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Adult human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by mitogen-activated protein kinase

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第13期275卷 9645-9652页

作者： Jaiswal, RK Jaiswal, N Bruder, SP Mbalaviele, G Marshak, DR Pittenger, MF Osiris Therapeut Inc Baltimore MD 21231 USA Johns Hopkins Univ Sch Med Baltimore MD 21205 USA

Adult human mesenchymal stem cells are primary, multipotent cells capable of differentiating to osteocytic, chondrocytic, and adipocytic lineages when stimulated under appropriate conditions. To characterize the molecular mechanisms that regulate osteogenic differentiation, we examined the contribution of mitogen-activated protein kinase family members, ERK, JNK, and p38, Treatment of these stem cells with osteogenic supplements resulted in a sustained phase of ERK activation from day 7 to day 11 that coincided with differentiation, before decreasing to basal levels. Activation of JNK occurred much later (day 13 to day 17) in the osteogenic differentiation process. This JNK activation was associated with extracellular matrix synthesis and increased calcium deposition, the two hallmarks of bone formation. Inhibition of ERK activation by PD98059, a specific inhibitor of the ERK signaling pathway, blocked the osteogenic differentiation in a dose-dependent manner, as did transfection with a dominant negative form of MAP kinase kinase (MEK-1), Significantly, the blockage of osteogenic differentiation resulted in the adipogenic differentiation of the stem cells and the expression of adipose-specific mRNAs peroxisome proliferator-activated receptor gamma 2, aP2, and lipoprotein lipase. These observations provide a potential mechanism involving MAP kinase activation in osteogenic differentiation of adult stem cells and suggest that commitment of hMSCs into osteogenic or adipogenic lineages is governed by activation or inhibition of ERK, respectively.

关键词：脂肪组织/细胞学碱基序列骨和骨组织/细胞学细胞分化细胞谱系 DNA引物丝裂原激活蛋白激酶类/拮抗剂和抑制剂丝裂原激活蛋白激酶类/代谢骨生成信号传导干细胞/细胞学成年人人类

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Proliferative response of different human osteoblast-like cell models to proinflammatory cytokines

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PEDIATRIC RESEARCH 2000年第2期48卷 163-168页

作者： Harbour, ME Gregory, JW Jenkins, KR Evans, BAJ Cardiff Univ Dept Child Hlth Cardiff CF14 4XN S Glam Wales

Children with inflammatory bowel disease are known to be at risk of osteopenia. The cause of this osteopenia is likely to be multifactorial, but the inflammatory process with its characteristic overproduction of cytokines has been implicated. To investigate this possible contribution of the disease activity to the development of osteopenia, we performed in vitro assays of the proliferation of osteoblast-like cells of differing origins in response to the inflammatory cytokines tumor necrosis factor-alpha and IL-1 beta. Osteoblast-like cells derived from pediatric bone explants, adherent stromal cells derived from bone marrow (osteoprogenitors), MG-63 osteosarcoma cells, and SV-40 virally transformed osteoprogenitor cells (HCCl) were studied. Tumor necrosis factor-ct stimulated the proliferation of cells in primary cultures (i.e. from explants and marrow samples) in a linear, dose-dependent manner. In contrast, inhibition of proliferation was observed with the established cell lines (MG-63 and HCCl). IL-1 beta stimulated proliferation of all cells apart from the immortalized human bone marrow cell line, HCCl, in which case potent inhibition was observed. We conclude that proinflammatory cytokines are potent regulators of osteoblast-like cell proliferation, and that the responses are specific to cell type. The opposite results obtained with established cell lines compared with the primary cultures suggest that careful consideration should be given to choosing the most suitable cell line for in vitro studies relating to in vivo mechanisms predisposing to osteopenia.

