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Sample records for bone morphogenetic protein

  1. Bone morphogenetic proteins: Periodontal regeneration

    Directory of Open Access Journals (Sweden)

    Subramaniam M Rao

    2013-01-01

    Full Text Available Periodontitis is an infectious inflammatory disease that results in attachment loss and bone loss. Regeneration of the periodontal tissues entails de novo formation of cementum, periodontal ligament, and alveolar bone. Several different approaches are currently being explored to achieve complete, reliable, and reproducible regeneration of periodontal tissues. The therapeutic management of new bone formation is one of the key issues in successful periodontal regeneration. Bone morphogenetic proteins form a unique group of proteins within the transforming growth factor superfamily of genes and have a vital role in the regulation in the bone induction and maintenance. The activity of bone morphogenetic proteins was first identified in the 1960s, but the proteins responsible for bone induction were unknown until the purification and cloning of human bone morphogenetic proteins in the 1980s, because of their osteoinductive potential. Bone morphogenetic proteins have gained a lot of interest as therapeutic agents for treating periodontal defects. A systematic search for data related to the use of bone morphogenetic proteins for the regeneration of periodontal defects was performed to recognize studies on animals and human (PUBMED, MEDLINE, COCHRANE, and Google search. All the studies included showed noticeable regeneration of periodontal tissues with the use of BMP.

  2. The classic: Bone morphogenetic protein.

    Science.gov (United States)

    Urist, Marshall R; Strates, Basil S

    2009-12-01

    This Classic Article is a reprint of the original work by Marshall R. Urist and Basil S. Strates, Bone Morphogenetic Protein. An accompanying biographical sketch of Marshall R. Urist, MD is available at DOI 10.1007/s11999-009-1067-4; a second Classic Article is available at DOI 10.1007/s11999-009-1069-2; and a third Classic Article is available at DOI 10.1007/s11999-009-1070-9. The Classic Article is copyright 1971 by Sage Publications Inc. Journals and is reprinted with permission from Urist MR, Strates BS. Bone morphogenetic protein. J Dent Res. 1971;50:1392-1406.

  3. Cross-talk between bone morphogenetic proteins and inflammatory pathways.

    Science.gov (United States)

    van der Kraan, Peter M; Davidson, Esmeralda N Blaney

    2015-11-23

    Pro-inflammatory cytokines and bone morphogenetic proteins are generally studied separately and considered to be elements of different worlds, immunology and developmental biology. Varas and colleagues report that these factors show cross-talk in rheumatoid arthritis synoviocytes. They show that pro-inflammatory cytokines not only stimulate the production of bone morphogenetic proteins but that these endogenously produced bone morphogenetic proteins interfere with the effects of pro-inflammatory cytokines on synoviocytes.

  4. Multifunctional Bone Morphogenetic Protein System in Endocrinology

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    Otsuka,Fumio

    2013-04-01

    Full Text Available New biological activities of bone morphogenetic proteins (BMPs in the endocrine system have recently been revealed. The BMP system is composed of approximately 30 ligands and preferential combinations of type I and type II receptors. The BMP system not only induces bone formation but also plays unique tissue-specific roles in various organs. For instance, the ovarian BMP system is a physiological inhibitor of luteinization in growing ovarian follicles. In the ovary, the expression of oocyte-derived BMP-15 is critical for female reproduction. In the pituitary, BMP-4 is a key player for initial development of the anterior pituitary, while it is also functionally involved in some differentiated pituitary tumors, including prolactinoma and Cushingʼs disease. In the adrenal glands, BMP-6 and BMP-4 modulate aldosterone and catecholamine production, respectively, which contributes to a functional interaction between the cortex and medulla. In the present review, recent advances in BMP biology in the field of endocrinology are described and the possibility for clinical application of BMP activity is discussed.

  5. Nuclear variants of bone morphogenetic proteins

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    Meinhart Christopher A

    2010-03-01

    Full Text Available Abstract Background Bone morphogenetic proteins (BMPs contribute to many different aspects of development including mesoderm formation, heart development, neurogenesis, skeletal development, and axis formation. They have previously been recognized only as secreted growth factors, but the present study detected Bmp2, Bmp4, and Gdf5/CDMP1 in the nuclei of cultured cells using immunocytochemistry and immunoblotting of nuclear extracts. Results In all three proteins, a bipartite nuclear localization signal (NLS was found to overlap the site at which the proproteins are cleaved to release the mature growth factors from the propeptides. Mutational analyses indicated that the nuclear variants of these three proteins are produced by initiating translation from downstream alternative start codons. The resulting proteins lack N-terminal signal peptides and are therefore translated in the cytoplasm rather than the endoplasmic reticulum, thus avoiding proteolytic processing in the secretory pathway. Instead, the uncleaved proteins (designated nBmp2, nBmp4, and nGdf5 containing the intact NLSs are translocated to the nucleus. Immunostaining of endogenous nBmp2 in cultured cells demonstrated that the amount of nBmp2 as well as its nuclear/cytoplasmic distribution differs between cells that are in M-phase versus other phases of the cell cycle. Conclusions The observation that nBmp2 localization varies throughout the cell cycle, as well as the conservation of a nuclear localization mechanism among three different BMP family members, suggests that these novel nuclear variants of BMP family proteins play an important functional role in the cell.

  6. Evaluation of heterotopic bone formation induced by squalane and bone morphogenetic protein composite.

    Science.gov (United States)

    Kawakami, T; Kawai, T; Takei, N; Kise, T; Eda, S; Urist, M R

    1997-04-01

    Bone morphogenetic protein is an important molecule whose bioactivity depends on the carrier. Squalane is used in the formulation of various kinds of cosmetics because it is easily emulsified and has the property of spreading well. Thus, squalane might be effective as a bone morphogenetic protein delivery system. As a test for this possibility, gelatin capsules containing squalane and bone morphogenetic protein (bovine derived partially purified) composite were implanted under the hind-quarter perimuscular membrane of ddY mice. Control capsules containing only bone morphogenetic protein were used for controls. The implants were radiographically and histologically examined at 1 to 4 weeks after the operation. According to the radiographic analysis, squalane and bone morphogenetic protein composite and bone morphogenetic protein only control specimens formed widespread heterotopic bone tissues. The amount of heterotopic bone formation in the composite experimental specimens was approximately 40% greater than that in the controls. Histologic examination of experimental and control specimens revealed varying amounts of perichondral ossification by 2 weeks. By 3 and 4 weeks, the bone deposits were colonized by hematopoietic bone marrow. Squalane was effective for the slow local release of bone morphogenetic protein. Furthermore, the squalane and bone morphogenetic protein composite was a reliable osteoinductive biomaterial.

  7. Squalane as a possible carrier of bone morphogenetic protein.

    Science.gov (United States)

    Kawakami, T; Uji, H; Antoh, M; Hasegawa, H; Kise, T; Eda, S

    1993-07-01

    Gelatin capsules containing squalane partially purified bone morphogenetic protein (BMP) complex were placed on the perimuscular membrane of rats. Two kinds of control, gelatin capsules containing only BMP and those bearing squalane only, were used. The embedded areas were histopathologically examined at 3 and 6 wk after the operation. The observations revealed that the squalane/BMP complex elicited wide heterotopic bone formation with bone marrow tissue, suggesting that squalane is a possible carrier of BMP for clinical applications.

  8. Bone graft substitutes and bone morphogenetic proteins for osteoporotic fractures: What is the evidence?

    NARCIS (Netherlands)

    E.M.M. van Lieshout (Esther); V. Alt (Volker)

    2016-01-01

    textabstractDespite improvements in implants and surgical techniques, osteoporotic fractures remain challenging to treat. Among other major risk factors, decreased expression of morphogenetic proteins has been identified for impaired fracture healing in osteoporosis. Bone grafts or bone graft substi

  9. Erythropoietin Modulates the Structure of Bone Morphogenetic Protein 2–Engineered Cranial Bone

    OpenAIRE

    Sun, Hongli; Jung, Younghun; Shiozawa, Yusuke; Taichman, Russell S.; Krebsbach, Paul H.

    2012-01-01

    The ideally engineered bone should have similar structural and functional properties to the native tissue. Although structural integrity is critical for functional bone regeneration, we know less about modulating the structural properties of the engineered bone elicited by bone morphogenetic protein (BMP) than efficacy and safety. Erythropoietin (Epo), a primary erythropoietic hormone, has been used to augment blood transfusion in orthopedic surgery. However, the effects of Epo on bone regene...

  10. Bone Morphogenetic Protein-2 Nonviral Gene Therapy in a Goat Iliac Crest Model for Bone Formation

    NARCIS (Netherlands)

    Loozen, Loek D.; van der Helm, Yvonne J. M.; Oner, F. Cumhur; Dhert, W.J.A.; Kruyt, Moyo C.; Alblas, Jacqueline

    2015-01-01

    Treatment and reconstruction of large bone defects, delayed unions, and nonunions is challenging and has resulted in an ongoing search for novel tissue-engineered therapies. Bone morphogenetic protein-2 (BMP-2) gene therapy is a promising strategy to provide sustained production of BMP-2 locally. Al

  11. Novel approaches to bone grafting: porosity, bone morphogenetic proteins, stem cells, and the periosteum.

    Science.gov (United States)

    Petrochenko, Peter; Narayan, Roger J

    2010-01-01

    The disadvantages involving the use of a patient's own bone as graft material have led surgeons to search for alternative materials. In this review, several characteristics of a successful bone graft material are discussed. In addition, novel synthetic materials and natural bone graft materials are being considered. Various factors can determine the success of a bone graft substitute. For example, design considerations such as porosity, pore shape, and interconnection play significant roles in determining graft performance. The effective delivery of bone morphogenetic proteins and the ability to restore vascularization also play significant roles in determining the success of a bone graft material. Among current approaches, shorter bone morphogenetic protein sequences, more efficient delivery methods, and periosteal graft supplements have shown significant promise for use in autograft substitutes or autograft extenders.

  12. Bone morphogenetic proteins: from structure to clinical use

    Directory of Open Access Journals (Sweden)

    Granjeiro J.M.

    2005-01-01

    Full Text Available Bone morphogenetic proteins (BMPs are multi-functional growth factors belonging to the transforming growth factor ß superfamily. Family members are expressed during limb development, endochondral ossification, early fracture, and cartilage repair. The activity of BMPs was first identified in the 1960s but the proteins responsible for bone induction were unknown until the purification and cloning of human BMPs in the 1980s. To date, about 15 BMP family members have been identified and characterized. The signal triggered by BMPs is transduced through serine/threonine kinase receptors, type I and II subtypes. Three type I receptors have been shown to bind BMP ligands, namely: type IA and IB BMP receptors and type IA activin receptors. BMPs seem to be involved in the regulation of cell proliferation, survival, differentiation and apoptosis, but their hallmark is their ability to induce bone, cartilage, ligament, and tendon formation at both heterotopic and orthotopic sites. This suggests that, in the future, they may play a major role in the treatment of bone diseases. Several animal studies have illustrated the potential of BMPs to enhance spinal fusion, repair critical-size defects, accelerate union, and heal articular cartilage lesions. Difficulties in producing and purifying BMPs from bone tissue have prompted the attempts made by several laboratories, including ours, to express these proteins in the recombinant form in heterologous systems. This review focuses on BMP structure, molecular mechanisms of action and significance and potential applications in medical, dental and veterinary practice for the treatment of cartilage and bone-related diseases.

  13. Harmine promotes osteoblast differentiation through bone morphogenetic protein signaling

    Energy Technology Data Exchange (ETDEWEB)

    Yonezawa, Takayuki [Department of Nutriproteomics, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Research Institute for Biological Functions, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Lee, Ji-Won [Research Institute for Biological Functions, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Hibino, Ayaka; Asai, Midori [Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Hojo, Hironori [Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Cha, Byung-Yoon [Research Institute for Biological Functions, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Teruya, Toshiaki [Research Institute for Biological Functions, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Faculty of Education, University of the Ryukyus, 1 Senbaru, Nishihara, Okinawa 903-0213 (Japan); Nagai, Kazuo [Research Institute for Biological Functions, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501 (Japan); Chung, Ung-Il [Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Yagasaki, Kazumi [Department of Nutriproteomics, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Division of Applied Biological Chemistry, Institute of Agriculture, Tokyo Noko University, 3-5-8 Saiwai, Fuchu, Tokyo 183-8509 (Japan); and others

    2011-06-03

    Highlights: {yields} Harmine promotes the activity and mRNA expression of ALP. {yields} Harmine enhances the expressions of osteocalcin mRNA and protein. {yields} Harmine induces osteoblastic mineralization. {yields} Harmine upregulates the mRNA expressions of BMPs, Runx2 and Osterix. {yields} BMP signaling pathways are involved in the actions of harmine. -- Abstract: Bone mass is regulated by osteoblast-mediated bone formation and osteoclast-mediated bone resorption. We previously reported that harmine, a {beta}-carboline alkaloid, inhibits osteoclast differentiation and bone resorption in vitro and in vivo. In this study, we investigated the effects of harmine on osteoblast proliferation, differentiation and mineralization. Harmine promoted alkaline phosphatase (ALP) activity in MC3T3-E1 cells without affecting their proliferation. Harmine also increased the mRNA expressions of the osteoblast marker genes ALP and Osteocalcin. Furthermore, the mineralization of MC3T3-E1 cells was enhanced by treatment with harmine. Harmine also induced osteoblast differentiation in primary calvarial osteoblasts and mesenchymal stem cell line C3H10T1/2 cells. Structure-activity relationship studies using harmine-related {beta}-carboline alkaloids revealed that the C3-C4 double bond and 7-hydroxy or 7-methoxy group of harmine were important for its osteogenic activity. The bone morphogenetic protein (BMP) antagonist noggin and its receptor kinase inhibitors dorsomorphin and LDN-193189 attenuated harmine-promoted ALP activity. In addition, harmine increased the mRNA expressions of Bmp-2, Bmp-4, Bmp-6, Bmp-7 and its target gene Id1. Harmine also enhanced the mRNA expressions of Runx2 and Osterix, which are key transcription factors in osteoblast differentiation. Furthermore, BMP-responsive and Runx2-responsive reporters were activated by harmine treatment. Taken together, these results indicate that harmine enhances osteoblast differentiation probably by inducing the expressions of

  14. The effect of bone morphogenetic protein-2 on osteosarcoma metastasis

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    Gill, Jonathan; Connolly, Patrick; Roth, Michael; Chung, So Hak; Zhang, Wendong; Piperdi, Sajida; Hoang, Bang; Yang, Rui; Guzik, Hillary; Gorlick, Richard; Geller, David S.

    2017-01-01

    Purpose Bone Morphogenetic Protein-2 (BMP-2) may offer the potential to enhance allograft-host osseous union in limb-salvage surgery following osteosarcoma resection. However, there is concern regarding the effect of locally applied BMP-2 on tumor recurrence and metastasis. The purpose of this project was to evaluate the effect of exogenous BMP-2 on osteosarcoma migration and invasion across a panel of tumor cell lines in vitro and to characterize the effect of BMP-2 on pulmonary osteosarcoma metastasis within a xenograft model. Experimental design The effect of BMP-2 on in vitro tumor growth and development was assessed across multiple standard and patient-derived xenograft osteosarcoma cell lines. Tumor migration capacity, invasion, and cell proliferation were characterized. In addition, the effect on metastasis was measured using a xenograft model following tail-vein injection. The effect of exogenous BMP-2 on the development of metastases was measured following both single and multiple BMP-2 administrations. Results There was no significant difference in migration capacity, invasion, or cell proliferation between the BMP-2 treated and the untreated osteosarcoma cell lines. There was no significant difference in pulmonary metastases between either the single-dose or multi-dose BMP-2 treated animals and the untreated control animals. Conclusions In the model systems tested, the addition of BMP-2 does not increase osteosarcoma proliferation, migration, invasion, or metastasis to the lungs. PMID:28264040

  15. Bone morphogenetic proteins in multiple sclerosis: Role in neuroinflammation.

    Science.gov (United States)

    Eixarch, Herena; Calvo-Barreiro, Laura; Montalban, Xavier; Espejo, Carmen

    2017-02-27

    Bone morphogenetic proteins (BMPs) are growth factors that represent the largest subgroup of signalling ligands of the transforming growth factor beta (TGF-β) superfamily. Their participation in the proliferation, survival and cell fate of several cell types and their involvement in many pathological conditions are now well known. BMP expression is altered in multiple sclerosis (MS) patients, suggesting that BMPs have a role in the pathogenesis of this disease. MS is a demyelinating and neurodegenerative autoimmune disorder of the central nervous system (CNS). MS is a complex pathological condition in which genetic, epigenetic and environmental factors converge, although its aetiology remains elusive. Multifunctional molecules, such as BMPs, are extremely interesting in the field of MS because they are involved in the regulation of several adult tissues, including the CNS and the immune system. In this review, we discuss the extensive data available regarding the role of BMP signalling in neuronal progenitor/stem cell fate and focus on the participation and expression of BMPs in CNS demyelination. Additionally, we provide an overview of the involvement of BMPs as modulators of the immune system, as this subject has not been thoroughly explored even though it is of great interest in autoimmune disorders. Moreover, we describe the data on BMP signalling in autoimmunity and inflammatory diseases, including MS and its experimental models. Thus, we aim to provide an integrated view of the putative role of BMPs in MS pathogenesis and to open the field for the further development of alternative therapeutic strategies for MS patients.

  16. Uncovering Molecular Bases Underlying Bone Morphogenetic Protein Receptor Inhibitor Selectivity.

    Directory of Open Access Journals (Sweden)

    Abdelaziz Alsamarah

    Full Text Available Abnormal alteration of bone morphogenetic protein (BMP signaling is implicated in many types of diseases including cancer and heterotopic ossifications. Hence, small molecules targeting BMP type I receptors (BMPRI to interrupt BMP signaling are believed to be an effective approach to treat these diseases. However, lack of understanding of the molecular determinants responsible for the binding selectivity of current BMP inhibitors has been a big hindrance to the development of BMP inhibitors for clinical use. To address this issue, we carried out in silico experiments to test whether computational methods can reproduce and explain the high selectivity of a small molecule BMP inhibitor DMH1 on BMPRI kinase ALK2 vs. the closely related TGF-β type I receptor kinase ALK5 and vascular endothelial growth factor receptor type 2 (VEGFR2 tyrosine kinase. We found that, while the rigid docking method used here gave nearly identical binding affinity scores among the three kinases; free energy perturbation coupled with Hamiltonian replica-exchange molecular dynamics (FEP/H-REMD simulations reproduced the absolute binding free energies in excellent agreement with experimental data. Furthermore, the binding poses identified by FEP/H-REMD led to a quantitative analysis of physical/chemical determinants governing DMH1 selectivity. The current work illustrates that small changes in the binding site residue type (e.g. pre-hinge region in ALK2 vs. ALK5 or side chain orientation (e.g. Tyr219 in caALK2 vs. wtALK2, as well as a subtle structural modification on the ligand (e.g. DMH1 vs. LDN193189 will cause distinct binding profiles and selectivity among BMP inhibitors. Therefore, the current computational approach represents a new way of investigating BMP inhibitors. Our results provide critical information for designing exclusively selective BMP inhibitors for the development of effective pharmacotherapy for diseases caused by aberrant BMP signaling.

  17. The effect of statins in colorectal cancer is mediated through the bone morphogenetic protein pathway

    NARCIS (Netherlands)

    Kodach, Liudmila L.; Bleuming, Sylvia A.; Peppelenbosch, Maikel P.; Hommes, Daniel W.; Van Den Brink, Gus R.; Hardwick, James C. H.

    2007-01-01

    Background & Aims: Epidemiological evidence suggests that statins prevent colorectal cancer (CRC), but the biological mechanism remains obscure. Statins induce bone morphogenetic protein (BMP) expression in bone cells. We have previously shown that BMPs act as tumor suppressors in CRC. We hypothesiz

  18. Erythropoietin modulates the structure of bone morphogenetic protein 2-engineered cranial bone.

    Science.gov (United States)

    Sun, Hongli; Jung, Younghun; Shiozawa, Yusuke; Taichman, Russell S; Krebsbach, Paul H

    2012-10-01

    The ideally engineered bone should have similar structural and functional properties to the native tissue. Although structural integrity is critical for functional bone regeneration, we know less about modulating the structural properties of the engineered bone elicited by bone morphogenetic protein (BMP) than efficacy and safety. Erythropoietin (Epo), a primary erythropoietic hormone, has been used to augment blood transfusion in orthopedic surgery. However, the effects of Epo on bone regeneration are not well known. Here, we determined the role of Epo in BMP2-induced bone regeneration using a cranial defect model. Epo administration improved the quality of BMP2-induced bone and more closely resembled natural cranial bone with a higher bone volume (BV) fraction and lower marrow fraction when compared with BMP2 treatment alone. Epo increased red blood cells (RBCs) in peripheral blood and also increased hematopoietic and mesenchymal stem cell (MSC) populations in bone marrow. Consistent with our previous work, Epo increased osteoclastogenesis both in vitro and in vivo. Results from a metatarsal organ culture assay suggested that Epo-promoted osteoclastogenesis contributed to angiogenesis because angiogenesis was blunted when osteoclastogenesis was blocked by alendronate (ALN) or osteoprotegerin (OPG). Earlier calcification of BMP2-induced temporary chondroid tissue was observed in the Epo+BMP group compared to BMP2 alone. We conclude that Epo significantly enhanced the outcomes of BMP2-induced cranial bone regeneration in part through its actions on osteoclastogenesis and angiogenesis.

  19. Enhanced Bone Morphogenetic Protein-2-Induced Ectopic and Orthotopic Bone Formation by Intermittent Parathyroid Hormone (1-34) Administration

    NARCIS (Netherlands)

    Kempen, Diederik H. R.; Lu, Lichun; Hefferan, Theresa E.; Creemers, Laura B.; Heijink, Andras; Maran, Avudaiappan; Dhert, Wouter J. A.; Yaszemski, Michael J.

    2010-01-01

    Bone morphogenetic proteins (BMPs) play a central role in local bone regeneration strategies, whereas the anabolic features of parathyroid hormone (PTH) are particularly appealing for the systemic treatment of generalized bone loss. The aim of the current study was to investigate whether local BMP-2

  20. Role of osteogenic protein-1/bone morphogenetic protein-7 in spinal fusion

    Directory of Open Access Journals (Sweden)

    Justin Munns

    2009-10-01

    Full Text Available Justin Munns, Daniel K Park, Kern SinghDepartment of Orthopedic Surgery, Rush University Medical Center, Chicago, Illinois, USAAbstract: Osteogenic protein-1 (OP-1, also known as bone morphogenetic protein-7 (BMP-7, is a protein in the TGF-β family of cellular proteins that has shown potential for application in patients undergoing spinal fusion due to its proven osteoinductive effects, particularly in patients with spondylolisthesis. OP-1 initiates numerous processes at the cellular level, acting on mesenchymal stem cells (MSCs, osteoblasts, and osteoclasts to stimulate bone growth. Animal studies of OP-1 have provided strong evidence for the ability of OP-1 to initiate ossification in posterolateral arthrodesis. Promising findings in early clinical trials with OP-1 prompted FDA approval for use in long bone nonunions in 2001 and subsequently for revision posterolateral arthrodesis in 2004 under a conditional Humanitarian Device Exemption. Larger clinical trials have recently shown no notable safety concerns or increases in adverse events associated with OP-1. However, a recent clinical trial has not conclusively demonstrated the noninferiority of OP-1 compared to autograft in revision posterolateral arthrodesis. The future of OP-1 application in patients with spondylolisthesis thus remains uncertain with the recent rejection of Premarket Approval (PMA status by the FDA (April 2009. Further investigation of its treatment success and immunological consequences appears warranted to establish FDA approval for its use in its current form.Keywords: osteogenic protein-1, bone morphogenetic protein-7, spinal fusion

  1. Stromal cell-derived factor-1 potentiates bone morphogenetic protein-2 induced bone formation.

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    Higashino, Kosaku; Viggeswarapu, Manjula; Bargouti, Maggie; Liu, Hui; Titus, Louisa; Boden, Scott D

    2011-02-01

    The mechanisms driving bone marrow stem cell mobilization are poorly understood. A recent murine study found that circulating bone marrow-derived osteoprogenitor cells (MOPCs) were recruited to the site of recombinant human bone morphogenetic protein-2 (BMP-2)-induced bone formation. Stromal cell-derived factor-1α (SDF-1α) and its cellular receptor CXCR4 have been shown to mediate the homing of stem cells to injured tissues. We hypothesized that chemokines, such as SDF-1, are also involved with mobilization of bone marrow cells. The CD45(-) fraction is a major source of MOPCs. In this report we determined that the addition of BMP-2 or SDF-1 to collagen implants increased the number of MOPCs in the peripheral blood. BMP-2-induced mobilization was blocked by CXCR4 antibody, confirming the role of SDF-1 in mobilization. We determined for the first time that addition of SDF-1 to implants containing BMP-2 enhances mobilization, homing of MOPCs to the implant, and ectopic bone formation induced by suboptimal BMP-2 doses. These results suggest that SDF-1 increases the number of osteoprogenitor cells that are mobilized from the bone marrow and then home to the implant. Thus, addition of SDF-1 to BMP-2 may improve the efficiency of BMPs in vivo, making their routine use for orthopaedic applications more affordable and available to more patients.

  2. Inflammatory Cytokines Stimulate Bone Morphogenetic Protein-2 Expression and Release from Pancreatic Beta Cells

    DEFF Research Database (Denmark)

    Urizar, Adriana Ibarra; Friberg, Josefine; Christensen, Dan Ploug;

    2016-01-01

    The proinflammatory cytokines interleukin-1 beta (IL-1β) and interferon gamma (IFN-γ) play important roles in the progressive loss of beta-cell mass and function during development of both type 1 and type 2 diabetes. We have recently showed that bone morphogenetic protein (BMP)-2 and -4...

  3. The bone morphogenetic protein pathway is active in human colon adenomas and inactivated in colorectal cancer

    NARCIS (Netherlands)

    Kodach, Liudmila L.; Bleurning, Sylvia A.; Musler, Alex R.; Peppelenbosch, Maikel R.; Hommes, Daniel W.; van den Brink, Gijs R.; van Noesel, Carel J. M.; Offerhaus, G. Johan A.; Hardwick, James C. H.

    2008-01-01

    BACKGROUND. Transforming growth factor beta (TGF beta) is important in colorectal cancer (CRQ progression. Bone morphogenetic proteins (BMPs), a subgroup within the TGF beta superfamily, recently also have been implicated in CRC, but their precise role in CRC has yet to be investigated. METHODS. The

  4. Bone morphogenetic protein signaling suppresses tumorigenesis at gastric epithelial transition zones in mice

    NARCIS (Netherlands)

    Bleuming, Sylvia A.; He, Xi C.; Kodach, Liudmila L.; Hardwick, James C.; Koopman, Frieda A.; ten Kate, Fiebo J.; van Deventer, Sander J. H.; Hommes, Daniel W.; Peppelenbosch, Maikel P.; Offerhaus, G. Johan; Li, Linheng; van den Brink, Gijs R.

    2007-01-01

    Bone morphogenetic protein (BMP) signaling is known to suppress oncogenesis in the small and large intestine of mice and humans. We examined the role of Bmpr1a signaling in the stomach. On conditional inactivation of Bmpr1a, mice developed neoplastic lesions specifically in the squamocolumnar and ga

  5. Cross talk between insulin and bone morphogenetic protein signaling systems in brown adipogenesis

    DEFF Research Database (Denmark)

    Zhang, Hongbin; Schulz, Tim J; Espinoza, Daniel O;

    2010-01-01

    Both insulin and bone morphogenetic protein (BMP) signaling systems are important for adipocyte differentiation. Analysis of gene expression in BMP7-treated fibroblasts revealed a coordinated change in insulin signaling components by BMP7. To further investigate the cross talk between insulin and...

  6. Bone morphogenetic protein-2 is a negative regulator of hepatocyte proliferation downregulated in the regenerating liver

    NARCIS (Netherlands)

    Xu, Cui-Ping; Ji, Wen-Min; van den Brink, Gijs R.; Peppelenbosch, Maikel P.

    2006-01-01

    AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on proliferation of human Huh7 hepatoma cells. METHODS: Fifty-four adult male Wistar rats we

  7. Bone morphogenetic protein-2-encapsulated grafted-poly-lactic acid-polycaprolactone nanoparticles promote bone repair.

    Science.gov (United States)

    Xu, Xiaojun; Yang, Jun; Ding, Lifeng; Li, Jianjun

    2015-01-01

    The aim of this study is to test the efficacy of a novel tissue-engineered bone in repairing bone defects, using poly-lactic-acid-polycaprolactone (PLA-PCL) scaffolding seeded with PEG-bone morphogenetic protein-2 (BMP-2)-transfected rBMSCs (rabbit bone marrow stromal cells). The rBMSCs were transfected with PEG/BMP-2 or liposome/BMP-2, and then implanted into a PLA-PCL tissue-engineered bone. The protein level of BMP-2 was assessed by Western blot analysis and immunohistochemistry. ELISA was used to measure the amount of BMP-2 secreted in the culture media. The mRNA level of BMP-2 and osteocalcin was assayed quantitatively by real-time PCR. The middle portion of the bilateral radius in New Zealand rabbits was excised and implanted with tissue-engineered bone, and the modified areas were monitored by X-ray, hematoxylin-eosin staining, and immunohistochemistry staining of BMP-2. PEG-BMP-2 nanoparticles (NPs) and BMP-2-loaded PEG-PLA-PCL tissue-engineered bones were successfully constructed. The novel PEG-PLA-PCL NPs/DNA complex was a superior option for transfecting BMP-2 in rBMSCs compared to normal liposomes Moreover, the mRNA level of osteocalcin and alkaline phosphatase activity was also elevated upon transfection of BMP-2-encapsulated NPs. In vivo implants with BMP-2-carried tissue-engineered bone exhibited dramatic augmentation of BMP-2 and effective bone formation in the rabbit ectopic model. The PEG-PLA-PCL NPs/BMP-2 complex had an advantageous effect on bone repair, which provided an important theoretic basis for potential clinical treatments.

  8. Oxidized alginate hydrogels for bone morphogenetic protein-2 delivery in long bone defects.

    Science.gov (United States)

    Priddy, Lauren B; Chaudhuri, Ovijit; Stevens, Hazel Y; Krishnan, Laxminarayanan; Uhrig, Brent A; Willett, Nick J; Guldberg, Robert E

    2014-10-01

    Autograft treatment of large bone defects and fracture non-unions is complicated by limited tissue availability and donor site morbidity. Polymeric biomaterials such as alginate hydrogels provide an attractive tissue engineering alternative due to their biocompatibility, injectability, and tunable degradation rates. Irradiated RGD-alginate hydrogels have been used to deliver proteins such as bone morphogenetic protein-2 (BMP-2), to promote bone regeneration and restoration of function in a critically sized rat femoral defect model. However, slow degradation of irradiated alginate hydrogels may impede integration and remodeling of the regenerated bone to its native architecture. Oxidation of alginate has been used to promote degradation of alginate matrices. The objective of this study was to evaluate the effects of alginate oxidation on BMP-2 release and bone regeneration. We hypothesized that oxidized-irradiated alginate hydrogels would elicit an accelerated release of BMP-2, but degrade faster in vivo, facilitating the formation of higher quality, more mature bone compared to irradiated alginate. Indeed, oxidation of irradiated alginate did accelerate in vitro BMP-2 release. Notably, the BMP-2 retained within both constructs was bioactive at 26days, as observed by induction of alkaline phosphatase activity and positive Alizarin Red S staining of MC3T3-E1 cells. From the in vivo study, robust bone regeneration was observed in both groups through 12weeks by radiography, micro-computed tomography analyses, and biomechanical testing. Bone mineral density was significantly greater for the oxidized-irradiated alginate group at 8weeks. Histological analyses of bone defects revealed enhanced degradation of oxidized-irradiated alginate and suggested the presence of more mature bone after 12weeks of healing.

  9. ANA deficiency enhances bone morphogenetic protein-induced ectopic bone formation via transcriptional events.

    Science.gov (United States)

    Miyai, Kentaro; Yoneda, Mitsuhiro; Hasegawa, Urara; Toita, Sayaka; Izu, Yayoi; Hemmi, Hiroaki; Hayata, Tadayoshi; Ezura, Yoichi; Mizutani, Shuki; Miyazono, Kohei; Akiyoshi, Kazunari; Yamamoto, Tadashi; Noda, Masaki

    2009-04-17

    Ectopic bone formation after joint replacement or brain injury in humans is a serious complication that causes immobility of joints and severe pain. However, mechanisms underlying such ectopic bone formation are not fully understood. Bone morphogenetic protein (BMPs) are defined as inducers of ectopic bone formation, and they are regulated by several types of inhibitors. ANA is an antiproliferative molecule that belongs to Tob/BTG family, but its activity in bone metabolism has not been known. Here, we examined the role of ANA on ectopic bone formation activity of BMP. In ANA-deficient and wild-type mice, BMP2 was implanted to induce ectopic bone formation in muscle. ANA deficiency increased mass of newly formed bone in vivo compared with wild-type based on 3D-muCT analyses. ANA mRNA was expressed in bone in vivo as well as in osteoblastic cells in vitro. Such ANA mRNA levels were increased by BMP2 treatment in MC3T3-E1 osteoblastic cells. Overexpression of ANA suppressed BMP-induced expression of luciferase reporter gene linked to BMP response elements in these cells. Conversely, ANA mRNA knockdown by small interference RNA enhanced the BMP-dependent BMP response element reporter expression. It also enhanced BMP-induced osteoblastic differentiation in muscle-derived C2C12 cells. Immunoprecipitation assay indicated that ANA interacts with Smad8. Thus, ANA is a suppressor of ectopic bone formation induced by BMP, and this inhibitory ANA activity is a part of the negative feedback regulation of BMP function.

  10. Salicylic Acid-Based Polymers for Guided Bone Regeneration Using Bone Morphogenetic Protein-2.

    Science.gov (United States)

    Subramanian, Sangeeta; Mitchell, Ashley; Yu, Weiling; Snyder, Sabrina; Uhrich, Kathryn; O'Connor, J Patrick

    2015-07-01

    Bone morphogenetic protein-2 (BMP-2) is used clinically to promote spinal fusion, treat complex tibia fractures, and to promote bone formation in craniomaxillofacial surgery. Excessive bone formation at sites where BMP-2 has been applied is an established complication and one that could be corrected by guided tissue regeneration methods. In this study, anti-inflammatory polymers containing salicylic acid [salicylic acid-based poly(anhydride-ester), SAPAE] were electrospun with polycaprolactone (PCL) to create thin flexible matrices for use as guided bone regeneration membranes. SAPAE polymers hydrolyze to release salicylic acid, which is a nonsteroidal anti-inflammatory drug. PCL was used to enhance the mechanical integrity of the matrices. Two different SAPAE-containing membranes were produced and compared: fast-degrading (FD-SAPAE) and slow-degrading (SD-SAPAE) membranes that release salicylic acid at a faster and slower rate, respectively. Rat femur defects were treated with BMP-2 and wrapped with FD-SAPAE, SD-SAPAE, or PCL membrane or were left unwrapped. The effects of different membranes on bone formation within and outside of the femur defects were measured by histomorphometry and microcomputed tomography. Bone formation within the defect was not affected by membrane wrapping at BMP-2 doses of 12 μg or more. In contrast, the FD-SAPAE membrane significantly reduced bone formation outside the defect compared with all other treatments. The rapid release of salicylic acid from the FD-SAPAE membrane suggests that localized salicylic acid treatment during the first few days of BMP-2 treatment can limit ectopic bone formation. The data support development of SAPAE polymer membranes for guided bone regeneration applications as well as barriers to excessive bone formation.

  11. Construction and characterization of a recombinant human adenovirus vector expressing bone morphogenetic protein 2

    OpenAIRE

    Zhang, Zheng; WANG, GUOXIAN; Li, Chen; Liu, Danping

    2013-01-01

    The aim of this study was to construct and characterize a novel recombinant human adenovirus vector expressing bone morphogenetic protein 2 (BMP2) and green fluorescent protein (GFP). The BMP2 gene in the plasmid pcDNA3-BMP2 was sequenced and the restriction enzyme recognition sites were analyzed. Following mutagenesis using polymerase chain reaction (PCR), the gene sequence after the translation termination codon was removed and new restriction sites were added. The mutated BMP2 gene (BMP2+ ...

  12. Comparative experiment of four different materials as carriers of Bone morphogenetic protein to repair long bone defect

    Institute of Scientific and Technical Information of China (English)

    WEI Kuan-hai; PEI Guo-xian; YANG Run-gong

    2001-01-01

    @@ OBJECTIVE To investigate the effects of four different materials as carriers of bone morphogenetic protein (BMP) to repair long bone defect. METHODS 12 mm radius bone defects were made. They were divided into 4 groups in random and repaired respectively with the vascular muscle flap combined with FS/BMP (group A), vascular muscle flap/BMP (group B), bloodless muscle flap/BMP (group C) and autolyzed antigen-extracted allogeneic bone (AAA)/BMP (group D).Their abilities of bone forming to repair bone defects were observed.

  13. Binding of integrin α1 to bone morphogenetic protein receptor IA suggests a novel role of integrin α1β1 in bone morphogenetic protein 2 signalling.

    Science.gov (United States)

    Zu, Yan; Liang, Xudong; Du, Jing; Zhou, Shuai; Yang, Chun

    2015-11-01

    Here, we observed that integrin α1β1 and bone morphogenetic protein receptor (BMPR) IA formed a complex and co-localised in several cell types. However, the molecular interaction between these two molecules was not studied in detail to date and the role of the interaction in BMPR signalling remains unknown; thus, these were investigated here. In a steered molecular dynamics (SMD) simulation, the observed development of the rupture force related to the displacement between the A-domain of integrin α1 and the extracellular domain of BMPR IA indicated a strong molecular interaction within the integrin-BMPR complex. Analysis of the intermolecular forces revealed that hydrogen bonds, rather than salt bridges, are the major contributors to these intermolecular interactions. By using Enzyme-linked immunosorbent assay (ELISA) and co-immunoprecipitation (co-IP) experiments with site-directed mutants, we found that residues 85-89 in BMPR IA play the most important role for BMPR IA binding to integrin α1β1. These residues are the same as those responsible for bone morphogenetic protein 2 (BMP-2)/BMPR IA binding. In our experiments, we also found that the interference of integrin α1β1 up regulated the level of phosphorylated Smad1, 5, 8, which is the downstream of BMP/BMPR signalling. Therefore, our results suggest that integrin α1β1/BMPR IA may block BMP-2/BMPR IA complex information and interfere with the BMP-2 signalling pathway in cells.

  14. Creating new functional biomaterials : construction and production of Bone Morphogenetic 2-ELP hybrid proteins

    OpenAIRE

    Silva, J. Azevedo; Machado, Raul; Reis, R.L.; Rodríguez-Cabello, José Carlos; Casal, Margarida

    2010-01-01

    Bone morphogenetic protein 2 (BMP-2) is a potent osteoinductive cytokine from the TGF-β superfamily that triggers the development of stem cells into osteoblasts. Its therapeutic interest has led to the development of various production systems for recombinant variables of BMP-2. Production has been achieved in expression systems ranging from animal cells to bacteria, but is always associated with three major drawbacks: low production rates (in animal cells), low activity (bacterial cells) and...

  15. Calcium Phosphate Scaffolds Combined with Bone Morphogenetic Proteins or Mesenchymal Stem Cells in Bone Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Han Sun

    2015-01-01

    Full Text Available Objective: The purpose of this study was to review the current status of calcium phosphate (CaP scaffolds combined with bone morphogenetic proteins (BMPs or mesenchymal stem cells (MSCs in the field of bone tissue engineering (BTE. Date Sources: Data cited in this review were obtained primarily from PubMed and Medline in publications from 1979 to 2014, with highly regarded older publications also included. The terms BTE, CaP, BMPs, and MSC were used for the literature search. Study Selection: Reviews focused on relevant aspects and original articles reporting in vitro and/or in vivo results concerning the efficiency of CaP/BMPs or CaP/MSCs composites were retrieved, reviewed, analyzed, and summarized. Results: An ideal BTE product contains three elements: Scaffold, growth factors, and stem cells. CaP-based scaffolds are popular because of their outstanding biocompatibility, bioactivity, and osteoconductivity. However, they lack stiffness and osteoinductivity. To solve this problem, composite scaffolds of CaP with BMPs have been developed. New bone formation by CaP/BMP composites can reach levels similar to those of autografts. CaP scaffolds are compatible with MSCs and CaP/MSC composites exhibit excellent osteogenesis and stiffness. In addition, a CaP/MSC/BMP scaffold can repair bone defects more effectively than an autograft. Conclusions: Novel BTE products possess remarkable osteoconduction and osteoinduction capacities, and exhibit balanced degradation with osteogenesis. Further work should yield safe, viable, and efficient materials for the repair of bone lesions.

  16. Calcium Phosphate Scaffolds Combined with Bone Morphogenetic Proteins or Mesenchymal Stem Cells in Bone Tissue Engineering

    Institute of Scientific and Technical Information of China (English)

    Han Sun; Hui-Lin Yang

    2015-01-01

    Objective:The purpose of this study was to review the current status of calcium phosphate (CaP) scaffolds combined with bone morphogenetic proteins (BMPs) or mesenchymal stem cells (MSCs) in the field of bone tissue engineering (BTE).Date Sources:Data cited in this review were obtained primarily from PubMed and Medline in publications from 1979 to 2014,with highly regarded older publications also included.The terms BTE,CaP,BMPs,and MSC were used for the literature search.Study Selection:Reviews focused on relevant aspects and original articles reporting in vitro and/or in vivo results concerning the efficiency of CaP/BMPs or CaP/MSCs composites were retrieved,reviewed,analyzed,and summarized.Results:An ideal BTE product contains three elements:Scaffold,growth factors,and stem cells.CaP-based scaffolds are popular because of their outstanding biocompatibility,bioactivity,and osteoconductivity.However,they lack stiffness and osteoinductivity.To solve this problem,composite scaffolds of CaP with BMPs have been developed.New bone formation by CaP/BMP composites can reach levels similar to those of autografts.CaP scaffolds are compatible with MSCs and CaP/MSC composites exhibit excellent osteogenesis and stiffness.In addition,a CaP/MSC/BMP scaffold can repair bone defects more effectively than an autograft.Conclusions:Novel BTE products possess remarkable osteoconduction and osteoinduction capacities,and exhibit balanced degradation with osteogenesis.Further work should yield safe,viable,and efficient materials for the repair of bone lesions.

  17. Comparison between heparin-conjugated fibrin and collagen sponge as bone morphogenetic protein-2 carriers for bone regeneration

    OpenAIRE

    Yang, Hee Seok; La, Wan-Geun; Cho, Yong-Min; Shin, Wangsoo; Yeo, Guw-Dong; Kim, Byung-Soo

    2012-01-01

    Bone morphogenetic protein-2 (BMP-2) is used to promote bone regeneration. However, the bone regeneration ability of BMP-2 relies heavily on the delivery vehicle. Previously, we have developed heparin-conjugated fibrin (HCF), a vehicle for long-term delivery of BMP-2 and demonstrated that long-term delivery of BMP-2 enhanced its osteogenic efficacy as compared to short-term delivery at an equivalent dose. The aim of this study was to compare the bone-forming ability of the BMP-2 delivered by ...

  18. Influence of bone morphogenetic protein type IA receptor conditional knockout in lens on expression of bone morphogenetic protein 4 in lens

    Institute of Scientific and Technical Information of China (English)

    Qi; Zhao; Jiang-Yue; Zhao; Jin-Song; Zhang

    2015-01-01

    AIM: To investigate the influence of bone morphogenetic protein type IA receptor [BMPR-IA(ALK3)] conditional knockout in lens on expression of bone morphogenetic protein 4(BMP4) in lens during the development of the vertebrate eye.METHODS: Cre-positive mice were mated with Crenegative mice to generate 50% Cre-positive(conditional knockout, CKO) 4 embryos, 8 eyes and 50% Cre-negative offspring(wild type, WT) 4 embryos, 8 eyes. The embryos were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to a thickness of 4 μm.Removal of paraffin wax and dehydrating for sections,and then the procedure of in situ hybridization was processed, BMP4 MK1784-m(BOSTER) was used, and observed the expression of BMP4 in the lens in experimental group and control group. We selected SPSS11.0 software for statistical analysis, P<0.05 showed that the difference was statistically significant.· RESULTS: Four embryos of each genotype were examined, totally we had 8 embryos, 16 eyes. We got the uniform outcomes in all the embryos. We found ALK3 was required during lens growing, but was not essential for the formation of lens. We observed that the expression of BMP4 in the lens was significantly reduced in all 8 ALK3 CKO lens, BMP4 expression was normal in all the 8 WT lens, P <0.01. This phenomenon became increasingly visible in accordance with embryo development. The most apparent alteration was present at stage E15.5.CONCLUSION: ALK3 is essential for lens growth. The influence of ALK3 on the expression of BMP4 is present during the development of mice lens.

  19. Bone Morphogenetic Protein 4 Signalling in Neural Stem and Progenitor Cells during Development and after Injury

    Directory of Open Access Journals (Sweden)

    Alistair E. Cole

    2016-01-01

    Full Text Available Substantial progress has been made in identifying the extracellular signalling pathways that regulate neural stem and precursor cell biology in the central nervous system (CNS. The bone morphogenetic proteins (BMPs, in particular BMP4, are key players regulating neuronal and glial cell development from neural precursor cells in the embryonic, postnatal, and injured CNS. Here we review recent studies on BMP4 signalling in the generation of neurons, astrocytes, and oligodendroglial cells in the CNS. We also discuss putative mechanisms that BMP4 may utilise to influence glial cell development following CNS injury and highlight some questions for further research.

  20. Bone morphogenetic protein 7 regulates reactive gliosis in retinal astrocytes and Müller glia

    OpenAIRE

    2014-01-01

    Purpose The focus of this study was to determine whether bone morphogenetic proteins (BMPs) trigger reactive gliosis in retinal astrocytes and/or Müller glial cells. Methods Retinal astrocytes and the Müller glial cell line MIO-M1 were treated with vehicle, BMP7, or BMP4. Samples from the treated cells were analyzed for changes in gliosis markers using reverse transcriptase – quantitative PCR (RT-qPCR) and western blotting. To determine potential similarities and differences in gliosis states...

  1. No effect of bone morphogenetic protein-7 (OP-1) on the incorporation of impacted bone grafts in a realistic acetabular model.

    NARCIS (Netherlands)

    Buma, P.; Arts, J.J.C.; Gardeniers, J.W.M.; Verdonschot, N.J.J.; Schreurs, B.W.

    2008-01-01

    Bone morphogenetic proteins (BMPs) accelerate bone repair in experimental and clinical conditions. Impacted Morsellized Cancellous Bone grafts (MCB) are successfully used to reconstruct bone defects after failed hip implants. The main question in this study was if BMP-7 (OP-1) mixed with MCB could a

  2. Caenorhabditis elegans SMA-10/LRIG is a conserved transmembrane protein that enhances bone morphogenetic protein signaling.

    Directory of Open Access Journals (Sweden)

    Tina L Gumienny

    2010-05-01

    Full Text Available Bone morphogenetic protein (BMP pathways control an array of developmental and homeostatic events, and must themselves be exquisitely controlled. Here, we identify Caenorhabditis elegans SMA-10 as a positive extracellular regulator of BMP-like receptor signaling. SMA-10 acts genetically in a BMP-like (Sma/Mab pathway between the ligand DBL-1 and its receptors SMA-6 and DAF-4. We cloned sma-10 and show that it has fifteen leucine-rich repeats and three immunoglobulin-like domains, hallmarks of an LRIG subfamily of transmembrane proteins. SMA-10 is required in the hypodermis, where the core Sma/Mab signaling components function. We demonstrate functional conservation of LRIGs by rescuing sma-10(lf animals with the Drosophila ortholog lambik, showing that SMA-10 physically binds the DBL-1 receptors SMA-6 and DAF-4 and enhances signaling in vitro. This interaction is evolutionarily conserved, evidenced by LRIG1 binding to vertebrate receptors. We propose a new role for LRIG family members: the positive regulation of BMP signaling by binding both Type I and Type II receptors.

  3. The daf-4 gene encodes a bone morphogenetic protein receptor controlling C. elegans dauer larva development.

    Science.gov (United States)

    Estevez, M; Attisano, L; Wrana, J L; Albert, P S; Massagué, J; Riddle, D L

    1993-10-14

    The bone morphogenetic protein (BMP) family is a conserved group of signalling molecules within the transforming growth factor-beta (TGF-beta) superfamily. This group, including the Drosophila decapentaplegic (dpp) protein and the mammalian BMPs, mediates cellular interactions and tissue differentiation during development. Here we show that a homologue of human BMPs controls a developmental switch in the life cycle of the free-living soil nematode Caenorhabditis elegans. Starvation and overcrowding induce C. elegans to form a developmentally arrested, third-stage dauer larva. The daf-4 gene, which acts to inhibit dauer larva formation and promote growth, encodes a receptor protein kinase similar to the daf-1, activin and TGF-beta receptor serine/threonine kinases. When expressed in monkey COS cells, the daf-4 receptor binds human BMP-2 and BMP-4. The daf-4 receptor is the first to be identified for any growth factor in the BMP family.

  4. Turning Bone Morphogenetic Protein 2 (BMP2) on and off in Mesenchymal Cells.

    Science.gov (United States)

    Rogers, Melissa B; Shah, Tapan A; Shaikh, Nadia N

    2015-10-01

    The concentration, location, and timing of bone morphogenetic protein 2 (BMP2, HGNC:1069, GeneID: 650) gene expression must be precisely regulated. Abnormal BMP2 levels cause congenital anomalies and diseases involving the mesenchymal cells that differentiate into muscle, fat, cartilage, and bone. The molecules and conditions that influence BMP2 synthesis are diverse. Understandably, complex mechanisms control Bmp2 gene expression. This review includes a compilation of agents and conditions that can induce Bmp2. The currently known trans-regulatory factors and cis-regulatory elements that modulate Bmp2 expression are summarized and discussed. Bone morphogenetic protein 2 (BMP2, HGNC:1069, GeneID: 650) is a classical morphogen; a molecule that acts at a distance and whose concentration influences cell behavior. In mesenchymal cells, the concentration of BMP2 influences myogenesis, adipogenesis, chondrogenesis, and osteogenesis. Because the amount, timing, and location of BMP2 synthesis influence the allocation of cells to muscle, fat, cartilage, and bone, the mechanisms that regulate the Bmp2 gene are crucial. Key early mesodermal events that require precise Bmp2 regulation include heart specification and morphogenesis. Originally named for its osteoinductive properties, healing fractures requires BMP2. The human Bmp2 gene also has been linked to osteoporosis and osteoarthritis. In addition, all forms of pathological calcification in the vasculature and in cardiac valves involve the pro-osteogenic BMP2. The diverse tissues, mechanisms, and diseases influenced by BMP2 are too numerous to list here (see OMIM: 112261). However, in all BMP2-influenced pathologies, changes in the behavior and differentiation of pluripotent mesenchymal cells are a recurring theme. Consequently, much effort has been devoted to identifying the molecules and conditions that influence BMP2 synthesis and the complex mechanisms that control Bmp2 gene expression. This review begins with an

  5. Successful treatment of a humeral capitulum osteonecrosis with bone morphogenetic protein-7 combined with autologous bone grafting.

    Science.gov (United States)

    Marsell, Richard; Hailer, Nils P

    2014-08-01

    We present the case of a 27-year-old female with subcortical osteonecrosis of the humeral capitulum. Percutaneous retrograde drilling of the lesion and application of recombinant human bone morphogenetic protein (BMP)-7 were combined with autologous bone grafting. At follow-up the patient was almost pain-free, had normalized her range of motion, and radiography showed consolidation of the lesion without any heterotopic bone formation. By timing surgery prior to subchondral collapse, biomechanical stability of the subchondral bone was maintained. To our knowledge, this is the first report on the treatment of an osteonecrosis in this location with a BMP, and this strategy could potentially be applied in other locations with juxta-articular osteonecrosis.

  6. Deletion of the sequence encoding the tail domain of the bone morphogenetic protein type 2 receptor reveals a bone morphogenetic protein 7-specific gain of function.

    Science.gov (United States)

    Leyton, Patricio A; Beppu, Hideyuki; Pappas, Alexandra; Martyn, Trejeeve M; Derwall, Matthias; Baron, David M; Galdos, Rita; Bloch, Donald B; Bloch, Kenneth D

    2013-01-01

    The bone morphogenetic protein (BMP) type II receptor (BMPR2) has a long cytoplasmic tail domain whose function is incompletely elucidated. Mutations in the tail domain of BMPR2 are found in familial cases of pulmonary arterial hypertension. To investigate the role of the tail domain of BMPR2 in BMP signaling, we generated a mouse carrying a Bmpr2 allele encoding a non-sense mediated decay-resistant mutant receptor lacking the tail domain of Bmpr2. We found that homozygous mutant mice died during gastrulation, whereas heterozygous mice grew normally without developing pulmonary arterial hypertension. Using pulmonary artery smooth muscle cells (PaSMC) from heterozygous mice, we determined that the mutant receptor was expressed and retained its ability to transduce BMP signaling. Heterozygous PaSMCs exhibited a BMP7‑specific gain of function, which was transduced via the mutant receptor. Using siRNA knockdown and cells from conditional knockout mice to selectively deplete BMP receptors, we observed that the tail domain of Bmpr2 inhibits Alk2‑mediated BMP7 signaling. These findings suggest that the tail domain of Bmpr2 is essential for normal embryogenesis and inhibits Alk2‑mediated BMP7 signaling in PaSMCs.

  7. SPECIFIC BINDING OF HUMAN BONE MORPHOGENETIC PROTEIN (2A) WITH MOUSE OSTEOBLASTIC CELLS

    Institute of Scientific and Technical Information of China (English)

    刘新平; 陈苏民; 陈南春; 高磊; 赵忠良

    1996-01-01

    Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM2I. Hulnata BMP2A C-terminal peptide displayed on the surface of the phage can bind specifically to the sttrface of mouse osteoblastie cell (MC3T3) membrane. ELISA assay showed a positive signal of the binding by using antibody against M13 phage gene 8 protein. After labeling with 3HTdR,the counts of the binding groups were 3 to 10 times higher than the control groups. It suggests that the'surface of MC3T3 cells exist the recepzor for hBMP2A.

  8. Water-dispersed bone morphogenetic protein nanospheres prepared by co-precipitation method

    Institute of Scientific and Technical Information of China (English)

    江兵兵; 高长有; 胡玲; 沈家骢

    2004-01-01

    A modified complex coacervation-co-precipitation method was used to prepare bone morphogenetic protein(BMP)-loaded nanospheres. Three natural polymers were used as packing materials to obtain nanoscale delivery device for BMP,in the presence of phosphatidylcholine functioning as stabilizer. Positively charged polysaccharide, N,N-diethylaminoethyl dex-tran (DEAE-dextran) tended to form stable, uniform and smaller size particles carrying BMP. Negatively charged bovine serumalbumin (BSA) induced precipitation of the produced BMP particles due to its weak interaction with BMP molecules, although itproduced nanosized BMP spheres. While collagen, a weakly positively charged protein shaped larger particles due to the stronginteraction among themselves. A mechanism of co-precipitation process was also deduced to depict the formation of stablenanospheres.

  9. Water-dispersed bone morphogenetic protein nanospheres prepared by co-precipitation method

    Institute of Scientific and Technical Information of China (English)

    江兵兵; 高长有; 胡玲; 沈家骢

    2004-01-01

    A modified complex coacervation-co-precipitation method was used to prepare bone morphogenetic protein (BMP)-loaded nanospheres. Three natural polymers were used as packing materials to obtain nanoscale delivery device for BMP,in the presence of phosphatidylcholine functioning as stabilizer. Positively charged polysaccharide, N,N-diethylaminoethyl dex-tran (DEAE-dextran) tended to form stable, uniform and smaller size particles carrying BMP. Negatively charged bovine serum albumin (BSA) induced precipitation of the produced BMP particles due to its weak interaction with BMP molecules, although it produced nanosized BMP spheres. While collagen, a weakly positively charged protein shaped larger particles due to the strong interaction among themselves. A mechanism of co-precipitation process was also deduced to depict the formation of stable nanospheres.

  10. Promoting lumbar spinal fusion by adenovirus-mediated bone morphogenetic protein-4 gene therapy

    Institute of Scientific and Technical Information of China (English)

    ZHAO Jian; ZHAO Dun-yan; SHEN Ai-guo; LIU Fan; ZHANG Feng; SUN Yu; WU Hong-fu; LU Chun-feng; SHI Hong-guang

    2007-01-01

    Objective: To determine whether an adenoviral construct containing bone morphogenetic protein-4 (BMP-4) gene can be used for lumbar spinal fusion. Methods: Twelve New Zealand white rabbits were randomly divided into two groups, 8 in the experimental group and 4 in the control group. Recombinant, replication-defective type 5 adenovirus with the cytomegalovirus (CMV) promoter and BMP-4 gene (Ad-BMP-4) was used. Another adenovirus constructed with the CMV promoter and β-galactosidase gene (Ad-β-gal) was used as control. Using collagen sponge as a carrier, Ad-BMP-4 (2.9×108 pfu/ml ) was directly implanted on the surface of L5-L6 lamina in the experimental group, while Ad-β-gal was implanted simultaneously in the control group. X-ray was obtained at 3, 6, and 12 weeks postoperatively to observe new bone formation. When new bone formation was identified, CT scans and three-dimensional reconstruction were obtained. After that, the animals were killed and underwent histological inspection.Results: In 12 weeks after operation, new bone formation and fusion were observed on CT scans in the experimental group, without the evidence of ectopic calcification in the canal. Negative results were found in the control group. Histological analysis demonstrated endochondral bone formation at the operative site and fusion at early stage was testified.Conclusions: In vivo gene therapy using Ad-BMP-4 for lumbar posterolateral spinal fusion is practicable and effective.

  11. The application of bone morphogenetic proteins to periodontal and peri-implant tissue regeneration: A literature review

    OpenAIRE

    Karuppanan P Sasikumar; Sugumari Elavarasu; Jayaprakash S Gadagi

    2012-01-01

    Progress in understanding the role of bone morphogenetic proteins (BMPs) in craniofacial and tooth development and the demonstration of stem cells in periodontal ligament have set the stage for periodontal regenerative therapy and tissue engineering. Furthermore, recent approval by the Food and Drug Administration of recombinant human BMPs for accelerating bone fusion in slow-healing fractures indicates that this protein family may prove useful in designing regenerative treatments in periodon...

