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In vitro and in vivo evidence for orphan nuclear receptor RORα function in bone metabolism

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第16期97卷 9197-+页

作者： Meyer, T Kneissel, M Mariani, J Fournier, B Novartis Pharma AG Therapeut Area Arthrit & Bone Metab CH-4002 Basel Switzerland Univ Paris 06 Inst Neurosci Lab Dev & Vieillissement Syst Nerveux F-75005 Paris 5 France

Bone is a major target site for steroid hormone action. Steroid hormones like cortisol, vitamin D, and estradiol are responsible for principal events associated with bone formation and resorption. Over the past decade, new members of the nuclear hormone gene family have been identified that lack known ligands. These orphan receptors can be used to uncover signaling molecules that regulate yet unidentified physiological networks. In the present study the function of retinoic acid receptor-related orphan receptor (ROR) alpha in bone metabolism has been examined. We showed that ROR alpha and ROR gamma, but not ROR beta, are expressed in mesenchymal stem cells derived from bone marrow. Interestingly, for ROR alpha We observed an increased messenger signal expression between control cells and cells undergoing osteogenic differentiation. Furthermore, the direct activation of mouse bone sialoprotein by ROR alpha, typically 7-fold, has been shown. In contrast, transient overexpression of RORa! overrides the activation of the osteocalcin promoter by 1 alpha,25-dihydroxyvitamin D-3. In addition, we have investigated bone mass parameters and bone geometry in the mouse mutant staggerer (sg/sg), a mouse strain that carries a deletion within the ROR alpha gene. Homozygote mutants have thin long bones compared with the heterozygote animals and wild-type littermates. More interestingly, the bones of the sg/sg animals are osteopenic as indicated by the comparison of bone mineral contents of sg/sg animals to the heterozygote and wild-type animals. We conclude that these in vitro and in vivo results suggest a function for ROR alpha in bone biology. ROR alpha most likely acts by direct modulation of a bone matrix component.

关键词：吸收测定法光子碱基序列骨发育骨和骨组织/代谢细胞培养的 DNA引物小鼠近交C57BL 受体胞质和核/生理学动物人类小鼠

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DEFECTIVE STIMULATION OF PROLIFERATION AND COLLAGEN BIOSYNTHESIS OF HUMAN BONE-CELLS BY SERUM FROM DIABETIC-PATIENTS

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ACTA ENDOCRINOLOGICA 1992年第6期127卷 509-514页

作者： BRENNER, RE RIEMENSCHNEIDER, B BLUM, W MORIKE, M TELLER, WM PIRSIG, W HEINZE, E UNIV ULM DEPT OTORHINOLARYNGOLW-7900 ULMGERMANY UNIV TUBINGEN CHILDRENS HOSPW-7400 TUBINGEN 1GERMANY

We studied the influence of fasting serum from nine insulin-dependent diabetic children and adolescents under insufficient metabolic control on normal human bone cells in vitro compared with serum from eight sex- and age-matched controls. Cell number 24 h after plating was significantly less under diabetic serum, indicating impaired cell attachment, spreading and initiation of cell proliferation. Cell number after five days was reduced by 1% diabetic serum, while higher serum concentrations had diverging effects on osteoblast proliferation. Collagen synthesis of human osteoblasts was significantly reduced by 8% diabetic serum compared to 8% control serum, while synthesis of non-collagenous proteins was not affected. Duration of diabetes (several weeks up to 12 years) had no influence on these parameters. The serum from one patient, which was studied a second time under excellent metabolic control three months later, however, had lost its inhibitory influence on collagen synthesis of osteoblasts. The pattern of the interstitial collagen types I, III and V was not altered by diabetic serum. These results indicate that defective regulation of proliferation and collagen synthesis of osteoblasts by components present in human diabetic serum may be an important factor in the development of diabetic osteopenia. The negative influence might be explained in part by reduced levels of IGF-I and elevated levels of IGF binding protein-I in the diabetic sera.

