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Biomedical subjects

G Vaes

Publications and source records attributed to G Vaes.

At least 37 records · Page 2Linked to original sources
PubMed1988

Cellular biology and biochemical mechanism of bone resorption. A review of recent developments on the formation, activation, and mode of action of osteoclasts.

The newest knowledge on the osteoclast allows us to consider bone resorption in a global perspective, as the resultant of three successive steps that may each be individually regulated by physiopathologic or pharmacologic agents. The first involves the formation of osteoclast progenitors in hematopoietic tissues followed by their vascular dissemination and the generation of resting preosteoclasts and osteoclasts in bone. The second consists in the activation of osteoclasts at the contact of mineralized bone. Osteoblasts appear to control this step by exposing the mineral to osteoclasts and preosteoclasts and/or by releasing a soluble factor that activates these cells. In a third step, activated osteoclasts resorb both the mineral and the organic of mineralized bone through the action of agents that they secrete in the segregated zone underlying their ruffled border. The mineral appears to be solubilized by hydrogen ions secreted by an ATP-driven proton pump located at that border and fed by protons generated from CO2 by carbonic anhydrase. The removal of organic matrix, which could be prepared by osteoblast collagenase at the level of nonmineralized bone surfaces, appears dependent on acid proteinases, particularly cysteine-proteinases, secreted, together with other lysosomal enzymes, in the acid microenvironment of the resorption zone.

Animals↗
PubMed1987

The effects of inhibitors of cysteine-proteinases and collagenase on the resorptive activity of isolated osteoclasts.

The effects of specific inhibitors of cysteine-proteinases ((Z-Phe-Ala-CHN2: benzyloxycarbonyl-phenyl-alanyl alanyl diazomethane and E-64: trans-epoxy-succinyl-L-leucylamido(4-guanidino)-butane) and collagenase and collagenase ((Cl-1: N-(3-N-benzyloxycarbonyl amino-1-R-carboxypropyl)-L-leucyl-O-methyl-L-tyrosine N-methylamide) have been tested on the osteoclastic resorption of dentine. Chick osteoclasts were cultured in the presence or absence of 12.5 microM Z-Phe-Ala-CHN2, 40 or 60 microM E-64, or 40 or 100 microM Cl-1 for 1 or 2 days. In addition, osteoclasts were cultured on oyster shell calcitostracum with or without 12.5 microM Z-Phe-Ala-CHN2. Specimens were studied by light microscopy to count cells and resorption features and by scanning electron microscopy (SEM) stereophotogrammetry for the measurement of the depths, plan-areas and volumes of resorption pits. The numbers, depths and volumes (but not the plan-areas) of the resorption pits in dentine were significantly reduced by Z-Phe-Ala-CHN2 and E-64. Thus, for a given plan-area, the volumes and the depths of resorption pits were smaller in these experimental groups compared with control dentine specimens. The overall inhibition of resorption was at least 75%. Cl-1 did not have this inhibitory effect on the numbers or sizes of resorption pits in dentine. When the oyster calcitostracum was used as a substrate for the osteoclasts, Z-Phe-Ala-CHN2 did not reduce the numbers or volumes of pits, but increased the plan-areas and prevented the formation of deeper pits.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Direct extraction and assay of bone tissue collagenase and its relation to parathyroid-hormone-induced bone resorption.

A method has been developed for the quantitative extraction of collagenase from as little as one 19-day-fetal-mouse calvarium. About 20-40 munits of collagenase are extracted per mg of tissue, all in a latent form that, after proper activation, shows the typical properties of mammalian collagenase. Culturing the calvaria for 2 days with parathyroid hormone (PTH) increases their procollagenase content up to 3-fold and induces bone resorption. Both PTH effects are prevented by cycloheximide, but not by indomethacin. Calcitonin inhibits resorption without affecting the PTH-induced procollagenase synthesis. The role of this synthesis is discussed in relation to the mechanisms of bone resorption.

Animals↗
PubMed1986

Inhibition of bone resorption in culture by (+)-catechin.

A pretreatment with (+)-catechin renders embryonic mouse calvaria in culture resistant to the action of bone resorbing agents, either parathyroid hormone (PTH), prostaglandin E2 or retinoic acid, and inhibits in a parallel way the enhanced excretion of N-acetyl-beta-glucosaminidase, a reference lysosomal enzyme, induced by these agents; it has, however, no effect on the small spontaneous leakage of lactate dehydrogenase from the explants. Moreover, the resorption induced in calvaria by a pretreatment with PTH or retinoic acid is inhibited by a further culture with catechin. This inhibition of bone resorption is discussed in relation with the collagen-stabilizing properties of (+)-catechin.

Animals↗
PubMed1985

A new synthetic inhibitor of mammalian tissue collagenase inhibits bone resorption in culture.