  12. Osteogenic potential of the human bone morphogenetic protein 2 gene activated nanobone putty

    Institute of Scientific and Technical Information of China (English)

    TIAN Xiao-bin; SUN Li; YANG Shu-hua; ZHANG Yu-kun; HU Ru-yin; FU De-hao

    2008-01-01

    Background Nanobone putty is an injectable and bioresorbable bone substitute. The neutral-pH putty resembles hard bone tissue, does not contain polymers or plasticizers, and is self-setting and nearly isothermic, properties which are helpful for the adhesion, proliferation, and function of bone cells. The aim of this study was to investigate the osteogenic potential of human bone morphogenetic protein 2(hBMP2)gene activated nanobone putty in inducing ectopic bone formation, and the effects of the hBMP2 gene activated nanobone putry on repairing bone defects. Methods Twenty four Kunming mice were randomly divided into two groups. The nanobone putty+hBMP2 plasmid was injected into the right thigh muscle pouches of the mice(experiment side). The nanobone putty+blank plasmid or nanobone putty was injected into the left thigh muscle pouches of the group 1(control side 1)or group 2(control side 2), respectively. The effects of ectopic bone formation were evaluated by radiography, histology, and molecular biology analysis at 2 and 4 weeks after operation. Bilateral 15 mm radial defects were made in forty-eight rabbits. These rabbits were randomly divided into three groups: Group A, nanobone putty+hBMP2 plasmid;Group B, putty+blank plasmid; Group C, nanobone putty only. Six rabbits with left radial defects served as blank controls. The effect of bone repairing was evaluated by radiography, histology, molecular biology, and biomechanical analysis at 4, 8, and 12 weeks after operation. Results The tissue from the experimental side of the mice expressed hBMP2. Obvious cartilage and island-distributed immature bone formation in implants of the experiment side were observed at 2 weeks after operation, and massive mature bone observed at 4 weeks. No bone formation was observed in the control side of the mice. The ALP activity in the experiment side of the mice was higher than that in the control side. The tissue of Group A rabbits expressed hBMP2 protein and higher ALP level

  13. Non-viral bone morphogenetic protein 2 transfection of rat dental pulp stem cells using calcium phosphate nanoparticles as carriers.

    NARCIS (Netherlands)

    Yang, X.; Walboomers, X.F.; Dolder, J. van den; Yang, F.; Bian, Z.; Fan, M.; Jansen, J.A.

    2008-01-01

    Calcium phosphate nanoparticles have shown potential as non-viral vectors for gene delivery. The aim of this study was to induce bone morphogenetic protein (Bmp)2 transfection in rat dental pulp stem cells using calcium phosphate nanoparticles as a gene vector and then to evaluate the efficiency and

  14. Heterotopic ossification after the use of recombinant human bone morphogenetic protein-7

    Science.gov (United States)

    Papanagiotou, Marianthi; Dailiana, Zoe H; Karachalios, Theophilos; Varitimidis, Sokratis; Hantes, Michael; Dimakopoulos, Georgios; Vlychou, Marianna; Malizos, Konstantinos N

    2017-01-01

    AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7 (rhBMP-7) for the treatment of nonunions. METHODS Bone morphogenetic proteins (BMPs) promote bone formation by auto-induction. Recombinant human BMP-7 in combination with bone grafts was used in 84 patients for the treatment of long bone nonunions. All patients were evaluated radiographicaly for the development of heterotopic ossification during the standard assessment for the nonunion healing. In all patients (80.9%) with radiographic signs of heterotopic ossification, a CT scan was performed. Nonunion site palpation and ROM evaluation of the adjacent joints were also carried out. Factors related to the patient (age, gender), the nonunion (location, size, chronicity, number of previous procedures, infection, surrounding tissues condition) and the surgical procedure (graft and fixation type, amount of rhBMP-7) were correlated with the development of heterotopic ossification and statistical analysis with Pearsons χ2 test was performed. RESULTS Eighty point nine percent of the nonunions treated with rhBMP-7, healed with no need for further procedures. Heterotopic bone formation occurred in 15 of 84 patients (17.8%) and it was apparent in the routine radiological evaluation of the nonunion site, in a mean time of 5.5 mo after the rhBMP-7 application (range 3-12). The heterotopic ossification was located at the femur in 8 cases, at the tibia in 6, and at the humerus in οne patient. In 4 patients a palpable mass was present and only in one patient, with a para-articular knee nonunion treated with rhBMP-7, the size of heterotopic ossification affected the knee range of motion. All the patients with heterotopic ossification were male. Statistical analysis proved that patient’s gender was the only important factor for the development of heterotopic ossification (P = 0.007). CONCLUSION Heterotopic ossification after the use of rhBMP-7 in nonunions was

  15. A new biocompatible delivery scaffold containing heparin and bone morphogenetic protein 2

    Directory of Open Access Journals (Sweden)

    Thanyaphoo Suphannee

    2016-09-01

    Full Text Available Silicon-substituted calcium phosphate (Si-CaP was developed in our laboratory as a biomaterial for delivery in bone tissue engineering. It was fabricated as a 3D-construct of scaffolds using chitosan-trisodium polyphosphate (TPP cross-linked networks. In this study, heparin was covalently bonded to the residual -NH2 groups of chitosan on the scaffold applying carbodiimide chemistry. Bonded heparin was not leached away from scaffold surfaces upon vigorous washing or extended storage. Recombinant human bone morphogenetic protein 2 (rhBMP-2 was bound to conjugated scaffolds by ionic interactions between the negatively charged SO42- clusters of heparin and positively charged amino acids of rhBMP-2. The resulting scaffolds were inspected for bone regenerative capacity by subcutaneous implanting in rats. Histological observation and mineralization assay were performed after 4 weeks of implantation. Results from both in vitro and in vivo experiments suggest the potential of the developed scaffolds for bone tissue engineering applications in the future.

  16. TiO2 nanotubes functionalized with regions of bone morphogenetic protein-2 increases osteoblast adhesion.

    Science.gov (United States)

    Balasundaram, Ganesan; Yao, Chang; Webster, Thomas J

    2008-02-01

    Titanium (Ti) and its alloys are widely used in orthopedic and dental applications. However, the native TiO2 layer is not bioactive enough to form a direct bond with bone, which sometimes translates into a lack of osseointegration into juxtaposed bone that might lead to long term implant failure. In this study, the 20 amino acid peptide sequence (the so-called "knuckle epitope") of bone morphogenetic protein-2 (BMP-2) was immobilized onto Ti nanotubes created by electrochemical anodization. Further, human osteoblast (bone-forming cell) responses to such anodic Ti oxides functionalized with the BMP-2 knuckle epitope was examined in vitro. Materials were characterized by scanning electron and atomic force microscopy. Results of this in vitro study continued to provide evidence of increased osteoblast adhesion on Ti anodized to possess nanotubes compared to unanodized Ti. However, for the first time, results also showed that the immobilization of the BMP-2 knuckle epitope onto Ti anodized to possess nanotubes increased osteoblast adhesion compared to non-functionalized anodized Ti, anodized Ti functionalized with amine (NH2) groups, and unanodized Ti after 4 h. Results also showed increased osteoblast adhesion on amine terminated anodized Ti compared to respective non-functionalized anodized Ti and unanodized Ti. In summary, results of this in vitro study provided evidence that Ti anodized to possess nanotubes and then further functionalized with the BMP-2 knuckle epitope should be further studied for improved orthopedic applications.

  17. Bone morphogenetic protein Smads signaling in mesenchymal stem cells affected by osteoinductive calcium phosphate ceramics.

    Science.gov (United States)

    Tang, Zhurong; Wang, Zhe; Qing, Fangzhu; Ni, Yilu; Fan, Yujiang; Tan, Yanfei; Zhang, Xingdong

    2015-03-01

    Porous calcium phosphate ceramics (CaP ceramics) could induce ectopic bone formation which was regulated by various signal molecules. In this work, bone marrow mesenchymal stem cells (MSCs) were cultured on the surface of osteoinductive hydroxyapatite (HA) and biphasic calcium phosphate (BCP) ceramics in comparison with control (culture plate) for up to 14 days to detect the signal molecules which might be affected by the CaP ceramics. Without adding osteogenic factors, MSCs cultured on HA and BCP both expressed higher Runx2, Osterix, collagen type I, osteopontin, bone sialoprotein, and osteocalcin at various stages compared with control, thus confirmed the osteoblastic differentiation of MSCs. Later study demonstrated the messenger RNA level of bone morphogenetic protein 2 (BMP2) and BMP4 were also significantly enhanced by HA and BCP. Furthermore, Smad1, 4, 5, and Dlx5, the main molecules in the BMP/Smads signaling pathway, were upregulated by HA and BCP. Moreover, the higher expression of Smads and BMP2, 4 in BCP over HA, corresponded to the better performance of BCP in stimulating in vitro osteoblastic differentiation of MSCs. This was in accordance with the better osteoinductivity of BCP over HA in vivo. Altogether, these results implied that the CaP ceramics may initiate the osteoblastic differentiation of MSCs by influencing the expression of molecules in BMP/Smads pathway.

  18. Expression and regulation of the decoy bone morphogenetic protein receptor BAMBI in the developing avian face.

    Science.gov (United States)

    Higashihori, Norihisa; Song, Yiping; Richman, Joy M

    2008-05-01

    Here, we examine the expression and regulation of the gene BAMBI, a kinase-deficient decoy receptor capable of interacting with type I bone morphogenetic protein (BMP) receptors in avian embryos. Initially, expression was limited to the endoderm during neurula and pharyngula stages. From embryonic day 3.5 (stage 20) and onward, BAMBI expression almost perfectly overlapped with known expression patterns for BMP4, particularly in the face and limbs. We performed bead implant experiments in the face to see which signals could be repressing or promoting expression of BAMBI. Our data point to retinoids and BMPs as being major positive regulators of BAMBI expression; however, fibroblast growth factor 2 acts to repress BAMBI. Furthermore, retinoic acid is likely to act directly on BAMBI as induction occurs in the presence of cycloheximide. The data suggested that BAMBI could be used to regulate Bmp signaling during tissue interactions that are an integral part of facial morphogenesis.

  19. Establishment and identification of fibroblast clones expressing human bone morphogenetic protein 2

    Institute of Scientific and Technical Information of China (English)

    Juan Wang; Weibin Sun; Chun Lu; Guixia Tang

    2005-01-01

    Objective:To establish fibroblasts stably expressing human bone morphogenetic protein 2 (hBMP2). Methods:Eukaryonic expression vector(pcDNA3.1-B2) was transduced into NIH3T3 cells using SofastTM, a new generation cationic polymer gene transfection reagent. The positive cell clones were selected with G418. The stable transfection and expression of BMP2 in the NIH3T3 cells were determined by RT-PCR and immunohistochemical stain. Results: BMP2 mRNA was transcripted and expressed in the transfected NIH3T3 cells. Conclusion: With positive compound transfection, outside human BMP2 gene can be successfully transducted into NIH3T3 cells, which is the key step to induce periodontal cells to osseous phenotypes.

  20. Combination of bone morphogenetic protein-2 plasmid DNA with chemokine CXCL12 creates an additive effect on bone formation onset and volume

    NARCIS (Netherlands)

    Wegman, F.; Poldervaart, M. T.; van der Helm, Y. J.; Oner, F. C.; Dhert, W. J.; Alblas, J.

    2015-01-01

    Bone morphogenetic protein-2 (BMP-2) gene delivery has shown to induce bone formation in vivo in cell-based tissue engineering. In addition, the chemoattractant stromal cell-derived factor-1α (SDF-1α, also known as CXCL12) is known to recruit multipotent stromal cells towards its release site where

  1. The effect of nicotine on osteoinduction by recombinant human bone morphogenetic protein 2.

    Science.gov (United States)

    Tamura, K; Togo, Y; Kaihara, S; Hussain, A; Takahashi, K; Bessho, K

    2014-08-01

    Nicotine, one of the constituents of tobacco, is known to have an adverse effect on human health. We sought to clarify the interaction between nicotine and recombinant human bone morphogenetic protein 2 (rhBMP-2) in terms of osteogenesis in vitro and osteoinduction in vivo. Nicotine did not inhibit or stimulate alkaline phosphatase (ALP) activity or the amount of osteocalcin in C2C12 cells in the presence of rhBMP-2 in vitro. Ectopic bone formation using a collagen sponge containing rhBMP-2 was evaluated with and without nicotine after 21 days using radiographic, histological, biochemical, and immunohistochemical analyses. ALP activity in the medium-dose group (2.2±0.9IU/mg protein; P=0.047) and the high-dose group (2.0±0.1IU/mg protein; P=0.03) was significantly lower than in the control group. The calcium content in the medium-dose group (35.4±12.9μg/mg tissue; P=0.0099) and high-dose group (34.8±10.5μg/mg tissue; P=0.006) was significantly lower than in the control group. The number of vascular endothelial growth factor-positive cells in the high-dose group (671.9±57.3cells/mm(2); P=0.03) was significantly lower than in the control group. Results showed that nicotine did not inhibit the stimulatory effect of rhBMP-2 in vitro, but a high dose of nicotine inhibited bone formation in vivo by adversely affecting vascularization.

  2. RETINOIC ACID DOWN-REGULATES BONE MORPHOGENETIC PROTEIN 7 EXPRESSION IN RAT WITH CLEFT PALATE

    Institute of Scientific and Technical Information of China (English)

    Lei Guo; Yu-yan Zhao; Shi-liang Zhang; Kui Liu; Xiao-yu Gao

    2008-01-01

    Objective To evaluate the effects of retinoic acid (RA) on expression of bone morphogenetic protein 7 (BMP-7)in rat fetus with cleft palate, and the effects of RA on proliferation and apoptosis of osteoblasts. Methods All-trans RA (ATRA) was used to induce congenital cleft palate in Wistar rat. BMP-7 mRNA expres-sion in maxillary bone tissue of fetal rats was measured by Northern blotting analysis. Flow cytometry and MTT assay were used to measure the apoptosis and proliferation of ATRA-treated MC-3T3-E1 cells. BMP-7 mRNA and protein ex-pressions in ATRA-treated MC-3T3-E1 cells were detected by RT-PCR and Western blotting analysis.Results ATRA could induce cleft palate of rat fetus. The incidence rate of cleft palate induced by 100 mg/kg AT-RA (45.5%) was significantly higher than 50 mg/kg ATRA (12.5%, P<0.05). BMP-7 mRNA expression de-creased in maxillary bone tissue of rat fetus with cleft palate. MC-3T3-E1 cells proliferation treated with 1 × 10-6 mol/L ATRA decreased by 60%, the cell apoptosis increased by 2 times. BMP-7 mR.NA and protein levels in MC-3T3-E1cells treated with 1 × 10-6 mol/L ATRA decreased by 60% and 80%, respectively, compared with ATRA-untreated ceils (P<0.05).Conclusions BMP-7 may play an important role in embryonic palate development RA may possess the ability to down-regulate cell proliferation through regulation of BMP-7 gene expression.

  3. Construction of Adeno-associated Virus System for Human Bone Morphogenetic Protein 7 Gene

    Institute of Scientific and Technical Information of China (English)

    Ke SONG; Nianjing RAO; Meiling CHEN; Yingguang CAO

    2008-01-01

    To construct the recombinant adeno-associated virus (rAAV) vector with human bone morphogenetic protein 7 (BMP7) and observe the BMP7 mRNA expression in vitro, BMP7 CDS se- quence was cloned into expression plasmid pAAV-MCS of AAV Helper Free System. The recombi- nant plasmid was identified with enzyme digestion and sequencing. The recombinant plasmid, pAAV-RC, pHelper were co-transfected into AAV-293 cells according to the calcium phosphate-based protocol. The viral stock was collected by 4 rounds of freeze/thaw. After purified and concentrated,the recombinant virus titer was determined by dot-blot assay. HEK293 cells were transfected with the recombinant virus at different MOI, and the expression of BMP7 mRNA was detected by RT-PCR. The results showed rAAV-BMP7 was constructed and packaged successfully. The physical particle titer was 2.5×1011 vector genomes/mL. There was different expression level of BMP7 mRNA after transfecton. These data suggested that recombinant AAV mediated a stable expression of hBMP7 mRNA in 293 cells. The AAV production method may pave the way of an effective strategy for the jaw bone defection around dental implants.

  4. The importance and the differences of bone morphogenetic proteins for osteoporotic hip fractures.

    Science.gov (United States)

    Dincel, V Ercan; Sepici-Dincel, Aylin

    2014-06-01

    Bone morphogenetic proteins (BMPs), major contributors to tissue repair, have become one of the most exciting fields in rheumatic and orthopaedic research. In our study we aimed to evaluate the relationship between osteoporotic hip fractures and the serum levels of BMPs to reveal their potential roles in the diagnosis of patients. The study group included 62 patients with osteoporotic hip fracture (Group 1; intertrochanteric fracture, Group 2; collum femoris fracture) and the control group. All fractures were due to low energy trauma, simple falls. For all subjects BMD measurements were in agreement for osteoporosis and no significant differences were observed between the two fracture groups. Biochemical markers; BMP-4 and BMP-7 (pg/mL) were determined by commercial Elisa kits from the serum samples. The mean and standard error values of serum samples for BMP-4 and BMP-7 in Group 1 (100.70 +/- 10.03, 74.41 +/- 6.31 respectively) and in Group 2 (112.34 +/- 11.52, 81.91 +/- 10.14 respectively) were not statistically different however for both groups only BMP-7 values increased statistically when compared to the control group. BMP-7 measurements may not only serve as potential biochemical markers for determining disease severity but also the increased levels, an osteogenic factor and bone stimulating agent in vivo, after trauma elevated levels are adaptive or protective and therefore may reduce the severity of the fracture.

  5. Effects of bone morphogenetic protein-7 stimulation on osteoblasts cultured on different biomaterials.

    Science.gov (United States)

    Açil, Yahya; Springer, Ingo N G; Broek, Vanessa; Terheyden, Hendrik; Jepsen, Søren

    2002-01-01

    The objective of the present study was to investigate the effects of an in vitro stimulation of human osteoblasts by recombinant human bone morphogenetic protein-7 (rhBMP-7) on the collagen types and the quantity of the collagen cross-links synthesized in a three-dimensional culture on various biomaterials for bone replacement. Trabecular bone chips were harvested from human iliac crests, and cell cultures were established at standard conditions. One hundred and fifty nanograms per milliliter of rhBMP-7 was added. For the second passage a cell scraper was used to bring the cells into suspension, and 100 microl osteoblasts (at a density of 3.3 x 10(5)) were transferred onto nine blocks of either Bio-Oss, Tutoplast, or PepGen p-15. Blocks incubated with cells that were not treated with rhBMP-7 served as controls. Cell colonization of the biomaterials was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) after a period of 2, 4, and 6 weeks. Throughout the experiment medium, supernatants were collected and collagen was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Finally, the collagen cross-link residues hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) were quantified by HPLC. Within 4 weeks the cells became confluent on all of the studied biomaterials. All samples synthesized bone specific LP and collagen type I. However, in rhBMP-7-stimulated samples, the amount of HP and LP found was increased by 45% compared to non-stimulated samples. Cell proliferation and collagen synthesis was similar on the different biomaterials, but was consistently reduced in specimen not stimulated with rhBMP-7. In vitro stimulation of osteoblasts on Bio-Oss, Tutoplast, or PepGen p-15 with rhBMP-7 and subsequent transplantation of the constructs might lead to an enhanced osseointegration of the biomaterials in vivo.

  6. The Effects of Bone Morphogenetic Protein 2 Gene Transfection on Fibroblasts

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    目的:探讨骨形成蛋白2(bone morphogenetic protein 2,BMP2)基因转染对成纤维细胞成骨表型的调控作用.方法:利用脂质体将包含BMP2 cDNA全长编码序列的表达载体转染至NIH3T3细胞,原位杂交和免疫组化分别检测BMP2在NIH3T3细胞内的稳定转染和表达,同时观察转染细胞的增殖能力及成骨标志物包括碱性磷酸酶(alkaline phosphatase,ALP)活力和骨钙素(osteocalcin,OC)含量的变化.结果:BMP2只在转染细胞内表达.与未转染细胞相比,BMP2基因转染细胞的增殖能力降低,而ALP活力和OC含量增加.结论:结果表明BMP2基因转染能够衣导成纤维细胞的体外成骨潜能.%Objective: To explore the regulatory effects of Bone Morphogenetic Protein 2 (BMP2) gene transfection on the phenotype of fibroblasts. Methods: A phagemid expression vector containing the full length of human BMP2 cDNA coding sequence was transfected into NIH3T3 cells by using LipofectAMINE. The stable transfection and expression of BMP2 in NIH3T3 cells were determined by in situ hybridization and immunohistochemistry, respectively. The proliferation and the markers for osteogenic features, including alkaline phosphatase (碱性磷酸酶, ALP) activity and osteocalcin (骨钙素, OC) production were also investigated in the transfected cells. Results: The results showed that BMP2 was only expressed in the transfected cells. Compared with the non-transfected cells, the BMP2 gene transfected cells showed decreased proliferative ability, but enhanced both ALP activity and OC production (P < 0.05). Conclusions: The results indicate that BMP2 gene transfection can induce the osteogenic potential of fibroblasts in vitro.

  7. Regulation of Oligodendrocyte Progenitor Cell Maturation by PPARδ: Effects on Bone Morphogenetic Proteins

    Directory of Open Access Journals (Sweden)

    Maria Vittoria Simonini

    2009-12-01

    Full Text Available In EAE (experimental autoimmune encephalomyelitis, agonists of PPARs (peroxisome proliferator-activated receptors provide clinical benefit and reduce damage. In contrast with PPARγ, agonists of PPARδ are more effective when given at later stages of EAE and increase myelin gene expression, suggesting effects on OL (oligodendrocyte maturation. In the present study we examined effects of the PPARδ agonist GW0742 on OPCs (OL progenitor cells, and tested whether the effects involve modulation of BMPs (bone morphogenetic proteins. We show that effects of GW0742 are mediated through PPARδ since no amelioration of EAE clinical scores was observed in PPARδ-null mice. In OPCs derived from E13 mice (where E is embryonic day, GW0742, but not the PPARγ agonist pioglitazone, increased the number of myelin-producing OLs. This was due to activation of PPARδ since process formation was reduced in PPARδ-null compared with wild-type OPCs. In both OPCs and enriched astrocyte cultures, GW0742 increased noggin protein expression; however, noggin mRNA was only increased in astrocytes. In contrast, GW0742 reduced BMP2 and BMP4 mRNA levels in OPCs, with lesser effects in astrocytes. These findings demonstrate that PPARδ plays a role in OPC maturation, mediated, in part, by regulation of BMP and BMP antagonists.

  8. Purification of bone morphogenetic protein-2 from refolding mixtures using mixed-mode membrane chromatography.

    Science.gov (United States)

    Gieseler, Gesa; Pepelanova, Iliyana; Stuckenberg, Lena; Villain, Louis; Nölle, Volker; Odenthal, Uwe; Beutel, Sascha; Rinas, Ursula; Scheper, Thomas

    2017-01-01

    In this study, we present the development of a process for the purification of recombinant human bone morphogenetic protein-2 (rhBMP-2) using mixed-mode membrane chromatography. RhBMP-2 was produced as inclusion bodies in Escherichia coli. In vitro refolding using rapid dilution was carried out according to a previously established protocol. Different membrane chromatography phases were analyzed for their ability to purify BMP-2. A membrane phase with salt-tolerant properties resulting from mixed-mode ligand chemistry was able to selectively purify BMP-2 dimer from refolding mixtures. No further purification or polishing steps were necessary and high product purity was obtained. The produced BMP-2 exhibited a biological activity of 7.4 × 10(5) U/mg, comparable to commercial preparations. Mixed-mode membrane chromatography can be a valuable tool for the direct purification of proteins from solutions with high-conductivity, for example refolding buffers. In addition, in this particular case, it allowed us to circumvent the use of heparin-affinity chromatography, thus allowing the design of an animal-component-free process.

  9. Imaging bone morphogenetic protein 7 induced cell cycle arrest in experimental gliomas.

    Science.gov (United States)

    Klose, Anke; Waerzeggers, Yannic; Monfared, Parisa; Vukicevic, Slobodan; Kaijzel, Eric L; Winkeler, Alexandra; Wickenhauser, Claudia; Löwik, Clemens W G M; Jacobs, Andreas H

    2011-03-01

    Bone morphogenetic protein 7 (BMP-7) belongs to the superfamily of transforming growth factor β-like cytokines, which can act either as tumor suppressors or as tumor promoters depending on cell type and differentiation. Our investigations focused on analyzing the effects of BMP-7 during glioma cell proliferation in vitro and in vivo. BMP-7 treatment decreased the proliferation of Gli36ΔEGFR-LITG glioma cells up to 50%through a cell cycle arrest in the G(1) phase but not by induction of apoptosis. This effect was mediated by the modulation of the expression and phosphorylation of cyclin-dependent kinase 2, cyclin-dependent kinase inhibitor p21, and downstream retinoblastoma protein. Furthermore, in vivo optical imaging of luciferase activity of Gli36ΔEGFR-LITG cells implanted intracranially into nude mice in the presence or absence of BMP-7 treatment corroborated the antiproliferative effects of this cytokine. This report clearly underlines the tumor-suppressive role of BMP-7 in glioma-derived cells. Taken together, our results indicate that manipulating the BMP/transforming growth factor β signaling cascade may serve as a new strategy for imaging-guided molecular-targeted therapy of malignant gliomas.

  10. Imaging Bone Morphogenetic Protein 7 Induced Cell Cycle Arrest in Experimental Gliomas

    Directory of Open Access Journals (Sweden)

    Anke Klose

    2011-03-01

    Full Text Available Bone morphogenetic protein 7 (BMP-7 belongs to the superfamily of transforming growth factor β-like cytokines, which can act either as tumor suppressors or as tumor promoters depending on cell type and differentiation. Our investigations focused on analyzing the effects of BMP-7 during glioma cell proliferation in vitro and in vivo. BMP-7 treatment decreased the proliferation of Gli36ΔEGFR-LITG glioma cells up to 50%through a cell cycle arrest in the G1 phase but not by induction of apoptosis. This effect was mediated by the modulation of the expression and phosphorylation of cyclin-dependent kinase 2, cyclin-dependent kinase inhibitor p21, and downstream retinoblastoma protein. Furthermore, in vivo optical imaging of luciferase activity of Gli36ΔEGFR-LITG cells implanted intracranially into nude mice in the presence or absence of BMP-7 treatment corroborated the antiproliferative effects of this cytokine. This report clearly underlines the tumor-suppressive role of BMP-7 in glioma-derived cells. Taken together, our results indicate that manipulating the BMP/transforming growth factor β signaling cascade may serve as a new strategy for imaging-guided molecular-targeted therapy of malignant gliomas.

  11. Bone morphogenetic protein-4 enhances vascular endothelial growth factor secretion by human retinal pigment epithelial cells.

    Science.gov (United States)

    Vogt, Rhonda R; Unda, Richard; Yeh, Lee-Chuan C; Vidro, Eileen K; Lee, John C; Tsin, Andrew T

    2006-08-01

    Retinal pigment epithelial (RPE) cells secrete vascular endothelial growth factor (VEGF), a cytokine known to promote angiogenesis. Results from RNase protection assays (RPAs) show that RPE from non-diabetic human donors and from adult retinal pigment epithelium-19 (ARPE-19) cells expressed significant bone morphogenetic protein-4 (BMP-4) message. In addition, ARPE-19 cells cultured in high glucose (25 mM), compared to those in physiological glucose (5.5 mM) released significantly more BMP-4 into the conditioned media (CM). However, the effect of BMP-4 on the release of VEGF by ARPE-19 cells has not been studied. Accordingly, ARPE-19 cells were treated with BMP-4 to determine VEGF secretion. BMP-4 and VEGF levels in the CM and cell lysates were measured by enzyme-linked immunosorbent assay (ELISA). Cells treated with exogenous BMP-4 had higher VEGF in the CM and this treatment effect was dose- and time-dependent, while cell lysates had low levels of VEGF. Addition of cycloheximide (CHX) or actinomycin-D (ACT) significantly reduced VEGF secretion from cells treated with BMP-4, suggesting that the BMP-4-induced secretion of VEGF requires new RNA and protein synthesis. Our results suggest that BMP-4 may play a role in the regulation of ocular angiogenesis associated with diabetic retinopathy (DR) by stimulating VEGF release from RPE cells.

  12. Immature muscular tissue differentiation into bone-like tissue by bone morphogenetic proteins in vitro, with ossification potential in vivo.

    Science.gov (United States)

    Hayashi, Tatsuhide; Kobayashi, Syuichiro; Asakura, Masaki; Kawase, Mayu; Ueno, Atsuko; Uematsu, Yasuaki; Kawai, Tatsushi

    2014-09-01

    The objective of this study was to induce bone formation from immature muscular tissue (IMT) in vitro, using bone morphogenetic proteins (BMPs) as a cytokine source and an expanded polytetrafluoroethylene (ePTFE) scaffold. In addition, cultured IMTs were implanted subcutaneously into Sprague-Dawley (SD) rats to determine their in vivo ossification potential. BMPs, extracted from bovine cortical bones, were applied to embryonic SD rat IMT cultures, before 2 weeks culture on ePTFE scaffolds. Osteoblast-like cells and osteoid tissues were partially identified by hematoxylin-eosin staining 2 weeks after culture. Collagen type I (Col-I), osteopontin (OP), and osteocalcin (OC) were detected in the osteoid tissues by immunohistochemical staining. OC gene expression remained low, but OP and Col-I were upregulated during the culture period. In vivo implanted IMTs showed slight radiopacity 1 week after implantation and strong radiopacity 2 and 3 weeks after implantation. One week after implantation, migration of numerous capillaries was observed and ossification was detected after 2 weeks by histological observation. These results suggest that IMTs are able to differentiate into bone-like tissue in vitro, with an ossification potential after implantation in vivo.

  13. Delivery Systems for Bone Morphogenetic Protein (BMP) for Repair of Battle Incurred Bone Injuries.

    Science.gov (United States)

    1987-11-01

    infections, congenital malformations that fail to heal are eligible for BMP treatment. I (my child/my ward) will be one of 50 patients to be treated with...Fusions in Dogs 6. Craniotomy Defects in Sheep t0 7. Craniotomy Defects in Monkeys 10 8. BMP Delivery System of Bone Matrix Non Collagenous 11 Proteins...effects. The most important and indispensptle substitutes for experiments in human beings are adult mongrel dogs, monkeys, and sheep . Experimental .S

  14. Effect of human bone morphogenetic protein 2 implant on tooth eruption in an experimental design.

    Science.gov (United States)

    Steinberg, B; Chiego, D J; Huizinga, P J; Wozney, J M; Wikesjö, U M

    1999-07-01

    This study evaluated the influence of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the development and eruption of the secondary dentition. Primary premolar tooth extraction sockets in 12 16-week-old felines were implanted with either rhBMP-2, in collagen sponge or with buffer/absorbable collagen sponge (ACS). Unoperated jaw quadrants served as controls. Experimental conditions were randomized between jaw quadrants in all animals. Two animals receiving rhBMP-2/ACS and buffer/ACS in two quadrants per implant were sacrificed at 4 weeks postsurgery. Ten animals receiving rhBMP-2/ACS (two quadrants), buffer/ACS implants (one quadrant), and one quadrant serving as an unoperated control were evaluated at 12 weeks postsurgery. Clinical assessments included healing, eruption patterns, and crown development. Radiographic assessments included tooth development, eruption patterns, and bone formation. Histological observations were also made from the 4-week animals. The secondary dentition remained unerupted at 4 weeks postsurgery. Histological analysis showed normal alveolar bone coronal to the erupting teeth in rhBMP-2/ACS-implanted quadrants. At 12 weeks postsurgery, all teeth were erupted without differences between quadrants. Clinically, the crowns of all teeth were normal. Radiographs suggested that teeth in rhBMP-2/ACS- and buffer/ACS-implanted jaw quadrants exhibited similar tooth development and eruption patterns as the normal control. The evidence from this study suggests that surgical implantation of rh-BMP-2/ACS in the pathway of the developing and erupting secondary dentition does not interfere with the normal development and eruption patterns of the teeth.

  15. Bone morphogenetic protein 6 polymorphisms are associated with radiographic progression in ankylosing spondylitis.

    Directory of Open Access Journals (Sweden)

    Young Bin Joo

    Full Text Available Nearly 25 genetic loci associated with susceptibility to ankylosing spondylitis (AS have been identified by several large studies. However, there have been limited studies to identify the genes associated with radiographic severity of the disease. Thus we investigated which genes involved in bone formation pathways might be associated with radiographic severity in AS.A total of 417 Korean AS patients were classified into two groups based on the radiographic severity as defined by the modified Stoke' Ankylosing Spondylitis Spinal Score (mSASSS system. Severe AS was defined by the presence of syndesmophytes and/or fusion in the lumbar or cervical spine (n = 195. Mild AS was defined by the absence of any syndesmophyte or fusion (n = 170. A total of 251 single nucleotide polymorphisms (SNPs within 52 genes related to bone formation were selected and genotyped. Odds ratios (OR and 95% confidence interval (95% CI were analysed by multivariate logistic regression controlling for age at onset of symptoms, sex, disease duration, and smoking status as covariates.We identified new loci of bone morphogenetic protein 6 (BMP6 associated with radiographic severity in patients with AS that passed false discovery rate threshold. Two SNPs in BMP6 were significantly associated with radiologic severity [rs270378 (OR 1.97, p = 6.74 × 10(-4 and rs1235192 [OR 1.92, p = 1.17 × 10(-3] adjusted by covariates.This is the first study to demonstrate that BMP6 is associated with radiographic severity in AS, supporting the role wingless-type like/BMP pathway on radiographic progression in AS.

  16. Recombinant human bone morphogenetic protein-2 suspended in fibrin glue enhances bone formation during distraction osteogenesis in rabbits

    Science.gov (United States)

    Li, Yunfeng; Li, Rui; Hu, Jing; Song, Donghui; Jiang, Xiaowen

    2016-01-01

    Introduction Bone morphogenetic protein-2 (BMP-2) has high potential for bone formation, but its in vivo effects are unpredictable due to the short life time. This study was designed to evaluate the effects of recombinant human (rh) BMP-2 suspended in fibrin on bone formation during distraction osteogenesis (DO) in rabbits. Material and methods The in vitro release kinetics of rhBMP-2 suspended in fibrin was tested using an enzyme-linked immunosorbent assay. Unilateral tibial lengthening for 10 mm was achieved in 48 rabbits. At the completion of osteodistraction, vehicle, fibrin, rhBMP-2 or rhBMP-2 suspended in fibrin (rhBMP-2 + fibrin) was injected into the center of the lengthened gap, with 12 animals in each group. Eight weeks later, the distracted callus was examined by histology, micro-CT and biomechanical testing. Radiographs of the distracted tibiae were taken at both 4 and 8 weeks after drug treatment. Results It was found that fibrin prolonged the life span of rhBMP-2 in vitro with sustained release during 17 days. The rhBMP-2 + fibrin treated animals showed the best results in bone mineral density, bone volume fraction, cortical bone thickness by micro-CT evaluation and mechanical properties by the three-point bending test when compared to the other groups (p < 0.05). In histological images, rhBMP-2 + fibrin treatment showed increased callus formation and better gap bridging compared to the other groups. Conclusions The results of this study suggest that fibrin holds promise to be a good carrier of rhBMP-2, and rhBMP-2 suspended in fibrin showed a stronger promoting effect on bone formation during DO in rabbits. PMID:27279839

  17. Stable expression and characterization of N-terminal tagged recombinant human bone morphogenetic protein 15

    Science.gov (United States)

    Li, Qinglei; Rajanahally, Saneal; Edson, Mark A.; Matzuk, Martin M.

    2009-01-01

    Oocyte-derived growth factors are critically involved in multiple ovarian processes via paracrine actions. Although recombinant proteins have been applied to dissect the physiological functions of these factors, variation of activities among different protein preparations remains an issue. To further elucidate the roles of one of these growth factors, bone morphogenetic protein 15 (BMP15), in mediating oocyte-regulated molecular and cellular events and to explore its potential clinical application, we engineered the human BMP15 sequence to efficiently produce bioactive recombinant human BMP15 (rhBMP15). The proteolytic cleavage site of the hBMP15 precursor was optimized to facilitate the production of the mature protein, and a FLAG-tag was placed at the N-terminus of the mature region to ease purification and avoid potential interference of the tag with the cystine knot structure. The rhBMP15 protein was purified using anti-FLAG M2 affinity gel. Our results demonstrated that the N-terminal tagged rhBMP15 was efficiently processed in HEK-293 cells. Furthermore, the purified rhBMP15 could activate SMAD1/5/8 and induce the transcription of genes encoding cumulus expansion-related transcripts (Ptx3, Has2, Tnfaip6 and Ptgs2), inhibitory SMADs (Smad6 and Smad7), BMP antagonists (Grem1 and Fst), activin/inhibin βA (Inhba) and βB (Inhbb) subunits, etc. Thus, our rhBMP15 containing a genetically modified cleavage sequence and an N-terminal FLAG-tag can be efficiently produced, processed and secreted in a mammalian expression system. The purified rhBMP15 is also biologically active and very stable, and can induce the expression of a variety of mouse granulosa cell genes. PMID:19651638

  18. The application of bone morphogenetic proteins to periodontal and peri-implant tissue regeneration: A literature review

    Directory of Open Access Journals (Sweden)

    Karuppanan P Sasikumar

    2012-01-01

    Full Text Available Progress in understanding the role of bone morphogenetic proteins (BMPs in craniofacial and tooth development and the demonstration of stem cells in periodontal ligament have set the stage for periodontal regenerative therapy and tissue engineering. Furthermore, recent approval by the Food and Drug Administration of recombinant human BMPs for accelerating bone fusion in slow-healing fractures indicates that this protein family may prove useful in designing regenerative treatments in periodontics. In the near term, these advances are likely to be applied to periodontal surgery; ultimately, they may facilitate approaches to regenerating whole lost periodontal structures.

  19. Bone Morphogenetic Proteins stimulate mammary fibroblasts to promote mammary carcinoma cell invasion.

    Directory of Open Access Journals (Sweden)

    Philip Owens

    Full Text Available Bone Morphogenetic Proteins (BMPs are secreted cytokines that are part of the Transforming Growth Factor β (TGFβ superfamily. BMPs have been shown to be highly expressed in human breast cancers, and loss of BMP signaling in mammary carcinomas has been shown to accelerate metastases. Interestingly, other work has indicated that stimulation of dermal fibroblasts with BMP can enhance secretion of pro-tumorigenic factors. Furthermore, treatment of carcinoma-associated fibroblasts (CAFs derived from a mouse prostate carcinoma with BMP4 was shown to stimulate angiogenesis. We sought to determine the effect of BMP treatment on mammary fibroblasts. A large number of secreted pro-inflammatory cytokines and matrix-metallo proteases (MMPs were found to be upregulated in response to BMP4 treatment. Fibroblasts that were stimulated with BMP4 were found to enhance mammary carcinoma cell invasion, and these effects were inhibited by a BMP receptor kinase antagonist. Treatment with BMP in turn elevated pro-tumorigenic secreted factors such as IL-6 and MMP-3. These experiments demonstrate that BMP may stimulate tumor progression within the tumor microenvironment.

  20. Effective Inhibition of Bone Morphogenetic Protein Function by Highly Specific Llama-Derived Antibodies.

    Science.gov (United States)

    Calpe, Silvia; Wagner, Koen; El Khattabi, Mohamed; Rutten, Lucy; Zimberlin, Cheryl; Dolk, Edward; Verrips, C Theo; Medema, Jan Paul; Spits, Hergen; Krishnadath, Kausilia K

    2015-11-01

    Bone morphogenetic proteins (BMP) have important but distinct roles in tissue homeostasis and disease, including carcinogenesis and tumor progression. A large number of BMP inhibitors are available to study BMP function; however, as most of these antagonists are promiscuous, evaluating specific effects of individual BMPs is not feasible. Because the oncogenic role of the different BMPs varies for each neoplasm, highly selective BMP inhibitors are required. Here, we describe the generation of three types of llama-derived heavy chain variable domains (VHH) that selectively bind to either BMP4, to BMP2 and 4, or to BMP2, 4, 5, and 6. These generated VHHs have high affinity to their targets and are able to inhibit BMP signaling. Epitope binning and docking modeling have shed light into the basis for their BMP specificity. As opposed to the wide structural reach of natural inhibitors, these small molecules target the grooves and pockets of BMPs involved in receptor binding. In organoid experiments, specific inhibition of BMP4 does not affect the activation of normal stem cells. Furthermore, in vitro inhibition of cancer-derived BMP4 noncanonical signals results in an increase of chemosensitivity in a colorectal cancer cell line. Therefore, because of their high specificity and low off-target effects, these VHHs could represent a therapeutic alternative for BMP4(+) malignancies.

  1. Bone morphogenetic protein receptor II regulates pulmonary artery endothelial cell barrier function.

    Science.gov (United States)

    Burton, Victoria J; Ciuclan, Loredana I; Holmes, Alan M; Rodman, David M; Walker, Christoph; Budd, David C

    2011-01-06

    Mutations in bone morphogenetic protein receptor II (BMPR-II) underlie most heritable cases of pulmonary arterial hypertension (PAH). However, less than half the individuals who harbor mutations develop the disease. Interestingly, heterozygous null BMPR-II mice fail to develop PAH unless an additional inflammatory insult is applied, suggesting that BMPR-II plays a fundamental role in dampening inflammatory signals in the pulmonary vasculature. Using static- and flow-based in vitro systems, we demonstrate that BMPR-II maintains the barrier function of the pulmonary artery endothelial monolayer suppressing leukocyte transmigration. Similar findings were also observed in vivo using a murine model with loss of endothelial BMPR-II expression. In vitro, the enhanced transmigration of leukocytes after tumor necrosis factor α or transforming growth factor β1 stimulation was CXCR2 dependent. Our data define how loss of BMPR-II in the endothelial layer of the pulmonary vasculature could lead to a heightened susceptibility to inflammation by promoting the extravasation of leukocytes into the pulmonary artery wall. We speculate that this may be a key mechanism involved in the initiation of the disease in heritable PAH that results from defects in BMPR-II expression.

  2. In anemia of multiple myeloma, hepcidin is induced by increased bone morphogenetic protein 2

    Science.gov (United States)

    Maes, Ken; Nemeth, Elizabeta; Roodman, G. David; Huston, Alissa; Esteve, Flavia; Freytes, Cesar; Callander, Natalie; Katodritou, Eirini; Tussing-Humphreys, Lisa; Rivera, Seth; Vanderkerken, Karin; Lichtenstein, Alan

    2010-01-01

    Hepcidin is the principal iron-regulatory hormone and a pathogenic factor in anemia of inflammation. Patients with multiple myeloma (MM) frequently present with anemia. We showed that MM patients had increased serum hepcidin, which inversely correlated with hemoglobin, suggesting that hepcidin contributes to MM-related anemia. Searching for hepcidin-inducing cytokines in MM, we quantified the stimulation of hepcidin promoter-luciferase activity in HuH7 cells by MM sera. MM sera activated the hepcidin promoter significantly more than did normal sera. We then examined the role of bone morphogenetic proteins (BMPs) and interleukin-6 (IL-6), the major transcriptional regulators of hepcidin. Mutations in both BMP-responsive elements abrogated the activation dramatically, while mutations in the IL-6–responsive signal transducer and activator of transcription 3-binding site (STAT3-BS) had only a minor effect. Cotreatment with anti–BMP-2/4 or noggin-Fc blocked the promoter induction with all MM sera, anti–IL-6 blocked it with a minority of sera, whereas anti–BMP-4, -6, or -9 antibodies had no effect. BMP-2–immunodepleted MM sera had decreased promoter stimulatory capacity, and BMP-2 concentrations in MM sera were significantly higher than in normal sera. Our results demonstrate that BMP-2 is a major mediator of the hepcidin stimulatory activity of MM sera. PMID:20679527

  3. Osteoinductivity assay of the variability of repeated extractions of bone morphogenetic proteins from bovine bone at different times

    Institute of Scientific and Technical Information of China (English)

    HU Zhen-ming 胡侦明; Sean AF Peel; Cameron ML Clokie

    2004-01-01

    Objective:To observe the activity of repeated extracts of bone matrix and the production of purified bone morphogenetic proteins (BMPs).Methods: BMPs were extracted 1- 4 times from fresh bovine cortical bone by the modified Urist's method, with each collected precipitate separated and lyophilized as partially purified BMPs. Another fresh bovine bone was extracted three times and the precipitates were mixed and lyophilized. Meanwhile, the alkaline phosphatase (ALP)activity was measured by an in vitro assay employing cultured C2C12 mouse myoblast cells through the osteoinductivity of bovine BMPs extracted four times at days 1, 4, 7, and 14, and the correlation between BMPs quantities and costing during extraction processes was analyzed.Results:The purified and the cost showed a positive correlation(r=0.969).To separate and lyophilize each collected precipitate as partially purified BMPs raised the cost,and mixed precipitates also cost much.ALPactivities of 1st and mixed extractions of BMPs were shown to be highly osteoinductive and keep a significantly high level(P<0.05-0.01)4 days after culturing compared with the 2nd,3rd and 4th extractions,especially the control group.However,the more times the extraction ws done,the less activity of BMPs was shown and more costing was.The x-ray and histological analysis also showed that the 1st extraction of BMPs induced more ossicles and new bone formation.Conclusions:The results indicated that BMPs enhanced the abilities of osteoinductiviyt in C2C12 culture in vitro.The first extraction of BMPsfrom bone is fitfull,4th extractions are unnecessary for they cost more and waste more time,say nothing of mixed extractions.

  4. Bone morphogenetic protein signaling protects against cerulein-induced pancreatic fibrosis.

    Directory of Open Access Journals (Sweden)

    Xuxia Gao

    Full Text Available Bone morphogenetic proteins (BMPs have an anti-fibrogenic function in the kidney, lung, and liver. However, their role in chronic pancreatitis (CP is unknown. The aim of this study was to define the anti-fibrogenic role of BMP signaling in the pancreas in vivo under CP induction. Mice with a deletion of BMP type II receptor (BMPR2(+/- were used in this study in comparison with wild-type mice. CP was induced by repetitive cerulein injection intraperitoneally for 4 weeks, and the severity of CP was evaluated. Pancreatic stellate cells (PSCs were isolated from the mice and treated with BMP2 and TGF-β in vitro, and extracellular matrix protein (ECM production was measured. Smad and mitogen-activated protein kinase (MAPK signaling was also evaluated. BMPR2(+/- mice revealed a greater pancreatic fibrosis, PSC activation and leukocyte infiltration after CP induction compared to wild-type mice (P<0.05. Under CP induction, phospho (pSmad1/5/8 was elevated in wild-type mice and this effect was abolished in BMPR2(+/- mice; pSmad2 and pp38(MAPK were further enhanced in BMPR2(+/- mice compared to wild-type mice (P<0.05. In vitro, BMP2 inhibited TGF-β-induced ECM protein fibronectin production in wild-type PSCs; this effect was abolished in BMPR2(+/- PSCs (P<0.05. In BMPR2(+/- PSCs, pSmad1/5/8 level was barely detectable upon BMP2 stimulation, while pSmad2 level was further enhanced by TGF-β stimulation, compared to wild-type PSCs (P<0.05. BMPR2/Smad1/5/8 signaling plays a protective role against cerulein-induced pancreatic fibrosis by inhibiting Smad2 and p38(MAPK signaling pathways.

  5. Use of bone morphogenetic proteins in mesenchymal stemcell stimulation of cartilage and bone repair

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    The extracellular matrix-associated bone morphogeneticproteins (BMPs) govern a plethora of biological processes.The BMPs are members of the transforming growthfactor-β protein superfamily, and they actively participateto kidney development, digit and limb formation,angiogenesis, tissue fibrosis and tumor development.Since their discovery, they have attracted attentionfor their fascinating perspectives in the regenerativemedicine and tissue engineering fields. BMPs havebeen employed in many preclinical and clinical studiesexploring their chondrogenic or osteoinductive potentialin several animal model defects and in human diseases.During years of research in particular two BMPs, BMP2and BMP7 have gained the podium for their use inthe treatment of various cartilage and bone defects.In particular they have been recently approved foremployment in non-union fractures as adjunct therapies.On the other hand, thanks to their potentialities inbiomedical applications, there is a growing interest instudying the biology of mesenchymal stem cell (MSC),the rules underneath their differentiation abilities, andto test their true abilities in tissue engineering. In fact,the specific differentiation of MSCs into targeted celltypelineages for transplantation is a primary goal of theregenerative medicine. This review provides an overviewon the current knowledge of BMP roles and signaling inMSC biology and differentiation capacities. In particularthe article focuses on the potential clinical use of BMPsand MSCs concomitantly, in cartilage and bone tissuerepair.

  6. Potential Roles of Bone Morphogenetic Protein (BMP-9 in Human Liver Diseases

    Directory of Open Access Journals (Sweden)

    Blanca Herrera

    2014-03-01

    Full Text Available Bone morphogenetic proteins (BMP-2 to BMP-15 belong to the Transforming Growth Factor (TGF-β superfamily and, besides their well-documented roles during embryogenesis and bone formation, some of them have recently been described to be involved in the pathogenesis of different organs, including the liver. The role of BMPs in liver damage responses including hepatocellular carcinoma (HCC development has only begun to be addressed and strong evidence supports the concept of a pro-tumorigenic role of BMP signaling in HCC cells. BMP-9 (also termed Growth and Differentiation Factor (GDF-2 represents the most recently discovered member of the BMP family. We have previously demonstrated that in HCC patient samples BMP-9 expression was positively associated with the tumor seize (“T stage” and that it enhanced cell migration and induced epithelial to mesenchymal transition (EMT in HCC cells in vitro. In another study we recently found that BMP-9 promotes growth in HCC cells, but not in non-transformed hepatocytes. Published as well as unpublished results obtained with primary hepatocytes support the concept of a dual function of BMP-9 in the liver: while in primary, non-malignant cells BMP-9 stabilizes the epithelial phenotype and inhibits proliferation, in HCC cells it induces cell growth and the acquisition of a migratory phenotype. In this review article we summarize current knowledge about BMPs in liver diseases, with special focus on the role of BMP-9 in HCC development and progression, that may provide new clues for a better understanding of the contribution of BMP-signaling to chronic liver diseases.

  7. Potential Roles of Bone Morphogenetic Protein (BMP)-9 in Human Liver Diseases

    Science.gov (United States)

    Herrera, Blanca; Dooley, Steven; Breitkopf-Heinlein, Katja

    2014-01-01

    Bone morphogenetic proteins (BMP-2 to BMP-15) belong to the Transforming Growth Factor (TGF)-β superfamily and, besides their well-documented roles during embryogenesis and bone formation, some of them have recently been described to be involved in the pathogenesis of different organs, including the liver. The role of BMPs in liver damage responses including hepatocellular carcinoma (HCC) development has only begun to be addressed and strong evidence supports the concept of a pro-tumorigenic role of BMP signaling in HCC cells. BMP-9 (also termed Growth and Differentiation Factor (GDF)-2) represents the most recently discovered member of the BMP family. We have previously demonstrated that in HCC patient samples BMP-9 expression was positively associated with the tumor seize (“T stage”) and that it enhanced cell migration and induced epithelial to mesenchymal transition (EMT) in HCC cells in vitro. In another study we recently found that BMP-9 promotes growth in HCC cells, but not in non-transformed hepatocytes. Published as well as unpublished results obtained with primary hepatocytes support the concept of a dual function of BMP-9 in the liver: while in primary, non-malignant cells BMP-9 stabilizes the epithelial phenotype and inhibits proliferation, in HCC cells it induces cell growth and the acquisition of a migratory phenotype. In this review article we summarize current knowledge about BMPs in liver diseases, with special focus on the role of BMP-9 in HCC development and progression, that may provide new clues for a better understanding of the contribution of BMP-signaling to chronic liver diseases. PMID:24670474

  8. Basic science and spine literature document bone morphogenetic protein increases cancer risk

    Directory of Open Access Journals (Sweden)

    Nancy E Epstein

    2014-01-01

    Full Text Available Background: Increasingly, clinical articles document that bone morphogenetic protein (BMP/INFUSE: Medtronic, Memphis, TN, USA and its derivatives utilized in spinal surgery increase the risk of developing cancer. However, there is also a large body of basic science articles that also document that various types of BMP and other members of the TGF-Beta (transforming growth factor beta family promote the growth of different types of cancers. Methods: This review looks at many clinical articles citing BMP/INFUSE′s role, largely "off-label", in contributing to complications encountered during spinal surgery. Next, however, specific attention is given to the clinical and basic science literature regarding how BMP and its derivatives (e.g. members of the TGF-beta family may also impact the development of breast and other cancers. Results: Utilizing BMP/INFUSE in spine surgery increased the risk of cancers/new malignancy as documented in several studies. For example, Carragee et al. found that for single-level instrumented posterolateral fusions (PLF using high-dose rhBMP-2 (239 patients vs. autograft (control group; n = 224, the risks of new cancers at 2 and 5 years postoperatively were increased. In laboratory studies, BMP′s along with other members of the TGF-Beta family also modulated/contributed to the proliferation/differentiation of breast cancer (e.g. bone formation/turnover, breast cancer-related solid tumors, and metastases, lung, adrenal, and colon cancer. Conclusions: BMP/INFUSE when utilized clinically in spinal fusion surgery appears to promote cancer at higher rates than observed in the overall population. Furthermore, BMP and TGF-beta are correlated with increased cancer growth both in the clinic and the laboratory.

  9. Mode of heparin attachment to nanocrystalline hydroxyapatite affects its interaction with bone morphogenetic protein-2.

    Science.gov (United States)

    Goonasekera, Chandhi S; Jack, Kevin S; Bhakta, Gajadhar; Rai, Bina; Luong-Van, Emma; Nurcombe, Victor; Cool, Simon M; Cooper-White, Justin J; Grøndahl, Lisbeth

    2015-12-16

    Heparin has a high affinity for bone morphogenetic protein-2 (BMP-2), which is a key growth factor in bone regeneration. The aim of this study was to investigate how the rate of release of BMP-2 was affected when adsorbed to nanosized hydroxyapatite (HAP) particles functionalized with heparin by different methods. Heparin was attached to the surface of HAP, either via adsorption or covalent coupling, via a 3-aminopropyltriethoxysilane (APTES) layer. The chemical composition of the particles was evaluated using X-ray photoelectron spectroscopy and elemental microanalysis, revealing that the heparin grafting densities achieved were dependent on the curing temperature used in the fabrication of APTES-modified HAP. Comparable amounts of heparin were attached via both covalent coupling and adsorption to the APTES-modified particles, but characterization of the particle surfaces by zeta potential and Brunauer-Emmett-Teller measurements indicated that the conformation of the heparin on the surface was dependent on the method of attachment, which in turn affected the stability of heparin on the surface. The release of BMP-2 from the particles after 7 days in phosphate-buffered saline found that 31% of the loaded BMP-2 was released from the APTES-modified particles with heparin covalently attached, compared to 16% from the APTES-modified particles with the heparin adsorbed. Moreover, when heparin was adsorbed onto pure HAP, it was found that the BMP-2 released after 7 days was 5% (similar to that from unmodified HAP). This illustrates that by altering the mode of attachment of heparin to HAP the release profile and total release of BMP-2 can be manipulated. Importantly, the BMP-2 released from all the heparin particle types was found by the SMAD 1/5/8 phosphorylation assay to be biologically active.

  10. Anodic oxidized nanotubular titanium implants enhance bone morphogenetic protein-2 delivery.

    Science.gov (United States)

    Bae, In-Ho; Yun, Kwi-Dug; Kim, Hyun-Seung; Jeong, Byung-Chul; Lim, Hyun-Pil; Park, Sang-Won; Lee, Kwang-Min; Lim, Young-Chai; Lee, Kyung-Ku; Yang, Yunzhi; Koh, Jeong-Tae

    2010-05-01

    Implant failure has been attributed to loosening of an implant from the host bone possibly due to poor osseointegration. One promising strategy for improving osseointegration is to develop a functional implant surface that promotes osteoblast differentiation. In this study, a titanium (Ti) surface was functionalized by an anodic oxidation process and was loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2). The following four groups of Ti surfaces were prepared: machined (M), anodized machined (MA), resorbable blast medium (RBM), and anodized RBM (RBMA). The surfaces were characterized by scanning electron microscopy and contact angle measurements. The results showed that a Ti oxide layer (TiO(2)) was observed in the anodized surfaces in the form of nanotubes, approximately 100 nm in diameter and 500 nm in length. The hydrophilic properties of the anodized surfaces were significantly improved. The adsorbed rhBMP-2 loaded on the nonanodized surfaces and lyophilized showed spherical particle morphology. However, the adsorbed rhBMP-2 showed a dispersed pattern over the anodized surfaces. The velocity of the rhBMP-2 released from the surfaces was measured to determine if the anodized surface can improve in delivery efficiency. The results showed that the release velocity of the rhBMP-2 from the anodized surfaces was sustained when compared with that of the nonanodized surfaces. In addition, the rhBMP-2 released from the surface was found to be bioactive according to the alkaline phosphatase activity and the level of calcium mineral deposition. These results suggest that the TiO(2) nanotubular structure formed by anodizing is a promising configuration for sustained rhBMP-2 delivery.