关键词：血蛋白质类/药理学血蛋白质类/生理学骨疾病代谢性/病因学骨和骨组织/细胞学骨和骨组织/代谢载体蛋白质类/血液细胞分裂细胞培养的胶原/生物合成糖尿病 1型/血液糖尿病 1型/代谢糖尿病 1型/病理生理学胰岛素样生长因子结合蛋白质类胰岛素样生长因子Ⅰ/分析胰岛素样生长因子Ⅱ/分析成骨细胞/细胞学成骨细胞/代谢放射免疫测定青少年成年人儿童儿童学龄前女(雌)性人类男(雄)性

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EFFECT OF EXOGENOUS LIPID ON LIPID-SYNTHESIS BY BONE AND BONE CELL-CULTURES

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JOURNAL OF DENTAL RESEARCH 1975年第1期54卷 131-139页

作者： SCHUSTER, GS DIRKSEN, TR HARMS, WS MED COLL GEORGIA SCH DENTDEPT ORAL BIOLAUGUSTAGA 30902 MED COLL GEORGIA SCH MEDDEPT CELL & MOLEC BIOLAUGUSTAGA

Newborn rat calvaria and isolated calvaria cells are capable of de novo lipid synthesis when grown in the presence or absence of exogenous lipid sources. Synthesis decreases when exogenous lipids are supplied. Several cholesterol precursors were found in these tissues and the presence of dihydrocholesterol was established for the first time.

关键词：乙酸盐类/代谢动物新生骨和骨组织/代谢二氧化碳/代谢细胞培养的胆固醇/代谢色谱法薄层脂肪酸类/药理学脂肪酸类非酯化/药理学脂类/分析脂类/生物合成颅骨/分析颅骨/代谢动物大鼠

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In vitro release of vancomycin and tobramycin from impregnated human and bovine bone grafts

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JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY 2000年第3期46卷 423-428页

作者： Winkler, H Janata, O Berger, C Wein, W Georgopoulos, A Danube Hosp Dept Orthopaed A-1220 Vienna Austria Danube Hosp Cent Bone Bank Vienna A-1220 Vienna Austria Danube Hosp Dept Infect Control A-1220 Vienna Austria Univ Vienna Dept Clin Pharmacol A-1090 Vienna Austria Univ Vienna Clin Dept Infect & Chemotherapy A-1090 Vienna Austria

In order to combine the effects of bone repair and eradication of infection, with both Gram-positive and Gram-negative pathogens, the behaviour of a compound of bone graft and antibiotics was investigated. Samples of human and bovine bone, cancellous and cortical, were processed and incubated with vancomycin and tobramycin, respectively. The compound was placed in 5% human albumin and the surrounding liquid was exchanged completely every 24 h. Concentrations of antibiotics in the fluid were measured over less than or equal to 28 days using high pressure liquid chromatography and a bioassay. All tested combinations eluted mainly in the initial phase with a logarithmic decrease over the testing period. The concentration of antibiotics in the albumin was well above the MIC for common pathogens throughout the investigation in all tested specimens. The highest initial concentrations were measured in the compound of bovine bone together with vancomycin (24395.8 +/- 1138.9 mg/L), decreasing to 9.02 +/- 1.3 mg/L after 11 exchanges. Human and bovine bone did not have significantly different properties. The storage capability of cortical bone was generally lower than that of cancellous bone. Tobramycin concentrations were significantly lower in the initial phase;however, it eluted move steadily and over a longer period, so that from day 6 onwards, its concentration was greater than that of vancomycin. After 28 days, the tobramycin concentration was 18.09 +/- 2.46 mg/L (bovine cancellous bone). In conclusion, bone, if processed adequately, is an excellent carrier for vancomycin and tobramycin. Cortical bone is as suitable as cancellous bone. The pharmacokinetics of human and bovine bone are comparable. Using an antibiotic-graft compound, eradication of pathogens and grafting of bony defects may be accomplished in a one-stage procedure.