关键词：骨肿瘤骨和骨组织/细胞学细胞分裂/药物作用细胞系细胞系转化细胞培养的白细胞介素1/药理学成骨细胞/细胞学成骨细胞/药物作用骨肉瘤干细胞肿瘤坏死因子α/药理学青少年儿童女(雌)性人类婴儿男(雄)性

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Osteobiology, strain, and microgravity: Part I. Studies at the cellular level

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CALCIFIED TISSUE INTERNATIONAL 2000年第1期67卷 2-9页

作者： Marie, PJ Jones, D Vico, L Zallone, A Hinsenkamp, M Cancedda, R Ctr Biotecnol Avanzate IST I-16132 Genoa Italy Lariboisiere Hosp INSERM U349 Paris France Univ Marburg Dept Expt Orthopaed & Biomech Marburg Germany Fac Med LBBTO INSERM E9901 St Etienne France Univ Bari Sch Med Inst Human Anat I-70121 Bari Italy Free Univ Brussels Erasme Hosp Dept Orthopaed Surg B-1050 Brussels Belgium Univ Genoa Dipartimento Oncol Biol & Genet Genoa Italy

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A fundamental transcription factor for bone and cartilage

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2000年第3期276卷 813-816页

作者： Komori, T Osaka Univ Sch Med Dept Mol Med Suita Osaka 5650871 Japan

Cbfa1-deficient mice were found to show a complete lack of bone formation owing to the maturational arrest of osteoblasts. Cbfa1 plays key roles in the determination of osteoblastic lineage from multipotential mesenchymal cells, their differentiation into mature osteoblasts, and transcriptional regulation of bone matrix-related genes Cbfa1 positively regulates chondrocyte maturation and osteoclast differentiation and is required for vascular invasion into cartilage. Therefore, complete elucidation of the function of Cbfa1 and its signaling would be of great benefit in understanding skeletogenesis. (C) 2000 Academic Press.

关键词：骨和骨组织/细胞学骨和骨组织/胚胎学骨和骨组织/代谢软骨/细胞学软骨/胚胎学软骨/代谢细胞分化细胞谱系软骨细胞/细胞学核心结合因子α1亚基基因表达调控发育期中胚层/细胞学小鼠基因敲除肿瘤蛋白质类成骨细胞/细胞学转录因子/缺乏转录因子/遗传学转录因子/生理学动物小鼠

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Influence of mechanical forces on cells and tissues

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BIORHEOLOGY 2000年第1-2期37卷 3-14页

作者： Stoltz, JF Dumas, D Wang, X Payan, E Mainard, D Paulus, F Maurice, G Netter, P Muller, S Fac Med Vandoeuvre Nancy CNRS UMR 7563 Equipe LEMTA F-54500 Vandoeuvre Les Nancy France Fac Med Vandoeuvre Nancy CNRS UMR 7561 F-54500 Vandoeuvre Les Nancy France

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Possible involvement of the p57^Kip2 gene in bone metabolism

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2000年第2期269卷 422-426页

作者： Urano, T Hosoi, T Shiraki, M Toyoshima, H Ouchi, Y Inoue, S Univ Tokyo Grad Sch Med Dept Geriatr Med Bunkyo Ku Tokyo 1138655 Japan Tokyo Metropolitan Geriatr Hosp Itabashi Ku Tokyo 173 Japan Res Inst & Practice Involut Dis Nagano 39981 Japan Tokyo Metropolitan Inst Med Sci Dept Tumor Biochem Bunkyo Ku Tokyo 1130021 Japan Japan Sci & Technol Corp CREST Yokohama Kanagawa Japan

We previously uncovered that growth stimulation of rat primary osteoblasts by transforming growth factor-beta 1 (TGF-beta 1) resulted in a dramatic decrease in p57(Kip2), a member of cyclin-dependent kinase (CDH) inhibitors, through the proteasomal degradation pathway (Urano et at, J. Biol Chem. 274, 12197-12200, 1999). Here we demonstrated that the amount of p57 protein increases markedly, when rat calvarial primary osteoblasts treated with 1,25-dihydroxyvitamin D3 transit from proliferation toward differentiation. Next, we have analyzed the association of four amino acids deletion polymorphism of p57 and bone mineral density (BMD). The p57 genotype was determined in 154 postmenopausal Japanese women. When we separated the subjects into two groups, one having one or two copies of deletion polymorphism and the other without the deletion, the former subjects had higher BMD (Z score of total body, 0.67 +/- 0.93 vs 0.23 +/- 0.90, mean +/- standard deviation;P = 0.021). Taken together, these findings suggest that the p57 regulated in the osteoblast proliferation and differentiation mag play a role in determination of bone mineral density and pathogenesis of osteoporosis. (C) 2000 Academic Press.