  11. Bone morphogenetic protein-2 gene controls tooth root development in coordination with formation of the periodontium

    Institute of Scientific and Technical Information of China (English)

    Audrey Rakian; Wu-Chen Yang; Jelica Gluhak-Heinrich; Yong Cui; Marie A Harris; Demitri Villarreal; Jerry Q Feng; Mary MacDougall; Stephen E Harris

    2013-01-01

    Formation of the periodontium begins following onset of tooth-root formation in a coordinated manner after birth. Dental follicle progenitor cells are thought to form the cementum, alveolar bone and Sharpey’s fibers of the periodontal ligament (PDL). However, little is known about the regulatory morphogens that control differentiation and function of these progenitor cells, as well as the progenitor cells involved in crown and root formation. We investigated the role of bone morphogenetic protein-2 (Bmp2) in these processes by the conditional removal of the Bmp2 gene using the Sp7-Cre-EGFP mouse model. Sp7-Cre-EGFP first becomes active at E18 in the first molar, with robust Cre activity at postnatal day 0 (P0), followed by Cre activity in the second molar, which occurs after P0. There is robust Cre activity in the periodontium and third molars by 2 weeks of age. When the Bmp2 gene is removed from Sp71 (Osterix1) cells, major defects are noted in root, cellular cementum and periodontium formation. First, there are major cell autonomous defects in root-odontoblast terminal differentiation. Second, there are major alterations in formation of the PDLs and cellular cementum, correlated with decreased nuclear factor IC (Nfic), periostin and a-SMA1 cells. Third, there is a failure to produce vascular endothelial growth factor A (VEGF-A) in the periodontium and the pulp leading to decreased formation of the microvascular and associated candidate stem cells in the Bmp2-cKOSp7-Cre-EGFP. Fourth, ameloblast function and enamel formation are indirectly altered in the Bmp2-cKOSp7-Cre-EGFP. These data demonstrate that the Bmp2 gene has complex roles in postnatal tooth development and periodontium formation.

  12. Sustained release of bone morphogenetic protein 2 via coacervate improves the osteogenic potential of muscle-derived stem cells.

    Science.gov (United States)

    Li, Hongshuai; Johnson, Noah Ray; Usas, Arvydas; Lu, Aiping; Poddar, Minakshi; Wang, Yadong; Huard, Johnny

    2013-09-01

    Muscle-derived stem cells (MDSCs) isolated from mouse skeletal muscle by a modified preplate technique exhibit long-term proliferation, high self-renewal, and multipotent differentiation capabilities in vitro. MDSCs retrovirally transduced to express bone morphogenetic proteins (BMPs) can differentiate into osteocytes and chondrocytes and enhance bone and articular cartilage repair in vivo, a feature that is not observed with nontransduced MDSCs. These results emphasize that MDSCs require prolonged exposure to BMPs to undergo osteogenic and chondrogenic differentiation. A sustained BMP protein delivery approach provides a viable and potentially more clinically translatable alternative to genetic manipulation of the cells. A unique growth factor delivery platform comprised of native heparin and a synthetic polycation, poly(ethylene argininylaspartate diglyceride) (PEAD), was used to bind, protect, and sustain the release of bone morphogenetic protein-2 (BMP2) in a temporally and spatially controlled manner. Prolonged exposure to BMP2 released by the PEAD:heparin delivery system promoted the differentiation of MDSCs to an osteogenic lineage in vitro and induced the formation of viable bone at an ectopic site in vivo. This new strategy represents an alternative approach for bone repair mediated by MDSCs while bypassing the need for gene therapy.

  13. Growth differentiation factor 9:bone morphogenetic protein 15 heterodimers are potent regulators of ovarian functions

    Science.gov (United States)

    Peng, Jia; Li, Qinglei; Wigglesworth, Karen; Rangarajan, Adithya; Kattamuri, Chandramohan; Peterson, Randall T.; Eppig, John J.; Thompson, Thomas B.; Matzuk, Martin M.

    2013-01-01

    The TGF-β superfamily is the largest family of secreted proteins in mammals, and members of the TGF-β family are involved in most developmental and physiological processes. Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), oocyte-secreted paralogs of the TGF-β superfamily, have been shown genetically to control ovarian physiology. Although previous studies found that GDF9 and BMP15 homodimers can modulate ovarian pathways in vitro, the functional species-specific significance of GDF9:BMP15 heterodimers remained unresolved. Therefore, we engineered and produced purified recombinant mouse and human GDF9 and BMP15 homodimers and GDF9:BMP15 heterodimers to compare their molecular characteristics and physiological functions. In mouse granulosa cell and cumulus cell expansion assays, mouse GDF9 and human BMP15 homodimers can up-regulate cumulus expansion-related genes (Ptx3, Has2, and Ptgs2) and promote cumulus expansion in vitro, whereas mouse BMP15 and human GDF9 homodimers are essentially inactive. However, we discovered that mouse GDF9:BMP15 heterodimer is ∼10- to 30-fold more biopotent than mouse GDF9 homodimer, and human GDF9:BMP15 heterodimer is ∼1,000- to 3,000-fold more bioactive than human BMP15 homodimer. We also demonstrate that the heterodimers require the kinase activities of ALK4/5/7 and BMPR2 to activate SMAD2/3 but unexpectedly need ALK6 as a coreceptor in the signaling complex in granulosa cells. Our findings that GDF9:BMP15 heterodimers are the most bioactive ligands in mice and humans compared with homodimers explain many puzzling genetic and physiological data generated during the last two decades and have important implications for improving female fertility in mammals. PMID:23382188

  14. Bone morphogenetic protein-2 is a negative regulator of hepatocyte proliferation downregulated in the regenerating liver

    Institute of Scientific and Technical Information of China (English)

    Cui-Ping Xu; Wen-Min Ji; Gijs R van den Brink; Maikel P Peppelenbosch

    2006-01-01

    AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on proliferation of human Huh7 hepatoma cells.METHODS: Fifty-four adult male Wistar rats were randomly divided into three groups: A normal control (NC) group, a partial hepatectomized (PH) group and a sham operated (SO) group. To study the effect of liver regeneration on BMP-2 expression, rats were sacrificed before and at different time points after PH or the sham intervention (6, 12, 24 and 48 h). For each time point, six rats were used in parallel. Expression and distribution of BMP-2 protein were determined in regenerating liver tissue by Western blot analysis and immunohistochemistry. Effects of BMP-2 on cell proliferation of human Huh7 hepatoma cell line were assessed using an MTT assay.RESULTS: In the normal liver strong BMP-2 expression was observed around the central and portal veins. The expression of BMP-2 decreased rapidly as measured by both immunohistochemistry and Western blot analysis.This decrease was at a maximum of 3.22 fold after 12 h and returned to normal levels at 48 h after PH. No significant changes in BMP-2 immunoreactivity were observed in the SO group. BMP-2 inhibited serum induced Huh7 cell proliferation.CONCLUSION: BMP-2 is expressed in normal adult rat liver and negatively regulates hepatocyte proliferation.The observed down regulation of BMP-2 following partial hepatectomy suggests that such down regulation may be necessary for hepatocyte proliferation.

  15. The Effects of Tocotrienol and Lovastatin Co-Supplementation on Bone Dynamic Histomorphometry and Bone Morphogenetic Protein-2 Expression in Rats with Estrogen Deficiency

    OpenAIRE

    Kok-Yong Chin; Saif Abdul-Majeed; Norazlina Mohamed; Soelaiman Ima-Nirwana

    2017-01-01

    Both tocotrienol and statins are suppressors of the mevalonate pathway. Supplementation of tocotrienol among statin users could potentially protect them against osteoporosis. This study aimed to compare the effects of tocotrienol and lovastatin co-supplementation with individual treatments on bone dynamic histomorphometric indices and bone morphogenetic protein-2 (BMP-2) gene expression in ovariectomized rats. Forty-eight female Sprague-Dawley rats were randomized equally into six groups. The...

  16. Novel Approaches to Bone Grafting: Porosity, Bone Morphogenetic Proteins, Stem Cells, and the Periosteum

    OpenAIRE

    Petrochenko, Peter; Narayan, Roger J.

    2010-01-01

    The disadvantages involving the use of a patient’s own bone as graft material have led surgeons to search for alternative materials. In this review, several characteristics of a successful bone graft material are discussed. In addition, novel synthetic materials and natural bone graft materials are being considered. Various factors can determine the success of a bone graft substitute. For example, design considerations such as porosity, pore shape, and interconnection play significant roles i...

  17. Inhibitory Smads and bone morphogenetic protein (BMP) modulate anterior photoreceptor cell number during planarian eye regeneration.

    Science.gov (United States)

    González-Sastre, Alejandro; Molina, Ma Dolores; Saló, Emili

    2012-01-01

    Planarians represent an excellent model to study the processes of body axis and organ re-specification during regeneration. Previous studies have revealed a conserved role for the bone morphogenetic protein (BMP) pathway and its intracellular mediators Smad1/5/8 and Smad4 in planarian dorsoventral (DV) axis re-establishment. In an attempt to gain further insight into the role of this signalling pathway in planarians, we have isolated and functionally characte-rized the inhibitory Smads (I-Smads) in Schmidtea mediterranea. Two I-Smad homologues have been identified: Smed-smad6/7-1 and Smed-smad6/7-2. Expression of smad6/7-1 was detected in the parenchyma, while smad6/7-2 was found to be ex-pressed in the central nervous system and the eyes. Neither single smad6/7-1 and smad6/7-2 nor double smad6/7-1,-2 silencing gave rise to any apparent disruption of the DV axis. However, both regenerating and intact smad6/7-2 (RNAi) planarians showed defects in eye morphogenesis and displayed small, rounded eyes that lacked the anterior subpopulation of photoreceptor cells. The number of pigment cells was also reduced in these animals at later stages of regeneration. In contrast, after low doses of Smed-bmp(RNAi), planarians regenerated larger eyes in which the anterior subpopulation of photoreceptor cells was expanded. Our results suggest that Smed-smad6/7-2 and Smed-bmp control the re-specification and maintenance of anterior photoreceptor cell number in S. mediterranea.

  18. Influence of bone morphogenetic protein-2 on spiral ganglion neurite growth in vitro.

    Science.gov (United States)

    Volkenstein, Stefan; Brors, D; Hansen, S; Minovi, A; Laub, M; Jennissen, H P; Dazert, S; Neumann, A

    2009-09-01

    Recombinant human bone morphogenetic protein-2 (rhBMP-2) is a growth factor of the transforming growth factor-beta superfamily. Members of this protein family are involved in the development of various mammalian tissues, including the inner ear. As their notations indicate, they also have well-known effects on bone formation and regeneration. In this study, we examined the influence of rhBMP-2 on spiral ganglion (SG) neurite growth in vitro and showed the presence of its most preferred receptor BMPR-IB in spiral ganglion cells both in vitro and in vivo. SG explants of postnatal day 4 rats were analysed for neurite length and number after organotypical cell culture for 72 h, fixation and immunolabeling. Different concentrations of rhBMP-2 were used in a serum-free culture media. Neurite growth was compared with control groups that lacked stimulative effects; with neutrophin-3 (NT-3), which is a well-established positive stimulus on neurite length and number; and with combinations of these parameters. The results display that neurite number and total neurite length per explant in particular concentrations of rhBMP-2 increased by a maximum factor of two, while the mean neurite length was not affected. NT-3 demonstrated a much more potent effect, delivering a maximum increase of a factor of five. Furthermore, a combination of both growth factors shows a predominant effect on NT-3. Immunohistological detection of BMPR-IB was successful both in cell culture explants and in paraffin-embedded sections of animals of different ages. The results show that rhBMP-2 is, among other growth factors, a positive stimulus for SG neurite growth in vitro. Most growth factors are unstable and cannot be attached to surfaces without loss of their biological function. In contrast, rhBMP-2 can be attached to metal surfaces without loss of activity. Our findings suggest in vivo studies and a future clinical application of rhBMP-2 in cochlear implant technology to improve the tissue

  19. Mutual regulation of growth hormone and bone morphogenetic protein system in steroidogenesis by rat granulosa cells.

    Science.gov (United States)

    Nakamura, Eri; Otsuka, Fumio; Inagaki, Kenichi; Miyoshi, Tomoko; Matsumoto, Yoshinori; Ogura, Kanako; Tsukamoto, Naoko; Takeda, Masaya; Makino, Hirofumi

    2012-01-01

    GH induces preantral follicle growth and differentiation with oocyte maturation. However, the effects of GH on ovarian steroidogenesis and the mechanisms underlying its effects have yet to be elucidated. In this study, we investigated the actions of GH on steroidogenesis by rat granulosa cells isolated from early antral follicles by focusing on the ovarian bone morphogenetic protein (BMP) system. We found that GH suppressed FSH-induced estradiol production with reduction in aromatase expression and, in contrast, GH increased FSH-induced progesterone level with induction of steroidogenic acute regulatory protein, side chain cleavage cytochrome P450, and 3β-hydroxysteroid dehydrogenase. The effects of GH on steroidogenesis by granulosa cells were enhanced in the presence of the BMP antagonist noggin. Coculture of GH with oocytes did not alter GH regulation of steroidogenesis. Steroid production induced by cAMP donors was not affected by GH treatment and the GH effects on FSH-induced steroid production were not accompanied by changes in cAMP synthesis, suggesting that GH actions were not directly mediated by the cAMP-protein kinase A pathway. GH exerted synergistic effects on MAPK activation elicited by FSH, which regulated FSH-induced steroidogenesis. In addition, GH-induced signal transducer and activator of transcription phosphorylation was involved in the induction of IGF-I expression. GH increased IGF-I, IGF-I receptor, and FSH receptor expression in granulosa cells, and inhibition of IGF-I signaling restored GH stimulation of FSH-induced progesterone production, suggesting that endogenous IGF-I is functionally involved in GH effects on progesterone induction. BMP inhibited IGF-I effects that increased FSH-induced estradiol production with suppression of expression of the GH/IGF-I system, whereas GH/IGF-I actions impaired BMP-Sma and Mad related protein 1/5/8 signaling through down-regulation of the expression of BMP receptors. Thus, GH acts to modulate estrogen

  20. Repair of radius defect with bone-morphogenetic-protein loaded hydroxyapatite/collagen-poly(L-lactic acid) composite

    Institute of Scientific and Technical Information of China (English)

    胡蕴玉; 张超; 吕荣; 徐建强; 李丹

    2003-01-01

    Objective: To explore the method to repair bone defect with bone-morphogenetic-protein loaded hydroxyapatite/collagen-poly(L-lactic acid) composite. Methods: 18 adult beagle dogs were randomly divided into 3 groups. In Group A, bone-morphogenetic-protein (BMP) loaded hydroxyapatite/collagen-poly(L-lactic acid) (HAC-PLA) scaffold was implanted in a 2 cm diaphyseal defect in the radius. In Group B, unloaded pure HAC-PLA scaffold was implanted in the defects. No material was implanted in Group C (control group). The dogs were sacrificed 6 months postoperatively. Features of biocompatibility, biodegradability and osteoinduction were evaluated with histological, radiological examinations and bone mineral density (BMD) measurements.Results: In Group A, the radius defect healed after the treatment with BMP loaded HAC-PLA. BMD at the site of the defect was higher than that of the contralateral radius. Fibrous union developed in the animals of the control group. Conclusions: BMP not only promotes osteogenesis but also accelerates degradation of the biomaterials. Optimized design parameters of a three-dimensional porous biomaterial would give full scope to the role of BMP as an osteoinductive growth factor.

  1. Nanofibrous yet injectable polycaprolactone-collagen bone tissue scaffold with osteoprogenitor cells and controlled release of bone morphogenetic protein-2

    Energy Technology Data Exchange (ETDEWEB)

    Subramanian, Gayathri; Bialorucki, Callan [Department of Bioengineering, College of Engineering, University of Toledo, Toledo, OH 43606 (United States); Yildirim-Ayan, Eda, E-mail: eda.yildirimayan@utoledo.edu [Department of Bioengineering, College of Engineering, University of Toledo, Toledo, OH 43606 (United States); Department of Orthopaedic Surgery, University of Toledo Medical Center, Toledo, OH 43614 (United States)

    2015-06-01

    In this work, we developed a nanofibrous, yet injectable orthobiologic tissue scaffold that is capable of hosting osteoprogenitor cells and controlling kinetic release profile of the encapsulated pro-osteogenic factor without diminishing its bioactivity over 21 days. This innovative injectable scaffold was synthesized by incorporating electrospun and subsequently O{sub 2} plasma-functionalized polycaprolactone (PCL) nanofibers within the collagen type-I solution along with MC3T3-E1 cells (pre-osteoblasts) and bone morphogenetic protein-2 (BMP2). Through changing the PCL nanofiber concentration within the injectable scaffolds, we were able to tailor the mechanical strength, protein retention capacity, bioactivity preservation, and osteoinductive potential of the scaffolds. The nanofibrous internal structure of the scaffold allowed us to use a low dose of BMP2 (200 ng/ml) to achieve osteoblastic differentiation in in vitro culture. The osteogenesis capacity of the injectable scaffolds were evaluated though measuring MC3T3-E1 cell proliferation, ALP activity, matrix mineralization, and early- and late-osteoblast specific gene expression profiles over 21 days. The results demonstrated that the nanofibrous injectable scaffold provides not only an osteoinductive environment for osteoprogenitor cells to differentiate, but also a suitable biomechanical and biochemical environment to act as a reservoir for osteogenic factors with controlled release profile. - Highlights: • Injectable nanofibrous scaffold with osteoprogenitor cells and BMP2 was synthesized. • PCL nanofiber concentration within collagen scaffold affected the BMP2 retention and bioactivity. • Optimal PCL concentration was identified for mechanical stability, injectability, and osteogenic activity. • Scaffolds exhibited long-term osteoinductive capacity for bone repair and regeneration.

  2. Improving Bone Formation in a Rat Femur Segmental Defect by Controlling Bone Morphogenetic Protein-2 Release

    Science.gov (United States)

    2011-04-01

    delivered on a collagen sponge (INFUSE Bone Graft; Medtronic) has been approved by FDA for posterior-lateral spine fusions, tibial fractures, and sinus...area was defined by drawing a quadrilateral area using the periosteal corners of the four host cortices as points of reference. The relative areas of...section of an FR +BMP scaffold in Figure 8 (the ap- proximate boundary of the implant is denoted by the box) shows a mature and fully bridged periosteal

  3. Activation of bone morphogenetic protein-6 gene transcription in MCF-7 cells by estrogen

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ming; YAN Ji-dong; HANG Lei; WANG Qing; L(U) Shu-jun; ZHANG Jie; ZHU Tian-hui

    2005-01-01

    Background Bone morphogenetic protein-6 (BMP-6) is closely correlated with tumor differentiation and skeletal metastasis. Estrogen is considered as a stimulant for the initiation and promotion of breast cancer. Previous studies demonstrated that 17β-estadiol (E2) can selectively increase the expression of BMP-6. This experiment is designed to detect the molecular mechanism of estrogen activating BMP-6 gene transcription in human estrogen receptor positive (ER+) breast cancer cell line MCF-7. Methods After the treatment of MCF-7 cells with E2 at different concentrations (10-11 mol/L, 10-9 mol/L, 10-7 mol/L), the BMP-6 expression level was examined through real-time polymerase chain reaction. Through restriction enzyme digestion, human BMP-6 1.2 kb long promoter, BMP-6 0.7 kb long promoter was cloned into pGL-3 basic vector; after the treatment with 10-7 mol/L E2, luciferase activities of the two promoters were detected. Site-directed mutagenesis was performed to obtain the mutant forms of estrogen response element half-site (1/2 ERE) element and Sp1 sites in the BMP-6 promoter, the activities of these mutant form promoters were detected following the methods mentioned above. Chromatin immunoprecipitation (ChIP) assay was also used to confirm the binding of estrogen receptor α (Erα) on BMP-6 promoter in the presence of E2. Results E2 dose dependently increased BMP-6 mRNA expression in human ER+ breast cancer cell line MCF-7. At a dose of 10-7 mol/L E2, human BMP-6 1.2 kb promoter activity was increased by 90% compared with the control group treated with ethanol (P<0.05). Both the 1/2 ERE response element mutant form and the Sp1 site mutant form of the BMP-6 promoter abolished the activation of the BMP-6 promoter's response to E2. Through ChIP assay, the binding of Erα on 1/2 ERE response element in BMP-6 promoter was further validated. Conclusion Estrogen induces BMP-6 expression in human ER+ breast cancer cell line MCF-7 through its receptor Erα binding on 1

  4. Preparation of recombinant human bone morphogenetic protein-2 loaded dextran-based microspheres and their characteristics

    Institute of Scientific and Technical Information of China (English)

    Fa-ming CHEN; Zhi-fen WU; Qin-tao WANG; Hong WU; Yong-jie ZHANG; Xin NIE; Yan JIN

    2005-01-01

    Aim: To prepare new pharmaceutical forms with sustained delivery properties of recombinant human bone morphogenetic protein-2 (rhBMP2) for tissue engineering and guided tissue regeneration (GTR) use. Methods: rhBMP2-1oaded dextranbased hydrogel microspheres (rhBMP2-MPs), which aimed to keep rhBMP2 bioactivity and to achieve long-term sustained release of rhBMP2, were prepared by double-phase emulsified condensation polymerization. The physical, chemical performances and biological characteristics of those microspheres were studied both in vitro and in vivo. Results: The microspheres' average diameter was 30.33±4.32 μm with 75.4% ranging from 20 μm to 40 μm and the drug loading and encapsulation efficiency were 7.82% and 82.25%, respectively. The rhBMP2-releasing profiles in vitro showed that rhBMP2 release could be maintained more than 10 d. The rhBMP2-MPs, with good swelling and biodegradation behavior,could be kept for 6 months at below 4 ℃ without significant characteristic change or bioactivity loss. Cytology studies showed that rhBMP2-MPs could promote the proliferation of periodontal ligament cells (PDLCs) approximately 10 d, while the bioactivity of concentrated rhBMP2 solution could keep no more than 3 d.Scanning electron microscope showed that rhBMP2-MPs could be enchased into the porous structure of calcium phosphate ceremic (CPC) and the eugonic growth of PDLCs in CPC/rhBMP2-MPs scaffolds. Animal experiments indicated that using CPC/rhBMP2-MPs scaffolds could gain more periodontal tissue regeneration than using rhBMP2 compound firsthand with CPC (CPC/rhBMP2). Conclusion:By encapsulating rhBMP2 into dextran-based microspheres, a small quantity of rhBMP2 could achieve equivalent effects to the concentrated rhBMP2 solution and at the same time, could prolong rhBMP2 retention both in vitro and in vivo.

  5. Bone Morphogenetic Protein (BMP-7 expression is decreased in human hypertensive nephrosclerosis

    Directory of Open Access Journals (Sweden)

    Cohen Clemens D

    2010-11-01

    Full Text Available Abstract Background Bone Morphogenetic Protein (BMP-7 is protective in different animal models of acute and chronic kidney disease. Its role in human kidneys, and in particular hypertensive nephrosclerosis, has thus far not been described. Methods BMP-7 mRNA was quantified using real-time PCR and localised by immunostaining in tissue samples from normal and nephrosclerotic human kidneys. The impact of angiotensin (AT-II and the AT-II receptor antagonist telmisartan on BMP-7 mRNA levels and phosphorylated Smad 1/5/8 (pSmad 1/5/8 expression was quantified in proximal tubular cells (HK-2. Functional characteristics of BMP-7 were evaluated by testing its influence on TGF-β induced epithelial-to-mesenchymal transition (EMT, expression of TGF-β receptor type I (TGF-βRI and phosphorylated Smad 2 (pSmad 2 as well as on TNF-α induced apoptosis of proximal tubular cells. Results BMP-7 was predominantly found in the epithelia of the distal tubule and the collecting duct and was less abundant in proximal tubular cells. In sclerotic kidneys, BMP-7 was significantly decreased as demonstrated by real-time PCR and immunostaining. AT-II stimulation in HK-2 cells led to a significant decrease of BMP-7 and pSmad 1/5/8, which was partially ameliorated upon co-incubation with telmisartan. Only high concentrations of BMP-7 (100 ng/ml were able to reverse TNF-α-induced apoptosis and TGF-β-induced EMT in human proximal tubule cells possibly due to a decreased expression of TGF-βRI. In addition, BMP-7 was able to reverse TGF-β-induced phosphorylation of Smad 2. Conclusions The findings suggest a protective role for BMP-7 by counteracting the TGF-β and TNF-α-induced negative effects. The reduced expression of BMP-7 in patients with hypertensive nephrosclerosis may imply loss of protection and regenerative potential necessary to counter the disease.

  6. Characterization and expression of bone morphogenetic protein 4 gene in postnatal pigs.

    Science.gov (United States)

    Li, Ming; Chen, Qixin; Sun, Guirong; Shi, Xiaowei; Zhao, Qiaohui; Zhang, Chi; Zhou, Jianshe; Qin, Nan

    2010-06-01

    Bone morphogenetic protein 4 (BMP4) is involved in animal embryonic development and reproductive physiology. The human and murine BMP4 genes have been isolated and characterized. The objectives of this study were to: (1) characterize the full mRNA and genomic sequence for porcine BMP4, and (2) examine BMP4 gene expression in 10 tissues of postnatal female pigs. Using RT-PCR, RACE and general PCR techniques, a 1,626 bp DNA including the full coding region of BMP4 was isolated and identified as a homologue of human BMP4 transcript variant (TV)-c. The porcine TV-c contained 3 exons and astride 3.6 kb in the isolated 7.8 kb porcine BMP4 genome. The In silicon cloning identified other three forms of mRNAs, including the homologues of human TV-1, TV-a and a novel variant related to human TV-3 (TV-3p). The porcine TV-c, TV-1 and TV-3p bear internal ribosome entry sites (IRES) in 5' untranslated region (UTR), while there are two ARE elements in the 3'UTR. The full genomic sequence of porcine BMP4 gene showed 81.38, 76.23 and 64.00% identity with that of bovine, human and murine, respectively. The expression of BMP4 mRNA was determined by RT-PCR in 7, 14, and 28 day old female piglets and non-gestational sows. The results showed that porcine BMP4 occurred in all 10 examined tissues (heart, lung, liver, kidney, ovary, spleen, spinal medulla, brain, duodenum and thymus). The mRNA expression levels were relatively higher in lung and kidney in 7 day old piglets, thymus in 14 day old piglets, and spleen in 28 day old piglets, respectively, while the higher expressions were detected in liver of non-gestational pigs (P < 0.05). Moreover, the mRNA amounts both in 7 day old piglets and sows were generally higher than those in 14 and 28 day old piglets in nearly all examined tissues, except in thymus. It is concluded that the structure of porcine BMP4 gene is highly conservative with other mammalian BMP4 genes, but some differences may present in the regulation of gene expression

  7. Stimulatory effect of puerarin on bone formation through co-activation of nitric oxide and bone morphogenetic protein-2/mitogen-activated protein kinases pathways in mice

    Institute of Scientific and Technical Information of China (English)

    SHEU Shiow-yunn; TSAI Chia-chung; SUN Jui-sheng; CHEN Ming-hong; LIU Man-hai; SUN Man-ger

    2012-01-01

    Background Estrogen deficiency results in loss of bone mass.Phytoestrogens are plant-derived non-steroidal compounds with estrogen-like activity that bind to estrogen receptors.The main aim of this study was to investigate the effect of the phytoestrogen puerarin on adult mouse osteoblasts.Methods Osteoblast cells were harvested from 8-month old female imprinting control region (ICR) mice.The effects of puerarin stimulation on the proliferation,differentiation and maturation of osteoblasts were examined.The production of nitric oxide (NO) and the expression of bone morphogenetic protein-2 (BMP-2),SMAD4,mitogen-activated protein kinases (MAPK),core binding factor α1/runt-related transcription factor 2 (Cbfa1/Runx2),osteoprotegerin (OPG),and receptor activator of NF-kB ligand (RANKL) genes were analyzed.The activation of signal pathways was further confirmed by specific pathway inhibitors.Results The osteoblast viability reached its maximum at 10-8 mol/L puerarin.At this concentration,puerarin increases the proliferation and matrix mineralization of osteoblasts and promotes NO synthesis.With 10-8 mol/L puerarin treatment,BMP-2,SMAD4,Cbfa1/Runx2,and OPG gene expression were up-regulated,while the RANKL gene expression is down-regulated.Concurrent treatment involving the (bone morphogenetic protein) BMP antagonist Noggin or the NOS inhibitor L-NAME diminishes puerarin induced cell proliferation,Alkaline phosphatase (ALP) activity,NO production,as well as the BMP-2,SMAD4,Cbfa1/Runx2,OPG,and RANKL gene expression.Conclusions In this in vitro study,we demonstrate that puerarin is a bone anabolic agent that exerts its osteogenic effects through the induction of BMP-2 and NO synthesis,subsequently regulating Cbfa1/Runx2,OPG,and RANKL gene expression.This effect may contribute to its induction of osteoblast proliferation and differentiation,resulting in bone formation.

  8. Expression of human bone morphogenetic protein (BMP-2 and BMP-4 genes in transgenic bovine fibroblasts Expressão dos genes bone morphogenetic protein (BMP-2 e BMP-4 em fibroblastos bovinos transgênicos

    Directory of Open Access Journals (Sweden)

    C. Oleskovicz

    2004-08-01

    Full Text Available cDNAs dos genes bone morphogenetic protein-2 (BMP-2 e bone morphogenetic protein-4 (BMP-4 foram sintetizados a partir de RNA total extraído de tecidos ósseos de pacientes que apresentavam trauma facial (fraturas do maxilar entre o 7º e o 10º dia pós-trauma e clonados num vetor para expressão em células mamíferas, sob controle do promotor de citomegalovírus (CMV. Os vetores contendo os genes BMP-2 e o BMP-4 foram utilizados para a transfecção de fibroblastos bovinos. mRNAs foram indiretamente detectados por RT-PCR nas células transfectadas. As proteínas BMP-2 e BMP-4 foram detectadas mediante análises de Western blot. Os resultados demonstram a possibilidade de produção desses fatores de crescimento celular em fibroblastos bovinos. Essas células poderão ser utilizadas como fontes doadoras de material genético para a técnica de transferência nuclear na geração de animais transgênicos.

  9. Bone morphogenetic proteins in inflammation, glucose homeostasis and adipose tissue energy metabolism

    DEFF Research Database (Denmark)

    Grgurevic, Lovorka; Christensen, Gitte Lund; Schulz, Tim J;

    2016-01-01

    Bore morphogenetic proteins (BMPs) are members of the transforming growth factor (TGF)-β superfamily, a group of secreted proteins that regulate embryonic development. This review summarizes the effects of BMPs on physiological processes not exclusively linked to the musculoskeletal system....... Specifically, we focus on the involvement of BMPs in inflammatory disorders, e.g. fibrosis, inflammatory bowel disease, anchylosing spondylitis, rheumatoid arthritis. Moreover, we discuss the role of BMPs in the context of vascular disorders, and explore the role of these signalling proteins in iron...... homeostasis (anaemia, hemochromatosis) and oxidative damage. The second and third parts of this review focus on BMPs in the development of metabolic pathologies such as type-2 diabetes mellitus and obesity. The pancreatic beta cells are the sole source of the hormone insulin and BMPs have recently been...

  10. Synergistic actions of olomoucine and bone morphogenetic protein-4 in axonal repair after acute spinal cord contusion

    Institute of Scientific and Technical Information of China (English)

    Liang Chen; Jianjun Li; Liang Wu; Mingliang Yang; Feng Gao; Li Yuan

    2014-01-01

    To determine whether olomoucine acts synergistically with bone morphogenetic protein-4 in the treatment of spinal cord injury, we established a rat model of acute spinal cord contusion by impacting the spinal cord at the T8 vertebra. We injected a suspension of astrocytes derived from glial-restricted precursor cells exposed to bone morphogenetic protein-4 (GDAsBMP) into the spinal cord around the site of the injury, and/or olomoucine intraperitoneally. Olomoucine effectively inhibited astrocyte proliferation and the formation of scar tissue at the injury site, but did not prevent proliferation of GDAsBMP or inhibit their effects in reducing the spinal cord lesion cavity. Furthermore, while GDAsBMP and olomoucine independently resulted in small improve-ments in locomotor function in injured rats, combined administration of both treatments had a signiifcantly greater effect on the restoration of motor function. These data indicate that the combined use of olomoucine and GDAsBMP creates a better environment for nerve regeneration than the use of either treatment alone, and contributes to spinal cord repair after injury.

  11. Enhanced healing of rabbit segmental radius defects with surface-coated calcium phosphate cement/bone morphogenetic protein-2 scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Yi; Hou, Juan; Yin, ManLi [Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Wang, Jing, E-mail: biomatwj@163.com [Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Liu, ChangSheng, E-mail: csliu@sh163.net [Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237 (China); Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China)

    2014-11-01

    Large osseous defects remain a difficult clinical problem in orthopedic surgery owing to the limited effective therapeutic options, and bone morphogenetic protein-2 (BMP-2) is useful for its potent osteoinductive properties in bone regeneration. Here we build a strategy to achieve prolonged duration time and help inducting new bone formation by using water-soluble polymers as a protective film. In this study, calcium phosphate cement (CPC) scaffolds were prepared as the matrix and combined with sodium carboxymethyl cellulose (CMC-Na), hydroxypropylmethyl cellulose (HPMC), and polyvinyl alcohol (PVA) respectively to protect from the digestion of rhBMP-2. After being implanted in the mouse thigh muscles, the surface-modified composite scaffolds evidently induced ectopic bone formation. In addition, we further evaluated the in vivo effects of surface-modified scaffolds in a rabbit radius critical defect by radiography, three dimensional micro-computed tomographic (μCT) imaging, synchrotron radiation-based micro-computed tomographic (SRμCT) imaging, histological analysis, and biomechanical measurement. The HPMC-modified CPC scaffold was regarded as the best combination for segmental bone regeneration in rabbit radius. - Highlights: • A simple surface-coating method was used to fabricate composite scaffolds. • Growth factor was protected from rapid depletion via superficial coating. • Significant promotion of bone regeneration was achieved. • HPMC-modification displayed optimal effect of bone regeneration.

  12. Structure of protein related to Dan and Cerberus: insights into the mechanism of bone morphogenetic protein antagonism.

    Science.gov (United States)

    Nolan, Kristof; Kattamuri, Chandramohan; Luedeke, David M; Deng, Xiaodi; Jagpal, Amrita; Zhang, Fuming; Linhardt, Robert J; Kenny, Alan P; Zorn, Aaron M; Thompson, Thomas B

    2013-08-06

    The bone morphogenetic proteins (BMPs) are secreted ligands largely known for their functional roles in embryogenesis and tissue development. A number of structurally diverse extracellular antagonists inhibit BMP ligands to regulate signaling. The differential screening-selected gene aberrative in neuroblastoma (DAN) family of antagonists represents the largest group of BMP inhibitors; however, little is known of how they mechanistically inhibit BMP ligands. Here, we present the structure of the DAN family member, protein related to Dan and Cerberus (PRDC), solved by X-ray crystallography. The structure reveals a growth factor-like appearance with an unexpected dimerization mechanism that is formed through extensive β strand contacts. Using site-directed mutagenesis coupled with in vitro and in vivo activity assays, we identified a BMP-binding epitope on PRDC. We also determined that PRDC binds heparin with high affinity and that heparin binding to PRDC interferes with BMP antagonism. These results offer insight for how DAN family antagonists functionally inhibit BMP ligands.

  13. Compound soft regenerated skull material for repairing dog skull defects using bone morphogenetic protein as an inductor and nanohydroxyapatite as a scaffold

    Institute of Scientific and Technical Information of China (English)

    Zhidong Shi; Mingwang Liu; Zhongzong Qin; Qinmei Wang; Ying Guo; Haiyong He; Zhonghe Yu

    2008-01-01

    BACKGROUND: In previous studies of skull defects and regeneration, bone morphogenetic protein as an inductor and nanohydroxyapatite as a scaffold have been cocultured with osteoblasts.OBJECTIVE: To verify the characteristics of the new skull regenerated material after compound soft regenerated skull material implantatiom.DESIGN, TIME AND SETTING: The self-control and inter-group control animal experiment was perfurmed at the Sun Yat-sen University, China from February to July 2007.MATERIALS: Twenty-tour healthy adult dogs of both genders weighing 15-20 kg were used in this study. Nanohydroxyapatite as a scaffold was cocultured with osteoblasts. Using demineralized canine bone matrix as a carrier, recombinant human bone morphogenetic protein-2 was employed to prepare compound soft regenerated skull material. Self-designed compound soft regenerated skull material was implanted in models of skull defects.METHODS: Animals were randomly assigned into two groups, Group A (n = 16) and Group B (n = 8).Bilateral 2.5-cm-diameter full-thickness parietal skull defects were made in all animals. In Group A, the right side was reconstructed with calcium alginate gel, osteoblasts, and nanomcter bone meal composite;the left side was reconstructed with calcium alginate gel, osteoblasts, nanometer bone meal and recombinant human bone morphogenetic protein-2 composite. In Group B, the right side was kept as a simple skull detect, and the left side was reconstructed with calcium alginate gel, osteoblasts, nanometer bone meal and recombinant human bone morphogenetic protein-2 composite.MAIN OUTCOME MEASURES: Bone regeneration and histopathological changes at the site of the skull defect were observed with an optical microscope and a scanning electron microscope after surgery.The ability to form bone was measured by alizarin red S staining. In vitro cultured osteoblasts were observed for morphology.RESULTS: One month following surgery, newly formed bone trabeculae mostly covered the

  14. Prolonged propagation of rat neural stem cells relies on inhibiting autocrine/paracrine bone morphogenetic protein and platelet derived growth factor signals

    Institute of Scientific and Technical Information of China (English)

    Yirui Sun; Liangfu Zhou; Xing Wu; Hua Liu; Qiang Yuan; Ying Mao; Jin Hu

    2011-01-01

    Continuous expansion of rat neural stem cell lines has not been achieved due to proliferation arrest and spontaneous differentiation in vitro. In the current study, neural precursor cells derived from the subventricular zone of adult rats spontaneously underwent astroglial and oligodendroglial differentiation after limited propagation. This differentiation was largely induced by autocrine or paracrine bone morphogenetic protein and platelet derived growth factor signals. The results showed that, by inhibiting bone morphogenetic protein and platelet derived growth factor signals, adult rat neural precursor cells could be extensively cultured in vitro as tripotent stem cell lines. In addition to adult rat neural stem cells, we found that bone morphogenetic protein antagonists can promote the proliferation of human neural stem cells. Therefore, the present findings illustrated the role of autocrine or paracrine bone morphogenetic protein and platelet derived growth factor signaling in determining neural stem cell self-renewal and differentiation. By antagonizing both signals, the long-term propagation of rat neural stem cell lines can be achieved.

  15. Bone morphogenetic protein 4 inhibits insulin secretion from rodent beta cells through regulation of calbindin1 expression and reduced voltage-dependent calcium currents

    DEFF Research Database (Denmark)

    Christensen, Gitte L.; Jacobsen, Maria L. B.; Wendt, Anna

    2015-01-01

    AIMS/HYPOTHESIS: Type 2 diabetes is characterised by progressive loss of pancreatic beta cell mass and function. Therefore, it is of therapeutic interest to identify factors with the potential to improve beta cell proliferation and insulin secretion. Bone morphogenetic protein 4 (BMP4) expression...

  16. Altered bone morphogenetic protein signalling in the Helicobacter pylori-infected stomach

    NARCIS (Netherlands)

    Bleuming, S. A.; Kodach, L. L.; Leon, M. J. Garcia; Richel, D. J.; Peppelenbosch, M. P.; Reitsma, P. H.; Hardwick, J. C.; van den Brink, G. R.

    2006-01-01

    Morphogens regulate epithelial cell fate decisions in the adult gastrointestinal tract. The authors hypothesized that influx of inflammatory cells into the lamina propria may disturb the normal expression gradients of morphogens (morphogenetic landscape) in gastrointestinal epithelia. Changes in the

  17. Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer

    Directory of Open Access Journals (Sweden)

    E Ferreira

    2012-07-01

    Full Text Available Transplantation of mesenchymal stem cells (MSCs with electrotransferred bone morphogenetic protein-2 (BMP-2 transgene is an attractive therapeutic modality for the treatment of large bone defects: it provides both stem cells with the ability to form bone and an effective bone inducer while avoiding viral gene transfer. The objective of the present study was to determine the influence of the promoter driving the human BMP-2 gene on the level and duration of BMP-2 expression after transgene electrotransfer into rat MSCs. Cytomegalovirus, elongation factor-1α, glyceraldehyde 3-phosphate dehydrogenase, and beta-actin promoters resulted in a BMP-2 secretion rate increase of 11-, 78-, 66- and 36-fold over respective controls, respectively. In contrast, the osteocalcin promoter had predictable weak activity in undifferentiated MSCs but induced the strongest BMP-2 secretion rates in osteoblastically-differentiated MSCs. Regardless of the promoter driving the transgene, a plateau of maximal BMP-2 secretion persisted for at least 21 d after the hBMP-2 gene electrotransfer. The present study demonstrates the feasibility of gene electrotransfer for efficient BMP-2 transgene delivery into MSCs and for a three-week sustained BMP-2 expression. It also provides the first in vitro evidence for a safe alternative to viral methods that permit efficient BMP-2 gene delivery and expression in MSCs but raise safety concerns that are critical when considering clinical applications.

  18. Ultra-low-dose recombinant human bone morphogenetic protein-2 for 3-level anterior cervical diskectomy and fusion.

    Science.gov (United States)

    Pourtaheri, Sina; Hwang, Ki; Faloon, Michael; Issa, Kimona; Mease, Samuel J; Mangels, Daniel; Sinha, Kumar; Emami, Arash

    2015-04-01

    This study evaluated the safety of 3-level anterior cervical diskectomy and fusion (ACDF) with ultra-low-dose recombinant bone morphogenetic protein-2 (rhBMP-2). Thirty-seven consecutive patients with cervical spondylotic myelopathy who were treated with 3-level ACDF and rhBMP-2 were evaluated. Complications such as airway or cervical swelling or hematoma were not observed. The rate of dysphagia was no different at 1, 2, and 6 months postoperatively compared with reports in the literature without rhBMP-2. There were significant improvements in VAS neck/arm pain, Oswestry Neck Disability Index, and cervical lordosis. The use of ultra-low-dose rhBMP-2 for 3-level ACDF may be efficacious for surgically addressing 3-level spondylotic myelopathy.

  19. Osseointegration of titanium implants by addition of recombinant bone morphogenetic protein 2 (rhBMP-2)

    Energy Technology Data Exchange (ETDEWEB)

    Lichtinger, T.K.; Mueller, R.T.; Schuermann, N.; Oldenburg, M. [Essen Univ. (Germany). Dept. of Orthopaedic Surgery; Wiemann, M. [Inst. of Physiology, Univ. of Essen (Germany); Chatzinikolaidou, M.; Jennissen, H.P. [Inst. of Physiological Chemistry, Univ. of Essen (Germany); Rumpf, H.M.

    2001-12-01

    The osseointegration of long-term implants is often incomplete such that gaps remain between the implant surface and the surrounding hard tissue. This study examines the effect of soluble recombinant human bone morphogenic protein 2 (rhBMP-2) on gap healing and osseous integration. The effect of a single, intraoperative application of soluble rhBMP-2 on the formation of new bone around titanium implants was studied. A total of 8 titanium-alloy cylinders (Ti-6Al-4V) with a plasma spray coating (TPS; 400 {mu}m thickness) were implanted into femoral condyles of mature sheep: rhBMP-2 solution (1 {mu}g) was pipetted into the 1 mm wide cleft around 4 implants; 4 further implants served as rhBMP-2-free controls. Two of these controls exhibited an additional calciumphosphate-coating. The cleft around the implants served as testing zone to study the formation of new bone by microradiographical and histological analyses. The follow-up periods were 4 and 9 weeks, respectively. A significant amount of new bone contacting the implants' surface was detected where rhBMP-2-solution had been used: In 50% a circumferential osseoinduction occurred within 4 weeks and a nearly complete osseointegration was observed after 9 weeks. In all cases bone formation was exaggerated and filled the spongiosa with compact bone. Time matched TPS-controls and controls with calciumphosphate coating showed no notable formation of new bone. The results suggest that a single administration of soluble rhBMP-2 into a bone cavity can augment bone formation and also osseointegration of titanium implants. Further investigations based on these findings are necessary to develop long-term implants (e.g. joint replacements) with rhBMP-2-biocoating for humans. (orig.)

  20. Bone Morphogenetic Protein-9 Enhances Osteogenic Differentiation of Human Periodontal Ligament Stem Cells via the JNK Pathway

    Science.gov (United States)

    Wang, Xingxing; Pang, Yanan; Yang, Su; Wei, Yibo; Gao, Haochen; Wang, Dalin; Cao, Zhizhong

    2017-01-01

    Bone morphogenetic protein-9 (BMP9) shows great osteoinductive potential in bone regeneration. Periodontal ligament stem cells (PDLSCs) with multi-differentiation capability and low immunogenicity are increasingly used as seed cells for periodontal regenerative therapies. In the present study, we investigated the potent osteogenic activity of BMP9 on human PDLSCs (hPDLSCs), in which the c-Jun N-terminal kinase (JNK) pathway is possibly involved. Our results showed that JNK inhibition by the specific inhibitor SP600125 or adenovirus expressing small interfering RNA (siRNA) targeting JNK (AdR-si-JNK) significantly decreased BMP9-induced gene and protein expression of early and late osteogenic markers, such as runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN), in hPDLSCs. We also confirmed the in-vivo positive effect of JNKs on ectopic bone formation induced by hPDLSCs injected into the musculature of athymic nude mice and BMP9 ex vivo gene delivery. For the cellular mechanism, we found that BMP9 activated the phosphorylation of JNKs and Smad2/3, and that JNKs may engage in cross-talk with the Smad2/3 pathway in BMP9-mediated osteogenesis. PMID:28052093

  1. Synergistic effects of dimethyloxallyl glycine and recombinant human bone morphogenetic protein-2 on repair of critical-sized bone defects in rats

    Science.gov (United States)

    Qi, Xin; Liu, Yang; Ding, Zhen-Yu; Cao, Jia-Qing; Huang, Jing-Huan; Zhang, Jie-Yuan; Jia, Wei-Tao; Wang, Jing; Liu, Chang-Sheng; Li, Xiao-Lin

    2017-02-01

    In bone remodeling, osteogenesis is closely coupled to angiogenesis. Bone tissue engineering using multifunctional bioactive materials is a promising technique which has the ability to simultaneously stimulate osteogenesis and angiogenesis for repair of bone defects. We developed mesoporous bioactive glass (MBG)-doped poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) composite scaffolds as delivery vehicle. Two bioactive molecules, dimethyloxalylglycine (DMOG), a small-molecule angiogenic drug, and recombinant human bone morphogenetic protein-2 (rhBMP-2), an osteoinductive growth factor, were co-incorporated into the scaffold. The synergistic effects of DMOG and rhBMP-2 released in the composite scaffolds on osteogenic and angiogenic differentiation of hBMSCs were investigated using real-time quantitative polymerase chain reaction and western blotting. Moreover, in vivo studies were conducted to observe bone regeneration and vascular formation of critical-sized bone defects in rats using micro-computed tomography, histological analyses, Microfil® perfusion, fluorescence labeling, and immunohistochemical analysis. The results showed that DMOG and rhBMP-2 released in the MBG-PHBHHx scaffolds did exert synergistic effects on the osteogenic and angiogenic differentiation of hBMSCs. Moreover, DMOG and rhBMP-2 produced significant increases in newly-formed bone and neovascularization of calvarial bone defects in rats. It is concluded that the co-delivery strategy of both rhBMP-2 and DMOG can significantly improve the critical-sized bone regeneration.

  2. Synergistic effects of dimethyloxallyl glycine and recombinant human bone morphogenetic protein-2 on repair of critical-sized bone defects in rats

    Science.gov (United States)

    Qi, Xin; Liu, Yang; Ding, Zhen-yu; Cao, Jia-qing; Huang, Jing-huan; Zhang, Jie-yuan; Jia, Wei-tao; Wang, Jing; Liu, Chang-sheng; Li, Xiao-lin

    2017-01-01

    In bone remodeling, osteogenesis is closely coupled to angiogenesis. Bone tissue engineering using multifunctional bioactive materials is a promising technique which has the ability to simultaneously stimulate osteogenesis and angiogenesis for repair of bone defects. We developed mesoporous bioactive glass (MBG)-doped poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) composite scaffolds as delivery vehicle. Two bioactive molecules, dimethyloxalylglycine (DMOG), a small-molecule angiogenic drug, and recombinant human bone morphogenetic protein-2 (rhBMP-2), an osteoinductive growth factor, were co-incorporated into the scaffold. The synergistic effects of DMOG and rhBMP-2 released in the composite scaffolds on osteogenic and angiogenic differentiation of hBMSCs were investigated using real-time quantitative polymerase chain reaction and western blotting. Moreover, in vivo studies were conducted to observe bone regeneration and vascular formation of critical-sized bone defects in rats using micro-computed tomography, histological analyses, Microfil® perfusion, fluorescence labeling, and immunohistochemical analysis. The results showed that DMOG and rhBMP-2 released in the MBG-PHBHHx scaffolds did exert synergistic effects on the osteogenic and angiogenic differentiation of hBMSCs. Moreover, DMOG and rhBMP-2 produced significant increases in newly-formed bone and neovascularization of calvarial bone defects in rats. It is concluded that the co-delivery strategy of both rhBMP-2 and DMOG can significantly improve the critical-sized bone regeneration. PMID:28230059

  3. Reinforcing effect of calcium sulfate cement bovine bone morphogenetic protein on vertebral in the rabbit model of osteoporosis

    Institute of Scientific and Technical Information of China (English)

    Jie Zhang; Yu-Ming Chen; Chen Sheng-Guo; Kaken Habaerxi; Shawuti Alimujiang; Yu Chen; Ming-Zhen Peng; Rong Yue; Yu-Lian Wu; De-Quan Wang

    2014-01-01

    Objective:To observe reinforcing effect of calcium sulfate cement(CSC) bovine bone morphogenetic protein(bBMP) on vertebral in the rabbit model of osteoporosis.Methods:A total of48NewZealand white rabbits were randomly divided into groupⅠ(blank control group), group Ⅱ(CSC injection group), group Ⅲ(CSC/bBMP injection group) and control group.White rabbit osteoporosis model was established rapidly by using castration method+methylprednisolone candidate.After modeling, groups Ⅱ, Ⅲ were given corresponding vertebral body injection material, and4 animals were sacrificed respectively at24 h,6 weeks,12 weeks after vertebral plasty.Tissue pathological status, vertebral mineral density and vertebral body bone mechanical strength were observed.Results:Vertebral body structure form was normal in the groups Ⅱand Ⅲ.Trabecular bone coarsens, connection and repair were observed in micro fracture and bone defects, bone trabecular connectivity was superior to group Ⅰ significantly; vertebral body compression strength in the groupⅠ was on the decline, vertebral compression strength in the groups Ⅱand Ⅲ was on the rise, the largest vertebra.PostoperativeBMC andBMD in groups Ⅱand Ⅲ were incresed, andsignificantly higher than group Ⅰ after6 weeks(P<0.05),BMC and BMD in group Ⅲ after12 weeks were higher than the other three groups.Conclusion:Compound bBMPCSC has good bone induction.It can improve the three-dimensional construction effect for osteoporosis vertebral trabecula, and can significantly improve the vertebral strength, as a vertebral packing material with good application prospect.

  4. The Effects of Tocotrienol and Lovastatin Co-Supplementation on Bone Dynamic Histomorphometry and Bone Morphogenetic Protein-2 Expression in Rats with Estrogen Deficiency.

    Science.gov (United States)

    Chin, Kok-Yong; Abdul-Majeed, Saif; Mohamed, Norazlina; Ima-Nirwana, Soelaiman

    2017-02-15

    Both tocotrienol and statins are suppressors of the mevalonate pathway. Supplementation of tocotrienol among statin users could potentially protect them against osteoporosis. This study aimed to compare the effects of tocotrienol and lovastatin co-supplementation with individual treatments on bone dynamic histomorphometric indices and bone morphogenetic protein-2 (BMP-2) gene expression in ovariectomized rats. Forty-eight female Sprague-Dawley rats were randomized equally into six groups. The baseline was sacrificed upon receipt. All other groups were ovariectomized, except for the sham group. The ovariectomized groups were administered orally daily with (1) lovastatin 11 mg/kg/day alone; (2) tocotrienol derived from annatto bean (annatto tocotrienol) 60 mg/kg/day alone; (3) lovastatin 11 mg/kg/day, and annatto tocotrienol 60 mg/kg/day. The sham and ovariectomized control groups were treated with equal volume of vehicle. After eight weeks of treatment, the rats were sacrificed. Their bones were harvested for bone dynamic histomorphometry and BMP-2 gene expression. Rats supplemented with annatto tocotrienol and lovastatin concurrently demonstrated significantly lower single-labeled surface, but increased double-labeled surface, mineralizing surface, mineral apposition rate and bone formation rate compared to individual treatments (p bone parameters. In conclusion, tocotrienol can augment the bone formation and mineralization in rats receiving low-dose statins. Supplementation of tocotrienol in statin users can potentially protect them from osteoporosis.

  5. Expression analysis of bone morphogenetic protein 4 between fat and lean birds in adipose tissue and serum.

    Science.gov (United States)

    Cheng, B H; Leng, L; Wu, M Q; Zhang, Q; Zhang, X Y; Xu, S S; Cao, Z P; Li, Y M; Luan, P; Li, H

    2016-07-01

    The objectives of the present study were to characterize the tissue expression of chicken (Gallus gallus) bone morphogenetic protein 4 (BMP4) and compare differences in its expression in abdominal fat tissue and serum between fat and lean birds and to determine a potential relationship between the expression of BMP4 and abdominal fat tissue growth and development. The results showed that chicken BMP4 messenger RNA (mRNA) and protein were expressed in various tissues, and the expression levels of BMP4 transcript and protein were relatively higher in adipose tissues. In addition, the mRNA and protein expression levels of BMP4 in abdominal fat tissue of fat males were lower than those of lean males at 1, 2, 5, and 7 wk of age (P fat males was lower than that of lean males at 7 wk of age (P fat deposition through differences in its expression level. The results of this study will provide basic molecular information for studying the role of BMP4 in the regulation of adipogenesis in avian species.

  6. Structure of neuroblastoma suppressor of tumorigenicity 1 (NBL1): insights for the functional variability across bone morphogenetic protein (BMP) antagonists.

    Science.gov (United States)

    Nolan, Kristof; Kattamuri, Chandramohan; Luedeke, David M; Angerman, Elizabeth B; Rankin, Scott A; Stevens, Mariana L; Zorn, Aaron M; Thompson, Thomas B

    2015-02-20

    Bone morphogenetic proteins (BMPs) are antagonized through the action of numerous extracellular protein antagonists, including members from the differential screening-selected gene aberrative in neuroblastoma (DAN) family. In vivo, misregulation of the balance between BMP signaling and DAN inhibition can lead to numerous disease states, including cancer, kidney nephropathy, and pulmonary arterial hypertension. Despite this importance, very little information is available describing how DAN family proteins effectively inhibit BMP ligands. Furthermore, our understanding for how differences in individual DAN family members arise, including affinity and specificity, remains underdeveloped. Here, we present the structure of the founding member of the DAN family, neuroblastoma suppressor of tumorigenicity 1 (NBL1). Comparing NBL1 to the structure of protein related to Dan and Cerberus (PRDC), a more potent BMP antagonist within the DAN family, a number of differences were identified. Through a mutagenesis-based approach, we were able to correlate the BMP binding epitope in NBL1 with that in PRDC, where introduction of specific PRDC amino acids in NBL1 (A58F and S67Y) correlated with a gain-of-function inhibition toward BMP2 and BMP7, but not GDF5. Although NBL1(S67Y) was able to antagonize BMP7 as effectively as PRDC, NBL1(S67Y) was still 32-fold weaker than PRDC against BMP2. Taken together, this data suggests that alterations in the BMP binding epitope can partially account for differences in the potency of BMP inhibition within the DAN family.

  7. Co-stimulation with bone morphogenetic protein-9 and FK506 induces remarkable osteoblastic differentiation in rat dedifferentiated fat cells.

    Science.gov (United States)

    Nakamura, Toshiaki; Shinohara, Yukiya; Momozaki, Sawako; Yoshimoto, Takehiko; Noguchi, Kazuyuki

    2013-10-18

    Dedifferentiated fat (DFAT) cells, which are isolated from mature adipocytes using the ceiling culture method, exhibit similar characteristics to mesenchymal stem cells, and possess adipogenic, osteogenic, chondrogenic, and myogenic potentials. Bone morphogenetic protein (BMP)-2 and -9, members of the transforming growth factor-β superfamily, exhibit the most potent osteogenic activity of this growth factor family. However, the effects of BMP-2 and BMP-9 on the osteogenic differentiation of DFAT remain unknown. Here, we examined the effects of BMP-2 and BMP-9 on osteoblastic differentiation of rat DFAT (rDFAT) cells in the presence or absence of FK506, an immunosuppressive agent. Co-stimulation with BMP-9 and FK506 induced gene expression of runx2, osterix, and bone sialoprotein, and ALP activity compared with BMP-9 alone, BMP-2 alone and BMP-2+FK506 in rDFAT cells. Furthermore, it caused mineralization of cultures and phosphorylation of smad1/5/8, compared with BMP-9 alone. The ALP activity induced by BMP-9+FK506 was not influenced by addition of noggin, a BMP antagonist. Our data suggest that the combination of BMP-9 and FK506 potently induces osteoblastic differentiation of rDFAT cells.

  8. Implanting hydroxyapatite-coated porous titanium with bone morphogenetic protein-2 and hyaluronic acid into distal femoral metaphysis of rabbits

    Institute of Scientific and Technical Information of China (English)

    PENG Lei; BIAN Wei-guo; LIANG Fang-hui; XU Hua-zi

    2008-01-01

    Objective: To assess the osseointegration capability of hydroxyapatite-coated porous titanium with bone morphogenetic protein-2 (BMP-2) and hyaluronic acid to repair defects in the distal femur metaphysis in rabbits. Methods: Porous titanium implants were made by sintering titanium powder at high temperature, which were coated with hydroxyapatite by alkali and heat treatment and with BMP-2 combined with bone regeneration materials. And hyaluronic acid was further used as delivery system to prolong the effect of BMP-2. The implants were inserted into the metaphysis of the distal femur of rabbits. The animals were killed at 6, 12 and 24 weeks to accomplish histological and biomechanical analyses. Results: According to the result of histological analysis, the osseointegration in BMP-2 group was better than that of the HA-coated porous titanium group. In push-out test, all the samples had bigger shear stress as time passed by. There was statistical difference between the two groups in 6 and 12 weeks but not in 24 weeks. Conclusion: Hydroxyapatite-coated porous titanium with BMP-2 and hyaluronic acid has a good effect in repairing defects of distal fumur in rabbits, which is a fine biotechnology for future clinical application.