A specific and potent synthetic inhibitor of mammalian tissue collagenase and related metallo-proteinases inhibits the collagen matrix resorption induced by parathyroid hormone (PTH) in cultured embryonic mouse calvaria. The inhibition is reversible, dose-dependent and virtually complete at 50 microM inhibitor concentration whereas that due to a less potent stereoisomer is much weaker. The PTH-enhanced secretion of calvarial lysosomal enzymes and the small spontaneous leakage of lactate dehydrogenase are not affected by the inhibitor. These results suggest that collagenase plays a critical role in bone resorption. Its role is discussed in relation to that of cysteine-proteinases that have also been implicated in this process.

Animals↗
PubMed1985

Bisphosphonates and bone resorption: effects on collagenase and lysosomal enzyme excretion.

When added to cultures of parathyroid hormone (PTH)-stimulated bones, dichloromethylenebisphosphonate (C12MBP) and 3-amino-1-hydroxypropydilene-1,1-bisphosphonate (AHPrBP) inhibit completely and in a parallel manner the development of resorption lacunae, the loss of calcium by the explants and their PTH-induced excretion of lysosomal hydrolases (beta-glucuronidase and N-acetyl-beta-glucosaminidase). The loss of collagen (hydroxyproline) by the bones is usually less inhibited than their loss of calcium and their heparin-induced excretion of collagenase is unaffected. To interpret these data, it is proposed that these bisphosphonates act more on the activity of osteoclasts, suppressing simultaneously their excretion of lysosomal enzymes and their erosion of mineralized bone matrix, than on that of other cell types (osteoblasts ?) responsible for collagenase production and the removal of uncalcified collagen.

Acetylglucosaminidase↗
PubMed1985

Partial characterization of the macrophage factor that stimulates fibroblasts to produce collagenase and to degrade collagen.

Rabbit bone marrow-derived macrophages in culture produce and release a soluble factor that activates collagenase secretion and collagen degradation by cultured skin fibroblasts from either rabbit, mouse or human origin. The factor is heat-labile and is inactivated by phenylglyoxal. After gel filtration, it is recovered in both an apparent high-Mr (67000-76000) and a low-Mr (9000-14000) form. Chromatography on cation exchangers suggests two molecular species with different charge properties. These characteristics are compatible with known properties of rabbit interleukin 1.

Animals↗
PubMed1984

In vivo and in vitro evidence for the involvement of cysteine proteinases in bone resorption.

The excretion of cathepsin B, a lysosomal cysteine proteinase, by parathyroid hormone-stimulated embryonic mouse calvaria in culture, correlates closely with the extent of bone resorption evaluated by the loss of hydroxyproline and calcium and by the extension of resorption lacunae. E-64, a specific inhibitor of cysteine proteinases, inhibits reversibly the resorption of cultured bones without affecting the hormone-induced secretion of lysosomal hydrolases. Given in vivo to rats, the proteinase inhibitors, E-64 and leupeptin, both induce a concomitant fall in the serum calcium level and in the urinary excretion of hydroxyproline. These results provide evidence that cysteine proteinases, possibly lysosomal cathepsins, are necessary for bone resorption.

Animals↗
PubMed1984

Degradation of cartilage proteoglycan and collagen by synovial cells. Stimulation by macrophages under activation by phagocytosis, lymphocyte factors, bacterial products or other inflammatory stimuli.

When cultured together with dead 35S-labelled cartilage discs or at the surface of [3H]proteoglycan/[14C]collagen-coated plates, synovial cells from either arthritic or normal rabbit joints digested both the proteoglycan and the collagen of the substrates after a lag-period of 1-2 days. These digestions were inversely related to the age (number of subculture passages) of the synovial cells and they could be modulated by serum components that were either inhibitory or stimulatory. They were dependent on a protein synthesis by the cells and were paralleled, in young cultures, by the release of collagenase and of a proteoglycan-degrading neutral proteinase. The co-culture of synovial cells with macrophages or their culture with macrophage-conditioned culture media caused a more rapid and more extensive degradation of collagen and proteoglycan due to the stimulation of the synovial cells by a nondialysable macrophage factor. The production of this synovial cell-activating 'matrix regulatory monokine' by the macrophage was enhanced by several immunological or inflammatory stimuli such as lymphocyte factors, phagocytosis, asbestos fibres, endotoxin, adjuvant muramyl dipeptide or chemotactic formyl-methionyl peptide, as well as by other membrane-active agents (phorbol myristate acetate, concanavalin A). It is presumed that these interactions are of importance in the development of cartilage destruction in rheumatoid and other chronic inflammatory arthritis.

Animals↗
PubMed1983

Co-operation between metastatic tumor cells and macrophages in the degradation of basement membrane (type IV) collagen.

The co-culture of mouse peritoneal macrophages and Lewis lung carcinoma cells induces the release of a metal-dependent type IV collagen-degrading proteinase which is not produced in detectable amounts by either cell type cultivated alone. Conditioned media of the co-cultures degrade both pepsin-extracted type IV collagen from human placenta and mouse type IV procollagen. Thus macrophages can interact with tumor cells to degrade basement membrane type IV collagen: this might be of importance to allow cancer invasion and metastasis.

Animals↗
PubMed1983

Collagen degradation by metastatic variants of Lewis lung carcinoma: cooperation between tumor cells and macrophages.