关键词：抗菌药/投药和剂量抗菌药/药代动力学骨移植骨和骨组织/药物作用骨和骨组织/代谢冷冻干燥法妥布拉霉素/投药和剂量妥布拉霉素/药代动力学万古霉素/投药和剂量万古霉素/药代动力学动物牛人类

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Osteomalacia due to vitamin D depletion: A neglected consequence of intestinal malabsorption

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AMERICAN JOURNAL OF MEDICINE 2000年第4期108卷 296-300页

作者： Basha, B Rao, DS Han, ZH Parfitt, AM Henry Ford Hlth Syst Div Bone & Mineral Bone & Mineral Metab Res Lab Ctr Bone & Joint Detroit MI 48202 USA Univ Arkansas Med Sci Div Endocrinol Little Rock AR 72205 USA

PURPOSE: Osteomalacia due to vitamin D depletion is believed to be rare in the United States because of the routine fortification of milk and other dairy products with vitamin D. We present a series of patients with histologically verified osteomalacia due to vitamin D depletion to emphasize the need for more careful and systematic surveillance of patients at risk of this metabolic bone disease. METHODS: Between 1989 and 1994, 17 patients with osteomalacia due to vitamin D depletion were seen in the Bone and Mineral Division of Henry Ford Health System, Detroit. All patients had a transiliac bone biopsy after in vivo double tetracycline labeling. Biochemical indexes of vitamin D nutritional status, parathyroid function, markers of bone turnover, and bone mineral density were assessed at the time of bone biopsy. The duration of symptoms, the lag between the cause of vitamin D depletion and the development of symptoms, and the radiologic findings were recorded. RESULTS: Osteomalacia was suspected by the referring physician in only 4 of the 17 patients, although a gastrointestinal disorder that can lend tc, vitamin D depletion was present in every patient. Thirteen of the patients had sustained at least one osteoporotic fracture (wrist, spine, or hip), and most had low appendicular and axial bone mineral density. All patients had one or more biochemical abnormalities consistent with vitamin D depletion. In 4 patients, a progressive rise in the serum alkaline phosphatase level was recorded but was not investigated until the patient presented with bone pain, muscle weakness, or fracture. CONCLUSIONS: Osteomalacia due to vitamin D depletion appears not to be suspected or diagnosed promptly in susceptible patients, perhaps because their physicians were not sufficiently aware of this condition. Am J Med. 2000;108:296 -300. (C) 2000 by Excerpta Medica, Inc.

关键词：碱性磷酸酶/血液骨密度骨和骨组织/代谢骨和骨组织/病理学钙/血液胃肠疾病/并发症胃肠疾病/外科学吸收不良综合征/并发症骨软化症/诊断骨软化症/病因学骨软化症/病理生理学绝经后期维生素D缺乏/血液维生素D缺乏/并发症成年人老年人女(雌)性人类男(雄)性中年人

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Short- and long-term effects of ibandronate treatment on bone turnover in Paget disease of bone

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CLINICAL CHEMISTRY 2000年第5期46卷 684-690页

作者： Woitge, HW Oberwittler, H Heichel, S Grauer, A Ziegler, R Seibel, MJ Heidelberg Univ Dept Internal Med Div Endocrinol & Metab D-69115 Heidelberg Germany

Background: In Paget disease of bone (PD), serum total alkaline phosphatase (TAP) is a valid marker of disease activity. The aim of the present longitudinal study was to compare TAP with new and potentially more specific markers of bone turnover in bisphosphonate-treated patients with PD. Methods: Twenty patients with active PD were studied before and after treatment with 2 mg of intravenous ibandronate over a period of 12 months. TAP (by colorimetry), serum bone-specific alkaline phosphatase (BAP;by enzyme immunoassay), serum osteocalcin (OC;by ELISA), serum bone sialoprotein (BSP;by RIA), and urinary total pyridinoline (PYD;by HPLC) and deoxypyridinoline (DPD;by HPLC) were measured as markers of bone turnover. Results: Before treatment, TAP, BAP, and BSP were increased in all 20 patients, whereas OC was increased in 10, PYD in 13, and DPD in 15 patients. Three months post treatment, nine patients showed normalized TAP values, and a greater than or equal to 25% re-increase (i.e., relapse) was observed in all patients after 12 months. A normalization of BAP was achieved in six patients only. No significant changes were found for OC. BSP was decreased significantly at 24 h, and DPD at 48 h post treatment. A normalization of BSP was found in 8, of PYD in 18, and of DPD in 16 cases. Both PYD and DPD increased significantly from 9 months post treatment onward. Conclusions: Most markers of bone turnover show similar long-term changes after treatment of active PD with ibandronate. With regard to cost-effectiveness and assay performance, TAP remains the marker of choice in therapeutic monitoring of PD. However, more specific markers may improve the biochemical assessment of PD in certain situations. (C) 2000 American Association for Clinical. Chemistry.