  9. Osteogenic efficiency of in situ gelling poloxamine systems with and without bone morphogenetic protein-2

    Directory of Open Access Journals (Sweden)

    A Rey-Rico

    2011-04-01

    Full Text Available In situ gelling solutions for minimally invasive local application of bone growth factors are attracting increasing attention as efficient and patient-friendly alternative to bone grafts and solid scaffolds for repairing bone defects. Poloxamines, i.e., X-shaped poly(ethylene oxide-poly(propylene oxide block copolymers with an ethylenediamine core (Tetronic®, were evaluated both as an active osteogenic component and as a vehicle for rhBMP-2 injectable implants. After cytotoxicity screening of various poloxamine varieties, Tetronic 908, 1107, 1301 and 1307 solutions were chosen as the most cytocompatible and their sol-to-gel transitions were rheologically characterized. Viscoelastic gels, formed at 37 ºC, sustained protein release under physiological-like conditions. Formulations of rhBMP-2 led to differentiation of mesenchymal stem cells to osteoblasts, quantified as alkaline phosphatase activity with a maximum at day 7, and to mineralized nodules. Interestingly, poloxamine solely gels led to an initial proliferation of the mesenchymal stem cells (first week, followed by differentiation to osteoblasts (second to third week. Histochemical analysis revealed that Tetronic 908 is only osteoinductive; Tetronic 1107 is mostly osteoinductive, although its use leads to a minor differentiation to adipocytes; Tetronic 1307, solely or loaded with rhBMP-2, causes differentiation of both osteoblasts and adipocytes. Enhanced expression levels of CBFA-1 and collagen type I were observed for Tetronic 908, 1107 and 1307, both solely and combined with rhBMP-2. The intrinsic osteogenic activity of poloxamines (not observed for Pluronic F127 offers novel perspectives for bone regeneration using minimally invasive procedures (i.e., injectable scaffolds and overcoming the safety and the cost/effectiveness concerns associated with large scale clinical use of recombinant growth factors.

  10. Depot injectable biodegradable nanoparticles loaded with recombinant human bone morphogenetic protein-2: preparation, characterization, and in vivo evaluation

    Directory of Open Access Journals (Sweden)

    Hassan AH

    2015-07-01

    Full Text Available Ali Habiballah Hassan,1 Khaled Mohamed Hosny,2,3 Zuahir A Murshid,1 Adel Alhadlaq,4 Ahmed Alyamani,5 Ghada Naguib6 1Department of Orthodontics, Faculty of Dentistry, 2Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 3Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Beni Suef University, Beni Suef, Egypt; 4Department of Pediatric Dentistry and Orthodontics, College of Dentistry, King Saud University, Riyadh, 5Department of Oral Surgery, 6Department of Restorative Dentistry, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia Objective: The aim of this study is to utilize the biocompatibility characteristics of biodegradable polymers, viz, poly lactide-co-glycolide (PLGA and polycaprolactone (PCL, to prepare sustained-release injectable nanoparticles (NPs of bone morphogenetic protein-2 (BMP-2 for the repair of alveolar bone defects in rabbits. The influence of formulation parameters on the functional characteristics of the prepared NPs was studied to develop a new noninvasive injectable recombinant human BMP-2 (rhBMP-2 containing grafting material for the repair of alveolar bone clefts.Materials and methods: BMP-2 NPs were prepared using a water-in-oil-in-water double-emulsion solvent evaporation/extraction method. The influence of molar ratio of PLGA to PCL on a suitable particle size, encapsulation efficiency, and sustained drug release was studied. Critical size alveolar defects were created in the maxilla of 24 New Zealand rabbits divided into three groups, one of them treated with 5 µg/kg of rhBMP-2 NP formulations.Results: The results found that NPs formula prepared using blend of PLGA and PCL in 4:2 (w/w ratio showed the best sustained-release pattern with lower initial burst, and showed up to 62.7% yield, 64.5% encapsulation efficiency, 127 nm size, and more than 90% in vitro release. So, this formula was selected for

  11. Effect of nano-hydroxyapatite/collagen composite and bone morphogenetic protein-2 on lumbar intertransverse fusion in rabbits

    Institute of Scientific and Technical Information of China (English)

    孙天胜; 关凯; 时述山; 朱兵; 郑永军; 崔福斋; 张伟; 廖素三

    2004-01-01

    Objective: To investigate the effect of nano-hydroxyapatite/collagen (nHA/collagen) composite as a graft extender and enhancer when combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) on lumbar intertransverse fusion in rabbits.Methods: Sixty-four adult female New Zealand white rabbits, aged 1 year and weighing 3.5-4.5 kg, underwent similar posterolateral intertransverse process arthrodesis and were randomly divided into 4 groups based on different grafts: autogenous cancellous bone alone (ACB group), nHA/collagen alone (HAC group), half autogenous cancellous bone and half nHA/collagen (ACB+HAC group) and nHA/collagen combined with rhBMP-2 (HAC+BMP group). The fusion masses were analyzed by manual palpation, radiography, biomechanical testing and histological examination. Results: Fusion was observed in 4 cases in the 6th week and in 5 cases in the 10th week after surgery in ACB group. No case showed fusion in HAC group. In ACB+HAC group, there was fusion in 3 cases in the 6th week and in 4 cases in the 10th week after surgery. In HAC+BMP group, fusion in 1 case was found in the 4th week, in 5 cases in the 6th week and in 6 cases in the 10th week after surgery. It suggested that ACB, ACB+HAC and HAC+BMP groups showed similar fusion ratio and mechanical strength in the 6th and 10th week after surgery. According to the microstructure analysis of the samples, nHA/collagen had no negative effect when implanted together with ilium autograft. In HAC+BMP group, new bone-like tissue was observed in the 2nd week postoperatively, and nearly all of the implanted composites were replaced by mature bone matrix and new bones in 10th week postoperatively.Conclusions: The nHA/collagen, especially combined with rhBMP-2, is a promising bone substitute, for it has quick biodegradation, fine bone-bending ability, and high osteoconductivity on posterolateral spinal fusion in rabbits.

  12. Human bone morphogenetic protein-2 gene transfer induces human mesenchymal stem cell proliferation and differentiation in vitro

    Institute of Scientific and Technical Information of China (English)

    李军; 范清宇; 钱济先; 马保安; 周勇; 张明华

    2004-01-01

    Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation.Methods: The BMP2 gene was cloned into a eukaryotic expression vector pcDNA3. Transfected the recombinant into hMSCs by liposome. Immunnohistochemistry and in situ hybridization methods were used to identify the expression of BMP2 mRNA and protein; ALP and Von Kossa stains were performed to identify the BMP2 gene differentiated effect on the hMSCs. Results: The pcDNA3/BMP2 fragments were as large as theory. BMP2 mRNA and protein were expressed and synthesized both in 48 h and 4 weeks after transfection, the ALP and Ca deposit exhibition, which marked the osteogenic lineage of hMSCs,were enhanced and sped. Conclusion: Transfection of pcDNA3/BMP2 is able to provide transient and persistent expression in hMSCs, and promote the MSCs differentiation to osteogenic lineage.

  13. [Effects of phosphatidylinositol-3 kinase/protein kinase b/bone morphogenetic protein-15 pathway on the follicular development in the mammalian ovary].

    Science.gov (United States)

    Wu, Yan-qing; Chen, Li-yun; Zhang, Zheng-hong; wang, Zheng-chao

    2013-04-01

    In mammals, ovarian follicle is made of an oocyte with its surrounding granulosa cells and theca cells. Follicular growth and development is a highly coordinated programmable process, which guarantees the normal oocyte maturation and makes it having the fertilizing capacity. The paracrine and autocrine between oocytes and granulosa cells are essential for the follicular development to provide a suitable microenvironment. Phosphatidylinositol-3 kinase /protein kinase B is one of these important regulatory signaling pathways during this developmental process, and bone morphogenetic protein-15 an oocyte-specific secreted signal molecule, which regulates the follicular development by paracrine in the mammalian ovary. The present article overviewed the role of phosphatidylinositol-3 kinase / protein kinase B signaling during the follicular development based on our previous investigation about protein kinase B /forkhead transcription factor forkhead family of transcription factors -3a, and then focused on the regulatory effects of bone morphogenetic protein-15, as a downstream signal molecule of phosphatidylinositol-3 kinase / forkhead family of transcription factors -3a pathway, on ovarian follicular development, which helped to further understand the molecular mechanism regulating the follicular development and to treat ovarian diseases like infertility.

  14. Lack of Association of Bone Morphogenetic Protein 2 Gene Haplotypes with Bone Mineral Density, Bone Loss, or Risk of Fractures in Men

    Directory of Open Access Journals (Sweden)

    Satya S. Varanasi

    2011-01-01

    Full Text Available Introduction. The association of bone morphogenetic protein 2 (BMP2 with BMD and risk of fracture was suggested by a recent linkage study, but subsequent studies have been contradictory. We report the results of a study of the relationship between BMP2 genotypes and BMD, annual change in BMD, and risk of fracture in male subjects. Materials and Methods. We tested three single-nucleotide polymorphisms (SNPs across the BMP2 gene, including Ser37Ala SNP, in 342 Caucasian Englishmen, comprising 224 control and 118 osteoporotic subjects. Results. BMP2 SNP1 (Ser37Ala genotypes were found to have similar low frequency in control subjects and men with osteoporosis. The major informative polymorphism, BMP2 SNP3 (Arg190Ser, showed no statistically significant association with weight, height, BMD, change in BMD at hip or lumbar spine, and risk of fracture. Conclusion. There were no genotypic or haplotypic effects of the BMP2 candidate gene on BMD, change in BMD, or fracture risk identified in this cohort.

  15. Bone Morphogenetic Protein-7 Ameliorates Cerebral Ischemia and Reperfusion Injury via Inhibiting Oxidative Stress and Neuronal Apoptosis

    Directory of Open Access Journals (Sweden)

    Haitao Pei

    2013-11-01

    Full Text Available Previous studies have indicated that bone morphogenetic protein-7 (BMP-7 is neuroprotective against cerebral ischemia/reperfusion (IR injury. The present study was undertaken to determine the molecular mechanisms involved in this effect. Adult male Wistar rats were subjected to 2 h of transient middle cerebral artery occlusion (MCAO, followed by 24 h of reperfusion. BMP-7 (10−4 g/kg or vehicle was infused into rats at the onset of reperfusion via the tail vein. Neurological deficits, infarct volume, histopathological changes, oxidative stress-related biochemical parameters, neuronal apoptosis, and apoptosis-related proteins were assessed. BMP-7 significantly improved neurological and histological deficits, reduced the infarct volume, and decreased apoptotic cells after cerebral ischemia. BMP-7 also markedly enhanced the activities of antioxidant enzymes superoxide dismutase (SOD and glutathione peroxidase (GSH-PX, and reduced the level of malondialdehyde (MDA in IR rats. In addition, Western blot analysis indicated that BMP-7 prevented cytochrome c release, inhibited activation of caspase-3, caspase-9 and caspase-8. Our data suggested that BMP-7 has protective effects against cerebral IR injury in rats, and the neuroprotective effects may be attributed to attenuating oxidative stress and inhibiting neuronal apoptosis.

  16. Different temporal patterns in the expressions of bone morphogenetic proteins and noggin during astroglial scar formation after ischemic stroke.

    Science.gov (United States)

    Shin, Jin A; Kang, Jihee Lee; Lee, Kyung-Eun; Park, Eun-Mi

    2012-05-01

    Bone morphogenetic proteins (BMPs) and their antagonists have roles in scar formation and regeneration after central nervous system injuries. However, temporal changes in their expression during astroglial scar formation in the ischemic brain are unknown. Here, we examined protein levels of BMP2, BMP7, and their antagonist noggin in the ischemic brain up to 4 weeks after experimental stroke in mice. BMP2 and BMP7 levels were increased from 1 to 4 weeks in the ischemic brain, and their expression was associated with astrogliosis. BMP7 expression was more intense and co-localized in reactive astrocytes in the ischemic subcortex at 1 week. Noggin expression began to increase after 2 weeks and was further increased at 4 weeks only in the ischemic subcortex, but the intensity was weak compared to the intensity of BMPs. Noggin was co-localized mainly in activated microglia. These findings show that expression of BMPs and noggin differed over time, in intensity and in types of cell, and suggest that BMPs and noggin have different roles in the processes of glial scar formation and neurorestoration in the ischemic brain.

  17. Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling.

    Science.gov (United States)

    Liao, Wei-Ju; Tsao, Ku-Chi; Yang, Ruey-Bing

    2016-03-01

    SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. We describe novel mechanisms in targeting S1 to the plasma membrane and demonstrate that N-glycans are required for S1 functions during primitive haematopoiesis in zebrafish.

  18. Brown fat determination and development from muscle precursor cells by novel action of bone morphogenetic protein 6.

    Directory of Open Access Journals (Sweden)

    Ankur Sharma

    Full Text Available Brown adipose tissue (BAT plays a pivotal role in promoting energy expenditure by the virtue of uncoupling protein-1 (UCP-1 that differentiates BAT from its energy storing white adipose tissue (WAT counterpart. The clinical implication of "classical" BAT (originates from Myf5 positive myoblastic lineage or the "beige" fat (originates through trans-differentiation of WAT activation in improving metabolic parameters is now becoming apparent. However, the inducers and endogenous molecular determinants that govern the lineage commitment and differentiation of classical BAT remain obscure. We report here that in the absence of any forced gene expression, stimulation with bone morphogenetic protein 6 (BMP6 induces brown fat differentiation from skeletal muscle precursor cells of murine and human origins. Through a comprehensive transcriptional profiling approach, we have discovered that two days of BMP6 stimulation in C2C12 myoblast cells is sufficient to induce genes characteristic of brown preadipocytes. This developmental switch is modulated in part by newly identified regulators, Optineurin (Optn and Cyclooxygenase-2 (Cox2. Furthermore, pathway analyses using the Causal Reasoning Engine (CRE identified additional potential causal drivers of this BMP6 induced commitment switch. Subsequent analyses to decipher key pathway that facilitates terminal differentiation of these BMP6 primed cells identified a key role for Insulin Like Growth Factor-1 Receptor (IGF-1R. Collectively these data highlight a therapeutically innovative role for BMP6 by providing a means to enhance the amount of myogenic lineage derived brown fat.

  19. In situ osteogenesis: regeneration of 10-cm mandibular defect in porcine model using recombinant human bone morphogenetic protein-2 (rhBMP-2) and Helistat absorbable collagen sponge.

    Science.gov (United States)

    Carstens, Michael H; Chin, Martin; Li, X Jian

    2005-11-01

    Traditional bone grafting relies upon the incorporation of a bone-cell bearing structure into a recipient site. The graft serves as a scaffold that is eventually replaced and remodeled. This process is known as osteoconduction. Recombinant human bone morphogenetic protein-2 (rhBMP-2) is commercially available as an acellular implant in which the protein is bound to an absorbable collagen sponge (ACS). The rhBMP-2/ACS implant converts undifferentiated mesenchymal stem cells into osteoblasts and promotes an intense local neovascular response. This process, known as osteoinduction, produces bone via membranous, chondroid, or endochondral ossification. The type of bone synthesis depends upon the mesenchymal substrate and the local cellular environment. Using this simple technique, bone defects can be resynthesized with good outcomes and a significant reduction in donor site morbidity. Repair of a critical-sized mandibular resection defect with ISO is described. Basic science concepts of rhBMP-2, relevant histopathologic findings, and clinical application are described.

  20. The Effects of Tocotrienol and Lovastatin Co-Supplementation on Bone Dynamic Histomorphometry and Bone Morphogenetic Protein-2 Expression in Rats with Estrogen Deficiency

    Science.gov (United States)

    Chin, Kok-Yong; Abdul-Majeed, Saif; Mohamed, Norazlina; Ima-Nirwana, Soelaiman

    2017-01-01

    Both tocotrienol and statins are suppressors of the mevalonate pathway. Supplementation of tocotrienol among statin users could potentially protect them against osteoporosis. This study aimed to compare the effects of tocotrienol and lovastatin co-supplementation with individual treatments on bone dynamic histomorphometric indices and bone morphogenetic protein-2 (BMP-2) gene expression in ovariectomized rats. Forty-eight female Sprague-Dawley rats were randomized equally into six groups. The baseline was sacrificed upon receipt. All other groups were ovariectomized, except for the sham group. The ovariectomized groups were administered orally daily with (1) lovastatin 11 mg/kg/day alone; (2) tocotrienol derived from annatto bean (annatto tocotrienol) 60 mg/kg/day alone; (3) lovastatin 11 mg/kg/day, and annatto tocotrienol 60 mg/kg/day. The sham and ovariectomized control groups were treated with equal volume of vehicle. After eight weeks of treatment, the rats were sacrificed. Their bones were harvested for bone dynamic histomorphometry and BMP-2 gene expression. Rats supplemented with annatto tocotrienol and lovastatin concurrently demonstrated significantly lower single-labeled surface, but increased double-labeled surface, mineralizing surface, mineral apposition rate and bone formation rate compared to individual treatments (p osteoporosis. PMID:28212283

  1. Improved Bone Formation in Osteoporotic Rabbits with the Bone Morphogenetic Protein-2 (rhBMP-2 Coated Titanium Screws Which Were Coated By Using Plasma Polymerization Technique

    Directory of Open Access Journals (Sweden)

    Salih Gulsen

    2014-06-01

    Full Text Available Delaying of bone fusion in osteoporotic patients underwent spinal stabilization surgery leads to screw loosening, and this causes pseudoarticulation, mobility and fibrosis at vertebral segments. To prevent these complications, the screws coated with recombinant bone morphogenetic protein-2 (rhBMP-2 could be used. To verify this hypothesis, we coated 5 Titanium screws with rhBMP-2 using plasma polymerization method, and also used 10 uncoated screws for making comparison between coated and uncoated screws in different groups. And 15 skeletally mature white New Zealand female rabbits were assigned into three different groups: Group 1(N = 5: No osteoporosis induction and insertion of uncoated Titanium screw into right sacrum of each rabbit in group 1; group 2 (N = 5: Osteoporosis induction and insertion of uncoated Titanium screw into right sacrum of each rabbit in group 2; group 3 (N = 5 rhBMP-2 coated Titanium screw inserted into right sacrum of each rabbit in group 3. In summary, using of these coated screws provides new bone formation, but causes less fibrosis and less inflammation than uncoated screws at the interface between the coated screw and bone. Then the plasma polymerization technique provides controlled releasing of rhBMP-2 from the screw to the bone tissue in osteoporotic rabbits.

  2. Aldosterone breakthrough caused by chronic blockage of angiotensin II type 1 receptors in human adrenocortical cells: Possible involvement of bone morphogenetic protein-6 actions

    OpenAIRE

    Otani, Hiroyuki; Otsuka, Fumio; Inagaki, Kenichi; Suzuki, Jiro; Miyoshi, Tomoko; KANO, YOSHIHIRO; GOTO, Junko; Ogura, Toshio; Makino, Hirofumi

    2008-01-01

    Circulating aldosterone concentrations occasionally increase after initial suppression with angiotensin II (Ang II) converting enzyme inhibitors or Ang II type 1 receptor blockers (ARBs), a phenomenon referred to as aldosterone breakthrough. However, the underlying mechanism causing the aldosterone breakthrough remains unknown. Here we investigated whether aldosterone breakthrough occurs in human adrenocortical H295R cells in vitro. We recently reported that bone morphogenetic protein (BMP)-6...

  3. Effect of transforming growth factor beta and bone morphogenetic proteins on rat hepatic stellate cell proliferation and trans-differentiation

    Institute of Scientific and Technical Information of China (English)

    Hong Shen; Guo-Jiang Huang; Yue-Wen Gong

    2003-01-01

    AIM: To explore different roles of TGF-β (transforming growth factor beta) and bone morphogenetic proteins (BMPs)in hepatic stellate cell proliferation and trans-differentiation.METHODS: Hepatic stellate cells were isolated from male Sprague-Dawley rats. Sub-cultured hepatic stellate cells were employed for cell proliferation assay with WST-1 reagent and Western blot analysis with antibody against smooth muscle alpha actin (SMA).RESULTS: The results indicated that TGF-β1 significantly inhibited cell proliferation at concentration as low as 0.1 ng/ml, but both BMP-2 and BMP-4 did not affect cell proliferation at concentration as high as 10 ng/ml. The effect on hepatic stellate cell trans-differentiation was similar between TGFβ1 and BMPs. However, BMPs was more potent at transdifferentiation of hepatic stellate cells than TGF-β1. In addition, we observed that TGF-β1 transient reduced the abundance of SMA in hepatic stellate cells.CONCLUSION: TGF-β may be more important in regulation of hepatic stellate cell proliferation while BMPs may be the major cytokines regulating hepatic stellate cell transdifferentiation.

  4. Coleusin factor, a novel anticancer diterpenoid, inhibits osteosarcoma growth by inducing bone morphogenetic protein-2-dependent differentiation.

    Science.gov (United States)

    Geng, Shuo; Sun, Bo; Lu, Ran; Wang, Jingze

    2014-06-01

    Coleusin factor is a diterpenoid compound isolated from the root of a tropical plant, Coleus forskohlii. Although Coleusin factor has been reported to suppress proliferation of and induce apoptosis in several types of cancer cells, the effects of Coleusin factor on osteosarcoma and the underlying mechanism are still not fully understood. In this study, we show that Coleusin factor treatment potently inhibits the growth of osteosarcoma cells associated with G(1) cell-cycle arrest. Interestingly, apoptosis and cell death are not induced. Instead, Coleusin factor causes osteosarcoma cells to exhibit typical properties of differentiated osteoblasts, including a morphologic alteration resembling osteoblasts, the expression of osteoblast differentiation markers, elevated alkaline phosphatase activity, and increased cellular mineralization. Coleusin factor treatment significantly increases the expression of bone morphogenetic protein-2 (BMP-2), a crucial osteogenic regulator, and runt-related transcription factor 2 (RUNX2), one of the key transcription factors of the BMP pathway. When BMP-2 signaling is blocked, Coleusin factor fails to inhibit cell proliferation and to induce osteoblast differentiation. Thus, upregulation of BMP-2 autocrine is critical for Coleusin factor to induce osteoblast differentiation and exert its anticancer effects on osteosarcoma. Importantly, administration of Coleusin factor inhibits the growth of osteosarcoma xenografted in nude mice without systemic or immunologic toxicity. Osteosarcoma is a highly aggressive cancer marked by the loss of normal differentiation. Coleusin factor represents a new type of BMP-2 inducer that restores differentiation in osteosarcoma cells. It may provide a promising therapeutic strategy against osteosarcoma with minimal side effects.

  5. EFFECTS OF TRANSFORMING GROWTH FACTOR β AND RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN 2 ON HUMAN PERIODONTAL LIGAMENT FIBROBLASTS

    Institute of Scientific and Technical Information of China (English)

    司晓辉; 刘正

    2001-01-01

    Objective To evaluate the effects of transforming growth factor β(TGF-β) and recombinant human bone morphogenetic protein 2 (rhBMP2) on human periodontal ligament fibroblasts ( HPDLFs ). Methods HPDLFs were done primary culture to detect the distinct concentrations of TGF-β and rhBMP2 on its proliferation, alkaline phosphatase (ALP) activity, osteocalcin ( OC) synthesis and formation of the mineralized nodules, respectively. Results TGF-β(5~100ng /ml) significantly stimulated the proliferation of HPDLFs. The ALP activity of HPDLFs was evaluated evidently by 5ng /ml TGF-β. TGF-β(0.5~100ng /ml) had no effects on OC synthesis and formation of the mineralized nodules of HPDLFs. rhBMP2 (0.25~2mg/ ml) had no rernarkable effect on the proliferation of HPDLFs. The ALP activity, OC synthesis and formation of the mineralized nodules of HPDLFs were significantly stimulated by 0.5~2mg/ml rhBMP2. Conclusion The effects of TGF-β and rhBMP2 on HPDLFs are dose-dependent. TGF-β can stimulate HPDLFs to express the early marker of osteoblastic phenotype , and it lacks the ability to promote maturation of the osteogenic phenotype. rhBMP2 can not only stimulate the expression but also promote the maturation of osteoblastic phenotype of HPDLFs.

  6. Upregulation of Bone Morphogenetic Protein-2 Synthesis and Consequent Collagen II Expression in Leptin-stimulated Human Chondrocytes.

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    Shun-Fu Chang

    Full Text Available Bone morphogenetic proteins (BMPs play positive roles in cartilage development, but they can barely be detected in healthy articular cartilage. However, recent evidence has indicated that BMPs could be detected in osteoarthritic and damaged cartilage and their precise roles have not been well defined. Extremely high amounts of leptin have been reported in obese individuals, which can be associated with osteoarthritis (OA development. The aim of this study was to investigate whether BMPs could be induced in human primary chondrocytes during leptin-stimulated OA development and the underlying mechanism. We found that expression of BMP-2 mRNA, but not BMP-4, BMP-6, or BMP-7 mRNA, could be increased in human primary chondrocytes under leptin stimulation. Moreover, this BMP-2 induction was mediated through transcription factor-signal transducer and activator of transcription (STAT 3 activation via JAK2-ERK1/2-induced Ser727-phosphorylation. Of note, histone deacetylases (HDACs 3 and 4 were both involved in modulating leptin-induced BMP-2 mRNA expression through different pathways: HDAC3, but not HDAC4, associated with STAT3 to form a complex. Our results further demonstrated that the role of BMP-2 induction under leptin stimulation is to increase collagen II expression. The findings in this study provide new insights into the regulatory mechanism of BMP-2 induction in leptin-stimulated chondrocytes and suggest that BMP-2 may play a reparative role in regulating leptin-induced OA development.

  7. Identification of bone morphogenetic protein 7 (BMP7) as an instructive factor for human epidermal Langerhans cell differentiation.

    Science.gov (United States)

    Yasmin, Nighat; Bauer, Thomas; Modak, Madhura; Wagner, Karin; Schuster, Christopher; Köffel, Rene; Seyerl, Maria; Stöckl, Johannes; Elbe-Bürger, Adelheid; Graf, Daniel; Strobl, Herbert

    2013-11-18

    Human Langerhans cell (LC) precursors populate the epidermis early during prenatal development and thereafter undergo massive proliferation. The prototypic antiproliferative cytokine TGF-β1 is required for LC differentiation from human CD34(+) hematopoietic progenitor cells and blood monocytes in vitro. Similarly, TGF-β1 deficiency results in LC loss in vivo. However, immunohistology studies revealed that human LC niches in early prenatal epidermis and adult basal (germinal) keratinocyte layers lack detectable TGF-β1. Here we demonstrated that these LC niches express high levels of bone morphogenetic protein 7 (BMP7) and that Bmp7-deficient mice exhibit substantially diminished LC numbers, with the remaining cells appearing less dendritic. BMP7 induces LC differentiation and proliferation by activating the BMP type-I receptor ALK3 in the absence of canonical TGF-β1-ALK5 signaling. Conversely, TGF-β1-induced in vitro LC differentiation is mediated via ALK3; however, co-induction of ALK5 diminished TGF-β1-driven LC generation. Therefore, selective ALK3 signaling by BMP7 promotes high LC yields. Within epidermis, BMP7 shows an inverse expression pattern relative to TGF-β1, the latter induced in suprabasal layers and up-regulated in outer layers. We observed that TGF-β1 inhibits microbial activation of BMP7-generated LCs. Therefore, TGF-β1 in suprabasal/outer epidermal layers might inhibit LC activation, resulting in LC network maintenance.

  8. Estimated glomerular filtration rate in sickle cell anemia is associated with polymorphisms of bone morphogenetic protein receptor 1B.

    Science.gov (United States)

    Nolan, Vikki G; Ma, Qianli; Cohen, Herbert T; Adewoye, Adeboye; Rybicki, Anne C; Baldwin, Clinton; Mahabir, Rhea N; Homan, Erica P; Wyszynski, Diego F; Fabry, Mary E; Nagel, Ronald L; Farrer, Lindsay A; Steinberg, Martin H

    2007-03-01

    Renal disease is common in sickle cell anemia. In this exploratory work, we used data from a longitudinal study of the natural history of sickle cell disease to examine the hypothesis that polymorphisms (SNPs) in selected candidate genes are associated with glomerular filtration rate (GFR). DNA samples and clinical and laboratory data were available for 1,140 patients with sickle cell anemia. GFR was estimated using the Cockcroft-Gault and Schwartz formulas for adults and children, respectively. We examined approximately 175 haplotype tagging (ht) SNPs in about 70 genes of the TGFbeta/BMP pathway for their association with GFR using linear regression. Four SNPs in BMPR1B, a bone morphogenetic protein (BMP) receptor gene, yielded statistically significant associations (P values ranging from 0.015 to 0.046). Three haplotypes in this gene were also associated with GFR. The TGF-beta/BMP pathway has been associated with the development of diabetic nephropathy, which has some features in common with sickle cell nephropathy. Our results suggest that, as with other subphenotypes of sickle cell disease, renal function may be genetically modulated.

  9. Bone morphogenetic protein antagonist noggin promotes skin tumorigenesis via stimulation of the Wnt and Shh signaling pathways.

    Science.gov (United States)

    Sharov, Andrey A; Mardaryev, Andrei N; Sharova, Tatyana Y; Grachtchouk, Marina; Atoyan, Ruzanna; Byers, H Randolph; Seykora, John T; Overbeek, Paul; Dlugosz, Andrzej; Botchkarev, Vladimir A

    2009-09-01

    Bone morphogenetic proteins (BMPs) play pivotal roles in the regulation of skin development. To study the role of BMPs in skin tumorigenesis, BMP antagonist noggin was used to generate keratin 14-targeted transgenic mice. In contrast to wild-type mice, transgenic mice developed spontaneous hair follicle-derived tumors, which resemble human trichofolliculoma. Global gene expression profiles revealed that in contrast to anagen hair follicles of wild-type mice, tumors of transgenic mice showed stage-dependent increases in the expression of genes encoding the selected components of Wnt and Shh pathways. Specifically, expression of the Wnt ligands increased at the initiation stage of tumor formation, whereas expression of the Wnt antagonist and tumor suppressor Wnt inhibitory factor-1 decreased, as compared with fully developed tumors. In contrast, expression of the components of Shh pathway increased in fully developed tumors, as compared with the tumor placodes. Consistent with the expression data, pharmacological treatment of transgenic mice with Wnt and Shh antagonists resulted in the stage-dependent inhibition of tumor initiation, and progression, respectively. Furthermore, BMP signaling stimulated Wnt inhibitory factor-1 expression and promoter activity in cultured tumor cells and HaCaT keratinocytes, as well as inhibited Shh expression, as compared with the corresponding controls. Thus, tumor suppressor activity of the BMPs in skin epithelium depends on the local concentrations of noggin and is mediated at least in part via stage-dependent antagonizing of Wnt and Shh signaling pathways.

  10. Heparan sulfate acts as a bone morphogenetic protein coreceptor by facilitating ligand-induced receptor hetero-oligomerization.

    Science.gov (United States)

    Kuo, Wan-Jong; Digman, Michelle A; Lander, Arthur D

    2010-11-15

    Cell surface heparan sulfate (HS) not only binds several major classes of growth factors but also sometimes potentiates their activities--an effect usually termed "coreception." A view that coreception is due to the stabilization of growth factor-receptor interactions has emerged primarily from studies of the fibroblast growth factors (FGFs). Recent in vivo studies have strongly suggested that HS also plays an important role in regulating signaling by the bone morphogenetic proteins (BMPs). Here, we provide evidence that the mechanism of coreception for BMPs is markedly different from that established for FGFs. First, we demonstrate a direct, stimulatory role for cell surface HS in the immediate signaling activities of BMP2 and BMP4, and we provide evidence that HS-BMP interactions are required for this effect. Next, using several independent assays of ligand binding and receptor assembly, including coimmunoprecipitation, cross-linking, and fluorescence fluctuation microscopy, we show that HS does not affect BMP binding to type I receptor subunits but instead enhances the subsequent recruitment of type II receptor subunits to BMP-type I receptor complexes. This suggests a view of HS as a catalyst of the formation of signaling complexes, rather than as a stabilizer of growth factor binding.

  11. Mutation in bone morphogenetic protein receptor-IB is associated with increased ovulation rate in Booroola Mérino ewes

    Science.gov (United States)

    Mulsant, Philippe; Lecerf, Frédéric; Fabre, Stéphane; Schibler, Laurent; Monget, Philippe; Lanneluc, Isabelle; Pisselet, Claudine; Riquet, Juliette; Monniaux, Danielle; Callebaut, Isabelle; Cribiu, Edmond; Thimonier, Jacques; Teyssier, Jacques; Bodin, Loys; Cognié, Yves; Chitour, Nour; Elsen, Jean-Michel

    2001-01-01

    Ewes from the Booroola strain of Australian Mérino sheep are characterized by high ovulation rate and litter size. This phenotype is due to the action of the FecBB allele of a major gene named FecB, as determined by statistical analysis of phenotypic data. By genetic analysis of 31 informative half-sib families from heterozygous sires, we showed that the FecB locus is situated in the region of ovine chromosome 6 corresponding to the human chromosome 4q22–23 that contains the bone morphogenetic protein receptor IB (BMPR-IB) gene encoding a member of the transforming growth factor-β (TGF-β) receptor family. A nonconservative substitution (Q249R) in the BMPR-IB coding sequence was found to be associated fully with the hyperprolificacy phenotype of Booroola ewes. In vitro, ovarian granulosa cells from FecBB/FecBB ewes were less responsive than granulosa cells from FecB+/FecB+ ewes to the inhibitory effect on steroidogenesis of GDF-5 and BMP-4, natural ligands of BMPR-IB. It is suggested that in FecBB/FecBB ewes, BMPR-IB would be inactivated partially, leading to an advanced differentiation of granulosa cells and an advanced maturation of ovulatory follicles. PMID:11320249

  12. Repair of rat cranial bone defect by using bone morphogenetic protein-2-related peptide combined with microspheres composed of polylactic acid/polyglycolic acid copolymer and chitosan.

    Science.gov (United States)

    Li, Jingfeng; Jin, Lin; Wang, Mingbo; Zhu, Shaobo; Xu, Shuyun

    2015-07-08

    The effects of the transplanted bone morphogenetic protein-2 (BMP2) -related peptide P24 and rhBMP2 combined with poly(lactic-co-glycolic acid) (PLGA)/chitosan (CS) microspheres were investigated in promoting the repair of rat cranial bone defect. Forty white rats were selected and equally divided into four groups (group A: 1 μg of rhBMP2/PLGA/CS composite; group B: 3 mg of P24/PLGA/CS composite; group C: 0.5 μg of rhBMP2 + 1.5 mg of P24/PLGA/CS composite; group D: blank PLGA/CS material), and rat cranial bone defect models with a diameter of 5 mm were established. The materials were transplanted to the cranial bone defects. The animals were sacrificed on weeks 6 and 12 post-operation. Radiographic examinations (x-ray imaging and 3D CT scanning) and histological evaluations were performed. The repaired areas of cranial bone defects were measured, and the osteogenetic abilities of various materials were compared. Cranial histology, imaging, and repaired area measurements showed that the osteogenetic effects at two time points (weeks 6 and 12) in group C were better than those in groups A and B. The effects in groups A and B were similar. Group D achieved the worst repair effect of cranial bone defects, where a large number of fibrous connective tissues were observed. The PLGA/CS composite microspheres loaded with rhBMP2 and P24 had optimal concrescence and could mutually increase their osteogenesis capability. rhBMP2 + P24/PLGA/CS composite is a novel material for bone defect repair with stable activity to induce bone formation.

  13. Linkage between stature and a region on chromosome 20 and analysis of a candidate gene, bone morphogenetic protein 2

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, D.B.; Ossowski, V.; Janssen, R.C.; Knowler, W.C.; Bogardus, C. [National Inst. of Health, Phoenix, AZ (United States)

    1995-12-04

    Sib-pair linkage analysis of the quantitative trait, stature, in over 500 Pima Indians indicates that a genetic determinant of governing stature is located on chromosome 20. Analysis of 10 short tandem repeat polymorphisms localized this linkage to a 3. cM region that includes D20S98 and D20S66. Using all possible sib-pair combinations, linkage was detected to both stature (P = 0.0001) and to leg length (P = 0.001), but not to sitting height. Single-strand conformational polymorphism analysis of exon 3 of the bone morphogenetic protein 2 (BMP2) gene, a candidate gene in this region, in genomic DNA of 20 of the tallest and 20 of the shortest individuals did not show any consistent differences associated with leg length or height. Sequence analysis of the region encoding the mature protein revealed a single nucleotide substitution, a T to G transversion, not detected by single-strand conformational polymorphism (SSCP) analysis. This transversion results in a conservative amino acid substitution of glycine for valine at codon 80 of BMP2. The frequency of this allele was 0.23 in the sample. No significant differences in height were noted in persons carrying either allele. This indicates that this structural alteration in the mature BMP2 protein does not contribute to the differences in stature observed in the Pima Indians, nor is this structural change in the mature protein likely to be responsible for the linkage observed with stature on chromosome 20. 33 refs., 2 figs., 2 tabs.

  14. Recombinant human bone morphogenetic protein-2 promotes the proliferation of mesenchymal stem cells in vivo and in vitro

    Institute of Scientific and Technical Information of China (English)

    LIU Shui-bing; HU Pei-zhen; HOU Ying; LI Peng; CAO Wei; TIAN Qiong

    2009-01-01

    Background Bone morphogenetic protein (BMP) is a member of the superfamily of transforming growth factor-β.Recent studies show that it is an indispensable factor in hematopoiesis.To better characterize the effect of recombinant human BMP (rhBMP)-2 in hematopoiesis,we set out to determine whether rhBMP-2 could promote the proliferation of mesenchymal stem cells (MSCs) and increase the levels of hematopoietic cytokines in MSCs.Methods 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino) carbonyl)-2H-tetrazolium hydroxide (XTT),real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) were used to deteMP-2 on the proliferation and hematopoietic cytokine levels of MSCs.In addition,MSCs marked with Hoechst33342 were transplanted into BALB/c mice by the intravenous route or intra-bone marrow transplantation,and cluster numbers were counted.Results The XTT test revealed that rhBMP-2 significantly induced proliferation of MSCs in doses ranging from 10 ng/ml to 0.1 mg/ml in a dose-dependent manner.The experiments in vivo showed that there were more clusters of donor cells in bone marrow,spleen,liver and lung of the BMP group than those in the control group after both intra-bone marrow transplantation (P<0.001,P <0.001,P <0.001,and P=0.001,respectively) and intravenous transplantation (P <0.001,P <0.001,and P <0.001 respectively).The results of real-time PCR and ELISA revealed that rhBMP-2 significantly increased mRNA expressions and protein levels of IL-6,IL-7,IL-11,G-CSF,M-CSF and SCF.Conclusions The treatment with rhBMP-2 promotes the proliferation of MSCs in vivo and in vitro and increases the levels of hematopoietic cytokines in MSCs,which may contribute to the improvement of hematopoietic function.

  15. Form-deprivation myopia induces decreased expression of bone morphogenetic protein-2, 5 in guinea pig sclera

    Institute of Scientific and Technical Information of China (English)

    Qing; Wang; Mei-Lan; Xue; Gui-Qiu; Zhao; Mei-Guang; Liu; Yu-Na; Ma; Yan; Ma

    2015-01-01

    AIM: To identify the presence of various bone morphogenetic proteins(BMPs) and their receptors in normal sclera of human, rat and guinea pigs, and to determine whether their expression changed with form-deprivation myopia(FDM) in guinea pig sclera.METHODS: The expression of BMPs and BMP receptors were detected using reverse transcription polymerase chain reaction(RT-PCR) and immunofluorescence. Two-week-old guinea pigs were monocularly form-deprived with a translucent lens. After fourteen days induction of FDM, total RNA was isolated and subjected to RT-PCR to examine the changes of BMPs and BMP receptors in tissues from the posterior sclera. Western blotting analysis was used to investigate their changes in protein levels.RESULTS: Human sclera expressed m RNAs for BMP-2,-4,-5,-7,-RIA,-RIB and BMP-RII. Conversely, rat sclera only expressed m RNA for BMP-7 and BMP-RIB,while the expression of BMPs and BMP receptors in guinea pigs were similar to that of humans. Human sclera also expresses BMP-2,-4,-5,-7 in protein level.Fourteen days after the induction of myopia, significant decreased expressions for BMP-2 and BMP-5 in the posterior sclera of FDM-affected eyes(P <0.05 vs internal control eyes).· CONCLUSION: Various BMPs were expressed in human and guinea pig sclera. In the posterior sclera,expressions of BMP-2 and BMP-5 significantly decreased in FDM eyes. This finding indicates that various BMPs as components of the scleral cytokines regulating tissue homeostasis and provide evidence that alterations in the expression of BMP-2 and BMP-5 are associated with sclera remodeling during myopia induction.

  16. The Effects of Tocotrienol and Lovastatin Co-Supplementation on Bone Dynamic Histomorphometry and Bone Morphogenetic Protein-2 Expression in Rats with Estrogen Deficiency

    Directory of Open Access Journals (Sweden)

    Kok-Yong Chin

    2017-02-01

    Full Text Available Both tocotrienol and statins are suppressors of the mevalonate pathway. Supplementation of tocotrienol among statin users could potentially protect them against osteoporosis. This study aimed to compare the effects of tocotrienol and lovastatin co-supplementation with individual treatments on bone dynamic histomorphometric indices and bone morphogenetic protein-2 (BMP-2 gene expression in ovariectomized rats. Forty-eight female Sprague-Dawley rats were randomized equally into six groups. The baseline was sacrificed upon receipt. All other groups were ovariectomized, except for the sham group. The ovariectomized groups were administered orally daily with (1 lovastatin 11 mg/kg/day alone; (2 tocotrienol derived from annatto bean (annatto tocotrienol 60 mg/kg/day alone; (3 lovastatin 11 mg/kg/day, and annatto tocotrienol 60 mg/kg/day. The sham and ovariectomized control groups were treated with equal volume of vehicle. After eight weeks of treatment, the rats were sacrificed. Their bones were harvested for bone dynamic histomorphometry and BMP-2 gene expression. Rats supplemented with annatto tocotrienol and lovastatin concurrently demonstrated significantly lower single-labeled surface, but increased double-labeled surface, mineralizing surface, mineral apposition rate and bone formation rate compared to individual treatments (p < 0.05. There was a parallel increase in BMP-2 gene expression in the rats receiving combined treatment (p < 0.05. The combination of annatto tocotrienol and lovastatin exerted either additively or synergistically on selected bone parameters. In conclusion, tocotrienol can augment the bone formation and mineralization in rats receiving low-dose statins. Supplementation of tocotrienol in statin users can potentially protect them from osteoporosis.

  17. Demineralized bone matrix combined bone marrow mesenchymal stem cells, bone morphogenetic protein-2 and transforming growth factor-β3 gene promoted pig cartilage defect repair.

    Directory of Open Access Journals (Sweden)

    Xin Wang

    Full Text Available OBJECTIVES: To investigate whether a combination of demineralized bone matrix (DBM and bone marrow mesenchymal stem cells (BMSCs infected with adenovirus-mediated- bone morphogenetic protein (Ad-BMP-2 and transforming growth factor-β3 (Ad-TGF-β3 promotes the repair of the full-thickness cartilage lesions in pig model. METHODS: BMSCs isolated from pig were cultured and infected with Ad-BMP-2(B group, Ad-TGF-β3 (T group, Ad-BMP-2 + Ad-TGF-β3(BT group, cells infected with empty Ad served as a negative group(N group, the expression of the BMP-2 and TGF-β3 were confirmed by immunofluorescence, PCR, and ELISA, the expression of SOX-9, type II collagen(COL-2A, aggrecan (ACAN in each group were evaluated by real-time PCR at 1w, 2w, 3w, respectively. The chondrogenic differentiation of BMSCs was evaluated by type II collagen at 21d with immunohistochemical staining. The third-passage BMSCs infected with Ad-BMP-2 and Ad-TGF-β3 were suspended and cultured with DBM for 6 days to construct a new type of tissue engineering scaffold to repair full-thickness cartilage lesions in the femur condyles of pig knee, the regenerated tissue was evaluated at 1,2 and 3 months after surgery by gross appearance, H&E, safranin O staining and O'driscoll score. RESULTS: Ad-BMP-2 and Ad-TGF-β3 (BT group infected cells acquired strong type II collagen staining compared with Ad-BMP-2 (B group and Ad-TGF-β3 (T group along. The Ad-BMP-2 and Ad-TGF-β3 infected BMSCs adhered and propagated well in DBM and the new type of tissue engineering scaffold produced hyaline cartilage morphology containing a stronger type II collagen and safranin O staining, the O'driscoll score was higher than other groups. CONCLUSIONS: The DBM compound with Ad-BMP-2 and Ad-TGF-β3 infected BMSCs scaffold has a good biocompatibility and could well induce cartilage regeneration to repair the defects of joint cartilage. This technology may be efficiently employed for cartilage lesions repair in vivo.

  18. Posterior maxillary sandwich osteotomy combined with sinus grafting with bone morphogenetic protein-2 for alveolar reconstruction for dental implants: report of four cases.

    Science.gov (United States)

    Jensen, Ole T; Cottam, Jared

    2013-01-01

    Four patients underwent posterior sandwich osteotomy combined with sinus floor grafting using bone morphogenetic protein-2 and other grafting materials. The patients were treated over a period of 4 years. Two to four implants were placed in each site subsequently. Of the 12 implants placed, none failed. Alveolar crest bone levels appeared to be stable over time, with an average vertical gain of about 5 mm. Overall vertical gain, including the sinus graft, exceeded 13 mm in all patients. The procedure appears to hold promise for combined vertical alveolar defects and prominent pneumatization of the posterior maxilla.

  19. Positive Selection in Bone Morphogenetic Protein 15 Targets a Natural Mutation Associated with Primary Ovarian Insufficiency in Human

    Science.gov (United States)

    Meslin, Camille; Monestier, Olivier; Di Pasquale, Elisa; Pascal, Géraldine; Persani, Luca; Fabre, Stéphane

    2013-01-01

    Bone Morphogenetic Protein 15 (BMP15) is a TGFβ-like oocyte-derived growth factor involved in ovarian folliculogenesis as a critical regulator of many granulosa cell processes. Alterations of the BMP15 gene have been found associated with different ovarian phenotypic effects depending on the species, from sterility to increased prolificacy in sheep, slight subfertility in mouse or associated with primary ovarian insufficiency (POI) in women. To investigate the evolving role of BMP15, a phylogenetic analysis of this particular TGFβ family member was performed. A maximum likelihood phylogenetic tree of several TGFβ/BMP family members expressed by the ovary showed that BMP15 has a very strong divergence and a rapid evolution compared to others. Moreover, among 24 mammalian species, we detected signals of positive selection in the hominidae clade corresponding to F146, L189 and Y235 residues in human BMP15. The biological importance of these residues was tested functionally after site directed-mutagenesis in a COV434 cells luciferase assay. By replacing the positively selected amino acid either by alanine or the most represented residue in other studied species, only L189A, Y235A and Y235C mutants showed a significant increase of BMP15 signaling when compared to wild type. Additionally, the Y235C mutant was more potent than wild type in inhibiting progesterone secretion of ovine granulosa cells in primary culture. Interestingly, the Y235C mutation was previously identified in association with POI in women. In conclusion, this study evidences that the BMP15 gene has evolved faster than other members of the TGFß family and was submitted to a positive selection pressure in the hominidae clade. Some residues under positive selection are of great importance for the normal function of the protein and thus for female fertility. Y235 represents a critical residue in the determination of BMP15 biological activity, thus indirectly confirming its role in the onset of POI in

  20. A novel link between the proteasome pathway and the signal transduction pathway of the Bone Morphogenetic Proteins (BMPs

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    Kim Richard H

    2002-06-01

    Full Text Available Abstract Background The intracellular signaling events of the Bone Morphogenetic Proteins (BMPs involve the R-Smad family members Smad1, Smad5, Smad8 and the Co-Smad, Smad4. Smads are currently considered to be DNA-binding transcriptional modulators and shown to recruit the master transcriptional co-activator CBP/p300 for transcriptional activation. SNIP1 is a recently discovered novel repressor of CBP/p300. Currently, the detailed molecular mechanisms that allow R-Smads and Co-Smad to co-operatively modulate transcription events are not fully understood. Results Here we report a novel physical and functional link between Smad1 and the 26S proteasome that contributes to Smad1- and Smad4-mediated transcriptional regulation. Smad1 forms a complex with a proteasome β subunit HsN3 and the ornithine decarboxylase antizyme (Az. The interaction is enhanced upon BMP type I receptor activation and occur prior to the incorporation of HsN3 into the mature 20S proteasome. Furthermore, BMPs trigger the translocation of Smad1, HsN3 and Az into the nucleus, where the novel CBP/p300 repressor protein SNIP1 is further recruited to Smad1/HsN3/Az complex and degraded in a Smad1-, Smad4- and Az-dependent fashion. The degradation of the CBP/p300 repressor SNIP1 is likely an essential step for Smad1-, Smad4-mediated transcriptional activation, since increased SNIP1 expression inhibits BMP-induced gene responses. Conclusions Our studies thus add two additional important functional partners of Smad1 into the signaling web of BMPs and also suggest a novel mechanism for Smad1 and Smad4 to co-modulate transcription via regulating proteasomal degradation of CBP/p300 repressor SNIP1.

  1. Bone morphogenetic protein 2 promotes transforming growth factor β3-induced chondrogenesis of human osteoarthritic synovium-derived stem cells

    Institute of Scientific and Technical Information of China (English)

    RUI Yun-feng; DU Lin; WANG You; WANG Yang; LUI Pauline po-yee; TANG Ting-ting; CHAN Kai-ming; DAI Ke-rong

    2010-01-01

    Background Synovium-derived stem cells (SDSCs) with higher chondrogenic potential are attracting considerable attention as a cell source for cartilage regeneration. We investigated the effect of bone morphogenetic protein 2 (BMP-2) on transforming growth factor beta3 (TGF-β3)-induced chondrogenesis of SDSCs isolated from human osteoarthritic synovium in a pellet culture system. Methods The clonogenicity, stem cell marker expression and multi-differentiation potential of isolated SDSCs were determined by colony forming unit assay, flow cytometry and specific staining including alizarin red S, Oil red O and alcian blue staining, respectively. SDSCs pellet was cultured in chondrogenic medium with or without TGF-β3 or/and BMP-2. At day 21, the diameter and the weight of the pellets were measured. Chondrogenic differentiation of SDSCs was evaluated by Safranin O staining, immunohistochemical staining of collagen type Ⅱ, sulfated glycosaminoglycan (sGAG) synthesis and mRNA expression of collagen type Ⅱ, aggrecan, SOX9, link-protein, collagen type X and BMP receptor Ⅱ. Results Cells isolated under the optimized culturing density (104/60 cm2) showed clonogenicity and multi-differentiation potential. These cells were positive (>99%) for CD44, CD90, CD105 and negative (<10%) for CD34 and CD71. SDSCs differentiated to a chondrocytic phenotype in chondrogenic medium containing TGF-β3 with or without BMP-2. Safranin O staining of the extracellular matrix was positive and the expression of collagen type Ⅱ was detected. Cell pellets treated with TGF-β3 and BMP-2 were larger in diameter and weight, produced more sGAGs, and expressed higher levels of collagen type Ⅱ and other chondrogenic markers, except COL10A1, than medium with TGF-β3 alone. Conclusions SDSCs could be isolated from human osteoarthritic synovium. Supplementation with BMP-2 significantly promoted the in vitro TGF-β3-induced chondrogenic differentiation of SDSCs.

  2. Comparative, osteochondral defect repair: Stem cells versus chondrocytes versus Bone Morphogenetic Protein-2, solely or in combination

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    R Reyes

    2013-07-01

    Full Text Available Full-thickness articular cartilage damage does not resolve spontaneously. Studies with growth factors, implantation of autologous chondrocytes and mesenchymal stem cells have led to variable, to some extent inconsistent, results. This work compares osteochondral knee-defect repair in rabbits upon implantation of a previously described alginate/(poly(lactic-co-glycolic acid (PLGA osteochondral scaffold in distinct conditions. Systems were either in vitro pre-cultured with a small number of allogeneic chondrocytes under fibroblast growth factor (FGF-2 stimulation or the same amount of allogeneic, marrow derived, mesenchymal stem cells (without any pre-differentiation, or loaded with microsphere-encapsulated bone morphogenetic protein (BMP-2 within the alginate layer, or holding combinations of one or the other cell type with BMP-2. The experimental limit was 12 weeks, because a foregoing study with this release system had shown a maintained tissue response for at least 24 weeks post-operation. After only 6 weeks, histological analyses revealed newly formed cartilage-like tissue, which resembled the adjacent, normal cartilage in cell as well as BMP-2 treated defects, but cell therapy gave higher histological scores. This advantage evened out until 12 weeks. Combinations of cells and BMP-2 did not result in any additive or synergistic effect. Equally efficient osteochondral defect repair was achieved with chondrocyte, stem cell, and BMP-2 treatment. Expression of collagen X and collagen I, signs of ongoing ossification, were histologically undetectable, and the presence of aggrecan protein indicated cartilage-like tissue. In conclusion, further work should demonstrate whether spatiotemporally controlled, on-site BMP-2 release alone could become a feasible therapeutic approach to repair large osteochondral defects.

  3. Management of subtrochanteric femur fractures with internal fixation and recombinant human bone morphogenetic protein-7 in a patient with osteopetrosis: a case report

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    Golden Robert D

    2010-05-01

    Full Text Available Abstract Introduction Osteopetrosis is a group of conditions characterized by defects in the osteoclastic function of the bone resulting in defective bone resorption. Clinically, the condition is characterized by a dense, sclerotic, deformed bone which, despite an increased density observable by radiography, often results in an increased propensity to fracture and delayed union. Case Presentation We report the case of a 27-year-old Asian man presenting with bilateral subtrochanteric femur fractures. He had a displaced right subtrochanteric femur fracture after a low-energy fall, which was treated surgically. The second fracture that our patient endured was diagnosed as a stress fracture ten weeks later when he complained of pain in the contralateral left thigh. By that time, the right-sided fracture exhibited no radiographic evidence of healing, and when the left-sided stress fracture was being treated surgically, bone grafting with recombinant human bone morphogenetic protein-7 was also performed on the right side. Conclusion While there are no data supporting the use of bone morphogenic proteins in the management of delayed healing in patients with osteopetrosis, no other reliable osteoinductive grafting options are available to treat this condition. Both fractures in our patient healed, but based on the serial radiographic assessment it is uncertain to what degree the recombinant human bone morphogenetic protein-7 may have contributed to the successful outcome. It may have also contributed to the formation of heterotopic bone around the fracture site. Further investigation of the effectiveness and indications of bone morphogenic protein use for the management of delayed fracture healing in patients with osteopetrosis is warranted.

  4. Bone morphogenetic protein 2 promotes osteogenesis of bone marrow stromal cells in type 2 diabetic rats via the Wnt signaling pathway.

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    Qian, Chao; Zhu, Chenyuan; Yu, Weiqiang; Jiang, Xinquan; Zhang, Fuqiang; Sun, Jian

    2016-11-01

    Type 2 diabetes mellitus impairs osteogenesis in bone marrow stromal cells (BMSCs). Bone morphogenetic protein 2 (BMP2) has been extensively applied for bone defect restoration and has been shown to activate the Wnt signaling pathway. The objective of this study was to investigate the effects of BMP2 on the cell proliferation and osteogenesis of type 2 diabetic BMSCs in rats and explore whether BMP2 induced osteogenesis via the stimulation of Wnt signaling pathway. The cell experiments were divided into DM (diabetic BMSCs), BMP25 (induced with 25ng/ml BMP2), BMP100 (induced with 100ng/ml BMP2) and BMP25 +XAV groups. All cells with or without the different concentrations of BMP2 were cultured under the same experimental conditions. The in vitro results indicated that BMP2 enhanced cell proliferation by 130%-157% and osteogenic differentiation by approximately two-fold in type 2 diabetic BMSCs. The expression levels of β-catenin, cyclin D1, Runx2 and c-myc related to the Wnt signaling pathway were also upregulated from 180% to 212% in BMP2-induced type 2 diabetic rat BMSCs, while the level of GSK3β decreased to 43%. In BMP2-induced type 2 diabetic BMSCs with calcium phosphate cement (CPC) scaffolds for osteoblast study in vivo, the appearance of newly formed bone dramatically increased to 175% compared with type 2 diabetic BMSCs. These data demonstrated that BMP2 enhanced bone regeneration in diabetic BMSCs by stimulating the Wnt signaling pathway with the accumulation of β-catenin and the depressed expression of GSK3β. Diabetic BMSCs associated with BMP2 might be a potential tissue-engineered construct for bone defects in type 2 diabetes mellitus.

  5. Differential effects of bone morphogenetic protein-2 and transforming growth factor-β1 on gene expression of collagen-modifying enzymes in human adipose tissue-derived mesenchymal stem cells

    NARCIS (Netherlands)

    Knippenberg, M.; Helder, M.N.; Doulabi, B.Z.; Bank, R.A.; Wuisman, P.I.J.M.; Klein-Nulend, J.

    2009-01-01

    Adipose tissue-derived mesenchymal stem cells (AT-MSCs) in combination with bone morphogenetic protein-2 (BMP-2) or transforming growth factor-β1 (TGF-β1) are under evaluation for bone tissue engineering. Posttranslational modification of type I collagen is essential for functional bone tissue with

  6. Expression of genes for bone morphogenetic proteins BMP-2, BMP-4 and BMP-6 in various parts of the human skeleton

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    Włodarski Krzysztof

    2007-12-01

    Full Text Available Abstract Background Differences in duration of bone healing in various parts of the human skeleton are common experience for orthopaedic surgeons. The reason for these differences is not obvious and not clear. Methods In this paper we decided to measure by the use of real-time RT-PCR technique the level of expression of genes for some isoforms of bone morphogenetic proteins (BMPs, whose role is proven in bone formation, bone induction and bone turnover. Seven bone samples recovered from various parts of skeletons from six cadavers of young healthy men who died in traffic accidents were collected. Activity of genes for BMP-2, -4 and -6 was measured by the use of fluorescent SYBR Green I. Results It was found that expression of m-RNA for BMP-2 and BMP-4 is higher in trabecular bone in epiphyses of long bones, cranial flat bones and corpus mandibulae then in the compact bone of diaphyses of long bones. In all samples examined the expression of m-RNA for BMP-4 was higher than for BMP-2. Conclusion It was shown that m-RNA for BMP-6 is not expressed in the collected samples at all. It is postulated that differences in the level of activation of genes for BMPs is one of the important factors which determine the differences in duration of bone healing of various parts of the human skeleton.

