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Resorption Pits (resorption + pit)
Selected AbstractsQuantitative and reliable in vitro method combining scanning electron microscopy and image analysis for the screening of osteotropic modulatorsMICROSCOPY RESEARCH AND TECHNIQUE, Issue 8 2006Gaël Grimandi Abstract The increased generation and up-regulated activity of bone resorbing cells (osteoclasts) play a part in the impairment of bone remodeling in many bone diseases. Numerous drugs (bisphosphonates, calcitonin, selective estrogen receptor modulators) have been proposed to inhibit this increased osteoclastic activity. In this report, we describe a pit resorption assay quantified by scanning electron microscopy coupled with image analysis. Total rabbit bone cells with large numbers of osteoclasts were cultured on dentin slices. The whole surface of the dentin slice was scanned and both the number of resorption pits and the total resorbed surface area were measured. Resorption pits appeared at 48 h and increased gradually up to 96 h. Despite the observation of a strong correlation between the total resorption area and the number of pits, we suggest that area measurement is the most relevant marker for osteoclastic activity. Osteotropic factors stimulating or inhibiting osteoclastic activity were used to test the variations in resorption activity as measured with our method. This reproducible and sensitive quantitative method is a valuable tool for screening for osteoclastic inhibitors and, more generally, for investigating bone modulators. Microsc. Res. Tech., 2006. © 2006 Wiley-Liss, Inc. [source] Glucocorticoids maintain human osteoclasts in the active mode of their resorption cycleJOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2010Kent Søe Abstract Osteoclasts are known to exert their resorptive activity through a so-called resorption cycle consisting of alternating resorption and migration episodes and resulting typically in the formation of increasing numbers of discrete round excavations on bone slices. This study shows that glucocorticoids deeply modify this resorptive behavior. First, glucocorticoids gradually induce excavations with a trenchlike morphology while reducing the time-dependent increase in excavation numbers. This indicates that glucocorticoids make osteoclasts elongate the excavations they initiated rather than migrating to a new resorption site, as in control conditions. Second, the round excavations in control conditions contain undegraded demineralized collagen as repeatedly reported earlier, whereas the excavations with a trenchlike morphology generated under glucocorticoid exposure appear devoid of leftovers of demineralized collagen. This indicates that collagenolysis proceeds generally at a lower rate than demineralization under control conditions, whereas collagenolysis rates are increased up to the level of demineralization rates in the presence of glucocorticoids. Taking these observations together leads to a model where glucocorticoid-induced increased collagenolysis allows continued contact of osteoclasts with mineral, thereby maintaining resorption uninterrupted by migration episodes and generating resorption trenches. In contrast, accumulation of demineralized collagen, as prevails in controls, acts as a negative-feedback loop, switching resorptive activity off and promoting migration to a new resorption site, thereby generating an additional resorption pit. We conclude that glucocorticoids change the osteoclastic resorption mode from intermittent to continuous and speculate that this change may contribute to the early bone fragilization of glucocorticoid-treated patients. © 2010 American Society for Bone and Mineral Research. [source] Indapamide, a Thiazide-Like Diuretic, Decreases Bone Resorption In VitroJOURNAL OF BONE AND MINERAL RESEARCH, Issue 2 2001Agnes Lalande Abstract We recently showed that indapamide (IDP), a thiazide-related diuretic, increases bone mass and decreases bone resorption in spontaneously hypertensive rats supplemented with sodium. In the present study, we evaluated the in vitro effects of this diuretic on bone cells, as well as those of hydrochlorothiazide (HCTZ), the reference thiazide, and acetazolamide (AZ), a carbonic anhydrase (CA) inhibitor. We showed that 10,4 M IDP and 10,4 M AZ, as well as 10,5 M pamidronate (APD), decreased bone resorption in organ cultures and in cocultures of osteoblast-like cells and bone marrow cells in the presence of 10,8 M 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. We investigated the mechanism of this antiresorptive effect of IDP; IDP decreased osteoclast differentiation as the number of osteoclasts developing in coculture of marrow and osteoblast-like cells was decreased markedly. We then investigated whether IDP affected osteoblast-like cells because these cells are involved in the osteoclast differentiation. Indeed, IDP increased osteoblast-like cell proliferation and alkaline phosphatase (ALP) expression. Nevertheless, it did not modify the colony-stimulating factor 1 (CSF-1) production by these cells. In addition, osteoblast-like cells expressed the Na+/Cl, cotransporter that is necessary for