts, osteoblasts) contribute individually to bone remodeling. Their cellular interactions control their cellular activities and the bone remodeling intensity. These interactions can be established either through a cell-cell contact, involving molecules of the integrin family, or by the release of many polypeptidic factors and/or their soluble receptor chains. These factors can act directly on osteogenic cells and their precursors to control differentiation, formation and functions (matrix formation, mineralization, resorption...). Here, we present the involvement of three groups of cytokines which seem to be of particular importance in bone physiology: interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) (TNF-alpha)/IL-1, and the more recently known triad osteoprotegerin (OPG)/receptor activator of NF-kappaB (RANK)/RANK ligand (RANKL). The interactions between these three groups are presented within the framework of bone resorption pathophysiology such as tumor associated osteolysis. The central role of the OPG/RANK/RANKL triad is pointed out

Proteoglycans on bone tumor development

Proteoglycans, extracellular matrix components, exert several activities on bone cells and seem crucial for maintaining an appropriate number of osteoblasts and osteoclasts. The overall data strengthen a pro-bone resorptive role for proteoglycans, through the control of osteoprotegerin availability and of receptor activator of NF-κB ligand bioactivity. In parallel, proteoglycans participate in the control of tumor development at different levels, including bone tumor development and bone metastases dissemination. This dual role makes them good candidates as regulatory molecules in the vicious cycle between tumor proliferation and bone resorption observed during tumor development in bone site. Knowledge of the biological roles of these molecules in cancer biology, tumor angiogenesis and metastasis has promoted the development of drugs targeting them

Proteases and bone remodelling

Bone remodelling is regulated by osteogenic cells which act individually through cellular and molecular interaction. These interactions can be established either through a cell–cell contact, involving molecules of the integrin family, or by the release of many polypeptidic factors and/or their soluble receptor chains. Proteolytic shedding of membrane-associated proteins regulates the physiological activity of numerous proteins. Proteases located on the plasma membrane, either as transmembrane proteins or anchored to cell-surface molecules, serve as activators or inhibitors of different cellular and physiological processes. This review will focus on the role of the proteases implicated in bone remodelling either through the proteolytic degradation of the extracellular matrix or through their relations with osteogenic factors. Their implication in bone tumor progression will be also considered

Effects of a sulfated exopolysaccharide produced by Altermonas infernus on bone biology

The growth and differentiation of bone cells is controlled by various factors which can be modulated by heparan sulphates. Here, we investigated the effects of an oversulphated “heparin-like” exopolysaccharide (OS-EPS) on bone. We compared the effect of this compound with that of a native exopolysaccharide (EPS). Long-term administration of OS-EPS causes cancellous bone loss in mice due, in part, to an increase in the number of osteoclasts lining the trabecular bone surface. No significant difference in cancellous bone volume was found between EPS-treated mice and age-matched control mice, underlying the importance of sulphatation in trabecular bone loss. However, the mechanism sustaining this osteoporosis was unclear. To clarify OS-EPS activities, we investigated the effect of OS-EPS in osteogenesis. Our results demonstrated that OS-EPS inhibited osteoclastogenesis in two cell models. By surface plasmon resonance technique we revealed that OS-EPS can constitute a hetero-molecular complex OS-EPS/RANKL/RANK and that RANK had a higher affinity for RANKL pre-incubated with OS-EPS than for RANKL alone which would be in favour of an increase in bone resorption. However, in vitro, OS-EPS inhibit the early steps of osteoclast precursor adhesion and therefore inhibits the step of cell fusion. In addition, we showed that OS-EPS reduces the proliferation and accelerates osteoblastic differentiation, leading to strong inhibition of mineralized nodule formation, which would be in favour of an increase in bone resorption. Taken together, these data show different levels of bone resorption regulation by exopolysaccharides, most of them leading to proresorptive effects

Effects of a sulfated exopolysaccharide produced by Altermonas infernus on bone biology. Glycobiology 2011, 21, 781–795. Samples Availability: Available from the authors. © 2011 by the authors; licensee MDPI

Abstract: The growth and differentiation of bone cells is controlled by various factors, which can be modulated by heparan sulfates. Here, we investigated the effects of an oversulfated exopolysaccharide (OS-EPS) on the bone. We compared the effect of this compound with that of a native EPS. Long-term administration of OS-EPS causes cancellous bone loss in mice due, in part, to an increase in the number of osteoclasts lining the trabecular bone surface. No significant difference in cancellous bone volume was found between EPS-treated mice and age-matched control mice, underlying the importance of sulfation in trabecular bone loss. However, the mechanism sustaining this osteoporosis was unclear. To clarify OS-EPS activities, we investigated the effect of OS-EPS on osteogenesis. Our results demonstrated that OS-EPS inhibited osteoclastogenesis in two cell models. Using the surface plasmon resonance technique, we revealed that OS-EPS can form a hetero-molecular complex OS-EPS/receptor activator of NF-κB ligand (RANKL)/RANK and that RANK had a higher affinity for RANKL pre-incubated with OS-EPS than for RANKL alone, which would be in favor of an increase in bone resorption. However, in vitro, OS-EPS inhibited the early steps of osteoclast precursor adhesion and therefore inhibited the cell fusion step. In addition, we showed that OS-EPS reduced proliferation and accelerated osteoblastic differentiation, leading to strong inhibition of mineralized nodule formation, which would be in favor of an increase in bone resorption. Taken together, these data show different levels of bone resorption regulation by EPSs, most of them leading to proresorptive effects

Potential synergies between matrix proteins and soluble factors on resorption and proteinase activities of rabbit bone cells

Human growth hormone (GH) has recently been found to stimulate osteoclastic resorption, cysteineproteinase and metalloproteinase activities (MMP-2 and MMP-9) in vitro via insulin-like growth factor-1 (IGF-1) produced by stromal cells. The present study investigated the effects of two extracellular matrix components (vitronectin and type-1 collagen) on hGHand hIGF-1-stimulated osteoclastic resorption and proteinase activities in a rabbit bone cell model. After 4 days of rabbit bone cell culture on dentin slices with vitronectin coating, hGH and hIGF-1 stimulated bone resorption and hIGF-1 upmodulated cysteine-proteinase activities. MMP-2 expression (but not resorption, cathepsin or MMP-9 activities) was upmodulated by hGH and hIGF-1 on dentin slices coated with type 1 collagen as compared to those without coating. Then, vitronectin was synergistic with hIGF-1 in the regulation of cysteine-proteinase production whereas collagen showed synergy with hGH and hIGF-1 in the regulation of MMP-2 production. Anti-avB3 totally abolished the effects of hGH and hIGF-1 on metalloproteinase release, but had no influence on cathepsin release. The results suggest that cysteine-proteinase modulation is not mediated by avB3 integrin (strongly expressed on osteoclastic surface) whereas the resorption process and metalloproteinase modulation are clearly'mediated by this integrin. Our finding about the collagen coating also suggests that hGH- and hIGF-1-stimulated MMP-2 activity are mediated, along with avB3 integrin, by another adhesion molecule