关键词：骨和骨组织/细胞学骨和骨组织/胚胎学骨和骨组织/代谢周期素依赖激酶抑制剂p57 核蛋白质类/遗传学成骨细胞/药物作用成骨细胞/代谢多态现象遗传绝经后期大鼠 Sprague-Dawley 维生素D/药理学老年人动物女(雌)性人类中年人大鼠

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Primary human bone cultures from older patients do not respond at continuum levels of in vivo strain magnitudes

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JOURNAL OF BIOMECHANICS 2000年第1期33卷 63-71页

作者： Stanford, CM Welsch, F Kastner, N Thomas, G Zaharias, R Holtman, K Brand, RA Univ Iowa Coll Dent Dows Inst Dent Res Iowa City IA 52242 USA Univ Iowa Coll Med Dept Orthopaed Surg Iowa City IA 52242 USA Univ Iowa Coll Engn Dept Ind Engn Iowa City IA 52242 USA Graz Univ Dept Orthopaed Surg A-8036 Graz Austria

Osteoporosis is characterized by excessive loss of bone mass, while exercise is believed to maintain or enhance bone mass. Since exercise marginally affects osteoporosis, we wondered whether bone cells from osteoporotic patients would fail to respond to strain. Primary human bone-like cultures were obtained from females over age 60 with hip arthroplasty procedures performed for either osteoporotic fracture (n = 8) or non-osteoporotic osteoarthrosis (n = 5). Cultures (96,000 cell/cm(2)) were strained in rectangular optically clear silastic wells. Three periods of uniaxial substratum strain (1000 mu-strain, 1 Hz, 10,000 cycles, sine wave) were provided every 24 h using a four-point bending, computer-controlled device. Results at a frequency of 1 Hz were compared to cultures exposed to 20 Hz with bone cells derived from one osteoarthritic subject. Alterations in protein level expression of bone-related proteins were determined using a semi-quantitative confocal approach along with enzyme (alkaline phosphatase) activity and enzyme mRNA copy number using cRNA RT-PCR. Strain did not alter levels of bone-related protein levels, enzyme activity, or steady slate copy number per cell in response to strain in either group. Strained cultures from osteoporotic patients exhibited little variation from unstrained controls, while individual cultures from osteoarthritic patients exhibited increases in one protein or the other. The results suggest that bone cells from older individuals may not be responsive to continuum levels of strain anticipated with vigorous activity. (C) 1999 Elsevier Science Ltd. All rights reserved.

关键词：衰老/生理学碱性磷酸酶/遗传学碱性磷酸酶/代谢碱基序列生物力学骨和骨组织/细胞学骨和骨组织/生理学细胞培养的胶原/代谢 DNA引物/遗传学骨关节炎/病理学骨关节炎/病理生理学骨结合素/代谢骨桥蛋白质骨质疏松绝经后/病理学骨质疏松绝经后/病理生理学 RNA 信使/遗传学 RNA 信使/代谢唾液糖蛋白类/代谢应力物理老年人老年人 80以上女(雌)性人类中年人

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The mechanism by which IGFBP-5 exerts anabolic effects in bone

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KIDNEY INTERNATIONAL 2000年第3期57卷 1209-1210页

