Ultrastructural evidence in vitro of osteoclast-induced degradation of calcium phosphate ceramic by simultaneous resorption and phagocytosis mechanisms

Osteoclasts are physiological polykaryons specialized in the resorption of calcified tissue. In the context of the clinical use of calcium-phosphate (Cap) ceramics as bone substitutes, this study used transmission electron microscopy to investigate the in vitro mechanisms of Cap ceramic degradation by osteoclastic cell types. Osteoclasts cultured on Cap ceramic developed typical ultrastructural features of bone osteoclasts, such as a polarized dome shape, a clear zone and a ruffled border. Modification of the shape and density of Cap crystals under the ruffled border indicated an acidic microenvironment. Moreover, osteoclasts were able to degrade ceramic by simultaneous resorption and phagocytosis mechanisms. Phagocytosis did not alter the ability of osteoclasts to resorb Cap ceramic. The phagocytosis mechanism consisted of three steps: crystal phagocytosis, disappearance of the endophagosome envelope membrane and fragmentation of phagocytosed crystals within the cytoplasm. The common mechanism of phagocytosis described here is similar to that observed with the monocyte/macrophage lineage, confirming that osteoclasts are part of the mononuclear phagocyte system. Osteoclasts are thus clearly involved in Cap degradation by means of resorption and phagocytosi

Bone remodelling and tumour grade modifications induced by interactions between bone and swarm rat chondrosarcoma

Chondrosarcoma is currently defined as a malignant cartilage tumour arising de novo or within a pre-existing benign cartilage tumour. Chondrosarcoma can be surgically resected, but all grades have significant rates of local recurrence. The purpose of the present study was to develop an animal intraosseous chondrosarcoma model simulating the progression of human chondrosarcoma and elucidating its behaviour and biology. An intraosseous Swarm rat model was designed to assess interactions between bone and chondrosarcoma. A comparison of tumour grading was carried out according to transplantation site. The effects of chondrosarcoma cells (SRC cells) on the mineralisation capacities of osteoblasts and on osteoclast differentiation were studied in relation to modifications observed in vivo at the cellular level. Transplantation of Swarm rat chondrosarcoma within bone marrow or contiguous to induced periosteal lesions led to extensive bone remodelling with trabecular bone rarefaction and periosteal apposition. Transplantation in close contact to bone but without any periosteal lesion had no effect on bone, suggesting that bone healing factors interact with tumour development. With the intramedullary model, the development of tumours of different grade confirms that bone environment is an important factor in malignancy. A decrease of bone nodule formation was noted after cocultures of SRC cells with rat bone marrow, but there was no modification of osteoclast differentiation after cultures of total rabbit bone cells with SRC cells. These data reveal the importance of interactions between bone environment and tumour in inducing bone remodelling and variations in tumour malignancy

Influence of metal ion solutions on rabbit osteoclast activities in vitro

The purpose of the present study was to compare the effects of various metal ions (aluminium, chromium, cobalt, gold, iron, strontium, titanium and vanadium) on rabbit osteoclast activities, with respect to their number, size, resorptive capacity and their capacity to release proteinases. Marked heterogeneous osteoclastic behaviour was observed early in culture with metal ions (24 h) in term of resorption parameters. In contrast, protease activities (cysteine-proteinase and metalloproteinase activities) were not modulated in our culture conditions. Aluminium, iron, gold and titanium reduced the number of osteoclasts significantly. Aluminium and gold had no effect on osteoclastmediated resorption on dentin-slices, although aluminium induced a greater number of very small lacunae. Titanium reduced only the mean surface area per lacunae, cobalt reduced the mean surface area of lacunae and increased their number, and iron reduced both parameters. Strontium had no effect on osteoclast formation and on total dentin slice surface resorbed. However, strontium increased the number of small lacunae formed on dentin-slices by osteoclasts. Chromium had no effect on osteoclast activities. These findings indicate that metal ions induce very early effects on osteoclasts, which can contribute to periprosthetic pathologies via different cellular mechanisms

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

The PHEMU09 catalogue and astrometric results of the observations of the mutual occultations and eclipses of the Galilean satellites of Jupiter made in 2009

Context. In 2009, the Sun and the Earth passed through the equatorial plane of Jupiter and therefore the orbital planes of its main satellites. It was the equinox on Jupiter. This occurrence made mutual occultations and eclipses between the satellites possible. Experience has shown that the observations of such events provide accurate astrometric data able to bring new information on the dynamics of the Galilean satellites. Observations are made under the form of photometric measurements, but need to be made through the organization of a worldwide observation campaign maximizing the number and the quality of the data obtained. Aims. This work focuses on processing the complete database of photometric observations of the mutual occultations and eclipses of the Galilean satellites of Jupiter made during the international campaign in 2009. The final goal is to derive new accurate astrometric data. Methods. We used an accurate photometric model of mutual events adequate with the accuracy of the observation. Our original method was applied to derive astrometric data from photometric observations of mutual occultations and eclipses of the Galilean satellites of Jupiter. Results. We processed the 457 lightcurves obtained during the international campaign of photometric observations of the Galilean satellites of Jupiter in 2009. Compared with the theory, for successful observations, the r.m.s. of O-C residuals are equal to 45.8 mas and 81.1 mas in right ascension and declination, respectively; the mean O-C residuals are equal to -2 mas and -9 mas in right ascension and declination, respectively, for mutual occultations; and -6 mas and +1 mas in right ascension and declination, respectively, for mutual eclipses. © ESO, 2014