the renal action of thiazide diuretics, but IDP inhibited bone resorption in mice lacking this cotransporter, so the inhibition of bone resorption and osteoclast differentiation did not involve this pathway. Thus, we hypothesized that IDP may act directly on cells of the osteoclast lineage. We observed that resorption pits produced by spleen cells cultured in the presence of soluble osteoclast differentiation factor (sODF) and CSF-1 were decreased by 10,4 M IDP as well as 10,5 M APD. In conclusion, in vitro IDP increased osteoblast proliferation and decreased bone resorption at least in part by decreasing osteoclast differentiation via a direct effect on hematopoietic precursors. [source] Deproteinized cancellous bovine bone (Bio-Oss®) as bone substitute for sinus floor elevationJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 3 2003A retrospective, histomorphometrical study of five cases Abstract Objectives: To study in detail the performance of deproteinized cancellous bovine bone (DPBB, Bio-Osso®) granules as a bone substitute, a histomorphometric was performed on five patients treated with DPBB for reconstruction of the severely atrophic maxilla. Material and Methods: DPBB was used as mixture with autogenous bone particles, in concentrations that increased from 20% to 100% DPBB, with the time of healing increasing accordingly from 5 to 8 months. A total of 20 vertical biopsies was taken at the time of fixture installation and used for histomorphometry as undecalcified Goldner stained sections. Results: The results show that in all cases, the DPBB granules had been interconnected by bridges of vital newly formed bone. The volume of bone in the grafted area correlated inversely with the concentration of DPBB grafted, and varied between 37% and 23%. However, the total volume of mineralized material (bone plus DPI3B granules) remained within the same range in all five patients (between 53% and 59%). The high values for osteoid and resorption surface, and the presence of tartrate-resistant acid phosphatase-positive multinucleated osteoclasts in resorption lacunae, indicated that bone remodeling was very active in all grafts. Osteoclasts were also observed in shallow resorption pits on DPBB surfaces. The percentage DPBB surface in contact with bone remained stable at about 35% and could not be related to the proportion of DPBB grafted. Conclusion: Although the number of patients examined was limited, the data suggest that deproteinized cancellous bovine bone, preferably combined with autogenous bone particles, is a suitable material for sinus floor elevation in the severely atrophic human maxilla. Zusammenfassung Ent-Proteinisierter spöngiöser boviner Knochen (Bio-Oss) als Knochenersatz zur Sinusboden-Elevation. Eine retrospektive histomorphometrische Studie an 5 Fällen. Ziele:Um das Verhalten Ent-Proteinisierten spöngiösen bovinen Knochengranulats (DPBB, Bio,Oss) als Knochenersatzmaterial detailliert zu studieren, wurde an 5 Patienten, die mit DPBB zur Rekonstruktion der stark atrophischen Maxilla behandelt wurden, eine histomorphometrische Studie durchgeführt. Material und Methoden:DPBB wurde als Mischung mit autogenen Knochenpartikeln in einer Konzentration von 20 , 100% DPBB verwendet. Die Heilungsdauer wurde entsprechend von 5 auf 8 Monate erhöht. Zum Zeitpunkt der Implantatsetzung wurden insgesamt 20 vertikale Biopsien entnommen und für die Histomorphometrie als nicht-entkalkte, Goldner-gefärbte Schnitte verwendet. Ergebnisse: Die Ergebnisse zeigten in allen Fällen, dass die DPBB-Granula über Brücken von vitalem neu gebildetem Knochen miteinander verbunden waren. Das Volumen des Knochens in dem transplantiertem Gebiet korrelierte umgekehrt mit der Konzentration der transpantierten DPBB und variierte zwischen 37% und 23%. Jedoch lag das Gesamtvolumen des mineralisierten Materials (Knochen+DPBB-Granula)bei allen 5 Patienten im selben Bereich (zwischen 53% und 59%). Die hohen Werte für Ostoid und Resorptionsflächen sowie die Anwesenheit von TRAP-positiven multinukleären Osteoklasten in Resorptionslakunen, zeigte, dass das Knochenremodelling in allen Transplantaten sehr aktiv war. Auch in flachen Resorpionsgrübchen auf der DPBB-Oberfläche wurden Osteoklasten beobachtet. Der Prozentsatz der DPBB-Oberfläche welche im Knochenkontakt war blieb mit etwa 35% stabil und korrelierte nicht mit dem Anteil des transplantierten DPBBs. Schlussfolgerung:Obwohl die untersuchte Patientenzahl sehr gering ist, lassen die Daten vermuten, dass Ent-Proteinisierter spöngiöser boviner Knochen, vorzugsweise mit autogenen Knochenpartikeln kombiniert, ein geeignetes Material für die Sinusboden-Elevation bei stark atrophischer Maxilla ist. Résumé Os bovin spongieux déprotéiné comme substitut osseux dans l'épaississement sinusal. Une étude histomorphométrique rétrospective de cinq cas Le but de cette étude a été d'analyser en détail les performances de l'os bovin spongieux déprotéiné (DPBB, Bio-Oss®) en granules comme substitut osseux par une analyse histomorphométrique effectuée chez cinq patients traités pour une reconstruction au niveau d'un maxillaire sévèrement atrophié. DPBB a été utilisé en mélange avec des particules osseuses autogènes