关键词：碱性磷酸酶/血液氨基酸类/血液骨和骨组织/酶学骨和骨组织/代谢色谱法高压液相二膦酸盐类/治疗应用酶联免疫吸附测定整合素结合涎蛋白变形性骨炎/药物疗法变形性骨炎/酶学变形性骨炎/代谢骨钙素/血液前瞻性研究放射免疫测定唾液糖蛋白类/血液老年人女(雌)性人类男(雄)性中年人

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Dietary supplementation of arachidonic acid is associated with higher whole body weight and bone mineral density in growing pigs

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PEDIATRIC RESEARCH 2000年第5期47卷 692-697页

作者： Weiler, HA Univ Manitoba Dept Food & Nutr Winnipeg MB R3T 2N2 Canada

Essential fatty acids are fundamental to normal growth and development, but North American formulas do not contain arachidonic (AA) and docosahexaenoic acid (DHA). The main objective of the present study was to determine whether addition of AA and DHA to formula elevates growth and bone mineralization in piglets. A secondary objective was to establish whether liver fatty acid composition is related to that of bone. Twelve 10-d-old male piglets were randomized to receive either a standard formula with an n-6:n-3 fatty acid ratio of 4.9:1.0 or the same formula made with an equal amount of fat but containing AA (0.5% wt/wt total fat) and DHA (0.1% wt/wt total fat) for 14 d. Piglets in the supplemented group had significantly (p < 0.05) higher weight and greater bone mineral density of the whole body, lumbar spine, and femur. No differences were observed in whole body length, calcium absorption, or biochemical markers of bone metabolism. Feeding AA resulted in lower linoleic acid (p < 0.05) and higher (p < 0.05) AA in liver total lipid (% wt/wt) and bone FFA (% wt/wt) but no change to DHA. Liver AA (% wt/wt total lipid) was positively related (p < 0.05) to growth, free AA (% wt/wt) in bone, bone mineral content, bone mineral density, and urinary prostaglandin E-2 but negatively related (p < 0.05) to free linoleic acid in bone. Inverse relationships were observed when liver linoleic acid was substituted for liver AA as the independent variable. These data indicate that feeding AA is associated with elevated weight and higher whole body and regional bone mineral density.

关键词：动物吮乳期花生四烯酸/投药和剂量身高/药物作用身高/生理学体重/药物作用骨密度/药物作用骨和骨组织/药物作用骨和骨组织/代谢胶原/血液胶原Ⅰ型营养保健品脂肪酸类/代谢肠/生理学肝/药物作用肝/代谢器官测量/生理学骨钙素/血液肽碎片/血液肽类/血液磷脂类/代谢前胶原/血液猪动物男(雄)性

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STUDIES ON REGULATION OF INSULIN-LIKE GROWTH-FACTOR BINDING-PROTEIN (IGFBP)-3 AND IGFBP-4 PRODUCTION IN HUMAN BONE-CELLS

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ACTA ENDOCRINOLOGICA 1992年第6期127卷 555-564页

作者： MOHAN, S STRONG, DD LEMPERT, UG TREMOLLIERES, F WERGEDAL, JE BAYLINK, DJ LOMA LINDA UNIV DEPT MEDLOMA LINDACA 92350 LOMA LINDA UNIV DEPT BIOCHEMLOMA LINDACA 92350 LOMA LINDA UNIV DEPT MICROBIOLLOMA LINDACA 92350 LOMA LINDA UNIV DEPT PHYSIOLLOMA LINDACA 92350