  7. Mutation analysis of exon1 of bone morphogenetic protein-15 gene in Iranian patients with polycystic ovarian syndrome

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    Mehdizadeh, Anahita; Sheikhha, Mohammad Hasan; Kalantar, Seyed Mehdi; Aali, Bibi Shahnaz; Ghanei, Azam

    2016-01-01

    Background: With the prevalence of 6-10%, polycystic ovarian syndrome (PCOS) is considered the most common endocrinological disorder affecting women in their reproductive age. It has been suggested that genetic factors participate in the development of PCOS. Follicular development has been considered as one of the impaired processes in PCOS. Bone morphogenetic protein-15 (BMP-15) gene is a candidate gene in follicular development and its variants may play role in pathogenesis of PCOS. Objective: To investigate whether BMP-15 gene mutations are present in Iranian women with PCOS. Materials and Methods: In this cross-sectional study 5 ml venous blood samples was taken from 70 PCOS women referring to Afzalipour Hospital, Kerman University of Medical Sciences, Kerman, Iran, between January to December 2014. Genomic DNA was extracted from the blood sample by salting out method. Then a set of PCR reactions for exon1 of BMP-15 gene was performed using specific primers followed by genotyping with direct sequencing. Results: Two different polymorphisms were found in the gene under study. In total 20 patients (28.6%) were heterozygote (C/G), and 2 patients (2.86%) were homozygous (G/G) for c.-9C>G in 5´UTR promoter region of BMP-15 gene (rs3810682). In addition, in the coding region of exon1, three patients (4.3%) were heterozygote (G/A) for c.A308G (rs41308602). Two PCOS patients (2.86%) appeared to have both c.-9C>G (C/G) and c.A308G (G/A) variants simultaneously. Conclusion: Our research detected two polymorphisms of BMP-15 gene among PCOS patients, indicating that even though it cannot be concluded that variants of BMP-15 gene are the principal cause of polycystic ovarian syndrome; they could be involved in pathogenic process in development of PCOS. PMID:27679828

  8. Bone morphogenetic protein 4 (BMP4) induces buffalo (Bubalus bubalis) embryonic stem cell differentiation into germ cells.

    Science.gov (United States)

    Shah, Syed Mohmad; Saini, Neha; Ashraf, Syma; Singh, Manoj Kumar; Manik, Radhey Sham; Singla, Suresh Kumar; Palta, Prabhat; Chauhan, Manmohan Singh

    2015-12-01

    The aim of the present study was to investigate the effect of Bone morphogenetic protein 4 (BMP4) stimulation on differentiation of buffalo embryonic stem (ES) cells into germ lineage cells. ES cells were subjected to in vitro differentiation in floating and adherent cultures, under different BMP4 concentrations (20, 50 and 100 ngml(-1)) for different culture intervals (4, 8 and 14 days). qPCR analysis revealed that BMP4 at a concentration of 50-100 ngml(-1) for a culture period of 14 days led to maximum induction of germ lineage genes like DAZL, VASA, PLZF (PGC-specific); SYCP3, MLH1, TNP1/2 and PRM2 (Meiotic genes); BOULE and TEKT1 (Spermatocyte markers); GDF9, ZP2 and 3 (Oocyte markers). The expression levels of all the genes were significantly higher under BMP4 differentiation as compared to BMP4 + NOGGIN and spontaneously differentiated cultures. Immunocytochemical analysis of embryoid bodies (EBs) and monolayer adherent cultures revealed expression of PGC- (c-KIT, DAZL and VASA); Meiotic- (SYCP3, MLH1 and PROTAMINE1); Spermatocyte- (ACROSIN and HAPRIN); and Oocyte- markers (GDF9 and ZP4). Western blotting was positive for VASA, GDF9 and ZP4. Oocyte-like structures (OLS) obtained in monolayer differentiated cultures harbored a big nucleus and a ZP4 coat. They showed embryonic development and progressed through 2-cell, 4-cell, 8-cell and blastocyst-like structures. Global DNA methylation analysis showed significantly (p < 0.05) decreased levels of 5-methyl-2-deoxycytidine in EBs obtained in optimum differentiation medium. The expression of meiotic markers coupled with expression of spermatocyte and oocyte markers is an indication of post-meiotic progression into spermatogenesis and oogenesis, respectively.

  9. Endoglin-mediated suppression of prostate cancer invasion is regulated by activin and bone morphogenetic protein type II receptors.

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    Michael J Breen

    Full Text Available Mortality from prostate cancer (PCa is due to the formation of metastatic disease. Understanding how that process is regulated is therefore critical. We previously demonstrated that endoglin, a type III transforming growth factor β (TGFβ superfamily receptor, suppresses human PCa cell invasion and metastasis. Endoglin-mediated suppression of invasion was also shown by us to be dependent upon the type I TGFβ receptor, activin receptor-like kinase 2 (ALK2, and the downstream effector, Smad1. In this study we demonstrate for the first time that two type II TGFβ receptors are required for endoglin-mediated suppression of invasion: activin A receptor type IIA (ActRIIA and bone morphogenetic protein receptor type II (BMPRII. Downstream signaling through these receptors is predominantly mediated by Smad1. ActRIIA stimulates Smad1 activation in a kinase-dependent manner, and this is required for suppression of invasion. In contrast BMPRII regulates Smad1 in a biphasic manner, promoting Smad1 signaling through its kinase domain but suppressing it through its cytoplasmic tail. BMPRII's Smad1-regulatory effects are dependent upon its expression level. Further, its ability to suppress invasion is independent of either kinase function or tail domain. We demonstrate that ActRIIA and BMPRII physically interact, and that each also interacts with endoglin. The current findings demonstrate that both BMPRII and ActRIIA are necessary for endoglin-mediated suppression of human PCa cell invasion, that they have differential effects on Smad1 signaling, that they make separate contributions to regulation of invasion, and that they functionally and physically interact.

  10. The hedgehog signal induced modulation of bone morphogenetic protein signaling: an essential signaling relay for urinary tract morphogenesis.

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    Ryuma Haraguchi

    Full Text Available BACKGROUND: Congenital diseases of the urinary tract are frequently observed in infants. Such diseases present a number of developmental anomalies such as hydroureter and hydronephrosis. Although some genetically-modified mouse models of growth factor signaling genes reproduce urinary phenotypes, the pathogenic mechanisms remain obscure. Previous studies suggest that a portion of the cells in the external genitalia and bladder are derived from peri-cloacal mesenchymal cells that receive Hedgehog (Hh signaling in the early developmental stages. We hypothesized that defects in such progenitor cells, which give rise to urinary tract tissues, may be a cause of such diseases. METHODOLOGY/PRINCIPAL FINDINGS: To elucidate the pathogenic mechanisms of upper urinary tract malformations, we analyzed a series of Sonic hedgehog (Shh deficient mice. Shh(-/- displayed hydroureter and hydronephrosis phenotypes and reduced expression of several developmental markers. In addition, we suggested that Shh modulation at an early embryonic stage is responsible for such phenotypes by analyzing the Shh conditional mutants. Tissue contribution assays of Hh-responsive cells revealed that peri-cloacal mesenchymal cells, which received Hh signal secreted from cloacal epithelium, could contribute to the ureteral mesenchyme. Gain- and loss-of-functional mutants for Hh signaling revealed a correlation between Hh signaling and Bone morphogenetic protein (Bmp signaling. Finally, a conditional ablation of Bmp receptor type IA (BmprIA gene was examined in Hh-responsive cell lineages. This system thus made it possible to analyze the primary functions of the growth factor signaling relay. The defective Hh-to-Bmp signaling relay resulted in severe urinary tract phenotypes with a decrease in the number of Hh-responsive cells. CONCLUSIONS/SIGNIFICANCE: This study identified the essential embryonic stages for the pathogenesis of urinary tract phenotypes. These results suggested that Hh

  11. Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10.

    Science.gov (United States)

    Susan-Resiga, Delia; Essalmani, Rachid; Hamelin, Josée; Asselin, Marie-Claude; Benjannet, Suzanne; Chamberland, Ann; Day, Robert; Szumska, Dorota; Constam, Daniel; Bhattacharya, Shoumo; Prat, Annik; Seidah, Nabil G

    2011-07-01

    Bone morphogenetic protein 10 (BMP10) is a member of the TGF-β superfamily and plays a critical role in heart development. In the postnatal heart, BMP10 is restricted to the right atrium. The inactive pro-BMP10 (∼60 kDa) is processed into active BMP10 (∼14 kDa) by an unknown protease. Proteolytic cleavage occurs at the RIRR(316)↓ site (human), suggesting the involvement of proprotein convertase(s) (PCs). In vitro digestion of a 12-mer peptide encompassing the predicted cleavage site with furin, PACE4, PC5/6, and PC7, showed that furin cleaves the best, whereas PC7 is inactive on this peptide. Ex vivo studies in COS-1 cells, a cell line lacking PC5/6, revealed efficient processing of pro-BMP10 by endogenous PCs other than PC5/6. The lack of processing of overexpressed pro-BMP10 in the furin- and PACE4-deficient cell line, CHO-FD11, and in furin-deficient LoVo cells, was restored by stable (CHO-FD11/Fur cells) or transient (LoVo cells) expression of furin. Use of cell-permeable and cell surface inhibitors suggested that endogenous PCs process pro-BMP10 mostly intracellularly, but also at the cell surface. Ex vivo experiments in mouse primary hepatocytes (wild type, PC5/6 knock-out, and furin knock-out) corroborated the above findings that pro-BMP10 is a substrate for endogenous furin. Western blot analyses of heart right atria extracts from wild type and PACE4 knock-out adult mice showed no significant difference in the processing of pro-BMP10, implying no in vivo role of PACE4. Overall, our in vitro, ex vivo, and in vivo data suggest that furin is the major convertase responsible for the generation of BMP10.

  12. Furin Is the Major Processing Enzyme of the Cardiac-specific Growth Factor Bone Morphogenetic Protein 10*

    Science.gov (United States)

    Susan-Resiga, Delia; Essalmani, Rachid; Hamelin, Josée; Asselin, Marie-Claude; Benjannet, Suzanne; Chamberland, Ann; Day, Robert; Szumska, Dorota; Constam, Daniel; Bhattacharya, Shoumo; Prat, Annik; Seidah, Nabil G.

    2011-01-01

    Bone morphogenetic protein 10 (BMP10) is a member of the TGF-β superfamily and plays a critical role in heart development. In the postnatal heart, BMP10 is restricted to the right atrium. The inactive pro-BMP10 (∼60 kDa) is processed into active BMP10 (∼14 kDa) by an unknown protease. Proteolytic cleavage occurs at the RIRR316↓ site (human), suggesting the involvement of proprotein convertase(s) (PCs). In vitro digestion of a 12-mer peptide encompassing the predicted cleavage site with furin, PACE4, PC5/6, and PC7, showed that furin cleaves the best, whereas PC7 is inactive on this peptide. Ex vivo studies in COS-1 cells, a cell line lacking PC5/6, revealed efficient processing of pro-BMP10 by endogenous PCs other than PC5/6. The lack of processing of overexpressed pro-BMP10 in the furin- and PACE4-deficient cell line, CHO-FD11, and in furin-deficient LoVo cells, was restored by stable (CHO-FD11/Fur cells) or transient (LoVo cells) expression of furin. Use of cell-permeable and cell surface inhibitors suggested that endogenous PCs process pro-BMP10 mostly intracellularly, but also at the cell surface. Ex vivo experiments in mouse primary hepatocytes (wild type, PC5/6 knock-out, and furin knock-out) corroborated the above findings that pro-BMP10 is a substrate for endogenous furin. Western blot analyses of heart right atria extracts from wild type and PACE4 knock-out adult mice showed no significant difference in the processing of pro-BMP10, implying no in vivo role of PACE4. Overall, our in vitro, ex vivo, and in vivo data suggest that furin is the major convertase responsible for the generation of BMP10. PMID:21550985

  13. Kartogenin, transforming growth factor-β1 and bone morphogenetic protein-7 coordinately enhance lubricin accumulation in bone-derived mesenchymal stem cells.

    Science.gov (United States)

    Liu, Chun; Ma, Xueqin; Li, Tao; Zhang, Qiqing

    2015-09-01

    Osteoarthritis, a common joint degeneration, can cause breakdown of articular cartilage with the presence of lubricin metabolic abnormalities. Lubricin is a multi-level chondroprotective mucinous glycoprotein in articular joints. Joint defect and infection is elevated and accompanied by accelerated cartilage lesions involving degradation and loss of lubricin. However, a novel, heterocyclic compound called kartogenin (KGN) was discovered to stimulate chondrogenic differentiation of bone-derived mesenchymal stem cells (BMSCs). And the synergistic effect of transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7) could provoke lubricin accumulation. This paper attempted to explore the connection between accumulation of lubricin and the effect of TGF-β1, BMP-7 and/or KGN. Hence, we investigated the expression and secretion of lubricin in BMSCs treated with different combinations of TGF-β1, BMP-7, and/or KGN. Using an in vitro BMSCs system, we observed the content of lubricin from BMSCs treated with TGF-β1, BMP-7, and KGN was the highest at both the protein level and the gene level. The accumulation of lubricin was enhanced coordinately by the increase of synthesis and decrease of degradation possibly via c-Myc and adamts5 pathway. These results further suggested that supplementation of the defect parts with lubricin by using growth factors and small molecules showed a promising potential on preventing joint deterioration in patients with acquired or genetic deficiency of lubricin in the future of regenerative medicine.

  14. Wnt5a signaling is a substantial constituent in bone morphogenetic protein-2-mediated osteoblastogenesis

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    Nemoto, Eiji, E-mail: e-nemoto@dent.tohoku.ac.jp [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Ebe, Yukari; Kanaya, Sousuke [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tsuchiya, Masahiro [Department of Aging and Geriatric Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Nakamura, Takashi [Department of Pediatric Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tamura, Masato [Department of Biochemistry and Molecular Biology, Hokkaido University Graduate School of Dentistry, Sapporo 060-8586 (Japan); Shimauchi, Hidetoshi [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)

    2012-06-15

    Highlights: Black-Right-Pointing-Pointer Wnt5a is identified in osteoblasts in tibial growth plate and bone marrow. Black-Right-Pointing-Pointer Osteoblastic differentiation is associated with increased expression of Wnt5a/Ror2. Black-Right-Pointing-Pointer Wnt5a/Ror2 signaling is important for BMP-2-mediated osteoblastic differentiation. Black-Right-Pointing-Pointer Wnt5a/Ror2 operates independently of BMP-Smad pathway. -- Abstract: Wnts are secreted glycoproteins that mediate developmental and post-developmental physiology by regulating cellular processes including proliferation, differentiation, and apoptosis through {beta}-catenin-dependent canonical and {beta}-catenin-independent noncanonical pathway. It has been reported that Wnt5a activates noncanonical Wnt signaling through receptor tyrosine kinase-like orphan receptor 2 (Ror2). Although it appears that Wnt5a/Ror2 signaling supports normal bone physiology, the biological significance of noncanonical Wnts in osteogenesis is essentially unknown. In this study, we identified expression of Wnt5a in osteoblasts in the ossification zone of the tibial growth plate as well as bone marrow of the rat tibia as assessed by immunohistochemistry. In addition, we show that osteoblastic differentiation mediated by BMP-2 is associated with increased expression of Wnt5a and Ror2 using cultured pre-osteoblasts, MC3T3-E1 cells. Silencing gene expression of Wnt5a and Ror2 in MC3T3-E1 cells results in suppression of BMP-2-mediated osteoblastic differentiation, suggesting that Wnt5a and Ror2 signaling are of substantial importance for BMP-2-mediated osteoblastic differentiation. BMP-2 stimulation induced phosphorylation of Smad1/5/8 in a similar fashion in both siWnt5a-treated cells and control cells, suggesting that Wnt5a was dispensable for the phosphorylation of Smads by BMP-2. Taken together, our results suggest that Wnt5a/Ror2 signaling appears to be involved in BMP-2-mediated osteoblast differentiation in a Smad independent

  15. Wnt5a signaling is a substantial constituent in bone morphogenetic protein-2-mediated osteoblastogenesis.

    Science.gov (United States)

    Nemoto, Eiji; Ebe, Yukari; Kanaya, Sousuke; Tsuchiya, Masahiro; Nakamura, Takashi; Tamura, Masato; Shimauchi, Hidetoshi

    2012-06-15

    Wnts are secreted glycoproteins that mediate developmental and post-developmental physiology by regulating cellular processes including proliferation, differentiation, and apoptosis through β-catenin-dependent canonical and β-catenin-independent noncanonical pathway. It has been reported that Wnt5a activates noncanonical Wnt signaling through receptor tyrosine kinase-like orphan receptor 2 (Ror2). Although it appears that Wnt5a/Ror2 signaling supports normal bone physiology, the biological significance of noncanonical Wnts in osteogenesis is essentially unknown. In this study, we identified expression of Wnt5a in osteoblasts in the ossification zone of the tibial growth plate as well as bone marrow of the rat tibia as assessed by immunohistochemistry. In addition, we show that osteoblastic differentiation mediated by BMP-2 is associated with increased expression of Wnt5a and Ror2 using cultured pre-osteoblasts, MC3T3-E1 cells. Silencing gene expression of Wnt5a and Ror2 in MC3T3-E1 cells results in suppression of BMP-2-mediated osteoblastic differentiation, suggesting that Wnt5a and Ror2 signaling are of substantial importance for BMP-2-mediated osteoblastic differentiation. BMP-2 stimulation induced phosphorylation of Smad1/5/8 in a similar fashion in both siWnt5a-treated cells and control cells, suggesting that Wnt5a was dispensable for the phosphorylation of Smads by BMP-2. Taken together, our results suggest that Wnt5a/Ror2 signaling appears to be involved in BMP-2-mediated osteoblast differentiation in a Smad independent pathway.

  16. Bone morphogenetic protein-1 and its related metalloproteinase%骨形态发生蛋白-1及其相关金属蛋白酶

    Institute of Scientific and Technical Information of China (English)

    陈冬瑛; 朱全胜; 丘钜世

    2004-01-01

    Bone morphogenetic protein-1(BMP-1) and its related molecules are members of metalloendoproteinase astacin family, including BMP-1, mTLD, mTLL-1 and mTLL-2. Even though all of them lack of the ability to induce bone or cartilage formation directly, they play key roles in numerable activities in ECM from embryo to adult, then affect the procedure and the result of osteogenesis and bone remodeling directly or indirectly. They are critical in maturation and deposition of some major collagen types, and in regulating the signaling of some growth factors in TGF-β superfamily by degradation of TGF-β inhibitor such as Chordin. The investigations about tissue distribution of BMP-1 and its related proteinases and also gene knock-out studies strongly indicate that they play key roles in osteogenesis and bone development.

  17. Profile of serum alkaline phosphatase after inoculation of mononuclear cells and bone morphogenetic protein in the repair of osteochondral defects in rabbits

    Directory of Open Access Journals (Sweden)

    Luiz Augusto de Souza

    2011-12-01

    Full Text Available In this study, serum alkaline phosphatase activity was measured in response to the repair of osteochondral defects in twenty-four New Zealand rabbits. The animals were divided into three groups: a control (GC, those treated with bone marrow mononuclear cells (GCM and those that received mononuclear cells with autologous bone morphogenetic protein (BMP + GCM. After exposing the trochlear groove of the left stifle joint, a wedge-shaped segment was removed. Later, the defect was filled with an osteochondral autograft preserved in 98% glycerin. For the GC group, only the bone graft was performed. For the GCM, in addition to the graft, 2x106 seed mononuclear cells were implanted. For the GCM + BMP, the same number of cells, associated with 1μg of bone morphogenetic protein, were intraarticularly administered. The osteoblastic response was measured by analyzing the serum alkaline phosphatase on day 0 (preoperative 3, 15, 30, and 45 after surgery, and by radiographic examinations. Analysis of variance in randomized blocks, factorial and Tukey’s test (p = 0.05 were made. The overall mean GCM was superior to the other groups and the highest rates were among the 15th and 45th days postoperatively. The discrepancy in values between individuals of the same group casts doubts on the veracity of the test.

  18. Inhibition of beta cell growth and function by bone morphogenetic proteins

    DEFF Research Database (Denmark)

    Bruun, Christine; Christensen, Gitte Lund; Jacobsen, Marie L B;

    2014-01-01

    : BMP2 and -4 were found to inhibit basal as well as growth factor-stimulated proliferation of primary beta cells from rats and mice. Bmp2 and Bmp4 mRNA and protein were expressed in islets and regulated by inflammatory cytokines. Neutralisation of endogenous BMP activity resulted in enhanced....../INTERPRETATION: These data show that BMP2 and -4 exert inhibitory actions on beta cells in vitro and suggest that BMPs exert regulatory roles of beta cell growth and function.......AIMS/HYPOTHESIS: Impairment of beta cell mass and function is evident in both type 1 and type 2 diabetes. In healthy physiological conditions pancreatic beta cells adapt to the body's increasing insulin requirements by proliferation and improved function. We hypothesised that during the development...

  19. Bone morphogenetic protein-5 (BMP-5 promotes dendritic growth in cultured sympathetic neurons

    Directory of Open Access Journals (Sweden)

    Higgins Dennis

    2001-09-01

    Full Text Available Abstract Background BMP-5 is expressed in the nervous system throughout development and into adulthood. However its effects on neural tissues are not well defined. BMP-5 is a member of the 60A subgroup of BMPs, other members of which have been shown to stimulate dendritic growth in central and peripheral neurons. We therefore examined the possibility that BMP-5 similarly enhances dendritic growth in cultured sympathetic neurons. Results Sympathetic neurons cultured in the absence of serum or glial cells do not form dendrites; however, addition of BMP-5 causes these neurons to extend multiple dendritic processes, which is preceded by an increase in phosphorylation of the Smad-1 transcription factor. The dendrite-promoting activity of BMP-5 is significantly inhibited by the BMP antagonists noggin and follistatin and by a BMPR-IA-Fc chimeric protein. RT-PCR and immunocytochemical analyses indicate that BMP-5 mRNA and protein are expressed in the superior cervical ganglia (SCG during times of initial growth and rapid expansion of the dendritic arbor. Conclusions These data suggest a role for BMP-5 in regulating dendritic growth in sympathetic neurons. The signaling pathway that mediates the dendrite-promoting activity of BMP-5 may involve binding to BMPR-IA and activation of Smad-1, and relative levels of BMP antagonists such as noggin and follistatin may modulate BMP-5 signaling. Since BMP-5 is expressed at relatively high levels not only in the developing but also the adult nervous system, these findings suggest the possibility that BMP-5 regulates dendritic morphology not only in the developing, but also the adult nervous system.

  20. Recombinant human bone morphogenetic protein 2-induced heterotopic ossification of the retroperitoneum, psoas muscle, pelvis and abdominal wall following lumbar spinal fusion

    Energy Technology Data Exchange (ETDEWEB)

    Shah, Raj K. [The George Washington University School of Medicine, Washington, DC (United States); Moncayo, Valeria M.; Pierre-Jerome, Claude; Terk, Michael R. [Emory University School of Medicine, Radiology Department, Musculoskeletal Division, Atlanta, GA (United States); Smitson, Robert D. [Emory University School of Medicine, Atlanta, GA (United States)

    2010-05-15

    A 45-year-old man presented with vertebral collapse at L5 as an initial manifestation of multiple myeloma and underwent spinal fusion surgery using recombinant human bone morphogenetic protein-2 (rhBMP-2). Subsequent computed tomography (CT) scans and X-rays revealed heterotopic ossification of the left psoas muscle, pelvis, and anterior abdominal wall. While the occurrence of heterotopic ossification has previously been reported when rhBMP-2 has been used for spinal fusion surgery, this case demonstrates that it can occur to a much greater degree than previously seen. (orig.)

  1. Bone morphogenetic protein-15 in follicle fluid combined with age may differentiate between successful and unsuccessful poor ovarian responders

    Directory of Open Access Journals (Sweden)

    Wu Yan-Ting

    2012-12-01

    Full Text Available Abstract Background The counselling of poor ovarian responders about the probability of pregnancy remains a puzzle for gynaecologists. The aim of this study was to optimise the management of poor responders by investigating the role of the oocyte-derived factor bone morphogenetic protein-15 (BMP-15 combined with chronological age in the prediction of the outcome of in-vitro fertilisation-embryo transfer (IVF-ET in poor responders. Methods A retrospective study conducted in a university hospital. A total of 207 poor ovarian responders who reached the ovum pick-up stage undergoing IVF/intracytoplasmic sperm injection (ICSI with three or fewer follicles no less than 14 mm on the day of oocyte retrieval were recruited from July 1, 2008 to December 31, 2009. Another 215 coinstantaneous cycles with normal responses were selected as controls. The BMP-15 levels in the follicular fluid (FF of the 207 poor responders were analysed by western blot. Based on the FF BMP-15 level and age, poor responders were sub-divided into four groups. The main outcome measures were the FF BMP-15 level, implantation rate, pregnancy rate, and live birth rate. Results The implantation rate (24.2% vs. 15.3%, chemical pregnancy rate (40% vs. 23.7%, clinical pregnancy rate (36.5% vs. 20.4% and live birth rate (29.4% vs. 15.1% in the high BMP-15 group were significantly higher than those in the low BMP-15 group. Furthermore, poor responders aged less than or equal to 35 years with a higher FF BMP-15 level had the best implantation, pregnancy and live birth rates, which were comparable with those of normal responders. Conclusions Our study suggests a potential role of BMP-15 in the prediction of the IVF outcome. A high FF BMP-15 combined with an age less than or equal to 35 years may be used as a potential indicator for repeating IVF cycles in poor ovarian responders.

  2. Expression of bone morphogenetic protein 7 in the cerebral cortex of rats after ischemic-hypoxic injury

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    BACKGROUND: Some researches demonstrate that exogenous bone morphogenetic protein 7 (BMP-7) can protect ischemic cerebral nerve tissue and promote recovery of motor energy function; however, there is lack of direct evidences of endogenous BMP-7 effect.OBJECTIVE: To observe the expression of endogenous BMP-7 in nerve tissue with ischemic-hypoxic injury and investigate the possible effects on damaged nerve tissue.DESIGN: Observational contrast animal study.SETTING: Department of Anatomy and Histoembryology, Peking University Health Science Center.MATERIALS: The experiment was carried out in the Nerve Researching Laboratory of Anatomy Department, Peking University Health Science Center from October 2006 to March 2007. A total of 25 adult male SD rats weighing 250 - 300 g and several newborn SD rats were selected from Experimental Animal Center, Peking University Health Science Center. Rabbit-anti-BMP-7 polyclonal antibody was provided by Wuhan Boster Company.METHODS: ① Adult rats were randomly divided into ischemia group (n =10), sham operation group (n =10) and normal group (n =5). Right external-internal carotid artery occlusion was used to infarct middle cerebral artery of adult rats in the ischemia group so as to copy focal cerebral infarction models. Line cork was inserted in crotch of internal and external carotid artery of adult rats in the sham operation group, while adult rats in the normal group were not given any treatments. ② Cerebral cortex of newborn rats was separated to obtain cell suspension. Cells which were cultured for 10 days were divided into control group and hypoxia/reoxygenation group. And then, cells in the hypoxia/reoxygenation group were cultured in hypoxic incubator for 4 hours and given reoxygenation for 24 hours.MAIN OUTCOME MEASURES: Immunohistochemical method was used to measure expression of BMP-7 in cerebral cortex at 24 hours after ischemia/reperfusion culture and in primary hypoxic culture.RESULTS: ① At 24 hours after

  3. The interactions between rat-adipose-derived stromal cells, recombinant human bone morphogenetic protein-2, and beta-tricalcium phosphate play an important role in bone tissue engineering.

    Science.gov (United States)

    E, Ling-Ling; Xu, Lu-Lu; Wu, Xia; Wang, Dong-Sheng; Lv, Yan; Wang, Jia-Zhu; Liu, Hong-Chen

    2010-09-01

    Cells, scaffolds, and growth factors are the three main factors for creating a stem-cell-based tissue-engineered construct, but the interactions between three factors are not very clear. We hereby explored the interactions between rat-adipose-derived stromal cells (rASCs), recombinant human bone morphogenetic protein-2 (rhBMP-2), and beta-tricalcium phosphate (beta-TCP) to provide evidence for their application in bone tissue engineering by evaluating the protein adsorption of beta-TCP, the cell attachment, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, mineral formation, calcium content, phosphonium content, cell vitality, gene expression, and implantation in the backs of severe combined immunodeficient mice of rhBMP-2 preinducing rASCs seeded onto beta-TCP. The results showed that beta-TCP could adsorb the proteins from the media. The attachment, proliferation, and osteogenic properties of rASCs were supported by beta-TCP, as revealed using scanning electron microscopy. Compared with rASCs cultured on the culture plate, rASCs cultured on beta-TCP had significantly higher ALP activity/protein, OCN content, and mineral formation. These values for rASCs cultured on beta-TCP with rhBMP-2 increased most significantly. The rhBMP-2 significantly increased the calcium content, phosphonium content, and ALP, type I collagen, and OCN mRNA levels of rASCs cultured on beta-TCP. The methylthiazol tetrazolium method revealed that the vitality of rASCs cultured on beta-TCP with or without rhBMP-2 for 4, 7, and 28 days in vitro was insignificantly different. After 8 and 12 weeks of implantation, each group displayed increased bone formation over the 12-week period. The percentage of the new bone formed areas for beta-TCP/rhBMP-2 and beta-TCP was not significantly different. This value for rASCs/beta-TCP construct was significantly higher than that for beta-TCP group, but the maximal and robust bone formation was presented in rASCs/beta-TCP with rhBMP-2

  4. Effects of Escherichia Coli-derived Recombinant Human Bone Morphogenetic Protein-2 Loaded Porous Hydroxyaptite-based Ceramics on Calvarial Defect in Rabbits

    Science.gov (United States)

    Kim, Shin-Young; Lee, Youngkyun; Seo, Seung-Jun; Lim, Jae-Hong

    2017-01-01

    Background Recombinant human bone morphogenetic proteins (rhBMPs) have been widely used in regenerative therapies to promote bone formation. The production of rhBMPs using bacterial systems such as Escherichia coli (E. coli) is estimated to facilitate clinical applications by lowering the cost without compromising biological activity. In clinical practice, rhBMP-2 and osteoconductive carriers (e.g., hydroxyapatite [HA] and bovine bone xenograft) are used together. This study examined the effect of E. coli-derived rhBMP-2 combined with porous HA-based ceramics on calvarial defect in rabbits. Methods Six adult male New Zealand white rabbits were used in this study. The experimental groups were divided into the following 4 groups: untreated (NC), bovine bone graft (BO), porous HA (HA) and porous HA with rhBMP-2 (HA-BMP). Four transosseous defects of 8 mm in diameter were prepared using stainless steel trephine bur in the frontal and parietal bones. Histological and histomorphometric analyses at 4 weeks after surgery revealed significant new bone formation by porous HA alone. Results HA-BMP showed significantly higher degree of bone formation compared with BO and HA group (Pceramics can promote new bone formation. PMID:28326298

  5. Increased osteoinductivity and mineralization by minimal concentration of bone morphogenetic protein-2 loaded onto biphasic calcium phosphate in a rabbit sinus

    Science.gov (United States)

    2016-01-01

    Purpose The purpose of the present study was to evaluate the effectiveness of a minimal concentration of bone morphogenetic protein-2 (BMP-2) in terms of quantitative and qualitative analyses of newly formed bone in a rabbit maxillary sinus model. Methods In 7 rabbits, sinus windows were prepared bilaterally. Biphasic calcium phosphate (BCP) loaded with 0.05 mg/mL BMP-2 was grafted into one sinus (the BMP group) and saline-soaked BCP was placed into the other (the control group) in each animal. The animals were allowed an 8-week healing period before being sacrificed. Specimens including the augmented area and surrounding tissues were then removed and evaluated both radiographically and histologically. Results There was a difference in the mineralization of new bone between the groups. In the BMP group, the greater part of the new bone consisted of mature lamellar bone with an evident trabecular pattern, whereas the control group showed mostly woven bone, consisting only partially of lamellar bone. Histometrically, the area of new bone was significantly greater (4.55±1.35 mm2 vs. 2.99±0.86 mm2) in the BMP group than in the control group (Pmineralization in a rabbit sinus model using a BCP carrier. PMID:27800217

  6. Bone morphogenetic protein 2-induced human dental pulp cell differentiation involves p38 mitogen-activated protein kinase-activated canonical WNT pathway

    Institute of Scientific and Technical Information of China (English)

    Jing Yang; Ling Ye; Tian-Qian Hui; Dong-Mei Yang; Ding-Ming Huang; Xue-Dong Zhou; Jeremy J Mao; Cheng-Lin Wang

    2015-01-01

    Both bone morphogenetic protein 2 (BMP2) and the wingless-type MMTV integration site (WNT)/b-catenin signalling pathway play important roles in odontoblast differentiation and dentinogenesis. Cross-talk between BMP2 and WNT/b-catenin in osteoblast differentiation and bone formation has been identified. However, the roles and mechanisms of the canonical WNT pathway in the regulation of BMP2 in dental pulp injury and repair remain largely unknown. Here, we demonstrate that BMP2 promotes the differentiation of human dental pulp cells (HDPCs) by activating WNT/b-catenin signalling, which is further mediated by p38 mitogen-activated protein kinase (MAPK) in vitro. BMP2 stimulation upregulated the expression of b-catenin in HDPCs, which was abolished by SB203580 but not by Noggin or LDN193189. Furthermore, BMP2 enhanced cell differentiation, which was not fully inhibited by Noggin or LDN193189. Instead, SB203580 partially blocked BMP2-induced b-catenin expression and cell differentiation. Taken together, these data suggest a possible mechanism by which the elevation of b-catenin resulting from BMP2 stimulation is mediated by the p38 MAPK pathway, which sheds light on the molecular mechanisms of BMP2-mediated pulp reparative dentin formation.

  7. Induction of bone formation in biphasic calcium phosphate scaffolds by bone morphogenetic protein-2 and primary osteoblasts.

    Science.gov (United States)

    Strobel, L A; Rath, S N; Maier, A K; Beier, J P; Arkudas, A; Greil, P; Horch, R E; Kneser, U

    2014-03-01

    Bone tissue engineering strategies mainly depend on porous scaffold materials. In this study, novel biphasic calcium phosphate (BCP) matrices were generated by 3D-printing. High porosity was achieved by starch consolidation. This study aimed to characterise the porous BCP-scaffold properties and interactions of osteogenic cells and growth factors under in vivo conditions. Five differently treated constructs were implanted subcutaneously in syngeneic rats: plain BCP constructs (group A), constructs pre-treated with BMP-2 (group B; 1.6 µg BMP-2 per scaffold), seeded with primary osteoblasts (OB) (group C), seeded with OB and BMP-2 (group D) and constructs seeded with OB and pre-cultivated in a flow bioreactor for 6 weeks (group E). After 2, 4 and 6 weeks, specimens were explanted and subjected to histological and molecular biological analyses. Explanted scaffolds were invaded by fibrovascular tissue without significant foreign body reactions. Morphometric analysis demonstrated significantly increased bone formation in samples from group D (OB + BMP-2) compared to all other groups. Samples from groups B-E displayed significant mRNA expression of bone-specific genes after 6 weeks. Pre-cultivation in the flow bioreactor (group E) induced bone formation comparable with group B. In this study, differences in bone distribution between samples with BMP-2 or osteoblasts could be observed. In conclusion, combination of osteoblasts and BMP-2 synergistically enhanced bone formation in novel ceramic scaffolds. These results provide the basis for further experiments in orthotopic defect models with a focus on future applications in orthopaedic and reconstructive surgery.

  8. Using poly(lactic-co-glycolic acid microspheres to encapsulate plasmid of bone morphogenetic protein 2/polyethylenimine nanoparticles to promote bone formation in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    Qiao C

    2013-08-01

    Full Text Available Chunyan Qiao,1,* Kai Zhang,2,* Han Jin,1 Leiying Miao,3 Ce Shi,1 Xia Liu,1 Anliang Yuan,1 Jinzhong Liu,1 Daowei Li,1 Changyu Zheng,4 Guirong Zhang,5 Xiangwei Li,1 Bai Yang,2 Hongchen Sun11Department of Pathology, School of Stomatology, Jilin University, Changchun, 2State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 3Institute and Hospital of Stomatology, Nanjing University Medical School, Nanjing, People's Republic of China; 4Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA; 5Department of Biochemistry, School of Basic Medicine, Jilin University, Changchun, People's Republic of China*These authors contributed equally to this workAbstract: Repair of large bone defects is a major challenge, requiring sustained stimulation to continually promote bone formation locally. Bone morphogenetic protein 2 (BMP-2 plays an important role in bone development. In an attempt to overcome this difficulty of bone repair, we created a delivery system to slowly release human BMP-2 cDNA plasmid locally, efficiently transfecting local target cells and secreting functional human BMP-2 protein. For transfection, we used polyethylenimine (PEI to create pBMP-2/PEI nanoparticles, and to ensure slow release we used poly(lactic-co-glycolic acid (PLGA to create microsphere encapsulated pBMP-2/PEI nanoparticles, PLGA@pBMP-2/PEI. We demonstrated that pBMP-2/PEI nanoparticles could slowly release from the PLGA@pBMP-2/PEI microspheres for a long period of time. The 3–15 µm diameter of the PLGA@pBMP-2/PEI further supported this slow release ability of the PLGA@pBMP-2/PEI. In vitro transfection assays demonstrated that pBMP-2/PEI released from PLGA@pBMP-2/PEI could efficiently transfect MC3T3-E1 cells, causing MC3T3-E1 cells to secrete human BMP-2 protein, increase calcium deposition and gene expressions of alkaline

  9. DSP-PP precursor protein cleavage by tolloid-related-1 protein and by bone morphogenetic protein-1.

    Science.gov (United States)

    Ritchie, Helena H; Yee, Colin T; Tang, Xu-Na; Dong, Zhihong; Fuller, Robert S

    2012-01-01

    Dentin sialoprotein (DSP) and phosphophoryn (PP), acidic proteins critical to dentin mineralization, are translated from a single transcript as a DSP-PP precursor that undergoes specific proteolytic processing to generate DSP and PP. The cleavage mechanism continues to be controversial, in part because of the difficulty of obtaining DSP-PP from mammalian cells and dentin matrix. We have infected Sf9 cells with a recombinant baculovirus to produce large amounts of secreted DSP-PP(240), a variant form of rat DSP-PP. Mass spectrometric analysis shows that DSP-PP(240) secreted by Sf9 cells undergoes specific cleavage at the site predicted from the N-terminal sequence of PP extracted from dentin matrix: SMQG(447)↓D(448)DPN. DSP-PP(240) is cleaved after secretion by a zinc-dependent activity secreted by Sf9 cells, generating DSP(430) and PP(240) products that are stable in the medium. DSP-PP processing activity is constitutively secreted by Sf9 cells, but secretion is diminished 3 days after infection. Using primers corresponding to the highly conserved catalytic domain of Drosophila melanogaster tolloid (a mammalian BMP1 homolog), we isolated a partial cDNA for a Spodopotera frugiperda tolloid-related-1 protein (TLR1) that is 78% identical to Drosophila TLR1 but only 65% identical to Drosophila tolloid. Tlr1 mRNA decreased rapidly in Sf9 cells after baculovirus infection and was undetectable 4d after infection, paralleling the observed decrease in secretion of the DSP-PP(240) processing activity after infection. Human BMP1 is more similar to Sf9 and Drosophila TLR1 than to tolloid, and Sf9 TLR1 is more similar to BMP1 than to other mammalian homologs. Recombinant human BMP1 correctly processed baculovirus-expressed DSP-PP(240) in a dose-dependent manner. Together, these data suggest that the physiologically accurate cleavage of mammalian DSP-PP(240) in the Sf9 cell system represents the action of a conserved processing enzyme and support the proposed role of BMP1 in

  10. DSP-PP precursor protein cleavage by tolloid-related-1 protein and by bone morphogenetic protein-1.

    Directory of Open Access Journals (Sweden)

    Helena H Ritchie

    Full Text Available Dentin sialoprotein (DSP and phosphophoryn (PP, acidic proteins critical to dentin mineralization, are translated from a single transcript as a DSP-PP precursor that undergoes specific proteolytic processing to generate DSP and PP. The cleavage mechanism continues to be controversial, in part because of the difficulty of obtaining DSP-PP from mammalian cells and dentin matrix. We have infected Sf9 cells with a recombinant baculovirus to produce large amounts of secreted DSP-PP(240, a variant form of rat DSP-PP. Mass spectrometric analysis shows that DSP-PP(240 secreted by Sf9 cells undergoes specific cleavage at the site predicted from the N-terminal sequence of PP extracted from dentin matrix: SMQG(447↓D(448DPN. DSP-PP(240 is cleaved after secretion by a zinc-dependent activity secreted by Sf9 cells, generating DSP(430 and PP(240 products that are stable in the medium. DSP-PP processing activity is constitutively secreted by Sf9 cells, but secretion is diminished 3 days after infection. Using primers corresponding to the highly conserved catalytic domain of Drosophila melanogaster tolloid (a mammalian BMP1 homolog, we isolated a partial cDNA for a Spodopotera frugiperda tolloid-related-1 protein (TLR1 that is 78% identical to Drosophila TLR1 but only 65% identical to Drosophila tolloid. Tlr1 mRNA decreased rapidly in Sf9 cells after baculovirus infection and was undetectable 4d after infection, paralleling the observed decrease in secretion of the DSP-PP(240 processing activity after infection. Human BMP1 is more similar to Sf9 and Drosophila TLR1 than to tolloid, and Sf9 TLR1 is more similar to BMP1 than to other mammalian homologs. Recombinant human BMP1 correctly processed baculovirus-expressed DSP-PP(240 in a dose-dependent manner. Together, these data suggest that the physiologically accurate cleavage of mammalian DSP-PP(240 in the Sf9 cell system represents the action of a conserved processing enzyme and support the proposed role of BMP

  11. Advances in Bone Morphogenetic Protein-Mediated Gene Therapy%骨形成蛋白介导的基因治疗研究进展

    Institute of Scientific and Technical Information of China (English)

    司晓辉; 杨连君; 冯志军; 刘艳琳; 吕晶; 张雪

    2011-01-01

    Gene therapy is a technique that draws on the introduction of new genes into cells for the purpose of treating disease by restoring or adding gene expression. Bone morphogenetic proteins (BMP) can be delivered in bone defect or fracture local site by means of gene therapy without using heterogeneous carrier. At present, there are two alternative approaches for gene therapy in vivo and ex vivo, which is useful in the healing of the bone and cartilage defect, spinal fusion, craniofacial and dental repair, tendon and ligament formation,and in the treatment of degenerative disc disease. In conclusion, BMP gene therapy is an efficient, economic and promising strategy.%基因治疗是指通过导入基因的功能片段改善机体生理状况或者治疗疾病.利用基因治疗可在骨缺损或骨折局部释放骨形成蛋白(bone morphogenetic proteins,BMP),并且无需异体载体,方法包括体内法和离体法.BMP基因治疗可以促进骨和软骨形成、脊柱融合、颌面骨和牙齿修复、肌腱韧带形成,此外对椎间盘退变性疾病也可采用BMP基因治疗.总之BMP基因治疗方法经济有效,是有前景的治疗手段.

  12. Crystallization and preliminary X-ray analysis of the complex of the first von Willebrand type C domain bound to bone morphogenetic protein 2

    Energy Technology Data Exchange (ETDEWEB)

    Qiu, Li-yan; Zhang, Jin-li [Lehrstuhl für Physiologische Chemie II, Theodor-Boveri-Institut (Biozentrum) der Universität Würzburg, Am Hubland, D-97074 Würzburg (Germany); Kotzsch, Alexander [Lehrstuhl für Molekulare Pflanzenphysiologie und Biophysik, Julius-von-Sachs Institut der Universität Würzburg, Julius-von-Sachs Platz 2, D-97082 Würzburg (Germany); Sebald, Walter [Lehrstuhl für Physiologische Chemie II, Theodor-Boveri-Institut (Biozentrum) der Universität Würzburg, Am Hubland, D-97074 Würzburg (Germany); Rudolf-Virchow-Zentrum (DFG Forschungszentrum) der Universität Würzburg, Versbacher Strasse 9, D-97070 Würzburg (Germany); Mueller, Thomas D., E-mail: mueller@botanik.uni-wuerzburg.de [Lehrstuhl für Molekulare Pflanzenphysiologie und Biophysik, Julius-von-Sachs Institut der Universität Würzburg, Julius-von-Sachs Platz 2, D-97082 Würzburg (Germany); Rudolf-Virchow-Zentrum (DFG Forschungszentrum) der Universität Würzburg, Versbacher Strasse 9, D-97070 Würzburg (Germany); Lehrstuhl für Physiologische Chemie II, Theodor-Boveri-Institut (Biozentrum) der Universität Würzburg, Am Hubland, D-97074 Würzburg (Germany)

    2008-04-01

    Crystals of the complex of the first von Willebrand type C domain (VWC1) of crossveinless 2 (CV2) bound to bone morphogenetic protein 2 (BMP2) exist in two tetragonal crystal forms belonging to either space group P4{sub 1}2{sub 1}2 or I4{sub 1}, with one complete BMP2 dimer and two CV2 VWC1 domains per asymmetric unit, and diffract to 2.6 Å resolution. Crossveinless 2 (CV2) is a member of the chordin family, a protein superfamily that modulates the activity of bone morphogenetic proteins such as BMP2. The BMPs represent a large group of secreted proteins that control many steps during embryonal development and in tissue and organ homeostasis in the adult organism. The gene encoding the first von Willebrand type C domain (VWC1) of CV2 was cloned, expressed in Escherichia coli and purified to homogeneity. The binary complex of CV2 VWC1 and BMP2 was purified and subjected to crystallization. Crystals of SeMet-labelled proteins were obtained in two different forms belonging to the tetragonal space groups P4{sub 1}2{sub 1}2 and I4{sub 1}, with unit-cell parameters a = b = 86.7, c = 139.2 Å and a = b = 83.7, c = 139.6 Å, respectively. Initial analysis suggests that a complete binary complex consisting of one BMP2 dimer bound to two CV2 VWC1 domains is present in the asymmetric unit.

  13. The use of recombinant human bone morphogenetic protein-2 for the treatment of a delayed union following femoral neck open-wedge osteotomy

    Directory of Open Access Journals (Sweden)

    Axel W.A. Baltzer

    2012-03-01

    Full Text Available Although the clinical potential of bone morphogenetic proteins (BMPs has been known for decades, their use in humans has only been approved for a limited number of orthopaedic conditions. Promising results in animals demonstrate the utility of BMP-2 in regional bone repair without using osteoconductors. To our knowledge, no comparable human case has been described. We report the case of a 50- year-old who suffered a femoral neck fracture. After 9 months of extensive treatment, he was still not pain-free. The following open-wedge osteotomy resulted in a therapy-resistant delayed union. We therefore conducted 4 computer tomography-guided injections of recombinant human (rh BMP-2 into the bone gap. No osteoconductor was employed. Six weeks later, there was a 55-60% defect filling. Followup examination showed a complete union of the bone defect. Our case report shows that in a complicated delayed union rhBMP-2 can be successfully used to induce bone formation without any osteoconductor.

  14. [Experimental study on application recombinant human bone morphogenetic protein 2(rhBMP-2)/poly-lactide-co-glycolic acid (PLGA)/fibrin sealant(FS) on repair of rabbit radial bone defect].

    Science.gov (United States)

    Fan, Zhongkai; Cao, Yang; Zhang, Zhe; Zhang, Mingchao; Lu, Wei; Tang, Lei; Yao, Qi; Lu, Gang

    2012-10-01

    This paper is aimed to investigate the repair of rabbit radial bone defect by the recombinant human bone morphogenetic protein 2/poly-lactideco-glycolic acid microsphere with fibrin sealant (rhBMP-2/PLGA/FS). The radial bone defect models were prepared using New Zealand white rabbits, which were randomly divided into 3 groups, experiment group which were injected with eMP-2/PLGA/FS at bone defect location, control group which were injected with FS at bone defect location, and blank control group without treatment. The ability of repairing bone defect was evaluated with X-ray radiograph. Bone mineral density in the defect regions was analysed using the level of ossification. The osteogenetic ability of repairing bone defect, the degradation of the material, the morphologic change and the bone formation were assessed by HE staining and Masson staining. The result showed that rhBMP-2/PLGA/FS had overwhelming superiority in the osteogenetic ability and quality of bone defect over the control group, and it could promote the repair of bone defect and could especially repair the radial bone defect of rabbit well. It may be a promising and efficient synthetic bone graft.

  15. Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication

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    Ma, W.H.; Liu, Y.J.; Wang, W.; Zhang, Y.Z. [The Third Hospital of Hebei Medical University, The Provincial Key Laboratory for Orthopedic Biomechanics of Hebei, Shijiazhuang, Hebei Province (China)

    2015-02-13

    Bone homeostasis seems to be controlled by delicate and subtle “cross talk” between the nervous system and “osteo-neuromediators” that control bone remodeling. The purpose of this study was to evaluate the effect of interactions between neuropeptides and human bone morphogenetic protein 2 (hBMP2) on human osteoblasts. We also investigated the effects of neuropeptides and hBMP2 on gap junction intercellular communication (GJIC). Osteoblasts were treated with neuropeptide Y (NPY), substance P (SP), or hBMP2 at three concentrations. At various intervals after treatment, cell viability was measured by the MTT assay. In addition, cellular alkaline phosphatase (ALP) activity and osteocalcin were determined by colorimetric assay and radioimmunoassay, respectively. The effects of NPY, SP and hBMP on GJIC were determined by laser scanning confocal microscopy. The viability of cells treated with neuropeptides and hBMP2 increased significantly in a time-dependent manner, but was inversely associated with the concentration of the treatments. ALP activity and osteocalcin were both reduced in osteoblasts exposed to the combination of neuropeptides and hBMP2. The GJIC of osteoblasts was significantly increased by the neuropeptides and hBMP2. These results suggest that osteoblast activity is increased by neuropeptides and hBMP2 through increased GJIC. Identification of the GJIC-mediated signal transduction capable of modulating the cellular activities of bone cells represents a novel approach to studying the biology of skeletal innervation.

  16. Recombinant Bone Morphogenetic Protein 2 Stimulates the Remodeling Chitosan-Based Porous Scaffold Into Hyaline-like Cartilage: Study in Heterotopic Implantation

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    Nurshat M. Gaifullin

    2013-09-01

    Full Text Available To study the morphology of remodeling the chitosan-based three-dimensional porous scaffold, containing bone morphogenetic protein-2 (BMP-2 for chondroinduction, the experiments with heterotopic implantation using 28 Wistar rats were carried out. Scaffolds with growth factor (n=12 or without it (n=12, against intact control (n=4 were implanted subcutaneously. Classical methods of histology and morphometry as well as immune histochemical markers (CD-68, CD-31, MMP-9, TIMP-1, and osteonectin expression, one used to investigate zone of remodeling in euthanized animals at 4 and 8 weeks after implantation. The BMP-2 application provides more intensive and rapid new cartilage formation from the scaffold matter. The additional chondroinductive effect proved more intensive settlement and proliferation of chondral cells in the regenerate, expression of chondral phenotype with the building the hyaline-like matrix, and the supporting necessary balance between the matrix metalloproteinases and their tissue inhibitors.

  17. Improved osseointegration of dental titanium implants by TiO2 nanotube arrays with recombinant human bone morphogenetic protein-2: a pilot in vivo study

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    Lee, Jae-Kwan; Choi, Dong-Soon; Jang, Insan; Choi, Won-Youl

    2015-01-01

    TiO2 nanotube arrays on the surface of dental implants were fabricated by two-step anodic oxidation. Their effects on bone-implant contact were researched by a pilot in vivo study. The implants were classified into four groups. An implant group with TiO2 nanotube arrays and recombinant human bone morphogenetic protein-2 (rhBMP-2) was compared with various surface implants, including machined surface, sandblasted large-grit and acid-etched surface, and TiO2 nanotube array surface groups. The diameter of the TiO2 nanotube window and TiO2 nanotube were ~70 nm and ~110 nm, respectively. The rhBMP-2 was loaded into TiO2 nanotube arrays and elution was detected by an interferometric biosensing method. A change in optical thickness of ~75 nm was measured by flow cell testing for 9 days, indicating elution of rhBMP-2 from the TiO2 nanotube arrays. For the in vivo study, the four groups of implants were placed into the proximal tibia of New Zealand White rabbits. In the implant group with TiO2 nanotube arrays and rhBMP-2, the bone-to-implant contact ratio was 29.5% and the bone volume ratio was 77.3%. Bone remodeling was observed not only in the periosteum but also in the interface between the bone and implant threads. These values were higher than in the machined surface, sandblasted large-grit and acid-etched surface, and TiO2 nanotube array surface groups. Our results suggest that TiO2 nanotube arrays could potentially be used as a reservoir for rhBMP-2 to reinforce osseointegration on the surface of dental implants. PMID:25709438

  18. In vivo local co-delivery of recombinant human bone morphogenetic protein-7 and pamidronate via poly-D, L-lactic acid

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    NYC Yu

    2010-12-01

    Full Text Available The effects of bone anabolic agents such as bone morphogenetic proteins (BMPs have the potential to be augmented by co-treatment with an anti-catabolic such as a bisphosphonate. We hypothesised that the effects of bisphosphonates on BMP-induced bone anabolism would be dose dependent, and we aimed to test this in a small animal model. Agents were delivered locally using a biodegradable poly-d, l-lactic-acid (PDLLA polymer delivery system. Recombinant human BMP-7 (25 µg was tested with a range of doses of the bisphosphonate pamidronate (0.02 mg, 0.2 mg and 2 mg local PAM; 0.3 mg/kg and 3 mg/kg thrice-weekly systemic PAM versus BMP-7 alone. Polymer pellets were surgically implanted in the hind limbs of female C57BL6/J mice (8-10 week and ectopic bone nodules were harvested at 3 and 8 weeks post-operatively. At 3 weeks, local low dose PAM (0.02 mg induced a 102% increase in rhBMP-7 induced bone volume (p<0.01 as measured by miroCT, and this was comparable to systemic PAM (0.3 mg/kg thrice-weekly. In contrast, local high dose PAM (2 mg resulted in a 97% decrease in bone volume (p<0.01. Radiography and histology indicated that the polymer vehicle was still largely present at 8 weeks indicating inefficient biodegradation. This is the first study to validate the utility of local co-delivery of BMP/bisphosphonate via biodegradable polymer and supports the continued refinement of more advanced bioresorbable delivery systems for clinical applications.

  19. Evaluation of an injectable silk fibroin enhanced calcium phosphate cement loaded with human recombinant bone morphogenetic protein-2 in ovine lumbar interbody fusion.

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    Gu, Yong; Chen, Liang; Yang, Hui-Lin; Luo, Zong-Ping; Tang, Tian-Si

    2011-05-01

    The objective of this study was to investigate the efficacy of an injectable calcium phosphate cement/silk fibroin/human recombinant bone morphogenetic protein-2 composite (CPC/SF/rhBMP-2) in an ovine interbody fusion model. Twenty-four mature sheep underwent anterior lumbar interbody fusion at the levels of L1/2, L3/4, and L5/6 with random implantation of CPC/SF, CPC/rhBMP-2, CPC/SF/rhBMP-2, or autogenous iliac bone. After the sheep were sacrificed, the fusion segments were evaluated by manual palpation, CT scan, undestructive biomechanical testing, undecalcified histology, and histomorphology. The fusion rates of CPC/SF/rhBMP-2 were 55.56% and 77.78% at 6 and 12 months, respectively. The fusion was superior to all the biomaterial grafts in stiffness, and reached the same stiffness as the autograft at 12 months. The new bone formation was less than autograft at 6 months, but similar with that at 12 months. However, the ceramic residue volume of CPC/SF/rhBMP-2 was significantly decreased compared with CPC/SF and CPC/rhBMP-2 at both times. The results indicated that CPC/SF/rhBMP-2 composite had excellent osteoconduction and osteoinduction, and balanced degradation and osteogenesis.

  20. Interplay between self-assembled structure of bone morphogenetic protein-2 (BMP-2) and osteoblast functions in three-dimensional titanium alloy scaffolds: Stimulation of osteogenic activity.

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    Nune, K C; Kumar, A; Murr, L E; Misra, R D K

    2016-02-01

    Three-dimensional cellular scaffolds are receiving significant attention in bone tissue engineering to treat segmental bone defects. However, there are indications of lack of significant osteoinductive ability of three-dimensional cellular scaffolds. In this regard, the objective of the study is to elucidate the interplay between bone morphogenetic protein (BMP-2) and osteoblast functions on 3D mesh structures with different porosities and pore size that were fabricated by electron beam melting. Self-assembled dendritic microstructure with interconnected cellular-type morphology of BMP-2 on 3D scaffolds stimulated osteoblast functions including adhesion, proliferation, and mineralization, with prominent effect on 2-mm mesh. Furthermore, immunofluorescence studies demonstrated higher density and viability of osteoblasts on lower porosity mesh structure (2 mm) as compared to 3- and 4-mm mesh structures. Enhanced filopodia cellular extensions with extensive cell spreading was observed on BMP-2 treated mesh structures, a behavior that is attributed to the unique self-assembled structure of BMP-2 that effectively communicates with the cells. The study underscores the potential of BMP-2 in imparting osteoinductive capability to the 3D printed scaffolds.

  1. The bone morphogenetic protein antagonist gremlin 1 is overexpressed in human cancers and interacts with YWHAH protein

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    Hur Soo

    2006-03-01

    Full Text Available Abstract Background Basic studies of oncogenesis have demonstrated that either the elevated production of particular oncogene proteins or the occurrence of qualitative abnormalities in oncogenes can contribute to neoplastic cellular transformation. The purpose of our study was to identify an unique gene that shows cancer-associated expression, and characterizes its function related to human carcinogenesis. Methods We used the differential display (DD RT-PCR method using normal cervical, cervical cancer, metastatic cervical tissues, and cervical cancer cell lines to identify genes overexpressed in cervical cancers and identified gremlin 1 which was overexpressed in cervical cancers. We determined expression levels of gremlin 1 using Northern blot analysis and immunohistochemical study in various types of human normal and cancer tissues. To understand the tumorigenesis pathway of identified gremlin 1 protein, we performed a yeast two-hybrid screen, GST pull down assay, and immunoprecipitation to identify gremlin 1 interacting proteins. Results DDRT-PCR analysis revealed that gremlin 1 was overexpressed in uterine cervical cancer. We also identified a human gremlin 1 that was overexpressed in various human tumors including carcinomas of the lung, ovary, kidney, breast, colon, pancreas, and sarcoma. PIG-2-transfected HEK 293 cells exhibited growth stimulation and increased telomerase activity. Gremlin 1 interacted with homo sapiens tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, eta polypeptide (14-3-3 eta; YWHAH. YWHAH protein binding site for gremlin 1 was located between residues 61–80 and gremlin 1 binding site for YWHAH was found to be located between residues 1 to 67. Conclusion Gremlin 1 may play an oncogenic role especially in carcinomas of the uterine cervix, lung, ovary, kidney, breast, colon, pancreas, and sarcoma. Over-expressed gremlin 1 functions by interaction with YWHAH. Therefore, Gremlin 1 and its binding

  2. Histogenesis and possible mechanism of chondroid changes in mixed tumour of the skin: immunohistochemical evaluation of bone morphogenetic protein, glycosaminoglycans, keratin, vimentin and neuronal markers.