Interactions between cancer cells and host macrophages might have important regulatory roles in controlling the expression of the metastatic phenotype, particularly by regulating the production of proteases necessary for tissue invasion. To investigate that possibility, mouse macrophages and Lewis lung carcinoma (LLC) cells from four clonal subpopulations with either low or high metastatic ability were cultured on [14C]collagen (type l)-coated plates. They did not degrade collagen when they were cultured independently on that substrate, but they were induced to do so when macrophages and cancer cells were cultured together. An increased production of neutral collagenase and other neutral protease activities was observed simultaneously. The degree of stimulation of collagen degradation varied according to the cancer cell subpopulation present in the cocultures. For a given LLC cell subpopulation, similar degrees of stimulation of collagen degradation were achieved with either bone marrow-derived or resident peritoneal macrophages, either syngeneic (from C57BL/6 mice) or allogeneic; lower stimulations were obtained with thioglycolate-elicited peritoneal macrophages. Macrophage-conditioned culture media could be substituted for living macrophages to stimulate collagen degradation or collagenase secretion by LLC cells, but LLC cell-conditioned media did not stimulate collagen degradation by macrophages. This suggests that, in the cocultures, collagen degradation is achieved mainly by the cancer cells, not by the macrophages, and that it is induced by a soluble factor, a monokine, produced by the macrophages. That factor might be identical to a recently identified rabbit monokine that stimulates fibroblasts or synovial cells to degrade collagen and proteoglycan and to activate plasminogen, because rabbit macrophage-conditioned media containing that monokine also stimulated collagen degradation by LLC cells.

Animals↗
PubMed1983

Metastatic heterogeneity of cells from Lewis lung carcinoma.

To allow investigations of the role of tumor cell proteases in invasion and metastasis, an attempt was made to obtain well-defined homogeneous populations of Lewis Lung carcinoma cells differing widely in their metastatic potential. From a single Lewis lung carcinoma, a parental line of cells was established and subsequently cloned so as to provide 18 clonal tumor cell lines. These clones differed in their ability to produce spontaneous, macroscopically visible metastases in the lung after i.m. inoculation into syngeneic C57BL/6 mice. Several of them were less metastatic than the parental line. The parental line expressed a metastatic behavior close to that of the high-metastatic cell subpopulations that it contained. There was, within certain limits, a good correlation between the potential for spontaneous lung metastases arising from a primary tumor and that for "artificial" lung colonies obtained after i.v. injection of the Lewis lung carcinoma cells. Although positively correlated with the growth rate of the tumor cells, the metastatic ability of the clones could not be considered as a mere reflection of the proliferation rates of the cells constituting the primary tumors. Differences in metastatic behavior observed among clones persisted in several cases after the cells had been maintained in culture for prolonged periods. However, this stability of the clones in vitro was not absolute. Indeed, some subclones isolated from the low-metastatic clone H122 displayed metastatic abilities which were lower than that of the parent clone. Furthermore, a significant increase in metastatic potential was once observed after a prolonged culture period of that same clone, H122. Thus, new metastatic phenotypes can emerged under in vitro culture conditions. However, the relative rarity of this event suggests that some metastatic heterogeneity already preexisted in vivo among the tumor cells.

Animals↗
PubMed1982

Degradation of collagen and proteoglycan by macrophages and fibroblasts. Individual potentialities of each cell type and cooperative effects through the activation of fibroblasts by macrophages.

Fibroblasts and macrophages of various sources (peritoneal, alveolar or bone marrow-derived), from either rabbit or mouse, were cultured, independently or together, at the surface of [3H]proteoglycan/[14C]collagen-coated plates to evaluate their capacities for proteoglycan and collagen degradation. The various macrophage populations differed widely in their potentialities for proteoglycan and particularly, for collagen degradation, native collagen being significantly degraded, in this model only by rabbit alveolar macrophages. Fibroblasts were as active in proteoglycan degradation as the most active macrophage preparations, but their potential for collagen degradation appeared much higher than that of macrophages. Moreover, all types of macrophages secreted a factor, a monokine, that activated collagen and proteoglycan degradation by fibroblasts. Thus, fibroblasts might well be a major effector cell, active in connective tissue degradations occurring under chronic inflammatory situations.

Animals↗
PubMed1981

A direct simultaneous plate assay of proteoglycan and collagen degradation by cells in culture and its application to synovial cells.

1. A radiochemical plate assay is presented that allows a simultaneous evaluation of the capacity of cells in culture to degrade proteoglycan and collagen. Its principle consists of monitoring the release of soluble radioactive degradation products from Multiwell culture plates coated with dried reconstituted 3H-labelled-proteoglycan/14C-labelled-collagen mixed gels. The plates can also be used for the assay of proteolytic activities within enzyme solutions. 2. When cultured on the plates, rabbit synovial cells degrade collagen and proteoglycan almost simultaneously, owing to the secretion of collagenase and of a proteoglycan-degrading metal-dependent neutral proteinase.

Animals↗