Previous studies have shown that the actions of IGF-II in bone are determined not only by its concentration, but also by the concentration of IGFBP-4 as well as other IGFBPs. In this study, we sought to determine by Western ligand blotting the effects of growth hormone, IGF-I and IGF-II on the production of IGFBP-3 and IGFBP-4 in TE89 human osteosarcoma cells and in untransformed normal human bone cells derived from rib. Human growth hormone at 10 mug/l decreased the amount of IGFBP-4 but had no effect on the IGFBP-3 level in the conditioned medium of low density cultures of TE89 cells and human bone cells derived from rib. Human growth hormone had no effect on IGFBP-3 or IGFBP-4 levels in the conditioned medium of high density human bone cell cultures. IGF-I and IGF-II, which increased human bone cell proliferation, decreased the level of IGFBP-4 (30% of control at 100 mug/IIGF-I and IGF-II) but increased the level of IGFBP-3 (3-10 fold at 100 mug/l IGF-I and IGF-II) after 48 h of treatment in the conditioned medium of both low and high density TE89 cell cultures. Similar changes in IGFBP-3 and IGFBP-4 levels were also seen in the conditioned medium of human bone cells derived from rib after treatment with IGF-I and IGF-II. Studies to determine the underlying molecular mechanisms by which IGF-II decreased the amount of IGFBP-4 in the conditioned medium revealed that IGF-II decreased the IGFBP-4 mRNA abundance and increased the IGFBP-3 mRNA abundance in human bone cells. Based on the above findings, we conclude that the production of both IGFBP-3 and IGFBP-4 is regulated in bone cells and that local and systemic agents may modulate the responsiveness of bone cells to IGFs by regulated secretion of IGFBP-3 and IGFBP-4.

关键词：碱基序列印迹法 RNA 印迹法蛋白质骨肿瘤/代谢骨肿瘤/病理学骨和骨组织/细胞学骨和骨组织/代谢骨和骨组织/超微结构载体蛋白质类/分析载体蛋白质类/遗传学载体蛋白质类/代谢细胞分裂细胞膜/化学细胞膜/超微结构细胞培养的培养基条件性/药理学基因表达调控生长激素/药理学胰岛素样生长因子结合蛋白质4 胰岛素样生长因子结合蛋白质类胰岛素样生长因子Ⅰ/药理学胰岛素样生长因子Ⅱ/药理学分子序列数据骨肉瘤/代谢骨肉瘤/病理学 RNA 信使/分析 RNA 信使/遗传学肿瘤细胞培养的人类

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Measurement of osteocalcin

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ANNALS OF CLINICAL BIOCHEMISTRY 2000年第4期37卷 432-446页

作者： Lee, AJ Hodges, S Eastell, R Univ Sheffield No Gen Hosp Div Clin Sci NGHT Med SectBone MEtab Grp Sheffield S5 7AU S Yorkshire England

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Effect of elcatonin on osteoinduction by recombinant human bone morphogenetic protein-2

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2000年第2期269卷 317-321页

作者： Okubo, Y Bessho, K Fujimura, K Kusumoto, K Ogawa, Y Iizuka, T Kyoto Univ Grad Sch Med Dept Oral & Maxillofacial Surg Sakyo Ku Kyoto 6068507 Japan Kansai Med Univ Dept Plast & Reconstruct Surg Moriguchi Osaka 5708507 Japan

To evaluate the effect of elcatonin on osteoinduction by recombinant human bone morphogenetic protein-2 (rhBMP-2), 5 mu g of rhBMP-2 was implanted into intramuscular sites of rats. For 14 days after the implantation, elcatonin was administered intraperitoneally with total dosage of 80 U, 8 U, and 0.8 U, respectively. For the control group, only physiological saline was administered. At 21 days after implantation, the area of the oval shadow in the radiologic findings depended on the elcatonin dose and the amount of trabecular bone and the number of osteoblasts observed in the histologic findings depended on the dosage of elcatonin, The values of ALP activity and Ca content also showed an elcatonin dose dependency. These results suggested that elcatonin is effective in enhancing osteoinduction by rhBMP-2 within the dose range of this study, and that elcatonin has an anabolic effect on osteoblasts in addition to an antiresorptive effect. (C) 2000 Academic Press.

关键词：碱性磷酸酶/代谢骨形态发生蛋白质类/药理学骨重建/药物作用骨和骨组织/解剖学和组织学骨和骨组织/酶学骨和骨组织/代谢骨和骨组织/放射摄影术降钙素/类似物和衍生物降钙素/药理学钙/代谢大鼠 Wistar 重组蛋白质类/药理学动物人类男(雄)性大鼠

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