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      An Oligodeoxynucleotide with Promising Modulation Activity for the Proliferation and Activation of Osteoblast

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          Abstract

          The paper explored the regulatory role of oligodeoxynucleotides (ODNs) with specific sequences in the proliferation and activation of osteoblast, using human osteoblast-like cell line MG 63 as the model. Through the administration of ODNs to MG 63 cells at a concentration of 1.0 μg/mL, ODN MT01 with positive effects on proliferation and activation of osteoblast was selected from 11 different ODNs by methyl thiazolyl tetrazolium (MTT) assay and alkaline phosphatase (ALP) activity measurement. To get a deeper insight into the molecular mechanism, effects of ODN MT01 treatment on the expression level of Sp7, runx-2, collagen-I, osteoprotegerin (OPG) and RANK ligand (RANKL) were determined using quantitative real time PCR and Western blotting. Remarkably, the mRNA and protein expression levels of Sp7, runx-2, collagen-I and OPG were improved after ODN MT01 treatment. Meanwhile, the protein expression level of RANKL was dramatically decreased. These results suggested that ODN MT01 had a significant impact in facilitating osteogenic proliferation and activation, and provided a direct evidence for the notion that single strand ODN could regulate the balance of bone formation and resorption, and thus was of great potential in the rebuilding of alveolar bone.

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          Most cited references33

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          The osteoblast: a sophisticated fibroblast under central surveillance.

          The study of the biology of osteoblasts, or bone-forming cells, illustrates how mammalian genetics has profoundly modified our understanding of cell differentiation and physiologic processes. Indeed, genetic-based studies over the past 5 years have revealed how osteoblast differentiation is controlled through growth and transcription factors. Likewise, the recent identification, using mutant mouse models, of a central component in the regulation of bone formation expands our understanding of the control of bone remodeling. This regulatory loop, which involves the hormone leptin, may help to explain the protective effect of obesity on bone mass in humans. In addition, it provides a novel physiologic concept that may shed light on the etiology of osteoporosis and help to identify new therapeutic targets.
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            Lymphoid follicle destruction and immunosuppression after repeated CpG oligodeoxynucleotide administration.

            DNA containing unmethylated cytidyl guanosyl (CpG) sequences, which are underrepresented in mammalian genomes but prevalent in prokaryotes, is endocytosed by cells of the innate immune system, including macrophages, monocytes and dendritic cells, and activates a pathway involving Toll-like receptor-9 (TLR9). CpG-containing oligodeoxynucleotides (CpG-ODN) are potent stimulators of innate immunity, and are currently being tested as adjuvants of antimicrobial, antiallergic, anticancer and antiprion immunotherapy. Little is known, however, about the consequences of repeated CpG-ODN administration, which is advocated for some of these applications. Here we report that daily injection of 60 microg CpG-ODN dramatically alters the morphology and functionality of mouse lymphoid organs. By day 7, lymphoid follicles were poorly defined; follicular dendritic cells (FDC) and germinal center B lymphocytes were suppressed. Accordingly, CpG-ODN treatment for > or =7 d strongly reduced primary humoral immune responses and immunoglobulin class switching. By day 20, mice developed multifocal liver necrosis and hemorrhagic ascites. All untoward effects were strictly dependent on CpG and TLR9, as neither the CpG-ODN treatment of Tlr9(-/-) mice nor the repetitive challenge of wild-type mice with nonstimulatory ODN (AT-ODN) or with the TLR3 agonist polyinosinic:cytidylic acid (polyI:C) were immunotoxic or hepatotoxic.
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              BMP-2-induced Osterix expression is mediated by Dlx5 but is independent of Runx2.

              BMP-2 stimulates the expression of three osteogenic master transcription factors: Runx2, Dlx5, and Osterix (Osx). However, the hierarchical regulatory relationships among them are not yet clearly understood. Osx was commonly stimulated in osteogenic and non-osteogenic cells in response to BMP-signaling, as Dlx5 was in our previous report. A cycloheximide experiment indicated that Osx expression by BMP-2 requires new protein synthesis. Even if Osx has been suggested as a downstream target of Runx2, the results of this study indicated that Osx expression was still induced by BMP-2 treatment in Runx2 null cells, but not induced by Runx2 overexpression in myogenic C2C12 cells. Instead, Osx expression by BMP-2 was completely abrogated by the antisense blocking of Dlx5. Depending upon the coincident expression pattern of Osx and Dlx5, and the blocking of Osx expression by the antisense Dlx5, BMP-2-induced Osx expression is mainly mediated not by Runx2, but by Dlx5.
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                Author and article information

                Journal
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                Molecular Diversity Preservation International (MDPI)
                1422-0067
                15 April 2011
                2011
                : 12
                : 4
                : 2543-2555
                Affiliations
                [1 ]Department of Orthodontics, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mails: fzy00@ 123456sina.com (Z.F.); 33817805@ 123456qq.com (J.W.)
                [2 ]Department of Periodontal, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mail: shenyuqin326@ 123456sina.com
                [3 ]Department of Molecular Biology, College of Basic Medicine, Jilin University, Changchun 130021, China; E-Mail: wlying@ 123456jlu.edu.cn
                [4 ]Department of Oral Cavity, The Second Affiliated Hospital of Shanxi Medical University, 382 Wuyi Road, Taiyuan 030001, China; E-Mail: chenglin6002@ 123456sina.com
                [5 ]Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, 2699 Qianjin Road, Changchun 130021, China; E-Mail: quanshun@ 123456jlu.edu.cn
                Author notes
                [* ]Authors to whom correspondence should be addressed; E-Mails: shiw@ 123456jlu.edu.cn (W.S.); xinhuasun8@ 123456163.com (X.S.); Tel.: +86-431-88796023; Fax: +86-431-88975348.
                Article
                ijms-12-02543
                10.3390/ijms12042543
                3127133
                21731457
                549cfddc-fb5e-4d41-80df-96374783b0b8
                © 2011 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

                History
                : 11 March 2011
                : 6 April 2011
                : 7 April 2011
                Categories
                Article

                Molecular biology
                oligodeoxynucleotide,activation,proliferation,bone remodeling,osteoblast
                Molecular biology
                oligodeoxynucleotide, activation, proliferation, bone remodeling, osteoblast

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