en concentrations augmentaient de 20 à 100% de DPBB, avec un temps de guérison augmentant paralèllement de cinq à huit mois. Vingt biopsies verticales ont ainsi été prélevées au moment du placement des implants et utilisées pour l'histomorphométrie sur coupes colorées Goldner non-décalcifiées. Les résultats ont indiqué que dans tous les cas les granules DPBB ont été interconnectées par des bridges d'os néoformé vivant. Le volume d'os dans l'aire greffée était en corrélation inverse avec les concentrations de DPBB greffées et variait entre 37 et 23 %. Cependant le volume total de matériel minéralisé (os+granules DPBB) restait dans les mêmes moyennes chez les cinq patients (entre 53 et 59 %). Les valeurs importantes pour la surface de résorption et ostéides, et la présence d'ostéoclastes multinucléaires positifs au TRAP dans les lacunes de résorption indiquaient que le remodelage osseux était très actif dans tous les greffons. Les ostéoclastes étaient également observés dans des petites crevasses de résorption étroites sur les surfaces DPBB. Le pourcentage DPBB en contact avec l'os demeurait stable à environ 35% et ne pouvait pas être mis en relation avec la proportion de DPBB greffé. Bien que le nombre de patients examinés aie été très limité, les données suggèrent que l'os bovin spongieux déprotéiné de préférence en combinaison avec des particules d'os autogène est un bon matériel pour l'épaississement du plancher sinusal en présence de maxillaire humain sévèrement atrophié. [source] Quantitative and reliable in vitro method combining scanning electron microscopy and image analysis for the screening of osteotropic modulatorsMICROSCOPY RESEARCH AND TECHNIQUE, Issue 8 2006Gaël Grimandi Abstract The increased generation and up-regulated activity of bone resorbing cells (osteoclasts) play a part in the impairment of bone remodeling in many bone diseases. Numerous drugs (bisphosphonates, calcitonin, selective estrogen receptor modulators) have been proposed to inhibit this increased osteoclastic activity. In this report, we describe a pit resorption assay quantified by scanning electron microscopy coupled with image analysis. Total rabbit bone cells with large numbers of osteoclasts were cultured on dentin slices. The whole surface of the dentin slice was scanned and both the number of resorption pits and the total resorbed surface area were measured. Resorption pits appeared at 48 h and increased gradually up to 96 h. Despite the observation of a strong correlation between the total resorption area and the number of pits, we suggest that area measurement is the most relevant marker for osteoclastic activity. Osteotropic factors stimulating or inhibiting osteoclastic activity were used to test the variations in resorption activity as measured with our method. This reproducible and sensitive quantitative method is a valuable tool for screening for osteoclastic inhibitors and, more generally, for investigating bone modulators. Microsc. Res. Tech., 2006. © 2006 Wiley-Liss, Inc. [source] Development of an ex vivo cellular model of rheumatoid arthritis: Critical role of cd14-positive monocyte/macrophages in the development of pannus tissueARTHRITIS & RHEUMATISM, Issue 9 2007Toshiko Nozaki Objective To establish an ex vivo cellular model of pannus, the aberrant overgrowth of human synovial tissue (ST). Methods Inflammatory cells that infiltrated pannus tissue from patients with rheumatoid arthritis (RA) were collected without enzyme digestion, and designated as ST-derived inflammatory cells. Single-cell suspensions of ST-derived inflammatory cells were cultured in medium alone. Levels of cytokines produced in culture supernatants were measured using enzyme-linked immunosorbent assay kits. ST-derived inflammatory cells were transferred into the joints of immunodeficient mice to explore whether these cells could develop pannus. CD14 and CD2 cells were depleted by negative selection. Results Culture of ST-derived inflammatory cells from 92 of 111 patients with RA resulted in spontaneous reconstruction of inflammatory tissue in vitro within 4 weeks. Ex vivo tissue contained fibroblasts, macrophages, T cells, and tartrate-resistant acid phosphatase,positive multinucleated cells. On calcium phosphate,coated slides, ST-derived inflammatory cell cultures showed numerous resorption pits. ST-derived inflammatory cell cultures continuously produced matrix metalloproteinase 9 and proinflammatory cytokines associated with osteoclastogenesis, such as tumor necrosis factor ,, interleukin-8, and macrophage colony-stimulating factor. More importantly, transferring ST-derived inflammatory cells into the joints of immunodeficient mice resulted in the development of pannus tissue and erosive joint lesions. Both in vitro development and in vivo development of pannus tissue by ST-derived inflammatory cells were inhibited by depleting CD14-positive, but not CD2-positive, cells from ST-derived inflammatory cells. Conclusion These findings suggest that overgrowth of inflammatory cells from human rheumatoid synovium simulates the development of pannus. This may prove informative in the screening of potential antirheumatic drugs. [source] | ||||||||||