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    Mori, M; Shrestha, P; Sakamoto, F; Yang, L J; Qin, C; Tsujimura, T

    1994-01-01

    The distribution of immunoreactivity of bone morphogenetic protein (BMP), the glycosaminoglycans chondroitin 4-sulphate (C4SPG), chondroitin 6-sulphate (C6SPG), dermatan sulphate (DSPG) and keratan sulphate proteoglycans (KSPG), cytokeratin (K8.12), vimentin, glial fibrillary acidic protein (GFAP), actin, desmin, S-100 protein and neuron-specific enolase (NSE) in mixed tumour of the skin was investigated using immunohistochemical methods using monoclonal (MoAb) and polyclonal antibodies (PoAb). A strong BMP immunoreactivity was found characteristically in outer tumour cells of tubuloductal structures and modified myoepithelial cells. Modified myoepithelial cells and chondroidally changed cells showed positive immunoreactivity for C4SPG, C6SPG and DSPG; and KSPG was more pronounced in the modified myoepithelial cells. Vimentin, S-100 protein, GFAP and NSE, but not actin and desmin, were distribute in the outer tumour cells and modified myoepithelial cells in chondroidally changed tissue. Two factors show that chondrogenesis in mixed tumour of the skin is associated with the modified myoepithelial cells through the activity of BMP and biosynthesis of glycosaminoglycans as matrix substance. First, outer or basal tumour cells in mixed tumour of the skin is characterized by the presence of positive immunoreactivity for BMP, KSPG, vimentin, cytokeratin K8.12, S-100 protein, GFAP and NSE, and second, there is a matrix of chondroidally changed tissue containing the reaction products of C4SPG, C6SPG, DSPF and KSPG.

  3. Effects of secretive bone morphogenetic protein 2 induced by gene transfection on the biological changes of NIH3T3 cells

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    SUN Wei-bin; WANG Juan; LU Chun; TANG Gui-xia

    2005-01-01

    Background Bone morphogenetic proteins (BMPs), which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes of NIH3T3 cells incubated with secretive BMP2 that was induced by gene transfection through transwell. Methods Eukaryonic expression vector (pcDNA3.1-B2) was transfered into NIH3T3 cells with SofastTM,a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expressions of BMP2 were determined by immunohistochemical stain and enzyme-linked immunosorbent assay. NIH3T3 cells were co-cultured with hBMP2 gene transfecting cells through transwell, and the ultrastructure, alkaline phosphatase activity and the expression of osteocalcin (the marker of osteogenetic differentiation) changes were observed. Results There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 cells. The ultrastructural changes, the high activity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells. Conclusion Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.

  4. Possible Involvement of Smad Signaling Pathways in Induction of Odontoblastic Properties in KN-3 Cells by Bone Morphogenetic Protein-2: A Growth Factor to Induce Dentin Regeneration

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    Ayako Washio

    2012-01-01

    Full Text Available We examined the effects of bone morphogenetic protein-2 (BMP-2 on growth, differentiation, and intracellular signaling pathways of odontoblast-like cells, KN-3 cells, to clarify molecular mechanisms of odontoblast differentiation during pulp regeneration process. After treatment with BMP-2, the cell morphology, growth, alkaline phosphatase (ALP activity, and the activation and expression of BMP-induced intracellular signaling molecules, such as Smad1/5/8 and Smad6/7, as well as activities of dentin sialoprotein (DSP and dentin matrix protein 1 (DMP1, were examined. BMP-2 had no effects on the morphology, growth, or ALP activity of KN-3 cells, whereas it induced the phosphorylation of Smad1/5/8 and expression of Smad6/7. BMP-2 also induced the expressions of DSP and DMP-1. Our results suggest that KN-3 cells may express an odontoblastic phenotype with the addition of BMP-2 through the activation of Smad signaling pathways.

  5. Wnt3a enhances bone morphogenetic protein 9-induced osteogenic differentiation of C3H10T1/2 cells

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xiao; LIN Liang-bo; XU Dao-jing; CHEN Rong-fu; TAN Ji-xiang; LIANG Xi; HU Ning

    2013-01-01

    Background Bone morphogenetic protein 9 (BMP9) and Wnt/β-catenin signaling pathways are able to induce osteogenic differentiation of mesenchymal stem cells (MSCs),but the role of Wnt/β-catenin signaling pathway in BMP9-induced osteogenic differentiation is not well understood.Thus,our experiment was undertaken to investigate the interaction between BMP9 and Wnt/β-catenin pathway in inducing osteogenic differentiation of MSCs.Methods C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP9,Wnt3a,and BMP9+Wnt3a.ALP,the early osteogenic marker,was detected by quantitative and staining assay.Later osteogenic marker,mineral calcium deposition,was determined by Alizarin Red S staining.The expression of osteopotin (OPN),osteocalcin (OC),and Runx2 was analyzed by Real time PCR and Western blotting.In vivo animal experiment was carried out to further confirm the role of Wnt3a in ectopic bone formation induced by BMP9.Results The results showed that Wnt3a enhanced the ALP activity induced by BMP9 and increased the expressions of OC and OPN,with increase of mineral calcium deposition in vitro and ectopic bone formation in vivo.Furthermore,we also found that Wnt3a increased the level of Runx2,an important nuclear transcription factor of BMP9.Conclusion Canonical Wnt/β-catenin signal pathway may play an important role in BMP9-induced osteogenic differentiation of MSCs,and Runx2 may be a linkage between the two signal pathways.

  6. Mechanical loading induced expression of bone morphogenetic protein-2,alkaline phosphatase activity,and collagen synthesis in osteoblastic MC3T3-E1 cells

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    LU Hong-fei; MAI Zhi-hui; XU Ye; WANG Wei; AI Hong

    2012-01-01

    Background Bone morphogenetic protein(BMP)-2,alkaline phosphatase(ALP),and collagen typeⅠ?are known to play a critical role in the process of bone remodeling.However,the relationship between mechanical strain and the expression of BMP-2,ALP,and COL-Ⅰ?in osteoblasts was still unknown.The purpose of this study was to investigate the effects of different magnitudes of mechanical strain on osteoblast morphology and on the expression of BMP-2,ALP,and COL-Ⅰ.Methods Osteoblast-like cells were flexed at four deformation rates(0,6%,12%,and 18% elongation).The expression of BMP-2 mRNA,ALP,and COL-Ⅰ?in osteoblast-like cells were determined by real-time quantitative reverse transcription polymerase chain reaction,respectively.The results were subjected to analysis of variance(ANOVA)using SPSS 13.0 statistical software.Results The cells changed to fusiform and grew in the direction of the applied strain after the mechanical strain was loaded.Expression level of the BMP-2,ALP,and COL-Ⅰ?increased magnitude-dependently with mechanical loading in the experimental groups,and the 12% elongation group had the highest expression(P<0.05).Conclusion Mechanical strain can induce morphological change and a magnitude-dependent increase in the expression of BMP-2,ALP,and COL-Ⅰ?mRNA in osteoblast-like cells,which might influence bone remodeling in orthodontic treatment.

  7. Magnesium modification up-regulates the bioactivity of bone morphogenetic protein-2 upon calcium phosphate cement via enhanced BMP receptor recognition and Smad signaling pathway.

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    Ding, Sai; Zhang, Jing; Tian, Yu; Huang, Baolin; Yuan, Yuan; Liu, Changsheng

    2016-09-01

    Efficient presentation of growth factors is one of the great challenges in tissue engineering. In living systems, bioactive factors exist in soluble as well as in matrix-bound forms, both of which play an integral role in regulating cell behaviors. Herein, effect of magnesium on osteogenic bioactivity of recombinant human bone morphogenetic protein-2 (rhBMP-2) was investigated systematically with a series of Mg modified calcium phosphate cements (xMCPCs, x means the content of magnesium phosphate cement wt%) as matrix model. The results indicated that the MCPC, especially 5MCPC, could promote the rhBMP-2-induced in vitro osteogenic differentiation via Smad signaling of C2C12 cells. Further studies demonstrated that all MCPC substrates exhibited similar rhBMP-2 release rate and preserved comparable conformation and biological activity of the released rhBMP-2. Also, the ionic extracts of MCPC made little difference to the bioactivity of rhBMP-2, either in soluble or in matrix-bound forms. However, with the quartz crystal microbalance (QCM), we observed a noticeable enhancement of rhBMP-2 mass-uptake on 5MCPC as well as a better recognition of the bound rhBMP-2 to BMPR IA and BMPR II. In vivo results demonstrated a better bone regeneration capacity of 5MCPC/rhBMP-2. From the above, our results demonstrated that it was the Mg anchored on the underlying substrates that tailored the way of rhBMP-2 bound on MCPC, and thus facilitated the recognition of BMPRs to stimulate osteogenic differentiation. The study will guide the development of Mg-doped bioactive bone implants for tissue regeneration.

  8. Comparative study of osteogenic potential of a composite scaffold incorporating either endogenous bone morphogenetic protein-2 or exogenous phytomolecule icaritin: an in vitro efficacy study.

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    Chen, S-H; Wang, X-L; Xie, X-H; Zheng, L-Z; Yao, D; Wang, D-P; Leng, Y; Zhang, G; Qin, L

    2012-08-01

    A local delivery system with sustained and efficient release of therapeutic agents from an appropriate carrier is desirable for orthopedic applications. Novel composite scaffolds made of poly (lactic-co-glycolic acid) with tricalcium phosphate (PLGA/TCP) were fabricated by an advanced low-temperature rapid prototyping technique, which incorporated either endogenous bone morphogenetic protein-2 (BMP-2) (PLGA/TCP/BMP-2) or phytomolecule icaritin (ICT) (PLGA/TCP/ICT) at low, middle and high doses. PLGA/TCP served as control. In vitro degradation, osteogenesis and release tests showed statistical differences among PLGA/TCP/ICT, PLGA/TCP and PLGA/TCP/BMP-2 groups, where PLGA/TCP/ICT had the desired slow release of bioactive icaritin in a dose-dependent manner, whereas there was almost no BMP-2 release from the PLGA/TCP/BMP-2 scaffolds. PLGA/TCP/ICT significantly increased more ALP activity, upregulated mRNA expression of osteogenic genes and enhanced calcium deposition and mineralization in rabbit bone marrow stem cells cultured on scaffolds compared with the other two groups. These results indicate the desired degradation rate, osteogenic capability and release property in PLGA/TCP/ICT composite scaffold, as icaritin preserved its bioactivity and structure after incorporation, while PLGA/TCP/BMP-2 did not show an initially expected osteogenic potential, owing to loss of the original bioactivity of BMP-2 during its incorporation and fabrication procedure. The results suggest that PLGA/TCP composite scaffolds incorporating osteogenic ICT might be a promising approach for bone tissue bioengineering and regeneration.

  9. Effect on cochlea function of guinea pig after controlled release recombinant human bone morphogenetic protein 2 transplanted into the middle ear

    Institute of Scientific and Technical Information of China (English)

    LI Xue-sheng; SUN Jian-jun; JIANG Wei; LIU Xiao

    2010-01-01

    Background The recombinant human bone morphogenetic protein 2 (rhBMP-2) has been used to induce osteogenesis in animals' middle ear and this technique is possible to be used to reconstruct the defects of ossicles. The side effects of the rhBMP-2 in middle ear should be observed before using in clinic. Thus we prepared the controlled release rhBMP-2 and implanted it into the acoustic bulla of guinea pigs. The effect on the cochlea was observed. Methods We prepared the acellular cancellous bone, accompanied with rhBMP-2. The material accompanied with rhBMP-2 was implanted into one acoustic bulla of the animal and the opposite side of the acoustic bulla was implanted with acellular cancellous bone without rhBMP-2. Totally 20 guinea pigs were undergone this procedure. After the operation, the auditory brainstem response (ABR) of the animals was tested according to the time sequence. Three months after the operation, the animals were sacrificed. The osteogenesis induced by rhBMP-2, the acoustic bulla and cochlea affected by rhBMP-2 were observed. The structures of hair cells were observed after silver nitrate staining. Results The animals were recovered soon after surgery. The hearing thresholds of the animals were declined slightly just after the surgery and come back completely after 3 months. Also, the bulla and cochlea were normal in shape. The osteogenesis occurred in the pore of the acellular cancellous bone with rhBMP-2. There was not any abnormal hyperplasia of bone in the bulla and cochlea. The articulation between the stapes and oval window was not merged. The shapes of the hair cells were normal and there was no obvious deletion of the hair cells compared with control group. Conclusions The controlled release rhBMP-2 transplanted into the middle ear could induce osteogenesis in the bulla of the animals. It did not affect the shape of the bulla and the hearing threshold of the animal, and did not induce the abnormal hyperplasia of bone in the bulla and might

  10. 大鼠腭中缝牵张成骨中骨形态发生蛋白7的作用%Role of bone morphogenetic protein-7 in the distraction osteogenesis of rat mid-palatal suture

    Institute of Scientific and Technical Information of China (English)

    李威; 李鹍; 王培军; 韩晶莹

    2012-01-01

    BACKGROUND: Experiments have confirmed that bone morphogenetic protein-7 can induce bone and cartilage formation; it can also promote bone formation in the process of distraction osteogenesis. OBJECTIVE: To explore the role of bone morphogenetic protein-7 in the suture distraction osteogenesis of rat mid-palatal suture and to detect the changes of bone morphogenetic protein-7 under the mechanical expansion force. METHODS: Expansionary force was applied on rats using a self-made expansing arch spring for binoculus to construct maxillary expansion rat model. Rat mid-palatal sutures were randomly collected on 1, 3, 5, 7, 9, 12 and 14 days after model construction; real-time PCR was performed to detect the changing regularities of the target genes. RESULTS AND CONCLUSION: The expression of the bone morphogenetic protein-7 mRNA in the rat mid-palatal sutures increased gradually on 3, 9, 12 and 14 days after model construction (P < 0.05). These findings demonstrate that bone morphogenetic protein-7 mRNA expression is significantly increased in the distraction osteogenesis process of rat mid-palatal sutures under the stimulus of mechanical force.%背景:已有实验证实,骨形态发生蛋白7可诱导骨组织和软骨组织的形成,在牵张成骨的过程中促进骨组织形成.目的:测定骨形态发生蛋白7在腭中缝牵张成骨中的作用及机械扩张力作用下的变化.方法:采用自制双眼扩弓簧对大鼠施加扩张力,建立上颌骨扩张大鼠模型.分别于建模后第1,3,5,7,9,12,14天随机选取大鼠取腭中缝组织,实时定量PCR法测定目的基因的变化规律.结果与结论:建模后第3,9,12,14天大鼠腭中缝组织骨形态发生蛋白7 Mrna的表达逐渐升高(P < 0.05).结果证实,大鼠腭中缝牵张成骨时,骨形态发生蛋白7 Mrna在机械力刺激下表达明显上调.

  11. A Crucial Role of Bone Morphogenetic Protein Signaling in the Wound Healing Response in Acute Liver Injury Induced by Carbon Tetrachloride

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    Nao Oumi

    2012-01-01

    Full Text Available Background. Acute liver injury induced by administration of carbon tetrachloride (CCl4 has used a model of wound repair in the rat liver. Previously, we reported transient expression of bone morphogenetic protein (Bmp 2 or Bmp4 at 6–24 h after CCl4 treatment, suggesting a role of BMP signaling in the wound healing response in the injured liver. In the present study, we investigated the biological meaning of the transient Bmp expression in liver injury. Methods. Using conditional knockout mice carrying a floxed exon in the BMP receptor 1A gene, we determined the hepatic gene expressions and proliferative activity following CCl4-treated liver. Results. We observed retardation of the healing response in the knockout mice treated with CCl4, including aggravated histological feature and reduced expressions of the albumin and Tdo2 genes, and a particular decrease in the proliferative activity shown by Ki-67 immunohistochemistry. Conclusion. Our findings suggest a crucial role of BMP signaling in the amelioration of acute liver injury.

  12. Noggin-Mediated Retinal Induction Reveals a Novel Interplay Between Bone Morphogenetic Protein Inhibition, Transforming Growth Factor β, and Sonic Hedgehog Signaling.

    Science.gov (United States)

    Messina, Andrea; Lan, Lei; Incitti, Tania; Bozza, Angela; Andreazzoli, Massimiliano; Vignali, Robert; Cremisi, Federico; Bozzi, Yuri; Casarosa, Simona

    2015-08-01

    It has long been known that the depletion of bone morphogenetic protein (BMP) is one of the key factors necessary for the development of anterior neuroectodermal structures. However, the precise molecular mechanisms that underlie forebrain regionalization are still not completely understood. Here, we show that Noggin1 is involved in the regionalization of anterior neural structures in a dose-dependent manner. Low doses of Noggin1 expand prosencephalic territories, while higher doses specify diencephalic and retinal regions at the expense of telencephalic areas. A similar dose-dependent mechanism determines the ability of Noggin1 to convert pluripotent cells in prosencephalic or diencephalic/retinal precursors, as shown by transplant experiments and molecular analyses. At a molecular level, the strong inhibition of BMP signaling exerted by high doses of Noggin1 reinforces the Nodal/transforming growth factor (TGF)β signaling pathway, leading to activation of Gli1 and Gli2 and subsequent activation of Sonic Hedgehog (SHH) signaling. We propose a new role for Noggin1 in determining specific anterior neural structures by the modulation of TGFβ and SHH signaling.

  13. Effects of Treatment with Bone Morphogenetic Protein 4 and Co-culture on Expression of Piwil2 Gene in Mouse Differentiated Embryonic Stem Cells

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    Mehdi Forouzandeh-Moghadam

    2009-01-01

    Full Text Available Background: Specific growth factors and feeder layers seem to have important roles in in vitroembryonic stem cells (ESCs differentiation. In this study,the effects of bone morphogenetic protein4 (BMP4 and mouse embryonic fibroblasts (MEFs co-culture system on germ cell differentiationfrom mouse ESCs were studied.Materials and Methods: Cell suspension was prepared from one-day-old embryoid body (EBand cultured for four days in DMEM medium containing 20% fetal bovine serum (FBS in thefollowing groups: simple culture (SC, simple culture with BMP4 (SCB, co-culture (CO-C andco-culture with BMP4 (CO-CB. Expression of piwi-like homolog 2 (Piwil2, the germ cell-specificgene, was evaluated in the different study groups by using quantitative real time polymerase chainreaction (RT-PCR. Testis was used as a positive control.Results: The maximum and minimum Piwil2 expression was observed in SC and SCB groups,respectively. A significant difference was observed in Piwil2 expression between SCB and otherstudy groups (p<0.05.Conclusion: The findings of this study showed that neither the addition of BMP4 in culture mediumnor the use of MEFs as a feeder layer have a positive effect on late germ cell induction from mouseESCs.

  14. Genetic variation in bone morphogenetic proteins and breast cancer risk in hispanic and non-hispanic white women: The breast cancer health disparities study.

    Science.gov (United States)

    Slattery, Martha L; John, Esther M; Torres-Mejia, Gabriela; Herrick, Jennifer S; Giuliano, Anna R; Baumgartner, Kathy B; Hines, Lisa M; Wolff, Roger K

    2013-06-15

    Bone morphogenetic proteins (BMP) are thought to be important in breast cancer promotion and progression. We evaluated genetic variation in BMP-related genes and breast cancer risk among Hispanic (2,111 cases, 2,597 controls) and non-Hispanic White (NHW) (1,481 cases, 1,586 controls) women who participated in the 4-Corner's Breast Cancer Study, the Mexico Breast Cancer Study and the San Francisco Bay Area Breast Cancer Study. BMP genes and their receptors evaluated include ACVR1, AVCR2A, ACVR2B, ACVRL1, BMP1, BMP2, BMP4, BMP6, BMP7, BMPR1A, BMPR1B, BMPR2, MSTN and GDF10. Additionally, 104 ancestral informative markers were assessed to discriminate between European and native American ancestry. The importance of estrogen on BMP-related associations was suggested through unique associations by menopausal status and estrogen (ER) and progesterone (PR) receptor status of tumors. After adjustment for multiple comparisons ACVR1 (8 SNPs) was modestly associated with ER+PR+ tumors [odds ratios (ORs) between 1.18 and 1.39 padj breast cancer in this admixed population.

  15. Bone morphogenetic protein antagonist gremlin 1 is widely expressed by cancer-associated stromal cells and can promote tumor cell proliferation

    Science.gov (United States)

    Sneddon, Julie B.; Zhen, Hanson H.; Montgomery, Kelli; van de Rijn, Matt; Tward, Aaron D.; West, Robert; Gladstone, Hayes; Chang, Howard Y.; Morganroth, Greg S.; Oro, Anthony E.; Brown, Patrick O.

    2006-10-01

    Although tissue microenvironments play critical roles in epithelial development and tumorigenesis, the factors mediating these effects are poorly understood. In this work, we used a genomic approach to identify factors produced by cells in the microenvironment of basal cell carcinoma (BCC) of the skin, one of the most common human cancers. The global gene expression programs of stromal cell cultures derived from human BCCs showed consistent, systematic differences from those derived from nontumor skin. The gene most consistently expressed at a higher level in BCC tumor stromal cells compared with those from nontumor skin was GREMLIN 1, which encodes a secreted antagonist of the bone morphogenetic protein (BMP) pathway. BMPs and their antagonists are known to play a crucial role in stem and progenitor cell biology as regulators of the balance between expansion and differentiation. Consistent with the hypothesis that BMP antagonists might have a similar role in cancer, we found GREMLIN 1 expression in the stroma of human BCC tumors but not in normal skin in vivo. Furthermore, BMP 2 and 4 are expressed by BCC cells. Ex vivo, BMP inhibits, and Gremlin 1 promotes, proliferation of cultured BCC cells. We further found that GREMLIN 1 is expressed by stromal cells in many carcinomas but not in the corresponding normal tissue counterparts that we examined. Our data suggest that BMP antagonists may be important constituents of tumor stroma, providing a favorable microenvironment for cancer cell survival and expansion in many cancers. cancer biology | stem cell regulation | tissue microenvironment | tumor stroma

  16. Correlation between Systemic Arterial Hypertension and Bone Morphogenetic Protein-2 in Central Obese Non-Diabetic Men with Evidence of Coronary Artery Calcification

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    Antonia Anna Lukito

    2011-12-01

    Full Text Available BACKGROUND: Previous studies have confirmed separately the relationship between obesity, insulin-resistance, hypertension and bone morphogenetic protein-2 (BMP-2 with coronary artery calcification, a parameter of subclinical atherosclerosis. It was also reported that BMPs may function as proinflammatory, prohypertensive and proatherogenic mediators. The study aimed to assess the correlation between systemic hypertension and BMP-2 plasma concentration in central-obese non-diabetic men with evidence of coronary artery calcification. METHODS: This was a cross sectional study on 60 central-obese non-diabetic men, of an average age of 55.2 years, with evidence of coronary calcification, who came for health check-up and met the inclusion criteria consecutively as defined by waist circumference >90 cm and fasting blood glucose <126 mg/dL. Coronary calcification was defined by coronary artery calcium (CAC score ≥10 Agatson-unit Dual Source 64 slice CT scan. RESULTS: There is positive correlation between hypertension and BMP-2 in central-obese non-diabetic men with evidence of coronary artery calcification. BMP-2 plasma concentration was higher in the hypertensive subjects. The correlation was stronger in younger (<55 years old subjects and subjects with insulin-resitance. KEYWORDS: hypertension, BMP-2, coronary calcification, central obesity, age, insulin resistance.

  17. Effects of simulated weightlessness on the kinase activity of MEK1 induced by bone morphogenetic protein-2 in rat osteosarcoma cells

    Science.gov (United States)

    Zhang, S.; Wang, B.; Cao, X. S.; Yang, Z.

    Objective The mRNA expression of alpha 1 chain of type I collagen COL-I alpha 1 in rat osteosarcoma ROS17 2 8 cells induced by bone morphogenetic protein-2 BMP-2 was reduced under simulated microgravity The protein kinase MEK1 of MAPK signal pathway plays an important role in the expression of COL-I alpha 1 mRNA The purpose of this study is to investigate the effects of simulated weightlessness on the activity of MEK1 induced by BMP-2 in ROS17 2 8 cells Methods ROS17 2 8 cells were cultured in 1G control and rotating clinostat simulated weightlessness for 24 h 48 h and 72 h BMP-2 500 ng ml was added into the medium 1 h before the culture ended There was a control group in which ROS17 2 8 cells were cultured in 1G condition without BMP-2 Then the total protein of cells was extracted and the expression of phosphated-ERK1 2 p-ERK1 2 protein was detected by means of Western Blotting to show the kinase activity of MEK1 Results There were no significant differences in the expression of total ERK1 2 among all groups The expression of p-ERK1 2 was unconspicuous in the control group without BMP-2 but increased significantly when BMP-2 was added P 0 01 The level of p-ERK1 2 in simulated weightlessness group was much more lower than that in 1G group in every time point P 0 01 The expression of p-ERK1 2 gradually decreased along with the time of weightlessness simulation P 0 01 Conclusions The kinase activity of MEK1 induced by BMP-2 in rat osteosarcoma cells was reduced under simulated weightlessness

  18. Osteogenic potential of icariin compared with recombinant human bone morphogenetic protein 2 in vitro: a preliminary study

    NARCIS (Netherlands)

    Zhang, X.; Liu, T.; Huang, Y.; Zheng, Y.; Liu, T.; Wismeijer, D.; Liu, Y.

    2015-01-01

    Icariin, the primary active ingredient of Herba Epimedii which has been used for decades to treat bone related maladies in China, has the ability to support bone regeneration. In this study, we investigated icariin's potential to stimulate osteogenesis using an in vitro studies to compare icariin's

  19. Human Articular Cartilage Progenitor Cells Are Responsive to Mechanical Stimulation and Adenoviral-Mediated Overexpression of Bone-Morphogenetic Protein 2.

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    Alexander J Neumann

    Full Text Available Articular cartilage progenitor cells (ACPCs represent a new and potentially powerful alternative cell source to commonly used cell sources for cartilage repair, such as chondrocytes and bone-marrow derived mesenchymal stem cells (MSCs. This is particularly due to the apparent resistance of ACPCs to hypertrophy. The current study opted to investigate whether human ACPCs (hACPCs are responsive towards mechanical stimulation and/or adenoviral-mediated overexpression of bone morphogenetic protein 2 (BMP-2. hACPCs were cultured in fibrin-polyurethane composite scaffolds. Cells were cultured in a defined chondro-permissive medium, lacking exogenous growth factors. Constructs were cultured, for 7 or 28 days, under free-swelling conditions or with the application of complex mechanical stimulation, using a custom built bioreactor that is able to generate joint-like movements. Outcome parameters were quantification of BMP-2 and transforming growth factor beta 1 (TGF-β1 concentration within the cell culture medium, biochemical and gene expression analyses, histology and immunohistochemistry. The application of mechanical stimulation alone resulted in the initiation of chondrogenesis, demonstrating the cells are mechanoresponsive. This was evidenced by increased GAG production, lack of expression of hypertrophic markers and a promising gene expression profile (significant up-regulation of cartilaginous marker genes, specifically collagen type II, accompanied by no increase in the hypertrophic marker collagen type X or the osteogenic marker alkaline phosphatase. To further investigate the resistance of ACPCs to hypertrophy, overexpression of a factor associated with hypertrophic differentiation, BMP-2, was investigated. A novel, three-dimensional, transduction protocol was used to transduce cells with an adenovirus coding for BMP-2. Over-expression of BMP-2, independent of load, led to an increase in markers associated with hypertropy. Taken together ACPCs

  20. Continuous release of bone morphogenetic protein-2 through nano-graphene oxide-based delivery influences the activation of the NF-κB signal transduction pathway

    Science.gov (United States)

    Zhong, Cheng; Feng, Jun; Lin, Xiangjin; Bao, Qi

    2017-01-01

    Graphene oxide (GO) has been used as a delivery vehicle for small molecule drugs and nucleotides. To further investigate GO as a smart biomaterial for the controlled release of cargo molecules, we hypothesized that GO may be an appropriate delivery vehicle because it releases bone morphogenetic protein 2 (BMP2). GO characterization indicated that the size distribution of the GO flakes ranged from 81.1 nm to 45,749.7 nm, with an approximate thickness of 2 nm. After BMP2 adsorption onto GO, Fourier-transformed infrared spectroscopy (FTIR) and thermal gravimetric analysis were performed. Compared to GO, BMP2-GO did not induce significant changes in the characteristics of the materials. GO continuously released BMP2 for at least 40 days. Bone marrow stem cells (BMSCs) and chondrocytes were treated with BMP2-GO in interleukin-1 media and assessed in terms of cell viability, flow cytometric characterization, and expression of particular mRNA. Compared to GO, BMP2-GO did not induce any significant changes in biocompatibility. We treated osteoarthritic rats with BMP2 and BMP2-GO, which showed significant differences in Osteoarthritis Research Society International (OARSI) scores (P<0.05). Quantitative assessment revealed significant differences compared to that using BMP2 and BMP2-GO (P<0.05). These findings indicate that GO may be potentially used to control the release of carrier materials. The combination of BMP2 and GO slowed the progression of NF-κB-activated degenerative changes in osteoarthritis. Therefore, we infer that our BMP2-GO strategy could alleviate the NF-κB pathway by inducing continuous BMP2 release.

  1. Chondrogenesis of periodontal ligament stem cells by transforming growth factor-β3 and bone morphogenetic protein-6 in a normal healthy impacted third molar

    Institute of Scientific and Technical Information of China (English)

    Sunyoung Choi; Tae-Jun Cho; Soon-Keun Kwon; Gene Lee; Jaejin Cho

    2013-01-01

    The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency. However, the proof of chondrogenic potential of the cells is scarce. Therefore, we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor (TGF)-β3 and bone morphogenetic protein (BMP)-6. After isolation of periodontal ligament stem cells (PDLSCs) from human periodontal ligament, the cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 20% fetal bovine serum (FBS). A mechanical force initiated chondrogenic differentiation of the cells. For chondrogenic differentiation, 10 μg ·L-1 TGF-β3 or 100 μg ·L-1 BMP-6 and the combination treating group for synergistic effect of the growth factors. We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay, histology, immunohistochemistry and genetic analysis. PDLSCs showed mesenchymal stem cell properties proved by FACS analysis. Glycosaminoglycans contents were increased 217% by TGF-β3 and 220% by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281% compared to control. The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls, but also TGF-β3 or BMP-6 single treatment dramatically. The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions. The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis, which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.

  2. Single nucleotide polymorphism of bone morphogenetic protein 4 gene: A risk factor of non-syndromic cleft lip with or without palate

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    Sathyaprasad Savitha

    2015-01-01

    Full Text Available Background: The bone morphogenetic protein (BMP signalling pathway is crucial in a number of developmental processes and is critical in the formation of variety of craniofacial elements including cranial neural crest, facial primordium, tooth, lip and palate. It is an important mediator in regulation of lip and palate fusion, cartilage and bone formation. Aim: To study the role of mutation of BMP4 genes in the aetiology of non-syndromic cleft lip with or without palate (NSCL ± P and identify it directly from human analyses. Materials and Methods: A case-control study was done to evaluate whether BMP4T538C polymorphism, resulting in an amino acid change of Val=Ala (V152A in the polypeptide, is associated with NSCL ± P in an Indian paediatric population. Genotypes of 100 patients with NSCL ± P and 100 controls (in whom absence of CL ± P was confirmed in three generations were detected using a polymerase chain reaction-restriction fragment length polymorphism strategy. Logistic regression was performed to evaluate allele and genotype association with NSCLP. Results: Results showed significant association between homozygous CC genotype with CL ± P (odds ratio [OR]-5.59 and 95% confidence interval [CI] = 2.85-10.99. The 538C allele carriers showed an increased risk of NSCL ± P as compared with 538 T allele (OR - 4.2% CI = 2.75-6.41. Conclusion: This study suggests an association between SNP of BMP4 gene among carriers of the C allele and increased risk for NSCLP in an Indian Population. Further studies on this aspect can scale large heights in preventive strategies for NSCLP that may soon become a reality.

  3. Tribulus terrestris Alters the Expression of Growth Differentiation Factor 9 and Bone Morphogenetic Protein 15 in Rabbit Ovaries of Mothers and F1 Female Offspring.

    Science.gov (United States)

    Abadjieva, Desislava; Kistanova, Elena

    2016-01-01

    Although previous research has demonstrated the key role of the oocyte-derived factors, bone morphogenetic protein (BMP) 15 and growth differentiation factor (GDF) 9, in follicular development and ovulation, there is a lack of knowledge on the impact of external factors, which females are exposed to during folliculogenesis, on their expression. The present study investigated the effect of the aphrodisiac Tribulus terrestris on the GDF9 and BMP15 expression in the oocytes and cumulus cells at mRNA and protein levels during folliculogenesis in two generations of female rabbits. The experiment was conducted with 28 New Zealand rabbits. Only the diet of the experimental mothers group was supplemented with a dry extract of T. terrestris for the 45 days prior to insemination. The expression of BMP15 and GDF9 genes in the oocytes and cumulus cells of mothers and F1 female offspring was analyzed using real-time polymerase chain reaction (RT-PCR). The localization of the GDF9 and BMP15 proteins in the ovary tissues was determined by immunohistochemical analysis. The BMP15 and GDF9 transcripts were detected in the oocytes and cumulus cells of rabbits from all groups. T. terrestris caused a decrease in the BMP15 mRNA level in the oocytes and an increase in the cumulus cells. The GDF9 mRNA level increased significantly in both oocytes and cumulus cells. The downregulated expression of BMP15 in the treated mothers' oocytes was inherited in the F1 female offspring born to treated mothers. BMP15 and GDF9 show a clearly expressed sensitivity to the bioactive compounds of T. terrestris.

  4. Tribulus terrestris Alters the Expression of Growth Differentiation Factor 9 and Bone Morphogenetic Protein 15 in Rabbit Ovaries of Mothers and F1 Female Offspring.

    Directory of Open Access Journals (Sweden)

    Desislava Abadjieva

    Full Text Available Although previous research has demonstrated the key role of the oocyte-derived factors, bone morphogenetic protein (BMP 15 and growth differentiation factor (GDF 9, in follicular development and ovulation, there is a lack of knowledge on the impact of external factors, which females are exposed to during folliculogenesis, on their expression. The present study investigated the effect of the aphrodisiac Tribulus terrestris on the GDF9 and BMP15 expression in the oocytes and cumulus cells at mRNA and protein levels during folliculogenesis in two generations of female rabbits. The experiment was conducted with 28 New Zealand rabbits. Only the diet of the experimental mothers group was supplemented with a dry extract of T. terrestris for the 45 days prior to insemination. The expression of BMP15 and GDF9 genes in the oocytes and cumulus cells of mothers and F1 female offspring was analyzed using real-time polymerase chain reaction (RT-PCR. The localization of the GDF9 and BMP15 proteins in the ovary tissues was determined by immunohistochemical analysis. The BMP15 and GDF9 transcripts were detected in the oocytes and cumulus cells of rabbits from all groups. T. terrestris caused a decrease in the BMP15 mRNA level in the oocytes and an increase in the cumulus cells. The GDF9 mRNA level increased significantly in both oocytes and cumulus cells. The downregulated expression of BMP15 in the treated mothers' oocytes was inherited in the F1 female offspring born to treated mothers. BMP15 and GDF9 show a clearly expressed sensitivity to the bioactive compounds of T. terrestris.

  5. Surface functionalization of nanoporous alumina with bone morphogenetic protein 2 for inducing osteogenic differentiation of mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Song, Yuanhui; Ju, Yang; Morita, Yasuyuki; Xu, Baiyao [Department of Mechanical Science and Engineering, Nagoya University, Nagoya 464-8603 (Japan); Song, Guanbin [Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044 (China)

    2014-04-01

    Many studies have demonstrated the possibility to regulate cellular behavior by manipulating the specific characteristics of biomaterials including the physical features and chemical properties. To investigate the synergistic effect of chemical factors and surface topography on the growth behavior of mesenchymal stem cells (MSCs), bone morphorgenic protein 2 (BMP2) was immobilized onto porous alumina substrates with different pore sizes. The BMP2-immobilized alumina substrates were characterized with scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Growth behavior and osteogenic differentiation of MSCs cultured on the different substrates were investigated. Cell adhesion and morphological changes were observed with SEM, and the results showed that the BMP2-immobilized alumina substrate was able to promote adhesion and spreading of MSCs. MTT assay and immunofluorescence staining of integrin β1 revealed that the BMP2-immobilized alumina substrates were favorable for cell growth. To evaluate the differentiation of MSCs, osteoblastic differentiation markers, such as alkaline phosphatase (ALP) activity and mineralization, were investigated. Compared with those of untreated alumina substrates, significantly higher ALP activities and mineralization were detected in cells cultured on BMP2-immobilized alumina substrates. The results suggested that surface functionalization of nanoporous alumina substrates with BMP2 was beneficial for cell growth and osteogenic differentiation. With the approach of immobilizing growth factors onto material substrates, it provided a new insight to exploit novel biofunctional materials for tissue engineering. - Highlights: • BMP2 was immobilized onto nanoporous alumina substrates with different pore sizes. • BMP2-immobilized substrates were able to promote adhesion and spreading of MSCs. • BMP2-immobilized substrates were favorable for cell growth of MSCs. • BMP2-immobilized substrates promoted osteogenic

  6. Does LED phototherapy influence the repair of bone defects grafted with MTA, bone morphogenetic proteins, and guided bone regeneration? A description of the repair process on rodents.

    Science.gov (United States)

    Pinheiro, Antonio L B; Soares, Luiz G P; Barbosa, Artur F S; Ramalho, Luciana M P; dos Santos, Jean N

    2012-09-01

    This work carried out a histological analysis on bone defects grafted (MTA) treated or not with LED, BMPs, and membrane (GBR). Benefits of their isolated or combined usage on bone repair were reported, but not their association. Ninety rats were divided into ten groups and each subdivided into three. Defects on G II and I were filled with the blood clot. G II was further LED irradiated. G III and IV were filled with MTA; G IV was further LED irradiated. In G V and VI, the defects were filled with MTA and covered with a membrane (GBR). G VI was further LED irradiated. In G VII and VIII, BMPs were added to the MTA and group VIII was further LED irradiated. In G IX and X, the MTA + BMP graft was covered with a membrane (GBR). G X was further LED irradiated. LED was applied over the defect at 48-h intervals and repeated for 15 days. Specimens were processed, cut, and stained with H&E and Sirius red and underwent histological analysis. The use of LED light alone dramatically reduced inflammation. However, its use on MTA associated with BMP and/or GBR increased the severity of the inflammatory reaction. Regarding bone reabsorption, the poorest result was seen when the LED light was associated with the MTA + BMP graft. In the groups Clot and MTA + GBR, no bone reabsorption was detectable. Increased collagen deposition was observed when the LED light was associated with the use of the MTA associated with BMP and/or GBR. Increased new bone formation was observed when the LED light was used alone or associated with the use of MTA + GBR, MTA + BMP, on association of MTA + BMP + GBR and when BMP was added to the MTA. Our results indicate that the use of LED light alone or in association with MTA, MTA + BMP, MTA + GBR, and MTA + BMP + GBR caused less inflammation, and an increase of both collagen deposition and bone deposition as seen on both histological and morphometric analysis.

  7. Effects of Roughly Focused Extracorporeal Shock Waves Therapy on the Expressions of Bone Morphogenetic Protein-2 and Osteoprotegerin in Osteoporotic Fracture in Rats

    Science.gov (United States)

    Huang, Hai-Ming; Li, Xiao-Lin; Tu, Shu-Qiang; Chen, Xiao-Feng; Lu, Chang-Chun; Jiang, Liang-Hua

    2016-01-01

    Background: Roughly focused extracorporeal shock waves therapy (ESWT) is characterized by a wide focal area, a large therapy zone, easy positioning, and less pain during treatment. The purpose of this study was to investigate the effects of roughly focused ESWT on the expression of osteoprotegerin (OPG) and bone morphogenetic protein-2 (BMP-2) in osteoporotic fractures in rats. Methods: Seventy-two female Sprague-Dawley (SD) rats, 3 months old, were divided into sham-operated group (n = 6) and an ovariectomized (OVX) group (n = 66). Sixty OVX SD rats were used as a model of double proximal tibial osteotomy and inner fixation. The osteotomy site in the left tibia was treated with roughly focused ESWT once at an energy density of 0.26 mJ/mm2, 60 doses/min, and 2000 pact quantities. The contralateral right tibia was left untreated and served as a control. Expression of OPG and BMP-2 in the callus of the osteoporotic fracture area was assessed using immunohistochemistry, real-time polymerase chain reaction (PCR), and Western blotting analysis. Results: Bone mineral density (BMD) at the proximal tibia, femur, and L5 spine was significantly reduced after ovariectomy. BMD of proximal tibia was 12.9% less in the OVX group than that in the sham-operated group. Meanwhile, bilateral oophorectomy resulted in a lower trabecular bone volume fraction (BV/TV) in the proximal tibia of the sham-OVX animals. Three months after bilateral oophorectomy, BV/TV was 14.29% of baseline BV/TV in OVX legs versus 45.91% in the sham-OVX legs (P shock wave treatment, paired with a much earlier (at 4 weeks) increase of BMP-2, and declined close to normal at 8 weeks. Conclusions: Roughly focused ESWT may promote the expression of OPG and BMP-2 in the osteoporotic fracture area in rats. BMP-2 and OPG may act synergistically and may lead to a significant enhancement of bone formation and remodeling. PMID:27779163

  8. Closure of 1.5-cm alveolar oral antral fistula with intra-alveolar sinus membrane elevation and bone morphogenetic protein-2/collagen graft followed by dental implant restoration: case report.

    Science.gov (United States)

    Cottam, Jared R; Jensen, Ole T; Beatty, Lucas; Ringeman, Jason

    2013-01-01

    Closure of a 1.5-cm oral antral fistula was done in combination with sinus floor and extraction socket grafting using recombinant human bone morphogenetic protein-2 within a collagen sponge matrix. The approach to the sinus was transalveolar, with elevation of the sinus membrane done through a molar extraction socket. Following graft placement, soft tissue repair was done with a buccal advancement flap. A dental implant was subsequently placed and restored. Peri-implant bone and implant stability were well maintained at the 1-year follow up examination.

  9. A Fusion between Domains of the Human Bone Morphogenetic Protein-2 and Maize 27 kD γ-Zein Accumulates to High Levels in the Endoplasmic Reticulum without Forming Protein Bodies in Transgenic Tobacco.

    Science.gov (United States)

    Ceresoli, Valentina; Mainieri, Davide; Del Fabbro, Massimo; Weinstein, Roberto; Pedrazzini, Emanuela

    2016-01-01

    Human Bone Morphogenetic Protein-2 (hBMP2) is an osteoinductive agent physiologically involved in bone remodeling processes. A commercialized recombinant hBMP2 produced in mammalian cell lines is available in different clinical applications where bone regeneration is needed, but widespread use has been hindered due to an unfavorable cost/effective ratio. Protein bodies are very large insoluble protein polymers that originate within the endoplasmic reticulum by prolamine accumulation during the cereal seed development. The N-terminal domain of the maize prolamin 27 kD γ-zein is able to promote protein body biogenesis when fused to other proteins. To produce high yield of recombinant hBMP2 active domain (ad) in stably transformed tobacco plants we have fused it to the γ-zein domain. We show that this zein-hBMP2ad fusion is retained in the endoplasmic reticulum without forming insoluble protein bodies. The accumulation levels are above 1% of total soluble leaf proteins, indicating that it could be a rapid and suitable strategy to produce hBMP2ad at affordable costs.

  10. A fusion between domains of the human bone morphogenetic protein-2 and maize 27 kD gamma-zein accumulates to high levels in the endoplasmic reticulum without forming protein bodies in transgenic tobacco

    Directory of Open Access Journals (Sweden)

    Valentina eCeresoli

    2016-03-01

    Full Text Available Human Bone Morphogenetic Protein-2 (hBMP2 is an osteoinductive agent physiologically involved in bone remodelling processes. A commercialized recombinant hBMP2 produced in mammalian cell lines is available in different clinical applications where bone regeneration is needed, but widespread use has been hindered due to an unfavorable cost/effective ratio. Protein bodies are very large insoluble protein polymers that originate within the endoplasmic reticulum by prolamine accumulation during the cereal seed development. The N-terminal domain of the maize prolamin 27 kD -zein is able to promote protein body biogenesis when fused to other proteins. To produce high yield of recombinant hBMP2 active domain (ad in stably transformed tobacco plants we have fused it to the γ-zein domain. We show that this zein-hBMP2ad fusion is retained in the endoplasmic reticulum without forming insoluble protein bodies. The accumulation levels are above 1% of total soluble leaf proteins, indicating that it could be a rapid and suitable strategy to produce hBMP2ad at affordable costs.

  11. Cultured Human Periosteum-Derived Cells Can Differentiate into Osteoblasts in a Perioxisome Proliferator-Activated Receptor Gamma-Mediated Fashion via Bone Morphogenetic Protein signaling.

    Science.gov (United States)

    Chung, Jin-Eun; Park, Jin-Ho; Yun, Jeong-Won; Kang, Young-Hoon; Park, Bong-Wook; Hwang, Sun-Chul; Cho, Yeong-Cheol; Sung, Iel-Yong; Woo, Dong Kyun; Byun, June-Ho

    2016-01-01

    The differentiation of mesenchymal stem cells towards an osteoblastic fate depends on numerous signaling pathways, including activation of bone morphogenetic protein (BMP) signaling components. Commitment to osteogenesis is associated with activation of osteoblast-related signal transduction, whereas inactivation of this signal transduction favors adipogenesis. BMP signaling also has a critical role in the processes by which mesenchymal stem cells undergo commitment to the adipocyte lineage. In our previous study, we demonstrated that an agonist of the perioxisome proliferator-activated receptor γ (PPARγ), a master regulator of adipocyte differentiation, stimulates osteoblastic differentiation of cultured human periosteum-derived cells. In this study, we used dorsomorphin, a selective small molecule inhibitor of BMP signaling, to investigate whether BMP signaling is involved in the positive effects of PPARγ agonists on osteogenic phenotypes of cultured human periosteum-derived cells. Both histochemical detection and bioactivity of ALP were clearly increased in the periosteum-derived cells treated with the PPARγ agonist at day 10 of culture. Treatment with the PPARγ agonist also caused an increase in alizarin red S staining and calcium content in the periosteum-derived osteoblasts at 2 and 3 weeks of culture. In contrast, dorsomorphin markedly decreased ALP activity, alizarin red S staining and calcium content in both the cells treated with PPARγ agonist and the cells cultured in osteogenic induction media without PPARγ agonist during the culture period. In addition, the PPARγ agonist clearly increased osteogenic differentiation medium-induced BMP-2 upregulation in the periosteum-derived osteoblastic cells at 2 weeks of culture as determined by quantitative reverse transcriptase polymerase chain reaction (RT-PCR), immunoblotting, and immunocytochemical analyses. Although further study will be needed to clarify the mechanisms of PPARγ-regulated osteogenesis

  12. Metabolic differences in bovine cumulus-oocyte complexes matured in vitro in the presence or absence of follicle-stimulating hormone and bone morphogenetic protein 15.

    Science.gov (United States)

    Sutton-McDowall, Melanie L; Mottershead, David G; Gardner, David K; Gilchrist, Robert B; Thompson, Jeremy G

    2012-10-01

    Bidirectional communication between cumulus cells and the oocyte is necessary to achieve oocyte developmental competence. The aim of the present study was to examine the effects of recombinant human bone morphogenetic protein 15 (rhBMP15) and follicle-stimulating hormone (FSH) supplementation on bovine cumulus-oocyte complex (COC) metabolism during maturation. Bovine COCs were matured in the presence of absence of FSH, rhBMP15, or both for 23 h. The addition of FSH and rhBMP15 increased blastocyst development (without rhBMP15 and FSH, 28.4% ± 7.4%; with FSH and rhBMP15, 51.5% ± 5.4%; P levels. rhBMP15 supplementation (regardless of FSH) significantly decreased ADP levels in COCs, leading to an increase in ATP:ADP ratios (P Indicators of mitochondrial activity and cellular REDOX, oxidized flavin adenine dinucleotide (FAD(++)) and reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), levels within the oocyte of COCs were significantly higher with rhBMP15 alone, whereas the presence of FSH diminished the rhBMP15 effect. Regardless of treatment, no changes in REDOX state (FAD(++):NAD(P)H). The significant increase in FAD(++) and NAD(P)H in COCs with rhBMP15 was mediated via cumulus cells, because no differences were found in denuded oocytes cultured in the presence or absence of FSH, rhBMP15, or both. The present study demonstrates that a principal metabolic consequence of FSH supplementation of COCs is to alter the glycolytic rate of cumulus cells, whereas that of rhBMP15 is to regulate oxidative phosphorylation in the oocyte, even though it acts via cumulus cells. These effects are tempered when FSH and rhBMP15 are present together but, nonetheless, yield the best oocyte developmental competence.

  13. Comparison of expression patterns of fibroblast growth factor 8, bone morphogenetic protein 4 and sonic hedgehog in jaw development of the house shrew, Suncus murinus.

    Science.gov (United States)

    Ogi, Hidenao; Tabata, Makoto J; Yamanaka, Atsushi; Yasui, Kinya; Uemura, Masanori

    2002-01-01

    To elucidate the mechanism underlying jaw development in mammals, we used a new laboratory animal, Suncus murinus (house shrew, an insectivore) as the subject for the investigation, because Suncus has all types of teeth (incisor, canine, premolar and molar) in its upper and lower jaws and is thought to be a good model animal having a general mammalian tooth pattern. At the start, by use of degenerate primers we cloned Suncus homologues of fibroblast growth factor 8 (sFgf8), bone morphogenetic protein 4 (sBmp4) and sonic hedgehog (sShh) genes from cDNA library derived from whole Suncus embryos at day 12 (E12). Thereafter, we examined the expression patterns of these genes in the jaw development of Suncus E11-16 embryos (for mouse E9.5-12 embryos). sFgf8 and sBmp4 were expressed in E11 but not in E15 and onward during orofacial development. sShh was expressed from E11 onward, and its expression was increased in the orofacial area. The expression pattern of sFgf8 in the maxillary and mandibular arches of E14 coincided with the area of the presumptive tooth arch. However, sShh and sBmp4 were expressed only in the outer area (= buccal/labial side) of presumptive tooth arch. Thus, these 3 genes showed specific expression pattern in jaw development of Suncus, and their distributions did not overlap each other except in a few regions. These findings suggest that sFgf8, sBmp4 and sShh have a specific function respectively during jaw development in Suncus murinus.

  14. Aldosterone breakthrough caused by chronic blockage of angiotensin II type 1 receptors in human adrenocortical cells: possible involvement of bone morphogenetic protein-6 actions.

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    Otani, Hiroyuki; Otsuka, Fumio; Inagaki, Kenichi; Suzuki, Jiro; Miyoshi, Tomoko; Kano, Yoshihiro; Goto, Junko; Ogura, Toshio; Makino, Hirofumi

    2008-06-01

    Circulating aldosterone concentrations occasionally increase after initial suppression with angiotensin II (Ang II) converting enzyme inhibitors or Ang II type 1 receptor blockers (ARBs), a phenomenon referred to as aldosterone breakthrough. However, the underlying mechanism causing the aldosterone breakthrough remains unknown. Here we investigated whether aldosterone breakthrough occurs in human adrenocortical H295R cells in vitro. We recently reported that bone morphogenetic protein (BMP)-6, which is expressed in adrenocortical cells, enhances Ang II- but not potassium-induced aldosterone production in human adrenocortical cells. Accordingly, we examined the roles of BMP-6 in aldosterone breakthrough induced by long-term treatment with ARB. Ang II stimulated aldosterone production by adrenocortical cells. This Ang II stimulation was blocked by an ARB, candesartan. Interestingly, the candesartan effects on Ang II-induced aldosterone synthesis and CYP11B2 expression were attenuated in a course of candesartan treatment for 15 d. The impairment of candesartan effects on Ang II-induced aldosterone production was also observed in Ang II- or candesartan-pretreated cells. Levels of Ang II type 1 receptor mRNA were not changed by chronic candesartan treatment. However, BMP-6 enhancement of Ang II-induced ERK1/2 signaling was resistant to candesartan. The BMP-6-induced Smad1, -5, and -8 phosphorylation, and BRE-Luc activity was augmented in the presence of Ang II and candesartan in the chronic phase. Chronic Ang II exposure decreased cellular expression levels of BMP-6 and its receptors activin receptor-like kinase-2 and activin type II receptor mRNAs. Cotreatment with candesartan reversed the inhibitory effects of Ang II on the expression levels of these mRNAs. The breakthrough phenomenon was attenuated by neutralization of endogenous BMP-6 and activin receptor-like kinase-2. Collectively, these data suggest that changes in BMP-6 availability and response may be involved

  15. T allele at site 6007 of bone morphogenetic protein-4 gene increases genetic susceptibility to ossification of the posterior longitudinal ligament in male Chinese Han population

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    MENG Xiang-long; WANG Hao; YANG Hui; HAI Yong; TIAN Bao-peng; LIN Xin

    2010-01-01

    Background Several candidate genes of ossification of the posterior longitudinal ligament (OPLL) susceptibility have been identified, but their polymorphisms account for only a small percent of the total variance. Bone morphogenetic protein-4 (BMP4) is a potent ectopic ossification inducing factor. BMP4 protein and mRNA are present in cells from OPLL patients, but not non-OPLL controls. A single nucleotide polymorphism of 6007C>T(rs17563) of BMP4 has been reported to affect bone density in postmenopausal women. Thus, BMP4 may function in OPLL development. Appropriately, the relationship between BMP4 polymorphisms and OPLL was investigated.Methods A case-control association study investigated the genetic etiology in 179 OPLL patients and 298 non-OPLL controls. Extent of OPLL was analyzed by radiologic examinations. Whether single nucleotide polymorphism (SNP) of -5826G>A(rs1957860) 5′ of the transcription start site and 6007C>T(rs17563) in exon 4 of the BMP4 gene were statistically associated with genetic susceptibility to OPLL in Chinese Han subjects was assessed.Results A significant statistical difference in genotype of 6007C>T polymorphism between male OPLL patients and male controls was evident, and the frequency of "TT" genotype in male OPLL patients was significantly higher than in male controls (P=0.039). The frequency of the "T" allele was also significantly higher in male OPLL subjects than in male controls (P=0.014, 0R=1.57). A significant difference was also observed between the 6007C>T polymorphism and the number of ossified cervical vertebrae in OPLL patients, while no statistical difference was apparent between the -5826G>A polymorphism and OPLL occurrence.Conclusions The T allele in the 6007C>T polymorphism may be a risk factor for male Han Chinese with ossification of the posterior longitudinal ligament in the cervical spine. Chinese Han male patients with CT and TT 6007C>T genotypes have a genetic susceptibility to OPLL and more extensive OPLL in

  16. 血管内皮生长因子和骨形态发生蛋白在骨组织工程中的作用%Vascular endothelial growth factor and bone morphogenetic protein in the bone tissue engineering

    Institute of Scientific and Technical Information of China (English)

    纪经涛; 胡永成; 夏群; 苗军; 陈晓鹏; 方程

    2015-01-01

    and progressive bone disorder are very common, and bone tissue engineering provides a new approach to bone defect repair. Growth factors related to bone tissue engineering bone have been reported a lot and have achieved some results. How to mimick the natural timing of different growth factors with different bioactivities has become the current hotspot in bone repair. OBJECTIVE: To review the new developments in vascular endothelial growth factor and bone morphogenetic protein in bone tissue engineering. METHODS: The first author searched CNKI (1990/2015) and Medline database (1990/2015) for related articles using the key words of “osteogenic factors, angiogenic factors, tissue engineering bone, bone repair, vascularization, vascular endothelial growth factor, bone morphogenetic protein, sequential release, seed cels, cytoskeleton” in Chinese and English, respectively. Mechanism of action and research direction about vascular endothelial growth factor and bone morphogenetic protein were summarized. RESULTS AND CONCLUSION:Totaly 313 papers were searched initialy, and finaly 87 papers were enroled in result analysis. The results show that different growth factors play different roles in bone repair. Vascularization and osteogenesis are the most important processes in bone repair. The osteogenic factors play an important role in maintaining bone structure and bone formation. The angiogenic factors can provide oxygen and nutrients for tissue growth, differentiation and functionalization. The combination of osteogenic and angiogenic factors has a better osteogenic effect than osteogenic or angiogenic factors used alone. However, the most important problem is how to control the exogenous osteogenesis and the release dosage of angiogenic factors in bone repair.