作者： Jehle, PM Mohan, S Ulm Germany and Loma Linda California USAMain navigationJournal homeAdvance online publicationAbout AOPCurrent issueArchiveABC of Epidemiology ArchiveTranslational Nephrology ArchiveBrowse by CategoryAbout SupplementsFocusesIn the PressOnline submissionFor authorsFor refereesContact editorial officeAbout the journalAbout the ISNFor librariansSubscribeAdvertising / SponsorshipAbout SupplementsReprints and permissionsContact NPGCustomer servicesSite featuresISN resourcesISN home pageChinese editionSpanish editionPortuguese editionContact usSociety newsNPG resourcesKidney International SupplementsKI Japanese home pageNature Reviews NephrologyNature Reviews Urology NPG Journalsby Subject AreaChemistry Chemistry Drug discoveryBiotechnologyMaterialsMethods & Protocols Clinical Practice & Research Cancer Cardiovascular medicine Dentistry Endocrinology Gastroenterology & HepatologyMethods & Protocols Pathology & Pathobiology Urology Earth & Environment Earth sciences Evolution & Ecology Life sciencesBiotechnology Cancer Development Drug discovery Evolution & Ecology Genetics Immunology Medical researchMethods & Protocols Microbiology Molecular cell biology Neuroscience Pharmacology Systems biology Physical sciencesPhysicsMaterials by A - Z IndexExtra navigation.ABSTRACT NAVIGATION - ABSTRACTPrevious | NextTable of contentsFull textDownload PDFSend to a friendRights and permissionsOrder Commercial ReprintsCrossRef lists 1 article citing this articleSave this linkExport citationPapers by JehleADVERTISEMENT TopThis journal is a member of and subscribes to the principles of the Committee on Publication Ethics.Kidney InternationalISSN: 0085-2538EISSN: 1523-1755? 2014 International Society of NephrologyAbout NPGContact NPGAccessibility statementHelpPrivacy policyUse of cookiesLegal noticeTermsNature jobsNature AsiaNature EducationRSS web feedsSearch:partner of AGORA HINARI OARE INASP ORCID CrossRef and COUNTER

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Dietary protein restriction lowers plasma insulin-like growth factor I (IGF-I), impairs cortical bone formation, and induces osteoblastic resistance to IGF-I in adult female rats

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ENDOCRINOLOGY 2000年第9期141卷 3149-3155页

作者： Bourrin, S Ammann, P Bonjour, JP Rizzoli, R Univ Hosp Geneva Dept Internal Med Div Bone Dis WHO Collaborating Ctr Osteoporosis & Bone Dis CH-1211 Geneva 14 Switzerland

Dietary protein deficiency, common in elderly, is associated with decreased areal bone mineral density and plasma insulin-like growth factor I (IGF-I). To investigate the early adaptation of bone cells to protein restriction, B-month-old female rats were pair-fed with isocaloric 15% (control) or 2.5% casein diets for 14 days. Animals were then treated daily with rhIGF-I/IGFBP-3 (1:4, 2.5 mg IGF-I/kg BW) or with vehicle for 10 days. After double-labeling, proximal metaphysis and mid-diaphysis of the tibia were analyzed histomorphometrically. Plasma ostcocalcin, IGF-I, and urinary deoxypyridinoline were quantified. After 14 days of protein restriction, significant drops in plasma osteocalcin (13%) and IGF-I (37%), in periosteal formation (83%) and mineral apposition (49%) rates are observed, indicating a decreased osteoblast recruitment and activity. In cancellous bone, a significant decrease in active eroded surfaces (27%) and osteoclast number (24%) indicates a transient depression of resorption. In rats fed the 15% casein diet, rhIGF-I/IGFBP-3 increases cancellous (42%) and periosteal (600%) formation rates, indicating an increased osteoblast recruitment. In protein-restricted rats, rhIGF-I/IGFBP-3 fails to increase cancellous or periosteal bone formation and plasma osteocalcin is significantly lower than in 15% casein+rhIGF-I/IGFBP-3 rats. Protein restriction induces osteoblast resistance to rhIGF-I/IGFBP-3 in both bone envelopes. Low plasma IGF-I and osteoblast resistance to IGF-I, may contribute to the impaired periosteal formation.

关键词：生物学标记体重/药物作用骨发育/生理学骨质吸收/病理学骨和骨组织/细胞学膳食膳食蛋白质类/投药和剂量抗药性胰岛素样生长因子结合蛋白质3/药理学胰岛素样生长因子Ⅰ/代谢胰岛素样生长因子Ⅰ/药理学器官测量/药物作用成骨细胞/药物作用大鼠 Sprague-Dawley 重组蛋白质类/药理学动物女(雌)性大鼠

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The mechanism of coupling: A role for the vasculature

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BONE 2000年第4期26卷 319-323页

作者： Parfitt, AM Univ Arkansas Med Sci Dept Internal Med Div Endocrinol Little Rock AR 72205 USA Univ Arkansas Med Sci Ctr Osteoporosis & Metab Bone Dis Little Rock AR 72205 USA

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