  17. Repetitive recombinant human bone morphogenetic protein 2 injections improve the callus microarchitecture and mechanical stiffness in a sheep model of distraction osteogenesis

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    Marc-Frederic Pastor

    2012-03-01

    Full Text Available Evidence suggests that recombinant human bone morphogenetic protein 2 (rhBMP-2 increases the mechanical integrity of callus tissue during bone healing. This effect may be either explained by an increase of callus formation or a modification of the trabecular microarchitecture. Therefore the purpose of the study was to evaluate the potential benefit of rhBMP-2 on the trabecular microarchitecture and on multidirectional callus stiffness. Further we asked, whether microarchitecture changes correlate with optimized callus stiffness. In this study a tibial distraction osteogenesis (DO model in 12 sheep was used to determine, whether percutaneous injection of rhBMP-2 into the distraction zone influences the microarchitecture of the bone regenerate. After a latency period of 4 days, the tibiae were distracted at a rate of 1.25 mm/day over a period of 20 days, resulting in total lengthening of 25 mm. The operated limbs were randomly assigned to one treatment groups and one control group: (A triple injection of rhBMP-2 (4 mg rhBMP-2/injection and (B no injection. The tibiae were harvested after 74 days and scanned by μCT (90 μm/voxel. In addition, we conducted a multidirectional mechanical testing of the tibiae by using a material testing system to assess the multidirectional strength. The distraction zones were tested for torsional stiffness and bending stiffness antero-posterior (AP and medio-lateral (ML direction, compression strength and maximum axial torsion. Statistical analysis was performed using multivariate analysis of variance (ANOVA followed by student’s t-test and Regression analysis using power functions with a significance level of P<0.05. Triple injections of rhBMP-2 induced significant changes in the trabecular architecture of the regenerate compared with the control: increased trabecular number (Tb.N. (treatment group 1.73 mm/1 vs. control group 1.2 mm/1, increased cortical bone volume fraction (BV/TV (treatment group 0.68 vs

  18. Bone Morphogenetic Protein (BMP-4 and BMP-7 regulate differentially Transforming Growth Factor (TGF-β1 in normal human lung fibroblasts (NHLF

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    Lloyd Clare M

    2010-06-01

    Full Text Available Abstract Background Airway remodelling is thought to be under the control of a complex group of molecules belonging to the Transforming Growth Factor (TGF-superfamily. The Bone Morphogenetic Proteins (BMPs belong to this family and have been shown to regulate fibrosis in kidney and liver diseases. However, the role of BMPs in lung remodelling remains unclear. BMPs may regulate tissue remodelling in asthma by controlling TGF-β-induced profibrotic functions in lung fibroblasts. Methods Cell cultures were exposed to TGF-β1 alone or in the presence of BMP-4 or BMP-7; control cultures were exposed to medium only. Cell proliferation was assessed by quantification of the incorporation of [3H]-thymidine. The expression of the mRNA encoding collagen type I and IV, tenascin C and fibronectin in normal human lung fibroblasts (NHLF was determined by real-time quantitative PCR and the main results were confirmed by ELISA. Cell differentiation was determined by the analysis of the expression of α-smooth muscle actin (α-SMA by western blot and immunohistochemistry. The effect on matrix metalloproteinase (MMP activity was assessed by zymography. Results We have demonstrated TGF-β1 induced upregulation of mRNAs encoding the extracellular matrix proteins, tenascin C, fibronectin and collagen type I and IV when compared to unstimulated NHLF, and confirmed these results at the protein level. BMP-4, but not BMP-7, reduced TGF-β1-induced extracellular matrix protein production. TGF-β1 induced an increase in the activity of the pro-form of MMP-2 which was inhibited by BMP-7 but not BMP-4. Both BMP-4 and BMP-7 downregulated TGF-β1-induced MMP-13 release compared to untreated and TGF-β1-treated cells. TGF-β1 also induced a myofibroblast-like transformation which was partially inhibited by BMP-7 but not BMP-4. Conclusions Our study suggests that some regulatory properties of BMP-7 may be tissue or cell type specific and unveil a potential regulatory role for

  19. Addition of bone morphogenetic protein type 2 to ascorbate and β-glycerophosphate supplementation did not enhance osteogenic differentiation of human adipose-derived stem cells

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    Ariadne Cristiane Cabral Cruz

    2012-12-01

    Full Text Available Bone morphogenetic protein type 2 (BMP-2 is a potent local factor, which promotes bone formation and has been used as an osteogenic supplement for mesenchymal stem cells. OBJECTIVES: This study evaluated the effect of a recombinant BMP-2 as well as the endogenous BMP-4 and BMP-7 in the osteogenic differentiation of adipose-derived stem cells (ASCs in medium supplemented with ascorbate and β-glycerophosphate. MATERIAL AND METHODS: Human ASCs were treated with osteogenic medium in the presence (ASCs+OM+BMP-2 or absence (ASCs+OM of BMP-2. The alkaline phosphatase (ALP activity was determined and the extracellular matrix mineralization was evaluated by Von Kossa staining and calcium quantification. The expressions of BMP-4, BMP-7, Smad1, Smad4, and phosphorylated Smad1/5/8 were analyzed by western blotting. Relative mRNA expressions of Smad1, BMP receptor type II (BMPR-II, osteonectin, and osteocalcin were evaluated by qPCR. Results: ASCs+OM demonstrated the highest expression of BMP-4 and BMP-7 at days 21 and 7, respectively, the highest levels of BMPR-II mRNA expression at day 28, and the highest levels of Smad1 mRNA at days 14 and 28. ASCs+OM+BMP-2 demonstrated the highest levels of Smad1 mRNA expression at days 1, 7, and 21, the highest expression of Smad1 at day 7, the highest expression of Smad4 at day 14, the highest ALP activity at days 14 and 21, and expression of phosphorylated Smad1/5/8 at day 7. ASCs+OM and ASCs+OM+BMP2 showed similar ALP activity at days 7 and 28, similar osteonectin and osteocalcin mRNA expression at all time periods, and similar calcium depositions at all time periods. CONCLUSIONS: We concluded that human ASCs expressed endogenous BMP-4 and BMP-7. Moreover, the supplementation of ASCs with BMP-2 did not increase the level of osteogenic markers in the initial (ALP activity, intermediate (osteonectin and osteocalcin, or final (calcium deposition phases, suggesting that the exogenous addition of BMP-2 did not improve

  20. Continuous release of bone morphogenetic protein-2 through nano-graphene oxide-based delivery influences the activation of the NF-κB signal transduction pathway

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    Zhong C

    2017-02-01

    Full Text Available Cheng Zhong,1 Jun Feng,2 Xiangjin Lin,1,* Qi Bao3,4,* 1Department of Orthopaedic, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, People’s Republic of China; 2Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA, 3Department of Plastic and Reconstructive Surgery, Second Affiliated Hospital, School of Medicine, 4Institute of Gastroenterology, Zhejiang University, Hangzhou, People’s Republic of China *These authors contributed equally to this work Abstract: Graphene oxide (GO has been used as a delivery vehicle for small molecule drugs and nucleotides. To further investigate GO as a smart biomaterial for the controlled release of cargo molecules, we hypothesized that GO may be an appropriate delivery vehicle because it releases bone morphogenetic protein 2 (BMP2. GO characterization indicated that the size distribution of the GO flakes ranged from 81.1 nm to 45,749.7 nm, with an approximate thickness of 2 nm. After BMP2 adsorption onto GO, Fourier-transformed infrared spectroscopy (FTIR and thermal gravimetric analysis were performed. Compared to GO, BMP2-GO did not induce significant changes in the characteristics of the materials. GO continuously released BMP2 for at least 40 days. Bone marrow stem cells (BMSCs and chondrocytes were treated with BMP2-GO in interleukin-1 media and assessed in terms of cell viability, flow cytometric characterization, and expression of particular mRNA. Compared to GO, BMP2-GO did not induce any significant changes in biocompatibility. We treated osteoarthritic rats with BMP2 and BMP2-GO, which showed significant differences in Osteoarthritis Research Society International (OARSI scores (P<0.05. Quantitative assessment revealed significant differences compared to that using BMP2 and BMP2-GO (P<0.05. These findings indicate that GO may be potentially used to control the release of carrier materials. The combination of BMP2 and GO slowed the

  1. The Expression of Bone Morphogenetic Protein 2 and Matrix Metalloproteinase 2 through Retinoic Acid Receptor Beta Induced by All-Trans Retinoic Acid in Cultured ARPE-19 Cells.

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    Zhenya Gao

    Full Text Available All-trans retinoic acid (ATRA plays an important role in ocular development. Previous studies found that retinoic acid could influence the metabolism of scleral remodeling by promoting retinal pigment epithelium (RPE cells to secrete secondary signaling factors. The purpose of this study was to investigate whether retinoic acid affected secretion of bone morphogenetic protein 2 (BMP-2 and matrix metalloproteinase 2 (MMP-2 and to explore the signaling pathway of retinoic acid in cultured acute retinal pigment epithelial 19 (ARPE-19 cells.The effects of ATRA (concentrations from 10-9 to 10-5 mol/l on the expression of retinoic acid receptors (RARs in ARPE-19 cells were examined at the mRNA and protein levels using reverse transcription-polymerase chain reaction (RT-PCR and western blot assay, respectively. The effects of treating ARPE-19 cells with ATRA concentrations ranging from 10-9 to 10-5 mol/l for 24 h and 48 h or with 10-6mol/l ATRA at different times ranging from 6h to 72h were assessed using real-time quantitative PCR (qPCR and enzyme-linked immunosorbent assay (ELISA. The contribution of RARβ-induced activation of ARPE-19 cells was confirmed using LE135, an antagonist of RARβ.RARβ mRNA levels significantly increased in the ARPE-19 cells treated with ATRA for 24h and 48h. These increases in RARβ mRNA levels were dose dependent (at concentrations of 10-9 to 10-5 mol/l with a maximum effect observed at 10-6 mol/l. There were no significant changes in the mRNA levels of RARα and RARγ. Western blot assay revealed that RARβ protein levels were increased significantly in a time-dependent manner in ARPE-19 cells treated with 10-6 mol/l ATRA from 12 h to 72 h, with a marked increase observed at 24 h and 48 h. The upregulation of RARβ and the ATRA-induced secretion in ARPE-19 cells could be inhibited by the RARβ antagonist LE135.ATRA induced upregulation of RARβ in ARPE-19 cells and stimulated these cells to secrete BMP-2 and MMP-2.

  2. Acerogenin A, a natural compound isolated from Acer nikoense Maxim, stimulates osteoblast differentiation through bone morphogenetic protein action

    Energy Technology Data Exchange (ETDEWEB)

    Kihara, Tasuku [Section of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo (Japan); Section of Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo (Japan); Global Center of Excellence (GCOE) Program, International Research Center for Molecular Science in Tooth and Bone Diseases, Tokyo Medical and Dental University, Tokyo (Japan); Ichikawa, Saki [Section of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo (Japan); Yonezawa, Takayuki; Lee, Ji-Won [Research Institute for Biological Functions, Chubu University, Kasugai, Aichi (Japan); Akihisa, Toshihiro [College of Science and Technology, Nihon University, Tokyo (Japan); Woo, Je Tae [Research Institute for Biological Functions, Chubu University, Kasugai, Aichi (Japan); Michi, Yasuyuki; Amagasa, Teruo [Section of Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo (Japan); Yamaguchi, Akira, E-mail: akira.mpa@tmd.ac.jp [Section of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo (Japan); Global Center of Excellence (GCOE) Program, International Research Center for Molecular Science in Tooth and Bone Diseases, Tokyo Medical and Dental University, Tokyo (Japan)

    2011-03-11

    Research highlights: {yields} Acerogenin A stimulated osteoblast differentiation in osteogenic cells. {yields} Acerogenin A-induced osteoblast differentiation was inhibited by noggin. {yields} Acerogenin A increased Bmp-2, Bmp-4 and Bmp-7 mRNA expression in MC3T3-E1 cells. {yields} Acerogenin A is a candidate agent for stimulating bone formation. -- Abstract: We investigated the effects of acerogenin A, a natural compound isolated from Acer nikoense Maxim, on osteoblast differentiation by using osteoblastic cells. Acerogenin A stimulated the cell proliferation of MC3T3-E1 osteoblastic cells and RD-C6 osteoblastic cells (Runx2-deficient cell line). It also increased alkaline phosphatase activity in MC3T3-E1 and RD-C6 cells and calvarial osteoblastic cells isolated from the calvariae of newborn mice. Acerogenin A also increased the expression of mRNAs related to osteoblast differentiation, including Osteocalcin, Osterix and Runx2 in MC3T3-E1 cells and primary osteoblasts: it also stimulated Osteocalcin and Osterix mRNA expression in RD-C6 cells. The acerogenin A treatment for 3 days increased Bmp-2, Bmp-4, and Bmp-7 mRNA expression levels in MC3T3-E1 cells. Adding noggin, a BMP specific-antagonist, inhibited the acerogenin A-induced increase in the Osteocalcin, Osterix and Runx2 mRNA expression levels. These results indicated that acerogenin A stimulates osteoblast differentiation through BMP action, which is mediated by Runx2-dependent and Runx2-independent pathways.

  3. Alterations in bone morphogenetic protein 15, growth differentiation factor 9, and gene expression in granulosa cells in preovulatory follicles of dairy cows given porcine LH.

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    Behrouzi, Amir; Colazo, Marcos Germán; Ambrose, Divakar Justus

    2016-04-15

    In a previous work, using porcine LH (pLH) in lieu of GnRH for synchronizing ovulation in dairy cows improved pregnancy rates without increasing plasma progesterone concentrations after ovulation. The LH profile is known to remain elevated above basal concentrations (≥1 ng/mL) for up to 20 hours in pLH-treated cows compared to less than 6 hours in GnRH-treated cows. Because LH triggers a cascade of signaling networks in the preovulatory follicle to promote final maturation and support oocyte competence, we hypothesized that dissimilar LH profiles will differentially regulate the intrafollicular factors and expression of downstream genes associated with improved oocyte competence. Specific objectives were to determine differences in the abundance of oocyte-secreted factors in the preovulatory follicular fluid and target genes in granulosa cells associated with oocyte competence, in response to exogenous porcine LH or GnRH-induced endogenous bovine LH exposure, in dairy cows. Follicular contents were aspirated by a transvaginal ultrasound-guided procedure from the preovulatory follicle of cyclic, nonlactating Holstein cows 21 ± 1 hour after administration of either pLH (25-mg) or GnRH (100-μg). Mature forms of bone morphogenetic protein 15, growth differentiation factor 9, and transforming growth factorβ1 were approximately 2-fold more abundant in pLH-treated cows which were exposed to an extended, low LH profile, than in GnRH-treated cows that had a short, high LH profile. The relative abundance of messenger RNA for cyclooxygenase-2, LH receptor, and progesterone receptor in granulosa cells, was about two-, eight-, and two-fold higher, respectively, in cows subjected to pLH than GnRH treatment. We infer that the improved pregnancy rate after pLH-induced ovulation reported previously, occurred through greater activation of intrafollicular transforming growth factor-β1 superfamily members, as these proteins promote cumulus expansion and oocyte competence.

  4. Hepatocyte growth factor activator inhibitor-1 is induced by bone morphogenetic proteins and regulates proliferation and cell fate of neural progenitor cells.

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    Raili Koivuniemi

    Full Text Available BACKGROUND: Neural progenitor cells (NPCs in the developing neuroepithelium are regulated by intrinsic and extrinsic factors. There is evidence that NPCs form a self-supporting niche for cell maintenance and proliferation. However, molecular interactions and cell-cell contacts and the microenvironment within the neuroepithelium are largely unknown. We hypothesized that cellular proteases especially those associated with the cell surface of NPCs play a role in regulation of progenitor cells in the brain. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we show that NPCs, isolated from striatal anlage of developing rat brain, express hepatocyte growth factor activator inhibitor-1 and -2 (HAI-1 and HAI-2 that are cell surface-linked serine protease inhibitors. In addition, radial glia cells derived from mouse embryonic stem cells also express HAI-1 and HAI-2. To study the functional significance of HAI-1 and HAI-2 in progenitor cells, we modulated their levels using expression plasmids or silencing RNA (siRNA transfected into the NPCs. Data showed that overexpression of HAI-1 or HAI-2 decreased cell proliferation of cultured NPCs, whilst their siRNAs had opposite effects. HAI-1 also influenced NPC differentiation by increasing the number of glial fibrillary acidic protein (GFAP expressing cells in the culture. Expression of HAI-1 in vivo decreased cell proliferation in developing neuroepithelium in E15 old animals and promoted astrocyte cell differentiation in neonatal animals. Studying the regulation of HAI-1, we observed that Bone morphogenetic protein-2 (BMP-2 and BMP-4 increased HAI-1 levels in the NPCs. Experiments using HAI-1-siRNA showed that these BMPs act on the NPCs partly in a HAI-1-dependent manner. CONCLUSIONS: This study shows that the cell-surface serine protease inhibitors, HAI-1 and HAI-2 influence proliferation and cell fate of NPCs and their expression levels are linked to BMP signaling. Modulation of the levels and actions of HAI-1

  5. Effects of Roughly Focused Extracorporeal Shock Waves Therapy on the Expressions of Bone Morphogenetic Protein-2 and Osteoprotegerin in Osteoporotic Fracture in Rats

    Institute of Scientific and Technical Information of China (English)

    Hai-Ming Huang; Xiao-Lin Li; Shu-Qiang Tu; Xiao-Feng Chen; Chang-Chun Lu; Liang-Hua Jiang

    2016-01-01

    Background:Roughly focused extracorporeal shock waves therapy (ESWT) is characterized by a wide focal area,a large therapy zone,easy positioning,and less pain during treatment.The purpose of this study was to investigate the effects of roughly focused ESWT on the expression of osteoprotegerin (OPG) and bone morphogenetic protein-2 (BMP-2) in osteoporotic fractures in rats.Methods:Seventy-two female Sprague-Dawley (SD) rats,3 months old,were divided into sham-operated group (n =6) and an ovariectomized (OVX) group (n =66).Sixty OVX SD rats were used as a model of double proximal tibial osteotomy and inner fixation.The osteotomy site in the left tibia was treated with roughly focused ESWT once at an energy density of 0.26 mJ/mm2,60 doses/min,and 2000 pact quantities.The contralateral right tibia was left untreated and served as a control.Expression of OPG and BMP-2 in the callus of the osteoporotic fracture area was assessed using immunohistochemistry,real-time polymerase chain reaction (PCR),and Western blotting analysis.Results:Bone mineral density (BMD) at the proximal tibia,femur,and L5 spine was significantly reduced after ovariectomy.BMD of proximal tibia was 12.9% less in the OVX group than that in the sham-operated group.Meanwhile,bilateral oophorectomy resulted in a lower trabecular bone volume fraction (BV/TV) in the proximal tibia of the sham-OVX animals.Three months after bilateral oophorectomy,BV/TV was 14.29% of baseline BV/TV in OVX legs versus 45.91% in the sham-OVX legs (P < 0.001).These data showed that the SD rats became a suitable model of osteoporosis,3 months after they were OVX.Immunohistochemical analysis showed higher levels of BMP-2 and OPG expression in the treatment group than those in the control group.Compared with the contralateral controls,decreased expression of OPG and BMP-2 at 3 days after roughly focused ESWT,followed by a later increase at 7 days,was indicated by real-time PCR and Western blotting analysis.The OPG

  6. Experience with bone morphogenetic protein-2 and interpositional grafting of edentulous maxillae: a comparison of Le Fort I downfracture to full-arch (horseshoe) segmental osteotomy done in conjunction with sinus floor grafting.

    Science.gov (United States)

    Jensen, Ole T; Cottam, Jared R; Ringeman, Jason L; Leopardi, Aldo; Butler, Brian; Laviv, Amir; Fleissig, Yoram; Casap, Nardy

    2013-01-01

    This paper is a retrospective report of the treatment of six patients with severely resorbed maxillae. Patients were treated, based on the amount of maxillary retrognathia, with either a Le Fort I downfracture or a "horseshoe" interpositional sandwich osteotomy, along with sinus elevation. Recombinant human bone morphogenetic protein-2 in an absorbable collagen sponge carrier was used for grafting in all patients, either alone or in combination with other grafting materials. Implants were placed and the patients were restored with fixed prostheses. Both grafting techniques are described, and the treated patients are presented.

  7. Elevated extracellular calcium increases expression of bone morphogenetic protein-2 gene via a calcium channel and ERK pathway in human dental pulp cells

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    Tada, Hiroyuki [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Nemoto, Eiji, E-mail: e-nemoto@umin.ac.jp [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Kanaya, Sousuke; Hamaji, Nozomu; Sato, Hisae; Shimauchi, Hidetoshi [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)

    2010-04-16

    Dental pulp cells, which have been shown to share phenotypical features with osteoblasts, are capable of differentiating into odontoblast-like cells and generating a dentin-like mineral structure. Elevated extracellular Ca{sup 2+}Ca{sub o}{sup 2+} has been implicated in osteogenesis by stimulating the proliferation and differentiation of osteoblasts; however, the role of Ca{sub o}{sup 2+} signaling in odontogenesis remains unclear. We found that elevated Ca{sub o}{sup 2+} increases bone morphogenetic protein (BMP)-2 gene expression in human dental pulp cells. The increase was modulated not only at a transcriptional level but also at a post-transcriptional level, because treatment with Ca{sup 2+} increased the stability of BMP-2 mRNA in the presence of actinomycin D, an inhibitor of transcription. A similar increase in BMP-2 mRNA level was observed in other human mesenchymal cells from oral tissue; periodontal ligament cells and gingival fibroblasts. However, the latter cells exhibited considerably lower expression of BMP-2 mRNA compared with dental pulp cells and periodontal ligament cells. The BMP-2 increase was markedly inhibited by pretreatment with an extracellular signal-regulated kinase (ERK) inhibitor, PD98059, and partially inhibited by the L-type Ca{sup 2+} channels inhibitor, nifedipine. However, pretreatment with nifedipine had no effect on ERK1/2 phosphorylation triggered by Ca{sup 2+}, suggesting that the Ca{sup 2+} influx from Ca{sup 2+} channels may operate independently of ERK signaling. Dental pulp cells do not express the transcript of Ca{sup 2+}-sensing receptors (CaSR) and only respond slightly to other cations such as Sr{sup 2+} and spermine, suggesting that dental pulp cells respond to Ca{sub o}{sup 2+} to increase BMP-2 mRNA expression in a manner different from CaSR and rather specific for Ca{sub o}{sup 2+} among cations.

  8. Use of recombinant human bone morphogenetic protein-2 as an adjunct for instrumented posterior arthrodesis in the occipital cervical region: An analysis of safety, efficacy, and dosing

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    D Kojo Hamilton

    2010-01-01

    Full Text Available Background: There have been few reports on the use of recombinant human bone morphogenetic protein (rhBMP-2 in posterior spine. However, no study has investigated the dosing, safety, and efficacy of its use in the posterior atlantoaxial, and/or craniovertebral junction. Recent case report of the cytokine-mediated inflammatory reaction, following off label use of rhBMP-2 as an adjunct for cervical fusion, particularly in complex cases, has increased concern about complications associated with the product. Objective: To assess the safety, efficacy, and dosing of rhBMP-2 as an adjunct for instrumented posterior atlantoaxial and/or craniovertebral junction arthrodesis. Materials and Methods: We included all patients treated by the senior author that included posterior atlantoaxial and/or craniovertebral junction instrumented fusion using rhBMP-2 from 2003 to 2008 with a minimum two year follow-up. Diagnosis, levels fused, rhBMP-2 dose, complications, and fusion were assessed. Results: Twenty three patients with a mean age of 60.9 years (range 4 - 89 years and an average follow-up of 45 months (range 27 to 84 months met inclusion criteria. The indications for surgery included, atlantoaxial instability (n = 16, basilar invagination (n = 6, and kyphoscoliosis (n = 1. The specific pathologic diagnosis included type 2 dens fracture (n = 7, complex C1 and C2 ring fracture (n = 2, chordoma (n = 2, degenerative/osteoporosis (n = 3, rheumatoid disease (n = 8, and pseudogout (n = 1. The average rhBMP-2 dose was 2.38 mg/level, with a total of 76 levels treated (average 3.3 levels, SD= 1.4 levels. There were no complications. During the most recent follow-up, all patients had achieved fusion. Conclusions: In a series of patients with complex pathology and/or rheumatoid arthritis, 100% fusion rate was achieved with adjunct use of rhBMP-2, with a safe and effective average rhBMP-2 dose of 2.38 mg per level.

  9. Adenovirus-mediated human bone morphogenetic protein 2 gene transfects bone marrow mesenchymal stem cells%腺病毒介导的人骨形态发生蛋白2基因转染骨髓间充质干细胞*☆

    Institute of Scientific and Technical Information of China (English)

    尹承慧; 邱俊钦; 曾昭勋; 陈宗雄

    2013-01-01

      背景:骨髓间充质干细胞作为骨、软骨创伤缺损及退变修复的种子细胞越来越受到关注。目的:分析人骨形态发生蛋白2基因转染对白色封闭群大鼠(SD 大鼠)骨髓间充质干细胞的影响。方法:分离纯化 SD 大鼠骨髓间充质干细胞并体外扩增,通过腺病毒载体介导人骨形态发生蛋白2基因转染骨髓间充质干细胞,分别通过荧光显微镜观察荧光表达情况及蛋白质水平来测定转染后人骨形态发生蛋白2的表达,碱性磷酸酶定量测定鉴定成骨活性及 MTT 法评估人骨形态发生蛋白2转染对骨髓间充质干细胞的影响。结果与结论:从 SD 大鼠骨髓提取物中分离培养的细胞形态为梭形,呈铺路石状、漩涡状生长,经流式细胞仪检测及多项分化能力鉴定符合骨髓间充质干细胞的特征;经转染人骨形态发生蛋白2基因后,骨髓间充质干细胞表达人骨形态发生蛋白2、碱性磷酸酶;MTT 法检测转染人骨形态发生蛋白2基因后,骨髓间充质干细胞增殖能力明显增强(P <0.05)。说明人骨形态发生蛋白2基因转染骨髓间充质干细胞后可以持续、高效表达人骨形态发生蛋白2和碱性磷酸酶,在体外明显促进骨髓间充质干细胞的增殖。%BACKGROUND: Bone marrow mesenchymal stem cel s as the seed cel s for repair of bone and cartilage trauma and degeneration have been paid increasing attention. OBJECTIVE: To investigative the effects of human bone morphogenetic protein 2 gene transfection on Sprague-Dawley rat bone marrow mesenchymal stem cel s. METHODS: Sprague-Dawley rat bone marrow mesenchyal stem cel s were in vitro isolated, purified and amplified. Adenovirus-mediated human bone morphogenetic protein 2 was transfected into bone marrow mesenchymal stem cel s. CD90 and CD45 expression levels were tested by flow cytometry. The successful y packaged virus was transfected into bone marrow mesenchymal

  10. Construction and soluble expression of recombinant Bone Morphogenetic Protein-2%骨形态发生蛋白-2基因工程菌的构建及可溶性研究

    Institute of Scientific and Technical Information of China (English)

    赵瑾; 陈洪; 林陈水

    2012-01-01

    骨形态发生蛋白是一类调节骨组织发育的生长因子,其中骨形态发生蛋白-2诱导成骨活性最强,在骨组织工程研究中最具研究意义.为获得能高效表达溶解性高的人骨形成蛋白-2 (BMP-2)的基因工程菌,用PCR方法扩增得到BMP-2的基因序列,直接将PCR产物连接到胞内融合表达型T载体质粒pMT-L上,构建包括麦芽糖结合蛋白(MBP)、连接肽、6个His、EKsite(Asp-AspAsp-Asp-Lys)和BMP-2的表达载体,转化E,coli DH5a,经抗性筛选和菌落PCR鉴定,抽提阳性克隆质粒转化表达宿主E.coli BL21(DE3),成功构建可在大肠杆菌细胞质内表达MBP-BMP-2融合蛋白的基因工程菌.工程菌经0.1 mmol/L IPTG诱导后,可获得表观分子量约为55 kD的以可溶形式表达的BMP-2融合蛋白.%Bone morphogenetic proteins are a group of growth {actors known for their ability to induce the formation of bone and cartilage. The bone morphogenetic protein-2 (BMP-2) displays a higher osteogenic activity than others, showing it has the most research significance in the bone tissue engineering. To obtain a high expression of soluble bone morphogenetic protein-2 (BMP-2) genetic engineering bacteria,the DNA sequence of Bone Morphogenetic Protein-2 (BMP-2) was amplified by polymerase chain reaction (PCR). Then, the produce of PCR was directly recombined to the intracellular fusing expressional T-Vector pMT-L, constructing the expression plasmid that containing maltose-binding protein(MBP), linker peptide, 6 His, EKsite (Asp-Asp-Asp-Asp-Lys) and BMP-2, then the recombinant plasmid transformed into E.coli DH5a. After a drug-resistant selection and the identification by colonies PCR, the interested plasmid was extracted and transformed into E. coli BL21 (DE3), the genetically engineered E. coli that can express MBP-BMP-2 fusion protein in the E. coli cytoplasm was successfully constructed. The results indicated that the genetically engineered bacteria expressed a large quantity soluble

  11. Bone morphogenetic protein-4 and transforming growth factor-beta1 mechanisms in acute valvular response to supra-physiologic hemodynamic stresses

    Institute of Scientific and Technical Information of China (English)

    Ling; Sun; Philippe; Sucosky

    2015-01-01

    AIM:To explore ex vivo the role of bone morphogenetic protein-4(BMP-4) and transforming growth factorbeta1(TGF-β1) in acute valvular response to fluid shear stress(FSS) abnormalities.METHODS:Porcine valve leaflets were subjected ex vivo to physiologic FSS,supra-physiologic FSS magnitude at normal frequency and supra-physiologic FSS frequency at normal magnitude for 48 h in a double-sided cone-and-plate bioreactor filled with standard culture medium. The role of BMP-4 and TGF-β1 in the valvular response was investigated by promoting or inhibiting the downstream action of those cytokines via culture medium supplementation with BMP-4 or the BMP antagonist noggin,and TGF-β1 or the TGF-β1 inhibitor SB-431542,respectively. Fresh porcine leaflets were used as controls. Each experimental group consisted of six leaflet samples. Immunostaining and immunoblotting were performed to assess endothelial activation in terms of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expressions,paracrine signaling in terms of BMP-4 and TGF-β1 expressions and extracellular matrix(ECM) remodeling in terms of cathepsin L,cathepsin S,metalloproteinases(MMP)-2 and MMP-9 expressions. Immunostained images were quantified by normalizing the intensities of positively stained regions by the number of cells in each image while immunoblots were quantified by densitometry. R E S U LT S :Regardless of the culture medium,physiologic FSS maintained valvular homeostasis. Tissue exposure to supra-physiologic FSS magnitude in standard medium stimulated paracrine signaling(TGF-β1:467% ± 22% vs 100% ± 6% in freshcontrols,BMP-4:258% ± 22% vs 100% ± 4% in fresh controls; P < 0.05) and ECM degradation(MMP-2:941% ± 90% vs 100% ± 19% in fresh controls,MMP-9:1219% ± 190% vs 100% ± 16% in fresh controls,cathepsin L:1187% ± 175% vs 100% ± 12% in fresh controls,cathepsin S:603% ± 88% vs 100% ± 13% in fresh controls; P < 0.05),while BMP-4 supplementation also promoted fibrosa

  12. Effects of bone morphogenetic protein-4 on spatial memory and cholinergic expression in the dentate gyrus after fornix-fimbria transection in rats

    Institute of Scientific and Technical Information of China (English)

    Lei Liu; Yilong Xue; Jingkun Pan; Yazhuo Hu; Yuhong Gao; Yun Luo

    2008-01-01

    BACKGROUND: Previous experiments have confirmed bone morphogenetic proteins (BMPs) upregulate cholinergic expression in neurons isolated from the embryonic rat hippocampus and cerebral cortex. Therefore, BMPs could be useful for treating Alzheimer's disease and other neurodegenerative diseases. OBJECTIVE: BMP-4 was infused into the hippocampal dentate gyrus of fornix-fimbria transected rats to test the effects of BMP-4 on cholinergic expression in dentate gyrus neurons, and to observe changes in spatial memory behavior. DESIGN: A randomized controlled animal experiment. SETTING: Department of Neurosurgery and Laboratory for Cell Biology, Institute of Geriatrics, General Hospital of Chinese PLA.MATERIALS: Twenty-seven healthy adult male Sprague Dawley (SD) rats, weighing 250-300 g, were provided by the Laboratory Animal Center of the General Hospital of Chinese PLA. Reagents: BMP-4 (B-2680, Sigma Company) and choline acetyl transferase (ChAT) antibody (AB5042, Chemicon Company) were used in this study. Equipments: a rat stereotaxic instrument (type: SN-2N, Narushige Group, Japan) and Image-prog-plus image analysis software (Media Cybernetics company, USA) were used in this study. The protocol was carried out in accordance with ethical guidelines for the use and care of animals.METHODS: This experiment was performed in the Institute of Geriatrics, General Hospital of Chinese PLA between July 2004 and March 2005. Rats were randomly divided into 4 groups: Alzheimer's disease group (n = 7), normal control group (n = 5), BMP-4-Alzheimer's disease group (n = 8), and model group (n = 7). In the Alzheimer's disease group, the left hippocampal fornix-fimbria of rats was transected to mimic Alzheimer's disease symptoms. In the BMP-4-Alzheimer's disease group, 1 μL BMP-4 (10 mg/L) was perfused into the left dentate gyrus with a microinjector at 1 μL/min. In the model group, 1 μL saline was perfused into the same position by the same method. Twenty-eight days after injection

  13. 骨形态发生蛋白2缓释载体的研究进展%Research Progress of Bone Morphogenetic Protein-2 Controlled-release Carrier

    Institute of Scientific and Technical Information of China (English)

    张以财; 焦力刚

    2012-01-01

    自体骨移植一直是骨修复的"金标准",但仍存在一些问题.异体骨移植同样存在着骨愈合缓慢及排斥反应等问题.随着组织工程学的发展,应用骨组织工程方法来修复骨缺损成为研究热点.骨组织工程主要包括支架材料、种子细胞、生长因子三个方面.骨形态发生蛋白2是目前最强的促骨生长因子,其在体内半衰期很短,必须依靠缓释载体才能发挥其较长效的促骨生长作用.%Autogenous bone graft has long been the " golden standard" of bone repair, while there are some remaining problems. Allograft also have many problems, such as slow bone healing and rejection etc. . With the development of tissue engineering, lots of eyes focus on bone tissue engineering to repair bone defects. There are three key points in bone tissue engineering namely scaffolds, seed cells and growth factor. Bone morphogenetic protein-2 is the most efficient factor to promote bone growth so far,but it has a very short half-time in vivo, which must rely on control-released carrier to fulfill its long-term bone growth-promoting effect.

  14. Ultrastructure of periosteal cells induced by bone morphogenetic protein 7 in vitro%骨形态发生蛋白7诱导骨膜细胞体外构骨的超微结构

    Institute of Scientific and Technical Information of China (English)

    廖家成; 贝抗胜; 连银川; 徐进文; 刘延晓; 黄晓瑜

    2014-01-01

    BACKGROUND:Study confirms that bone morphogenetic protein can induce osteogenesis;however the ultrastructure of periosteal cells induced by bone morphogenetic protein-7 remains poorly reported. OBJECTIVE:To study the bioactivity and ultrastructure of periosteal cells induced by bone morphogenetic protein-7 in vitro. METHODS:The primary periosteal cells isolated from adult tibial bone were in vitro cultured, and then divided into experimental group and control group. In the experimental group, cells were cultured with bone morphogenetic protein-7 and culture adjuvant;while cells in the control group were only cultured with the adjuvant. Three samples in each group were tested at 5, 10, 15 days, respectively. The general structure of cultured cells was observed using von Kossa staining, and the ultrastructure was observed under transmission electron microscopy. RESULTS AND CONCLUSION:The periosteal cells in the two groups grew wel in vitro, showing uniform morphology. Early cells were spindle-shaped, with strong three-dimensional sense and ful transparency;mitotic cells were short columnar or cubic shaped, there were a lot of rough endoplasmic reticulum and Golgi complex in osteoblasts under electron microscope. Later stage of cells developed from long fusiform into wide shuttle and irregular shape, there were a large number of matrix vesicles within the cells under the electron microscope. The membrane coating, alkaline phosphatase and calcium-binding protein in the cytoplasm, as wel as calcium crystals were found. The osteogenesis basement and lateral sides appeared projections, which were connected with adjacent bone cells. Induction of bone morphogenetic protein-7 in vitro promotes the osteoblasts proliferation, division and bone formation speed. The results suggest that bone morphogenetic protein-7 can significantly enhance the proliferation ability of osteoblasts in vitro.%背景:骨形态发生蛋白被证实有成骨诱导作用,然

  15. Investigation of gene expressions in differentiated cell derived bone marrow stem cells during bone morphogenetic protein-4 treatments with Fourier transform infrared spectroscopy

    Science.gov (United States)

    Zafari, Jaber; Jouni, Fatemeh Javani; Ahmadvand, Ali; Abdolmaleki, Parviz; Soodi, Malihe; Zendehdel, Rezvan

    2017-02-01

    A model was set up to predict the differentiation patterns based on the data extracted from FTIR spectroscopy. For this reason, bone marrow stem cells (BMSCs) were differentiated to primordial germ cells (PGCs). Changes in cellular macromolecules in the time of 0, 24, 48, 72, and 96 h of differentiation, as different steps of the differentiation procedure were investigated by using FTIR spectroscopy. Also, the expression of pluripotency (Oct-4, Nanog and c-Myc) and specific genes (Mvh, Stella and Fragilis) were investigated by real-time PCR. However, the expression of genes in five steps of differentiation was predicted by FTIR spectroscopy. FTIR spectra showed changes in the template of band intensities at different differentiation steps. There are increasing changes in the stepwise differentiation procedure for the ratio area of CH2, which is symmetric to CH2 asymmetric stretching. An ensemble of expert methods, including regression tree (RT), boosting algorithm (BA), and generalized regression neural network (GRNN), was the best method to predict the gene expression by FTIR spectroscopy. In conclusion, the model was able to distinguish the pattern of different steps from cell differentiation by using some useful features extracted from FTIR spectra.

  16. Bone morphogenetic protein-2 (BMP-2 and transforming growth factor-β1 (TGF-β1 alter connexin 43 phosphorylation in MC3T3-E1 Cells

    Directory of Open Access Journals (Sweden)

    Rudkin George H

    2001-07-01

    Full Text Available Abstract Background Bone morphogenetic proteins (BMPs and transforming growth factor-βs (TGF-βs are important regulators of bone repair and regeneration. BMP-2 and TGF-β1 have been shown to inhibit gap junctional intercellular communication (GJIC in MC3T3-E1 cells. Connexin 43 (Cx43 has been shown to mediate GJIC in osteoblasts and it is the predominant gap junctional protein expressed in these murine osteoblast-like cells. We examined the expression, phosphorylation, and subcellular localization of Cx43 after treatment with BMP-2 or TGF-β1 to investigate a possible mechanism for the inhibition of GJIC. Results Northern blot analysis revealed no detectable change in the expression of Cx43 mRNA. Western blot analysis demonstrated no significant change in the expression of total Cx43 protein. However, significantly higher ratios of unphosphorylated vs. phosphorylated forms of Cx43 were detected after BMP-2 or TGF-β1 treatment. Immunofluorescence and cell protein fractionation revealed no detectable change in the localization of Cx43 between the cytosol and plasma membrane. Conclusions BMP-2 and TGF-β1 do not alter expression of Cx43 at the mRNA or protein level. BMP-2 and TGF-β1 may inhibit GJIC by decreasing the phosphorylated form of Cx43 in MC3T3-E1 cells.

  17. Effect of recombinant human bone morphogenetic protein 2/poly-lactide-co-glycolic acid (rhBMP-2/PLGA) with core decompression on repair of rabbit femoral head necrosis

    Institute of Scientific and Technical Information of China (English)

    Zhao-Xun Pan; Hong-Xin Zhang; Ye-Xin Wang; Long-Di Zhai; Wei Du

    2014-01-01

    Objective:To observe the effect of recombinant human bone morphogenetic protein 2/poly-lactide-co-glycolic acid (rhBMP-2/PLGA) with core decompression on repair of rabbit femoral head necrosis. Methods: Bilateral femoral head necrosis models of rabbit were established by steroid injection. A total of 48 rabbits (96 femoral head necrosis) were randomly divided into 4 groups: Group A, control group with12 rabbits, 24 femoral head necrosis;Group B, treated with rhBMP-2/PLGA implantation after core depression, with 12 rabbits, 24 femoral head necrosis;Group C, treated with rhBMP-2 implantation after core depression, with 12 rabbits, 24 femoral head necrosis;Group D treated with core depression group without implantation, with 12 rabbits, 24 femoral head necrosis. All animals were sacrificed after 12 weeks. The ability of repairing bone defect was evaluated by X-ray radiograph. Bone mineral density analysis of the defect regions were used to evaluate the level of ossification. The morphologic change and bone formation was assessed by HE staining. The angiogenesis was evaluated by VEGF immunohistochemistry. Results: The osteogenetic ability and quality of femoral head necrosis in group B were better than those of other groups after 12 weeks by X-ray radiograph and morphologic investigation. And the angiogenesis in group B was better than other groups. Group C had similar osteogenetic quality of femoral head necrosis and angiogenesis with group D. Conclusions:The treatment of rhBMP-2/PLGA implantation after core depression can promote the repair of rabbit femoral head necrosis. It is a promising and efficient synthetic bone material to treat the femoral head necrosis.

  18. The regulation of bone morphogenetic protein 2 on osseointegration under hypoxia%低氧条件下骨形态发生蛋白2对骨整合的调控

    Institute of Scientific and Technical Information of China (English)

    廖晓妤; 李迎春

    2012-01-01

    Under the condition of hypoxia, bone loss would happen, and bone heal would be delayed, even nonunion, and osseointegration would be delayed after implant surgery. Bone morphogenetic protein (BMP) 2 is the central regulator of osteoblast differentiation and participates in the process of bone and cartilage formation whether it is in an individual or combined way. In this article, the bone reaction, the influence of BMP2 on osteoblast and osteoclast differentiation, as well as the interaction between BMP2 and vascular endothelial growth factor and the mechanism of BMP2 regulation of osseointegration under hypoxia are reviewed.%在低氧状态下,骨量丢失、骨组织再生延迟,骨组织损伤后愈合缓慢甚至不愈合,种植手术后骨整合延迟;而骨形态发生蛋白(BMP)2为成骨细胞分化的中心调控因子,无论是单一的还是组合的BMP2皆参与了软骨形成和骨形成过程.本文就低氧环境下骨组织的反应,BMP2对骨整合过程中成骨细胞和破骨细胞的影响,骨整合过程中BMP2与血管内皮生长因子间的相互作用,低氧作用于BMP2的机制等研究进展作一综述.

  19. Oxygen-glucose deprivation preconditioning protects neurons against oxygen-glucose deprivation/reperfusion induced injury via bone morphogenetic protein-7 mediated ERK, p38 and Smad signalling pathways.

    Science.gov (United States)

    Guan, Junhong; Du, Shaonan; Lv, Tao; Qu, Shengtao; Fu, Qiang; Yuan, Ye

    2016-01-01

    Bone morphogenetic protein (BMP)-7 mediated neuroprotective effect of cerebral ischemic preconditioning (IPC) has been studied in an ischemic animal model, but the underlying cellular mechanisms have not been clearly clarified. In this study, primary cortical neurons and the SH-SY5Y cell line were used to investigate the role of BMP-7 and its downstream signals in the neuroprotective effects of oxygen-glucose deprivation preconditioning (OGDPC). Immunocytochemistry was used to detect the expression of neurofilament in neurons. MTT and lactate dehydrogenase activity assays were used to measure the cytotoxicity. Western blot was used to detect the protein expression of BMP-7 and downstream signals. BMP inhibitor, mitogen-activated protein kinase inhibitors, Smad inhibitor and siRNA of Smad 1 were used to investigate the role of corresponding signalling pathways in the OGDPC. Results showed that OGDPC-induced overexpression of BMP-7 in primary cortical neurons and SH-SY5Y cells. Both of endogenous and exogenous BMP-7 could replicate the neuroprotective effects seen in OGDPC pretreatment. In addition, extracellular regulated protein kinases, p38 and Smad signalling pathway were found to be involved in the neuroprotective effects mediated by OGDPC via BMP-7. This study primarily reveals the cellular mechanisms of the neuroprotection mediated by OGDPC, and provides evidence for better understanding of this intrinsic factor against ischemia.

  20. Effects of LED phototherapy on bone defects grafted with MTA, bone morphogenetic proteins, and guided bone regeneration in a rodent model: a description of the bone repair by light microscopy

    Science.gov (United States)

    Pinheiro, Antonio Luiz B.; Aciole, Gilberth T. S.; Soares, Luiz G. P.; Correia, Neandder A.; N. dos Santos, Jean

    2011-03-01

    We carried out a histological analysis on surgical bone defects grafted or not with MTA, treated or not with LED, BMPs and GBR. We have used several models to assess the effects of laser on bone. Benefits of the isolated or combined use them on bone healing has been suggested. There is no previous report on their association with LED light. 90 rats were divided into 10 groups. On Groups II and I the defect were filled with the clot. On Group II, were further irradiated. On groups III-VI, defect was filled with MTA + Collagen gel (III); animals of group IV were further irradiated. On groups V and VI, the defects filled with the MTA were covered with a membrane. Animals of Group VI were further irradiated. On Groups VII and VIII a pool of BMPs was added to the MTA and was further irradiated. On groups IX and X, the MTA + BMP graft was covered with a membrane. On group X, the defect was further irradiated. LED (λ850 +/- 10nm, 150mW, A= 0.5cm2, 54s, 0.3W/cm2, 16 J/cm2) was applied at 48 h intervals during 15 days. Specimens were taken, processed, cut and stained with H&E and Sirius red and underwent histological analysis. The results showed that MTA seemed not being affected by LED light. However, its use positively affected healing around the graft. It is concluded that MTA is not affected by the LED light due to it characteristics, but beneficial results with LED usage was found.

  1. Histological observation on transforming growth factor- β compounded with bone morphogenetic protein as a pulp capping material%TGF-β复合BMP盖髓的组织学观察

    Institute of Scientific and Technical Information of China (English)

    李国华; 赵京宁; 王学英; 金岩; 牛忠英

    2000-01-01

    目的:用β转化生长因子复合骨形成蛋白(TGF-β/BMP)覆盖狗暴露牙髓,观察TGF-β/BMP的盖髓效果。方法:用活体狗牙作直接盖髓实验,并用组织学方法评价盖髓效果。结果:术后8周,TGF-β/BMP能够诱导牙髓成纤维细胞、内皮细胞增殖并抑制炎症反应,提高牙髓修复能力,有完整的牙本质桥形成。结论:TGF-β/BMP能促进狗牙髓组织修复。%AIM: Observation of the effects of transforming growth factor- β compounded with bone morphogenetic protein (TGF- β/BMP) on the exposed pulp of dogs. METHODS: The teeth of dogs were used for direct pulp capping experiment and the results were evaluated histopathologically. RESULt: Eight weeks postoperatively TGF- β/BMP was able to induce the proliferations of the pulp cells (fibroblasts)and endothelial cell, inhibit the inflammatory reaction of pulp, and possessed the ability to enhance the pulp repairing, complete dentin bridge was induced. CONCLUSION:TGF- β/BMP can promote the regeneration of dog's pulp tissue.

  2. Multiple roles of Activin/Nodal, bone morphogenetic protein, fibroblast growth factor and Wnt/β-catenin signalling in the anterior neural patterning of adherent human embryonic stem cell cultures

    Science.gov (United States)

    Lupo, Giuseppe; Novorol, Claire; Smith, Joseph R.; Vallier, Ludovic; Miranda, Elena; Alexander, Morgan; Biagioni, Stefano; Pedersen, Roger A.; Harris, William A.

    2013-01-01

    Several studies have successfully produced a variety of neural cell types from human embryonic stem cells (hESCs), but there has been limited systematic analysis of how different regional identities are established using well-defined differentiation conditions. We have used adherent, chemically defined cultures to analyse the roles of Activin/Nodal, bone morphogenetic protein (BMP), fibroblast growth factor (FGF) and Wnt/β-catenin signalling in neural induction, anteroposterior patterning and eye field specification in hESCs. We show that either BMP inhibition or activation of FGF signalling is required for effective neural induction, but these two pathways have distinct outcomes on rostrocaudal patterning. While BMP inhibition leads to specification of forebrain/midbrain positional identities, FGF-dependent neural induction is associated with strong posteriorization towards hindbrain/spinal cord fates. We also demonstrate that Wnt/β-catenin signalling is activated during neural induction and promotes acquisition of neural fates posterior to forebrain. Therefore, inhibition of this pathway is needed for efficient forebrain specification. Finally, we provide evidence that the levels of Activin/Nodal and BMP signalling have a marked influence on further forebrain patterning and that constitutive inhibition of these pathways represses expression of eye field genes. These results show that the key mechanisms controlling neural patterning in model vertebrate species are preserved in adherent, chemically defined hESC cultures and reveal new insights into the signals regulating eye field specification. PMID:23576785

  3. 骨形态发生蛋白7诱导骨膜细胞碱性磷酸酶的表达*%Bone morphogenetic protein-7 induces the expression of alkaline phosphatase in periosteal cells

    Institute of Scientific and Technical Information of China (English)

    廖家成; 贝抗胜; 连银川

    2013-01-01

      背景:关于骨形态发生蛋白7作为刺激因子诱导细胞成骨的报道目前较少见。目的:观察骨膜细胞经骨形态发生蛋白7诱导后碱性磷酸酶的表达。  方法:取材于成人胫骨骨膜,常规细胞培养法行骨膜细胞体外培养,分为实验组和对照组,分别加入骨形态发生蛋白7加成骨细胞培养辅助剂和单纯成骨细胞培养辅助剂,相差显微镜观察骨膜细胞形态特征及超微结构。每组分别在第7,14,21天设3个时间点,每个时间点设3个样本,采用碱性磷酸酶试剂盒法检测成骨细胞特异性标志物碱性磷酸酶表达情况。  结果与结论:骨膜细胞经分组培养后,第7天时,实验组和对照组骨膜细胞均有明显增殖,碱性磷酸酶的可被检测出,但量不多,细胞外形为梭形,实验组比对照组检测的碱性磷酸酶数量稍多;第14天时,实验组及对照组骨膜细胞均显著增殖,细胞外形由梭形变为宽梭形,实验组比对照组检测的碱性磷酸酶数量明显增多。第21天时,实验组及对照组骨膜细胞均增殖,其中实验组细胞增殖明显,细胞外形为宽梭形,实验组比对照组检测的碱性磷酸酶数量显著增多。经过统计学分析由骨形态发生蛋白7诱导的骨膜细胞的成骨标志物碱性磷酸酶阳性率明显高于对照组(P OBJECTIVE:To study the expression of alkaline phosphatase of periosteal cel s after induced by bone morphogenetic protein-7 in vitro. METHODS:Periosteal cel s were obtained from adult tibial periosteum, and then the periosteal cel s were cultured by routine method in vitro. The cel s were divided into experimental group and control group, and then cultured with bone morphogenetic protein-7 plus osteoblast culture adjuvants and simple osteoblast culture adjuvants, respectively. The phase contrast microscope was used to observe the morphology and ultrastructure of periosteal

  4. Effects of Roughly Focused Extracorporeal Shock Waves Therapy on the Expressions of Bone Morphogenetic Protein-2 and Osteoprotegerin in Osteoporotic Fracture in Rats

    Directory of Open Access Journals (Sweden)

    Hai-Ming Huang

    2016-01-01

    Conclusions: Roughly focused ESWT may promote the expression of OPG and BMP-2 in the osteoporotic fracture area in rats. BMP-2 and OPG may act synergistically and may lead to a significant enhancement of bone formation and remodeling.

  5. Application of bone morphogenetic proteins in mandibular fast distraction osteogenesis%骨形态形成蛋白在下颌骨快速牵张成骨中的应用

    Institute of Scientific and Technical Information of China (English)

    刘浩; 王敏

    2012-01-01

    BACKGROUND: Distraction osteogenesis has became an effective method in treatment of different kinds of craniofacialdeformities and bone defects. However, the major disadvantage of this method is the long distraction and consolidation period,which may lead to severe complications during the distraction process.OBJECTIVE: To summarize the research progress of the application of bone morphogenetic proteins (BMPs) in fast distractionosteogenesis.METHODS: A computer-based online search in Pubmed database was performed to select the reviews and papers related toBMPs and distraction osteogenesis from 1989 to 2011.RESULTS AND CONCLUSION: A total of 32 literatures on the the application of BMPs in fast distraction osteogenesis wereincluded. BMPs have a strong osteogenic activity and capability of promoting regeneration and remodeling of bone. The presentresearch shows that BMPs can be used to accelerate new bone formation and shorten the treatment period. However, the clinicalapplication of BMPs needs further research.%背景:牵张成骨已经成为治疗不同类型颅面畸形和骨缺损的有效的方法,但是牵张成骨的主要缺点是牵张期和稳定期比较长,可能导致牵张过程中严重的并发症.目的:总结骨形态形成蛋白在快速牵张成骨过程中作用的研究现状.方法:电子检索计算机Pubmed 数据库(1989/2011)收录的骨形态形成蛋白和牵张成骨相关综述和论文报告.结果与结论:共纳入骨形态形成蛋白在快速牵张成骨中的作用相关文献32 篇.骨形态形成蛋白具有很强的成骨活性,能促进骨再生和骨改建.目前的研究显示应用骨形态形成蛋白能加快牵张成骨过程中新骨形成和缩短治疗的疗程.但是骨形态形成蛋白应用于临床还需要进一步的研究.

  6. In vivo bacterial morphogenetic protein interactions

    NARCIS (Netherlands)

    van der Ploeg, R.; den Blaauwen, T.; Meghea, A.

    2012-01-01

    This chapter will discuss none-invasive techniques that are widely used to study protein-protein interactions. As an example, their application in exploring interactions between proteins involved in bacterial cell division will be evaluated. First, bacterial morphology and cell division of the rod-s

  7. Profiling bone morphogenetic proteins and transforming growth factor-βs by hTGF-β3 pre-treated coral-derived macroporous bioreactors: the power of one.

    Science.gov (United States)

    Ripamonti, Ugo; Dix-Peek, Thérèse; Parak, Ruqayya; Milner, Brenda; Duarte, Raquel

    2015-05-01

    To study the expression profile of bone morphogenetic proteins and transforming growth factor-βs (BMPs and TGFβs), coral-derived calcium carbonate-based macroporous bioreactors with limited conversion to hydroxyapatite (7% HA/CC) were pre-loaded with and without 250 μg hTGF-β3 and implanted in the rectus abdominis of 3 non-human primates Papio ursinus euthanized on day 60. To investigate the required dose of hNoggin, a BMPs antagonist that controls the induction of bone formation, 7% HA/CC were pre-loaded with 150 μg hNoggin, with 125 μg hTGF-β3/150 μg hNoggin, with or without 125 μg hTGF-β3 and implanted in the r. abdominis of 3 additional animals euthanized on day 90. Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) evaluated the expression' profile of BMP-2, BMP-3, BMP-4, BMP-6, BMP-7 and TGF-β1, -β2, and -β3 in tissue generating bioreactors as well as in the adjacent r. abdominis muscle. On day 60, 250 μg hTGF-β3 induced bone formation at the periphery of the implanted bioreactors only. On day 90, 125 μg hTGF-β3/treated bioreactors showed the induction of bone formation throughout the macroporous spaces. Untreated bioreactors induced bone, 4.11% vs. 2.00% on days 60 and 90, respectively. In hTGF-β3/treated bioreactors, BMP-2 and BMP-3 were up-regulated at both time periods, both in the homogenized constructs and in the adjacent r. abdominis muscle whilst BMP-4 in the homogenized construct only. In untreated 7% HA/CC constructs, BMP-2 was up-regulated in the macroporous construct only. On day 60, 250 μg hTGF-β3/treated and untreated macroporous constructs showed up-regulation of TGF-β1 with a six fold increase vs. TGF-β1 expression in adjacent muscle of untreated constructs. TGF-β2 was down regulated in both untreated and 250 μg hTGF-β3/treated bioreactors. On day 60, 250 μg hTGF-β3/treated bioreactors showed TGF-β3 expression in untreated, treated and adjacent muscle tissues. On day 90, BMP-2 was up

  8. RESEARCH PROGRESS OF BONE MORPHOGENETIC PROTEIN AND LIABILITY OF OSSIFICATION OF POSTERIOR LONGITUDINAL LIGAMENT%BMP与后纵韧带骨化症易患性的研究进展

    Institute of Scientific and Technical Information of China (English)

    方钊; 孙天威; Sandip Kumar Yadav

    2012-01-01

    目的 综述BMP与后纵韧带骨化症(ossification of the posterior longitudinal ligament,OPLL)易患性的研究进展.方法 查阅近年国内外有关BMP与OPLL易患性的相关参考文献并进行总结.结果 BMP基因的单核苷酸多态性(single nueleotide polymorphisms,SNPs)产生微效累加效应增加对OPLL的易感性,多种环境因素可通过增加BMPs基因的表达,促进OPLL的发生、发展.结论 BMPs易感基因的SNPs可促进个体对OPLL的易感性,SNPs的累加效应及与环境因素的相互作用则可影响个体对OPLL易患性的高低.%Objective To review the research progress of bone morphogenetic protein (BMP) and the liability of ossification of the posterior longitudinal ligament (OPLL). Methods Recent literature concerning BMP and the liability of OPLL was reviewed, analysed, and summarized. Results The single nucleotide polymorphisms (SNPs) of BMP gene may produce a minor cumulative effect and increase individual susceptibility to OPLL. A variety of environmental factors can promote the occurrence and development of OPLL by increasing the expression of BMP gene. Conclusion The SNPs of BMP gene may increase individual susceptibility to OPLL. However, interaction of cumulative effect of the SNPs and environmental factors can promote the liability to OPLL.

  9. Trends in Bone Morphogenetic Protein Usage since the U.S. Food and Drug Administration Advisory in 2008: What Happens to Physician Practices When the Food and Drug Administration Issues an Advisory?

    Science.gov (United States)

    Mckie, Janay; Qureshi, Sheeraz; Iatridis, James; Egorova, Natalia; Cho, Samuel; Hecht, Andrew

    2014-06-01

    Study Design Retrospective cross-sectional study of spinal procedures from 2002 to 2010 using the Nationwide Inpatient Sample database. Objective To determine the patterns of bone morphogenetic protein (BMP) usage in fusion surgery before and after the U.S. Food and Drug Administration (FDA) 2008 advisory for the anterior cervical spine to understand how advisories affect U.S. physician practices. Methods Procedures were identified through International Classification of Diseases, Ninth Revision procedure codes and were plotted over time based on fusion procedure type, site, and area of fusion. U.S. national trends were approximated by polynomial regression analysis. Results The majority of the data trends of BMP usage reflect a second-order polynomial model. BMP usage in anterior cervical spine fusion procedures plateaued during the fourth quarter of 2007. The most apparent change in trend was noted in BMP usage pre- and postadvisory in the analysis of anterior cervical spine fusions. BMP percentage of use decreased in this area by 5% from the time of the FDA advisory to the fourth quarter of 2010. Conclusions The decrease in BMP usage in anterior cervical spinal fusion procedures coincided with the timing of the FDA advisory. The fact that BMP continued to be used in cervical spine fusion procedures, even at lower rates, despite the advisory, may reflect the availability of new clinical information that could lessen complications (i.e., lower BMP dose, perioperative steroids, BMP containment). Furthermore, factors like the natural ceiling effect of use or demand for new technology, complications, prohibitive institutional costs, access to information, and insurance compensation may have all contributed to the BMP usage trends observed.

  10. Adenovirus-mediated siRNA targeting TNF-α and overexpression of bone morphogenetic protein-2 promotes early osteoblast differentiation on a cell model of Ti particle-induced inflammatory response in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Guo, H.H.; Yu, C.C.; Sun, S.X. [Affiliated Hospital of Ningxia Medical University, Department of Orthopedic Surgery, Yinchuan (China); Ma, X.J. [Ningxia Medical Autonomous Region of the First People' s Hospital, Department of Orthopedic Surgery, Yinchuan (China); Yang, X.C.; Sun, K.N.; Jin, Q.H. [Affiliated Hospital of Ningxia Medical University, Department of Orthopedic Surgery, Yinchuan (China)

    2013-10-02

    Wear particles are phagocytosed by macrophages and other inflammatory cells, resulting in cellular activation and release of proinflammatory factors, which cause periprosthetic osteolysis and subsequent aseptic loosening, the most common causes of total joint arthroplasty failure. During this pathological process, tumor necrosis factor-alpha (TNF-α) plays an important role in wear-particle-induced osteolysis. In this study, recombination adenovirus (Ad) vectors carrying both target genes [TNF-α small interfering RNA (TNF-α-siRNA) and bone morphogenetic protein 2 (BMP-2)] were synthesized and transfected into RAW264.7 macrophages and pro-osteoblastic MC3T3-E1 cells, respectively. The target gene BMP-2, expressed on pro-osteoblastic MC3T3-E1 cells and silenced by the TNF-α gene on cells, was treated with titanium (Ti) particles that were assessed by real-time PCR and Western blot. We showed that recombinant adenovirus (Ad-siTNFα-BMP-2) can induce osteoblast differentiation when treated with conditioned medium (CM) containing RAW264.7 macrophages challenged with a combination of Ti particles and Ad-siTNFα-BMP-2 (Ti-ad CM) assessed by alkaline phosphatase activity. The receptor activator of nuclear factor-κB ligand was downregulated in pro-osteoblastic MC3T3-E1 cells treated with Ti-ad CM in comparison with conditioned medium of RAW264.7 macrophages challenged with Ti particles (Ti CM). We suggest that Ad-siTNFα-BMP-2 induced osteoblast differentiation and inhibited osteoclastogenesis on a cell model of a Ti particle-induced inflammatory response, which may provide a novel approach for the treatment of periprosthetic osteolysis.

  11. Adenovirus-mediated siRNA targeting TNF-α and overexpression of bone morphogenetic protein-2 promotes early osteoblast differentiation on a cell model of Ti particle-induced inflammatory response in vitro.

    Science.gov (United States)

    Guo, H H; Yu, C C; Sun, S X; Ma, X J; Yang, X C; Sun, K N; Jin, Q H

    2013-10-01

    Wear particles are phagocytosed by macrophages and other inflammatory cells, resulting in cellular activation and release of proinflammatory factors, which cause periprosthetic osteolysis and subsequent aseptic loosening, the most common causes of total joint arthroplasty failure. During this pathological process, tumor necrosis factor-alpha (TNF-α) plays an important role in wear-particle-induced osteolysis. In this study, recombination adenovirus (Ad) vectors carrying both target genes [TNF-α small interfering RNA (TNF-α-siRNA) and bone morphogenetic protein 2 (BMP-2)] were synthesized and transfected into RAW264.7 macrophages and pro-osteoblastic MC3T3-E1 cells, respectively. The target gene BMP-2, expressed on pro-osteoblastic MC3T3-E1 cells and silenced by the TNF-α gene on cells, was treated with titanium (Ti) particles that were assessed by real-time PCR and Western blot. We showed that recombinant adenovirus (Ad-siTNFα-BMP-2) can induce osteoblast differentiation when treated with conditioned medium (CM) containing RAW264.7 macrophages challenged with a combination of Ti particles and Ad-siTNFα-BMP-2 (Ti-ad CM) assessed by alkaline phosphatase activity. The receptor activator of nuclear factor-κB ligand was downregulated in pro-osteoblastic MC3T3-E1 cells treated with Ti-ad CM in comparison with conditioned medium of RAW264.7 macrophages challenged with Ti particles (Ti CM). We suggest that Ad-siTNFα-BMP-2 induced osteoblast differentiation and inhibited osteoclastogenesis on a cell model of a Ti particle-induced inflammatory response, which may provide a novel approach for the treatment of periprosthetic osteolysis.

  12. Association between two polymorphisms of the bone morpho-genetic protein-2 gene with genetic susceptibility to ossification of the posterior longitudinal ligament of the cervical spine and its severity

    Institute of Scientific and Technical Information of China (English)

    WANG Hao; YANG Zhao-hui; LIU Dong-mei; WANG Ling; MENG Xiang-long; TIAN Bao-peng

    2008-01-01

    Background Ossification of the posterior longitudinal ligament (OPLL) has a strong genetic background. Previous studies have shown that bone morphogenetic protein-2 (BMP2) and BMP2 mRNA are expressed in ossifying matrix and chondrocytes adjacent to cartilaginous areas of OPLL tissues and mesenchymal cells with fibroblastic features in the immediate vicinity of the cartilaginous areas. It is suggested that BMP2 plays different roles in the different stages of development of OPLL. However, it remains unknown which factors induce ligament cells to produce BMP2.Methods OPLL patients (n=192) and non-OPLL controls (n=304) were studied. Radiographs of the cervical spine were analyzed for extent of OPLL. We investigated whether single nucleotide polymorphisms of exons 3(-726) TIC and 3(-583) A/G in the BMP2 gene are statistically associated with genetic susceptibility to OPLL in Chinese Han subjects.Results There was no statistical difference between the occurrence of exons 3(-726) TIC and 3(-583) A/G and the occurrence of OPLL in the cervical spine. However, there was a significant association between occurrence of exon 3(-726) TIC polymorphism and occurrence of OPLL in males of cases and controls in the cervical spine. In addition, no significant association was found between the exons 3(-726) TIC and 3(-583) A/G with number of ossified cervical vertebrae in OPLL patients.Conclusions Exon 3(-583) A/G polymorphism in BMP2 gene is not associated with the occurrence and the extent of OPLL in the cervical spine. Chinese Han male patients with TC and CC genotypes in exon 3(-726) T/C have genetic susceptibility to OPLL but not to more extensive OPLL in the cervical spine.

  13. Advance on Relationship between Obesity and Bone Morphogenetic Protein-7%骨形态发生蛋白7与肥胖关系的研究进展

    Institute of Scientific and Technical Information of China (English)

    汪小娟

    2011-01-01

    Obesity is a common risk factor for many metabolic diseases,such as coronary heart disease, hypertension, type 2 diabetes and hyperlipemia. Obesity is also closely related to insulin resistance and metabolic syndrome. Bone morphogenetic protein-7( BMP-7 )is a kind of obesity-related active factor recently discovered. It can specifically activate brown fat-related regulatory factor,by promoting brown fat cell differentiation to control the body energy balance. Therefore,understandings about the role of BMP-7 in the physiological and pathological processes of obesity help to reveal the molecular mechanisms of the above multi-gene disease.%肥胖是冠心病、高血压、2型糖尿病、高脂血症等许多代谢性疾病的共同危险因子,与胰岛素抵抗、代谢综合征密切相关.骨形态发生蛋白7(BMP-7)是最近发现的与肥胖相关的一种活性因子,可以特异性地激活棕色脂肪调控因子的表达,通过促进棕色脂肪细胞的分化来调控体内能量平衡.因此,对BMP-7的深入研究,了解其在肥胖的生理和病理过程中的作用,有助于揭示上述多基因疾病的分子生物学机制.

  14. 重组人骨形成蛋白-4的大规模制备%The Large Scale Preparation of Recombinant Human Bone Morphogenetic Protein-4 Mature Peptide

    Institute of Scientific and Technical Information of China (English)

    王涛; 陈苏民; 陈南春; 赵伟钦; 张晓楠

    2006-01-01

    含有能够高表达重组人骨形成蛋白成熟肽(recombinant human bone morphogenetic protein 4 mature peptide,rhBMP-4m)质粒的工程菌株,导入15 L NBS发酵罐中进行恒溶氧高密度发酵和诱导表达,测定发酵菌液OD600值为30.8.离心收集菌体,PAGE电泳后光密度扫描表明hBMP-4m占细菌总蛋白量的42%.悬浮所收集的菌体,裂菌,洗涤4次后的包涵体中hBMP-4m占蛋白量的83.2%.预先取少量样品进行探索实验后,全部包涵体用8 mol尿素缓冲液溶解,上SP-Sepharose FF阳离子柱,以0.35mol NaCl洗脱,获得纯度为96%的hBMP-4m.其收获量为1.34 g/L发酵液;收得率为36.4%.结果表明:使用这套工艺流程大规模制备hBMP-4m,能够得到较好的收得率、较高的收获量和纯度.

  15. In vivo bacterial morphogenetic protein interactions

    OpenAIRE

    van der Ploeg, R.; den Blaauwen, T.; Meghea, A.

    2012-01-01

    This chapter will discuss none-invasive techniques that are widely used to study protein-protein interactions. As an example, their application in exploring interactions between proteins involved in bacterial cell division will be evaluated. First, bacterial morphology and cell division of the rod-shaped bacterium Escherichia coli will be introduced. Next, three bacterial two-hybrid methods and three Förster resonance energy transfer detection methods that are frequently applied to detect int...

  16. Lumbar interbody fusion with porous biphasic calcium phosphate enhanced by recombinant bone morphogenetic protein-2/silk fibroin sustained-released microsphere: an experimental study on sheep model.

    Science.gov (United States)

    Chen, Liang; Liu, Hai-Long; Gu, Yong; Feng, Yu; Yang, Hui-Lin

    2015-03-01

    Biphasic calcium phosphate (BCP) has been investigated extensively as a bone substitute nowadays. However, the bone formation capacity of BCP is limited owing to lack of osteoinduction. Silk fibroin (SF) has a structure similar to type I collagen, and could be developed to a microsphere for the sustained-release of rhBMP-2. In our previous report, bioactivity of BCP could be enhanced by rhBMP-2/SF microsphere (containing 0.5 µg rhBMP-2) in vitro. However, the bone regeneration performance of the composite in vivo was not investigated. Thus, the purpose of this study was to evaluate the efficacy of BCP/rhBMP-2/SF in a sheep lumbar fusion model. A BCP and rhBMP-2/SF microsphere was developed, and then was integrated into a BCP/rhBMP-2/SF composite. BCP, BCP/rhBMP-2 and BCP/rhBMP-2/SF were implanted randomly into the disc spaces of 30 sheep at the levels of L1/2, L3/4 and L5/6. After sacrificed, the fusion segments were evaluated by manual palpation, CT scan, biomechanical testing and histology at 3 and 6 months, respectively. The composite demonstrated a burst-release of rhBMP-2 (39.1 ± 2.8 %) on the initial 4 days and a sustained-release (accumulative 81.3 ± 4.9 %) for more than 28 days. The fusion rates, semi-quantitative CT scores, fusion stiffness in bending in all directions and histologic scores of BCP/rhBMP-2/SF were significantly greater than BCP and BCP/rhBMP-2 at each time point, respectively (P sheep using BCP constructs.

  17. Changes of the intensity of morphogenetic process in the bone skeleton under lowering of gravitational loading

    Science.gov (United States)

    Vasilievna Rodionova, Natalia; Zolotova-Haidamaka, Nadezhda

    The development of long skeleton bones and reconstruction of bone structures in ontogenesis during adaptive remodeling are performed due to a combination of the bone apposition and bone resorption processes. With the use of radioactive markers of specific biosyntheses -3H- thymidine and 3H-glycine we studied the dynamics and peculiarities of these processes under modeling microgravity conditions by unloading the hind limbs of young white rats (tail suspension method) during 28 days. The radionuclides were administered in a single dose at the end of the experiment and the biomaterial was taken 1, 24, 48, 120 and 192 h. after injection. In histoautographs the counts were made of a nuclei labeling index (3H-thymidine), of the number of silver grains over the cells and in the forming bone matrix in growth and remodeling zones of the femoral bone (3H-glycine). The tendency for a reduction of a labeling index in the 3H-thymidine-labeled osteogenic cells in the periost and endost has been established. The dynamics of labeled cells following various intervals after 3H-thymidine injection testifies to a delay in the rates of osteoblasts' differentiation and their transformation to osteocytes in the experiment animals. 3H-glycine is assimilated by osteogenic cells 30 min after the radionuclide injection and following 24 h. it is already incorporated into the forming bone matrix. As a result an appositional bone addition by 192 h. the silver grains are registered in the bone matrix as "labeling lines". A lower 3H-glycine uptake by the osteogenic cells and bone matrix as compared with a control is indicative of a decrease of the osteoplastic process under hypokinesia, particulary in the periost. At the same time the resorption and remodeling bone zones reveal regions of an intensive 3H-glycine uptake after 1 and 24 h. We associate this latter fact with an activation of collagen proteins in the differentiating fibroblasts (instead of osteoblasts) in these locations. This is

  18. Autoradiographic studies of the intensity of morphogenetic processes in the bone skeleton under modeling microgravity

    Science.gov (United States)

    Rodionova, N. V.; Zolotova-Haidamaka, N. V.; Nithevich, T. P.

    In ontogenesis the development of long skeleton bones and reconstruction of bone structures during adaptive remodeling are performed due to a combination of the bone apposition and bone resorption processes. With the use of radioactive markers of specific biosyntheses -3H-thymidine and 3H-glycine we studied the dynamics and peculiarities of these processes under hypokinesia by unloading the hind limbs of young white rats (tail suspension method) during 28 days. The radionuclides were administered in a single dose at the end of the experiment and the biomaterial was taken 1, 24, 48, 120 and 192 h. after injection. In histoautographs the counts were made of a nuclei labeling index (3H-thymidine), of the number of silver grains over the cells and in the forming bone matrix in growth and remodeling zones of the femoral bone (3H-glycine). The tendency for a reduction of a labeling index in the 3H-thymidine-labeled osteogenic cells in the periost and endost has been established. The dynamics of labeled cells following various intervals after 3H-thymidine injection testifies to a delay in the rates of osteoblasts' differentiation and their transformation to osteocytes in the experiment animals. 3H-glycine is assimilated by osteogenic cells 30 min after the radionuclide injection and following 24 h. it is already incorporated into the forming bone matrix. As a result an appositional bone addition by 192 h. the silver grains are registered in the bone matrix as "labeling lines". A lower 3H-glycine uptake by the osteogenic cells and bone matrix as compared with a control is indicative of a decrease of the osteoplastic process under hypokinesia, particulary in the periost. At the same time the resorption and remodeling bone zones reveal regions of an intensive 3H-glycine uptake after 1 and 24 h. We associate this latter fact with an activation of collagen proteins in the differentiating fibroblasts (instead of osteoblasts) in these locations. This is confirmed by our previous

  19. 钛网结合异体骨移植联合人骨形成蛋白与自体骨移植在种植前牙槽嵴骨增量中的应用对比%Comparison of Application of Titanium Mesh Combined with Allograft Combined with Human Bone Morphogenetic Protein and Autogenous Bone Transplantation in Alveolar Bone Augmentation

    Institute of Scientific and Technical Information of China (English)

    崔延军; 王红光; 程汇

    2015-01-01

    Objective To compare the application of titanium mesh combined with allograft combined with human bone morphogenetic protein and autogenous bone transplantation in alveolar bone augmentation.Methods A total of 46 cases receiving anterior implant plus alveolar bone increment admitted to Tianmen City First People′s Hospital from Jan.2008 to Dec.2011 were randomly divided into two groups according to random-number table method,group A(n =23) using titanium mesh combined with allograft combined with human bone morphogenetic protein,while group B (n =23) using autogenous bone graft.The alveolar ridge width,the incremental effects and postoperative bone implant stability were compared before and after surgery.Results Postoperative average alveolar ridge width of group A was significantly higher than that of group B [(6.9 ±0.5) mm vs (5.6 ±0.3) mm,P <0.01]; the cure rate of group A was 94.1%,of group B was 60.0%,and the clinical efficacy of group A was better than group B with statistically significant difference (P <0.05); no-loosening rate after 4 months implant was 91.2% in group A,50.6% in group B,the sta-bility of group A was much better than group B,with statistically significant difference(P <0.05).Conclu-sion Titanium mesh combined with allogeneic bone graft combined with human bone morphogenetic protein can significantly increase patient alveolar bone mass and help to stabilize the implant after surgery .%目的:比较钛网结合异体骨移植联合人骨形成蛋白与自体骨移植在种植前牙槽嵴骨增量中的应用效果。方法将2008年1月至2011年12月天门市第一人民医院收治的46例前牙种植并行牙槽嵴骨增量的患者依据随机数字表法分为两组:A 组23例,采用钛网结合异体骨移植联合人骨形成蛋白的方法;B 组23例,采用自体骨移植的方法。比较两组患者手术前后牙槽嵴宽度、骨增量效果及术后种植体稳定性。结果术后 A 组牙槽嵴

  20. Improved Osteointegration of Ti-6Al-4V-implants of Different Surface Texture by the Use of Bone Morphogenetic Protein-3 (BMP-3)

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Half of altogether 60 cylindrical implant devices mode of titanium-aluminum-vanadium alloy( Ti-6Al-4 V) were plasma-sprayed with a hydroxyapatite-coating and the other half had a corundum blasted porous surface. 15 implants of each group of the titanium test implants were coated with 230μgporcine, high-purified BMP- 3-precipitate per implant. In each case a BMP- 3-coated and an uncoated control-device were implanted into the femoral part of the patellofemoral joint of the right and left leg of 30 adult giant rabbits. Histomorphological and histomorphometrical we found in both groups with BMP- 3-coated test devices an improved osteointegration. Statistical evaluation using the t-test for matched samples showed 5 weeks after surgery a significant higher volume of new formed bone of the BMP- 3-coated corundum-blasted or hydroxyapatite-coated Ti-6Al-4 V test devices compared to the non-coated controls of the same type (p < 0.01, t-test for matched samples). In both implatt groups with BMP-coating a synergetic effect was verifiable although the bone ongrowth in the hydroxyapatite coated implants was more extensive than in the corundum blasted implants. Light microscopy demonstrated osteointegration without connective tissue membrane around the surface of the implants. Our results indicate that composite metal implants, as used in endoprosthetics and implantology , are suitable carriers for BMP- 3 and improved fixation of the implants can be achieved. The hydroxyapatite surface is superior to the corundum-blasted surface with regards to the observed parameters because of its pronounced bioactivity and its osteoconductive characteristics.

  1. Induction of chondro-, osteo- and adipogenesis in embryonic stem cells by bone morphogenetic protein-2: Effect of cofactors on differentiating lineages

    Directory of Open Access Journals (Sweden)

    zur Nieden Nicole I

    2005-01-01

    Full Text Available Abstract Background Recently, tissue engineering has merged with stem cell technology with interest to develop new sources of transplantable material for injury or disease treatment. Eminently interesting, are bone and joint injuries/disorders because of the low self-regenerating capacity of the matrix secreting cells, particularly chondrocytes. ES cells have the unlimited capacity to self-renew and maintain their pluripotency in culture. Upon induction of various signals they will then differentiate into distinctive cell types such as neurons, cardiomyocytes and osteoblasts. Results We present here that BMP-2 can drive ES cells to the cartilage, osteoblast or adipogenic fate depending on supplementary co-factors. TGFβ1, insulin and ascorbic acid were identified as signals that together with BMP-2 induce a chondrocytic phenotype that is characterized by increased expression of cartilage marker genes in a timely co-ordinated fashion. Expression of collagen type IIB and aggrecan, indicative of a fully mature state, continuously ascend until reaching a peak at day 32 of culture to approximately 80-fold over control values. Sox9 and scleraxis, cartilage specific transcription factors, are highly expressed at very early stages and show decreased expression over the time course of EB differentiation. Some smaller proteoglycans, such as decorin and biglycan, are expressed at earlier stages. Overall, proteoglycan biosynthesis is up-regulated 7-fold in response to the supplements added. BMP-2 induced chondrocytes undergo hypertrophy and begin to alter their expression profile towards osteoblasts. Supplying mineralization factors such as β-glycerophosphate and vitamin D3 with the culture medium can facilitate this process. Moreover, gene expression studies show that adipocytes can also differentiate from BMP-2 treated ES cells. Conclusions Ultimately, we have found that ES cells can be successfully triggered to differentiate into chondrocyte-like cells

  2. Bases teóricas y aplicación clínica de las proteínas morfogenéticas óseas en cirugía maxilofacial Base theories and the clinical application of bone morphogenetic proteins in maxillofacial surgery

    Directory of Open Access Journals (Sweden)

    C.M. Ardila Medina

    2009-06-01

    Full Text Available Uno de los grandes avances en la neoformación ósea ha sido la identificación de factores de crecimiento importantes para ella como son las proteinas morfogenéticas óseas (PMO que regulan la diferenciación ósea y cartilaginosa in vivo. La depuración, clonación genética y expresión de las PMO han establecido las bases para el análisis celular y molecular del desarrollo y la regeneración ósea. El estudio genético de las PMO señala que son esenciales para la función normal animal y en la osteogénesis postfetal es importante en el desarrollo embrionario orgánico, esquelético y de los tejidos dentales y craneofaciales. La disponibilidad de las PMO proporciona retos y oportunidades para mejorar los conocimientos que regulan la regeneración ósea con el fin de optimizar los resultados en el paciente.One of the fundamental advances in bone neoformation has been the identification of important growth factors like the bone morphogenetic proteins that regulate live cartilage and bone differentiation. The cleansing, genetic cloning and expression of recombinant human bone morphogenetic proteins (BMP have laid the basis for cellular and molecular analysis of bone development and regeneration. The genetic study of the BMPs indicates that they are essential to the normal development and function of animals. BMP post-natal bone development is also very important in embryonic organic, skeletal, craniofacial and dental tissue development. The availability of BMPs provides several challenges and opportunities to improve insights into the mechanisms that regulate the regeneration of bone for optimal outcome in the patient.

  3. Sustained Release of Bone Morphogenetic Protein 2 via Coacervate improves Muscle Derived Stem Cell Mediated Cartilage Regeneration in MIA-induced Osteoarthritis

    Science.gov (United States)

    Hicks, Justin James; Rocha, Jorge Luis; Li, Hongshuai; Huard, Johnny; Wang, Yadong; Hogan, MaCalus Vinson

    2016-01-01

    Objectives: Individuals who participate in sports have an increased risk of osteoarthritis (OA), characterized by articular cartilage degeneration. Currently, there is no cure for OA with treatment aimed at symptom relief and improved function. Muscle-derived stem cells (MDSCs) have been shown to exhibit long-term proliferation, high self-renewal, and multipotent differentiation capabilities in vitro. Previously, we have demonstrated that murine MDSCs retrovirally transduced to express chondrogenic proteins (BMPs) differentiate into chondrocytes and enhance cartilage repair in vivo. Direct injection of therapeutic proteins can promote cartilage healing; however, they have relatively short half-lives requiring muitiple injections of high dosages. This presents a challenge in terms of maintaining adequate local BMP levels and could negatively affect both injured and normal structures and lead to side effects such as osteophyte formation. Gene therapy is a promising approach that addresses this problem; however, its utilization in clinical applications is much further down the road. In order to circumvent viral transduction of cells for cartilage regeneration, we developed a unique growth factor delivery platform comprised of native heparin and a synthetic polycation, poly(ethylene argininylaspartate diglyceride) (PEAD) incorporated with BMP2 (BMP2 coacervate). In this study, we show that sustained delivery of BMP2 via a BMP2 coacervate can induce the differentiation of MDSCs to a chondrocyte lineage for in vivo cartilage regeneration and healing in a Monoiodoacetate (MIA)-induced osteoarthritis model. Methods: mMDSCs were isolated from muscle biopsies via a modified pre-plated technique. The BMP2 coacervates were prepared as previously described. The release profiles of BMP2 coacervate were tested by ELISA. The chondrogenic effects that delivery of BMP2 had on MDSCs were evaluated by RT-PCR. The efficacy of MDSC with BMP2 coacervate were evaluated in vivo in a MIA

  4. 自体髂骨与胶原-羟基磷灰石复合重组人骨形成蛋白2修复大鼠单侧腭裂%Autogenous iliac boneversuscollagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite material for repair of unilateral cleft palate in rats

    Institute of Scientific and Technical Information of China (English)

    沈悦; 马海英; 张彦升; 王娟; 时炳正

    2015-01-01

      结果与结论:随时间的推移,碱性磷酸酶活性逐渐升高,抗酒石酸酸性磷酸酶活性逐渐降低,实验组碱性磷酸酶活性始终高于对照组(P OBJECTIVE:To detect the healing effect of autologous bone graft versus colagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite graft in a rat model of unilateral complete cleft palate. METHODS: Firstly, we established the artificial unilateral complete cleft palate models in 32 Sprague-Dawley rats, and then the established animal models were randomly divided into control group and experimental group. The autologous iliac bone was transplanted into the fissures of control group, and the experimental group received colagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite graft. After that, the activities of serum alkaline phosphatase and tartrate-resistant acid phosphatase, bone mineral densities in the neonatal palate, expressions of osteocalcin, osteoprotegerin, core binding factor, and osteoclast differentiation factor were detected at 1, 2, 3, 4 weeks after treatment. RESULTS AND CONCLUSION: Over time, the alkaline phosphatase activity increased gradualy, while tartrate-resistant acid phosphatase activity decreased. Compared with the control group, the alkaline phosphatase activity was always higher (P < 0.05,P < 0.01) but the tartrate-resistant acid phosphatase activity was lower in the experimental group (P < 0.05,P < 0.01); the bone mineral density increased in both groups, but it was always higher in the experimental group than the control group (P < 0.01). The expression levels of osteocalcin, osteoprotegerin, and core-binding factor gene gradualy rose in both groups, but they were always higher in the experimental group than the control group; in contrast, the expression of osteoclast differentiation factor was decreased in both groups, and it was lower in the experimental group than the control group. These findings indicate that

  5. Improvement of ovarian response and oocyte quality of aged female by administration of bone morphogenetic protein-6 in a mouse model

    Directory of Open Access Journals (Sweden)

    Park Seung S

    2012-12-01

    Full Text Available Abstract Background Advancing female age remains a difficult problem in infertility treatment. Ovarian angiogenesis plays an important role in follicular development and the activation of ovarian angiogenesis has been emerged as a new strategy for the improvement of age-related decline of oocyte quality. BMP-6 affect gonadotropin signals in granulosa cells and it promotes normal fertility by enabling appropriate response to LH and normal oocyte quality. BMP-6 has a potential role in regulation of angiogenesis and regulates the expression of inhibitor of DNA-binding proteins (Ids. Ids involved in the control and timing of follicle selection and granulosa cells differentiation. Especially, Id-1 is well-characterized target of BMP-6 signaling. Therefore, this study investigated whether co-administration of BMP-6 during superovulation process improves ovarian response, oocyte quality and expression of Id-1 and vascular endothelial growth factor (VEGF in the ovary of aged female using a mouse model. Methods Aged C57BL/6 female mice (26–31 weeks old were superovulated by injection with 0.1 mL of 5 IU equine chorionic gonadotropin (eCG containing recombinant mouse BMP-6 at various doses (0, 0.01, 0.1, 1, and 10 ng, followed by injection with 5 IU human chorionic gonadotropin (hCG 48 h later. Then, the mice were immediately paired with an individual male. The aged control group was superovulated without BMP-6. Young mice of 6–9 weeks old were superovulated without BMP-6 as a positive control for superovulation and in vitro culture of embryos. Eighteen hours after hCG injection, zygotes were retrieved and cultured for 4 days. Both ovaries of each mouse were provided in the examination of ovarian expression of Id-1 and VEGF by reverse transcriptase-polymerase chain reaction, western blot, and immunohistochemistry. Results Administration of 0.1 ng BMP-6 significantly increased the number and blastocyst formation rate of oocytes ovulated and ovarian

  6. A single nucleotide polymorphism in the human bone morphogenetic protein-2 gene (109T>G) affects the Smad signaling pathway and the predisposition to ossification of the posterior longitudinal ligament of the spine

    Institute of Scientific and Technical Information of China (English)

    YAN Liang; CHANG Zhen; LIU Yang; LI Yi-bing; HE Bao-rong; HAO Ding-jun

    2013-01-01

    Background Although various systemic and local factors such as abnormal carbohydrate or calcium metabolism,aging,and hormonal disturbances have been suggested as causes of ossification of the posterior longitudinal ligament (OPLL),the etiology of OPLL is not fully understood.The purpose of this study was to investigate whether bone morphogenetic protein (BMP)-2 is a candidate gene to modify the susceptibility of OPLL and the mechanism of signal transduction in ossification.Methods A total of 420 OPLL patients and 506 age-and sex-matched controls were studied.The complete coding sequence of the human BMP-2 gene was analyzed using polymerase chain reaction (PCR) and direct sequencing.All single nucleotide polymorphisms (SNPs) were detected and genotyped.BMP-2 expression vectors containing positive polymorphisms were constructed and transfected into the C3H10T1/2 cells.The expression of BMP-2 and the Smad signal pathway in positive cell clones were detected by Western blotting.The alkaline phosphatase (ALP) activity was determined using quantitative detection kits.Results The frequencies for the 109T>G and 570A>T polymorphisms were different between the case and control groups.The "TG" genotype in 109T>G polymorphism is associated with the occurrence of OPLL,the frequency of the "G"allele is significantly higher in patients with OPLL than in control subjects (P <0.001).The "AT" genotype in 570A>T polymorphism is associated with the occurrence of OPLL,the frequency of the "T" allele is significantly higher in patients with OPLL than in control subjects (P=0.005).Western blotting analysis revealed that the expression of P-Smad1/5/8protein transfected by wild-type or mutant expression vectors were significantly higher than control groups (P <0.05),but there was no statistical difference in each experimental group (P >0.05).The expression of Smad4 protein transfected by wild-type or mutant expression vectors was significantly higher than control groups (P

  7. 原核表达的重组人骨形态发生蛋白7增高牙槽嵴的实验%Experimental study on recombinant human bone morphogenetic protein-7 expressed in prokaryocyte augmenting alveolar ridge

    Institute of Scientific and Technical Information of China (English)

    肖水清; 卞翠荣

    2006-01-01

    BACKGROUND: After loss of teeth, the dynamic equilibrium between osteoblast and osteoclast in alveolar bone is destroyed because of systematic and local factors, and residual ridge resorption and atrophy occur irreversibly, which result in the loss of massive alveolar bone. Bone morphogenetic protein-7 (BMP-7) exerts an important effect in the development and traumatic repair of bone and tooth.OBJECTIVE: To investigate the effect of human recombinant bone morphogenetic protein-7 (rhBMP-7) on new bone formation of alveolar ridge and absorption of alveolar ridge.DESIGN: Control experiment.SETTING: Department of Orthodontics, Jinan Stomatological Hospital;Shangdong Academy of Medical Sciences.MATERIALS: This study was conducted at the Shangdong Academy of Medical Sciences from June 2003 to December 2004. Totally 28 New Zealand white rabbits, of either gender, weighing 2 kg, were used in this study.METHODS: Animal extracted wound models were created on the rabbits.The mandibular left and right central incisor of the rabbits were removed.Two carriers containing BMP7 complex (40 μg/each) were implanted into the mandibular right central incisor (experimental group), two empty carriers containing only phosphate buffer solution(PBS) (40 μg/each) were implanted into the mandibular left central incisor (control group). The animals were sacrificed at postoperative 2, 4, 8 and 12 weeks. The specimen from the operated regions were harvested and observed by scanning electron microscope, and calcium content and alkaline phosphatase (ALP) activity were also measured.MAIN OUTCOME MEASURES: ① Result of scanning electron microscope (SEM). ② ALP activity and calcium content.RESULTS: ① The bone wound healing was 4-6 weeks earlier in the experimental group than in the control group . ② The ALP activity was significantly higher in the experimental group than in the control group [week 2:(38.191±5.384, (19.821±2.084) μkat/g;week 4: (160.815±9.669), (126.709±1.634)

  8. 骨缺损再生修复中人骨形成蛋白2基因的克隆和序列分析%Cloning and sequencing of human bone morphogenetic protein 2 gene in the regeneration and repair of bone defects

    Institute of Scientific and Technical Information of China (English)

    王德利; 阮狄克; 马巍; 张惠中; 范清宇

    2004-01-01

    目的:克隆人骨形成蛋白 2 (hBMP2)基因全长,用于临床难治性骨折和骨缺损的再生修复.方法:以成骨肉瘤细胞总 RNA为模板,应用反转录-聚合酶链反应法(RT-PCR)克隆 hBMP2的 cDNA全长;将获得的基因插入 pGEM-T-Easy载体质粒,并转化至大肠杆菌后挑选阳性克隆,利用限制性内切酶酶切分析鉴定重组质粒.结果:通过质粒酶切分析和序列测定,获得的基因为 hBMP2全长 DNA序列.结论:克隆获得 hBMP2的基因,为其进一步开发利用提供了前提条件.%AIM:To clone and sequence the bone morphogenetic protein 2 gene(hBMP2) for its clinical application in the regeneration and repair of failed bone fracture and defects. METHODS:The human genomic RNA was extracted from the human osteosarcome cells as templates,and the gene of bone morphogenetic protein 2(BMP2) was cloned by using reverse transcription-polymerase chain reaction(RT-PCR) and T-vector cloning method.The positive clone was screened and identified by the restriction enzymes, and then the cloned amplified fragment was sequenced and analyzed. RESULTS:DNA sequence comparison for the cloned gene of BMP2 with the GenBank(M22489) sequence demonstrated that both sequences were identical, 1190bp length. CONCLUSION:Cloning the BMP2 gene from the human gene has paved the way for further study on gene therapy of bone fracture.

  9. Preparation and performance of recombinant human bone morphogenetic protein-2-poly(hydroxybutyrate-co-hydroxyoctanoate) nanospheres%聚羟基丁酸-羟基辛酸共聚酯载生长因子的缓释纳米微球

    Institute of Scientific and Technical Information of China (English)

    王晓东; 张永红

    2014-01-01

    BACKGROUND:Bone morphogenetic protein-2 can increase the production of chondrocytes and progenitor cel matrix, enhance tissue inhibitor of metaloproteinase-1, sox9, type II colagen and aggrecan expressions, with the induction of mesenchymal cel migration, proliferation, and differentiation, leading to cartilage and bone formation. OBJECTIVE:Using the biodegradable copolymer of poly(hydroxybutyrate-co-hydroxyoctanoate)-based materials to fabricate recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres and to investigate its morphology, particle distribution, drug loading, encapsulation efficiency, in vitrorelease time, and bioactivity. METHODS:The recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres were prepared by multiple emulsion volatilizing method. Porcine chondrocytes were isolated and culturedin vitro. There were three groups: group 1, no drugs were added into the medium serving as control group; group 2, 20 μg/L recombinant human bone morphogenetic-2 was added; group 3, 20 μg/L recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres were added. The effectiveconcentration was 55 μg/L for recombinant human bone morphogenetic-2 in group 2, and 100 μg/L for recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres in group 3. Proliferative capacity of chondrocytes was detected using 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay.In vitrorelease profile and biological activity were observed under simulated in vivo conditions. RESULTS AND CONCLUSION: The surface of nanospheres was smooth and rounded, and the nanosheres had uniform size and particle size of 231-415 nm. Under scanning electron microscope, the average particle size was 323 nm. Microsphere encapsulation efficiency and drug loading were (79.63±0.16)% and (1

  10. Nano-hydroxyapatite/collagen composited with recombinant human bone morphogenetic protein-2 and titanium membrane in repairing peripheral bone defects of instant dental implants%胶原基纳米骨复合重组人骨形成蛋白2及钛膜修复即刻钛种植体周围骨缺损

    Institute of Scientific and Technical Information of China (English)

    刘冰; 陈鹏; 王忠义; 柯杰; 李晓华; 汪正文

    2009-01-01

    BACKGROUND:Recently,with the rapid development of material science and bioscience,the technology of dental implant has made great progress,especially the immediate implant technology.But the size and shape of implant are usually not fit for tooth extraction wound,so it is an important factor that leads to failure when implant and tooth extraction wound can not form close tangency.Guided bone regeneration or bone grafting materials are usually used to solve this problem.OBJECTIVE:To study the effects of nano-hydroxyapatite/collogen (nHAC) with recombinant human bone morphogenetic protein-2(rhBMP-2) and titanium (Ti) membrane on repairing peripheral bone defects of instant implant.DESIGN,TIME AND SETTING:A randomized,controlled animal study was performed at the Central Laboratory,the Fourth Military Medical University of Chinese PLA between January 2005 and January 2006.MATERIALS:Ti screw implants (diameter 2 mm,length 10 mm,and pitch 0.4 mm) without the part that went through gum were offered by Nonferrous Metal Academy in Baoji,China.The nonabsorbable Ti membranes (2 cm×2 cm) were offered by Zhongbang Biomaterial Limited Company in Xi'an,China.The nHAC materials were gifted by professor Cui Fu-zhai from Material Science and Engineering Department of Tsinghua University and fabricated into 0.5 mm×0.5 mm×0.5 mm small blocks.rhBMP-2 was offered by the Academy of Military Medical Sciences in Beijing,China.rhBMP-2 was dissolved with hydrochloric carbamidine and then nHAC was immersed in it.Vacuumization,freeze-drying,and Ekibon degermation were followed.Each gram of nHAC compounds required approximately 1 mg rhBMP-2.METHODS:Four healthy purebred male dogs were included in this study.According to the methods to repair bone defects rhBMP-2+Ti membrane,nHAC composited with rhBMP-2 was implanted,covering Ti membrane.Six defects were made on the mandible on each side.MAIN OUTCOME MEASURES:At 6 and 12 weeks after implantation,new bone formation and the correlation of new

  11. 骨形态发生蛋白-2与碱性成纤维细胞生长因子在异位和原位成骨中的作用%Response of bone morphogenetic protein-2 and basic fibroblast growth factor in bone marrow stromal cells in ectopic and in situ bone formation

    Institute of Scientific and Technical Information of China (English)

    王磊; 章燕; 游素兰; 谭鸾君; 黄远亮

    2012-01-01

    Objective We ascertained the effect of bone morphogenetic protein-2 (BMP-2) and basic fibroblast growth factor (bFGF) by a series of experiments: Proliferation and differentiation of bone marrow stromal cells (BMSCs) in vitro, ectopic and in situ bone formation and loaded porous calcium phosphate cement (CPC) on the repair of bone defects around dental implants. Methods BMSCs from Beagle dogs were cultured in vitro with basic culture medium containing BMP-2, bFGF, and BMP-2+bFGF. Proliferation and differentiation of BMSCs were quantified using methyl thiazolyl tetrazolium (MTT) and alkaline phosphatase GVLP) test. The CPC seeded with BMSCs and BMP-2, bFGF, com-bined BMP-2 with bFGF were implanted subcutaneously into nude rats in ectopic bone formation, and were implanted into critical-sized bone defects of Beagle dogs in the in situ bone formation. The hone formation was detected by his-tology examination and quantified using an image analysis system. Polychrome sequential fluorescent labels and fluores-cence histological examinations of undecalcified sections were performed post-operatively. Results It was determined that BMP-2+bFGF promoted BMSCs statistically significant proliferation and differentiation compared to either BMP-2 or bFGF in vitro. The CPC with BMP-2+bFGF group yielded more bone than those with either BMP-2 or bFGF in ectopic bone formation test. The percentages of newly ectopic formed bone were higher in the BMP-2+bFGF group(48/79%±1131%) than those in other groups (BMP-2 group, 30.71%±10.85%; bFGF group, 27.33%±9.67%; and the control group, 10.65%±6.05%). Undecalcified sections showed that new bone was actively formed in the BMP-2+bFGF group after 12 weeks in the in situ bone formation test. The bone mineralization apposition ate (MAR) was better in the BMP-2+bFGF group than in other groups (P<0.01). Conclusion BMP-2 combined with bFGF are more effective than one alone in promoting the formation of new bone.%目的 通过骨髓基质细

  12. 胶原蛋白膜包裹藻酸钙凝胶/骨髓基质细胞/BMP-2复合体修复兔桡骨缺损%Collagen membrane wraps the complex of calcium alginate gelatin, bone marrow stromal cells and bone morphogenetic protein to repair radius defect of rabbits

    Institute of Scientific and Technical Information of China (English)

    郑旺; 高丽娜; 李西成; 张隆; 田志

    2012-01-01

    [目的]探讨在外源性生长因子(BMP -2)作用下,使用胶原蛋白膜包裹经体外培养扩增的骨髓基质干细胞(BMSCs)与藻酸钙凝胶形成的复合物修复骨缺损的可行性.[方法]选用成年新西兰白兔36只.手术造成两侧桡骨标准缺损后,左侧植入复合体,右侧空白对照.取自体BMSCs,经体外分离培养扩增后,与BMP -2、藻酸钙凝胶及胶原蛋白膜形成复合体修复骨缺损,于2、4、8、12周时行大体、X线片、免疫组织化学观察.[结果]整个过程中可见实验组缺损区新生骨组织增殖明显、持续时间较长,且无过量的结缔组织生长;X线早期可见连续性骨痂通过缺损区,分布均匀;免疫组织化学可见多个成骨中心,骨小梁排列有序,成熟骨替代完全,BMP分布持续时间长,且在新骨组织中分布范围大.[结论]复合体能修复骨缺损,并且能够阻挡周围软组织进入缺损区,为新骨生长提供空间;同时作为BMP的载体,能减少外溢,提高其疗效.%[Objective]To investigate whether the expanded bone marrow stromal cells(BMSCs) in vitro mixed with calcium alginate gelatin and wrapped by collag#n membrane can repair bone defects with the addition of exogenous bone morphoge-netic protein -2. [Method]The experiment was done on 36 adult New Zealand rabbits. After the experimental model of standard radius bone defect had been made by operation, the complex implanting on the left radius defect area and nothing on the right as blank contrast. Autologous bone marrow stem cells cultured in vitro were mixed with BMP - 2, calcium alginate gelatin and collagen membrane to repair bone defect. The reconstructed tissues were observed by gross, X-ray, immunohistochemistry at 2,4,8 and 12 weeks. [ Result] The experimental group showed that new bone growth tissues proliferated distinctly in bone defect area, and lasted longer. There were no excessive connective in defect area. Early X-ray observation revealed that

  13. 大鼠钝性脊髓损伤后BMPR Ia型受体的表达%Expression of bone morphogenetic protein receptor lA in rats after contusive spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    李华凤; 江兴华; 邹定全; 曹启林; 吕静; 李媛; 张慧芳; 王亚平

    2011-01-01

    目的 观察脊髓损伤后BMPR Ia型受体的表达.方法 运用免疫组织化学方法,首先检测3种BMP受体(BMPR)Ia、Ib、II型在正常成体大鼠脊髓中的表达分布.运用大鼠钝性脊髓损伤模型,以150 kdyn的撞击力直接撞击脊髓,观察动物撞击后1、3、7、14、30、60 d后脊髓中BMPR Ia的表达改变.结果 在正常成年大鼠脊髓中,BMPR Ia、II型受体主要在少突胶质细胞、灰质神经元中表达,在部分星形胶质细胞和大多数小胶质细胞中表达.灰质神经元中未检测到Ib型受体的表达或表达很低.脊髓损伤后,BMPR Ia在星形胶质细胞中表达激剧增加,高表达可持续至损伤后1个月;脊髓损伤诱导脊髓小胶质细胞活化,活化的小胶质细胞中表达BMPR Ia明显增加.结论 大鼠脊髓损伤后,诱导BMPR Ia受体在星形胶质细胞、小胶质细胞激剧增加,BMPR Ia高表达提示BMP信号在胶质细胞的重要病理生理作用,这一发现为进一步研究BMP信号的功能作用提供基础.%Objective To observe the expression pattern of bone morphogenetic protein receptor IA (BMPR I A) in rats after contusive spinal cord injury. Methods The expressions of BMPR IA, IB, and II were detected by immunochemistry in the spinal cord of normal adult rats, and the expression of BMPR IA was detected in the infinite horizons impactor model at 1, 3, 7, 14, 30, and 60 days after spinal cord injury. Results In the spinal cord of normal adult rats, BMPR IA and II were expressed predominantly in the oligodentrocytes and neurons in the grey matter, and also in some astrocytes and numerous microglia cells. Only a low level of BMPR IB expression was detected in the neurons of the grey matter. After spinal cord injury, the expression of BMP IA markedly increased with sustained strong expression in the astrocytes till one month after the injury; its expression was also increased obviously in the microglia cells activated by the injury. Conclusion The

  14. 前列腺癌组织BMP2表达及其与DIF-1的相关性%EXPRESSION OF BONE MORPHOGENETIC PROTEIN 2 AND ITS ASSOCIATION WITH DIFFERENTIATION INHIBITING FACTOR 1 IN PROSTATE CANCER

    Institute of Scientific and Technical Information of China (English)

    刘玉波; 于小玲; 赵辉

    2011-01-01

    Objective To investigate the expression of bone morphogenetic protein 2 (BMP2) in prostate cancer (PCa)and benign prostatic hyperplasia (BPH), and its association with cell differentiation, and differentiation inhibiting factor 1 (DIF-1). Methods Expressions of BMP2 and DIF-1 PCa and BPH were detected using immunohistochemical streptavidin peroxidase (SP) method. The positive expressions were analyzed using image analysis software. Linear correlation analysis was applied to analyze the correlation of BMP2 with DIF-1. Results In BPH, weak or negative expression of both BMP2 and DIF-1 was observed;In PCa, both BMP2 and DIF-1 showed positive expression. In PCa, the expressions of BMP2 and DIF-1 were positively correlated with Gleason score (rs=O.61, 0.63;P<0.01), and BMP2 was positively correlated with DIF-1 (r=0.92,P<0.01). Conclusion In prostate cancer, there is a correlation between the expressions of BMP2 and DIF-1, their expressions are closely related with the extent of malignancy, which indicates that both parameters are involved in the occurrence and development of this malignant tumor.%目的 探讨骨形态发生蛋白2(BMP2)在前列腺癌(PCa)及前列腺增生(BPH)组织中的表达及其与细胞分化的关系,并进一步分析其与分化抑制因子1(DIF-1)的相关性.方法 采用免疫组化SP法,分别检测PCa和BPH组织中BMP2和DIF-1的表达.采用图像分析软件分析阳性表达的灰度和面积,并采用直线相关方法分析BMP2和DIF-1的相关性.结果 BPH组织中BMP2和DIF-1呈阴性或弱阳性表达;PCa组织中BMP2和DIF-1均呈阳性表达.在PCa组织中.BMP2和DIF-1的表达水平与Gleason评分呈正相关(r=0.61、0.63,P<0.01);BMP2与DIF-1的表达也呈正相关(r=0.92,P<0.01).结论 在PCa组织中BMP2与DIF-1的表达存在相关性,且二者的表达均与PCa的恶性程度密切相关,提示二者可能与PCa的发生发展有关.

  15. The study of bone repair effect of recombinant human bone morphogenetic protein-2 combined with artificial bone substitute materials in alveolar defect around the dental implant%骨形态发生蛋白2复合人工骨替代材料修复犬种植体周骨缺损的研究

    Institute of Scientific and Technical Information of China (English)

    张敬阳; 黄翠

    2014-01-01

    Objective To evaluate the bone repair capacity of β-tricalciumphosphate (β-TCP) or Bio-Oss combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) in Beagle dogs.Methods Ten healthy adult Beagle dogs were used in this study.The mandibular premolars of the dogs were extracted firstly.Three months later,dental implants were placed in the mandibular,and a 5 mm× 4 mm× 3 mm bone defect area was prepared on the buccal side of each implant.The bone defect areas were filled with β-TCP,rhBMP-2/β-TCP compound,Bio-Oss or rhBMP-2/Bio-Oss compound respectively.The bone defect areas were filled nothing as the blank.The dogs were sacrificed after 4 or 12 weeks.Samples were prepared for the gross observation,X-ray analysis and histomorphology analysis.Results The new bone formation was observed in all experiment groups.After 4 weeks,the bone density of materials combined with rhBMP-2 groups [(36.70 ± 1.73) % and (38.50 ± 1.83) %] were higher than the raw materials groups [(30.50 ± 1.81)% and (31.48 ± 1.86)%].After 12 weeks,the bone density of materials combined with rhBMP-2 groups [(52.84 ± 1.85) % and (54.85 ± 1.87) %] were also higher than the raw materials groups [(43.65 ± 1.81) % and (44.94 ± 1.88) %].And new bone formation rate of the materials combined with rhBMP-2 groups were higher than the raw materials groups at each evaluation time.Conclusion rhBMP-2 can promote the bone regeneration activity of the artificial bone substitute materials effectively.%目的 观察重组人骨形态发生蛋白-2(rhBMP-2)复合β-磷酸三钙(β-TCP)及无机牛骨(Bio-Oss)对骨缺损修复能力的影响.方法 拔除10条Beagle犬下颌前磨牙,3个月骨质愈合后预备种植窝并制备5 mm×4 mm×3 mm骨缺损区,植入种植体,并分别在骨缺损区填入β-TCP、rhBMP-2/β-TCP复合物、Bio-Oss或rhBMP-2/Bio-Oss复合物;空白对照组不充填.术后4、12周处死犬,行大体观察、X线观察和组织形态

  16. The contrastive research on the osteogenesis effects of recombinant human bone morphogenetic protein-2, platelet-rich fibrin and autologous bone compositeding with coralline hydroxyapatite respectively%rhBMP-2、PRF和自体骨分别复合珊瑚羟基磷灰石成骨效能的比较研究

    Institute of Scientific and Technical Information of China (English)

    宋亚平; 徐世同; 杨淑娟; 张彩美; 黄丞蔚; 刘虎

    2014-01-01

    目的:通过建立动物骨缺损模型,比较重组人骨形成蛋白-2(rhBMP-2)与珊瑚羟基磷灰石(CHA)复合物、富血小板纤维蛋白(PRF)与珊瑚羟基磷灰石复合物、自体骨与珊瑚羟基磷灰石复合物以及单纯珊瑚羟基磷灰石这四种骨移植材料在骨缺损中的成骨效能。方法:在比格犬双侧胫骨干骺端制备四个相同的骨缺损区,在缺损区分别植入rhBMP-2/CHA、 PRF/CHA、自体骨/CHA及CHA (对照);3个月后处死动物,行大体标本观察;拍牙科CT,观察各植骨区骨密度情况;制作石蜡切片、 HE染色,比较各植骨区骨组织学特点及新骨形成量。结果:大体标本见四组骨缺损间隙均完全关闭。 X线示自体骨/CHA组和PRF/CHA组骨密度较致密, rhBMP-2/CHA组致密性低于前两者, CHA组未见明显骨致密影。 HE切片见四组新生骨与宿主骨连接紧密,新生骨小梁不规则,粗细不一,排列无序;复合型骨移植材料的新生骨小梁比对照组更密集、粗大,连续性更好;四组植骨区成骨量比较:自体骨/CHA组>PRF/CHA组>rhBMP-2/CHA组>CHA组。结论:复合型骨移植材料成骨效应明显优于单纯珊瑚羟基磷灰石;三种复合型材料中自体骨/CHA成骨效应最好,其次为PRF/CHA, rhBMP-2/CHA最差。%Objective: By establishing bone defects animal model, the osteogenesis effects of four groups bone transplantation materials that they are compound of reco mbinant human bone morphogenetic protein-2 (rhBMP -2), coralline hydroxyapatite (CHA), compound of platelet -rich fibrin and coralline hydroxyapatite, compound of autologous bone and coralline hydroxyapatite and pure coralline hydroxyapatite were compared by repairing bone defects. Methods: Four same bone defects were prepared on each side of tibial metaphyseal of Beagles , rhBMP-2/CHA, PRF/CHA, autologous bone/CHA and pure CHA were respectively implanted into the bone defects. The Beagles

  17. ECTOPIC OSTEOGENESIS OF BONE MARROW STROMAL CELLS INDUCED BY BONE MORPHOGENETIC PROTEIN%骨髓基质干细胞在骨形成蛋白诱导下异位成骨及应用于人造骨的实验研究

    Institute of Scientific and Technical Information of China (English)

    董书堃; 文剑明; 毕?; 董愉

    2001-01-01

    Objective To investigate the ectopic osteogenesis of bone marrow stromal cells (MSC) induced by bone morphogenetic protein(BMP) in vitro and in vivo, providing the experimental evidence for making an artificial bone with its own capacity of bone formation. Methods MSC were separated and cultured from bone marrow of Wistar rats, MSC were co-cultured with BMP in vitro (cultured in plate and diffuse chamber). Artificial coral hydroxyapatites (CHA) with MSC and BMP were implanted into dorsal muscles of Wistar rats, their bone formation were observed by morphological examination, histochemistry and immunohistochemistry. Results Only cartilaginous matrix were produced by MSC in vitro (cultured in plate and diffuse chamber), and both cartilaginous and bone matrix production within the combined grafts were seen. The bone formation of experimental groups (CHA+BMP+MSC) was stronger than that of control A(CHA+MSC) and control B(CHA). Conclusion It may be possible to produce an artificial bone with its own capacity of bone formation by combined graft (CHA+BMP+MSC). There may be multiple factors as well as BMP inducing bone formation both in the whole body and the location of the implantation. Further research on these factors will have the significance for making the ideal artificial bone.%目的 研究经分离培养的骨髓基质干细胞(MSC)在骨形成蛋白(BMP)诱导下在体内外的异位成骨效应,为研制一种本身具有成骨能力的人造骨材料提供实验依据。方法 分离、培养Wistar大鼠MSC,体外实验将MSC+BMP分别在培养板和弥散小室内培养,体内实验将MSC+BMP与人工珊瑚骨(CHA)制成复合移植体,种植在大鼠背肌内,用形态学、组织化学和免疫组化方法观察其成骨效应。结果 MSC在体外(培养板和弥散小室)只形成软骨基质,在体内复合移植体(CHA+BMP+MSC)形成骨质,其成骨效应比对照组A(CHA+MSC)和对照组B(CHA)强。结论 复合移植体(CHA

  18. 骨形成蛋白与肌瓣修复骨缺损的研究%Repair of Bone Defects of Tibia With Rotated Muscle Flap Reacted With Bovine Bone Morphogenetic Protein in Sheep

    Institute of Scientific and Technical Information of China (English)

    毕林祥; 陈燕; 李军; 林振福; 刘瑞萱

    1994-01-01

    The experiment was designed to simulate the clinical bone defects.All experiments were performed on twelve immature sheep.Biolateral bone defects were created in the upperdiaphyseal shaft of tibia.One lateral defect served as the experiment,the other as the control.The musclitibilis anterior rotational flap and 50mg of bBMP/iNCP consisted of No.5 ultra gelatin capsule were implanted into the experimental defects,the rotational muscle flap was implanted into the control one.Observations showed that the new bone was identified along the edge of the defects,as well as the central island' of new formed bone in two-week experimental defect after operation.Four weeks after surgery repair were complete in experimental bone defects.In controls,two weeks after operation,new bone was only identified along the edges of the defects and the quantity of new bone was lower than those in experiments.%采用羊胫骨缺损作为模型,分别植入肌瓣+骨形成蛋白和肌瓣.植入后第1,2,3,4周分批将羊处死,并进行X线摄片和病理组织学检查.结果表明:植入肌瓣+骨形成蛋白组术后第2周即在缺损边缘及中央出现大量新骨,于第4周基本由新骨充填;而单纯肌瓣植入组由缺损边缘少量新骨生长,术后第4周缺损由新骨部分充填.说明植入骨形成蛋白+肌瓣能够较快地修复骨缺损.

  19. 新伤续断汤对成骨细胞增殖及骨形态发生蛋白的影响%Effects of Xinshang Xuduan Decoction on Osteoblast Proliferation and Bone Morphogenetic Protein in Vitro

    Institute of Scientific and Technical Information of China (English)

    张佩; 董路珏; 李义凯

    2016-01-01

    【目的】探讨新伤续断汤(由骨碎补、续断、地鳖虫等组成)对大鼠成骨细胞增殖以及骨形态发生蛋白2(BMP-2)表达的影响。【方法】采用血清药理学方法制备新伤续断汤含药血清,与α-MEM培养基配成细胞培养液。按酶序列消化法从SD大鼠乳鼠颅骨获取、培养成骨细胞,以碱性磷酸酯酶染色和钙结节染色鉴定细胞。取第3~4代细胞用细胞增殖—毒性(CCK-8)法检测经体积分数为5%、10%、20%新伤续断汤含药血清处理24、48、72 h后的细胞增殖情况。用含体积分数10%的新伤续断汤、续断、骨碎补、空白对照含药血清分别培养细胞,采用微量酶标法检测碱性磷酸酶的活性,酶联免疫吸附法检测BMP-2的表达。【结果】新伤续断汤对成骨细胞的影响呈时间、浓度上的依赖性,其中浓度为10%时效果明显。体积分数10%的新伤续断汤、骨碎补、续断含药血清的成骨细胞碱性磷酸酶活性较空白对照组显著升高,差异有统计学意义(P<0.01);7 d时,新伤续断汤组、骨碎补组与空白对照组比较, BMP-2表达显著升高,差异有统计学意义(P<0.05)。【结论】新伤续断汤能促进大鼠成骨细胞碱性磷酸酶活性及BMP-2的表达,其中单味药骨碎补可能发挥重要作用,其促进骨折愈合机制可能与提高细胞的成骨功能有关。%Objective To investigate the effects of Xinshang Xuduan Decoction(XXD, mainly composed of Rhizoma Drynariae, Radix Dipsaci, and Eupolyphaga seu Steleophaga) on the proliferation and differentiation of rat osteoblasts and bone morphogenetic protein 2(BMP-2)expression in the osteoblasts . Methods The animal serum containing XXD was prepared by serum pharmacological method and then was mixed together with α-MEM for the cell culture. Osteoblasts were isolated from the skull bone of SD neonatal rats by collagenase digestion and were identified by

  20. Zellbiologie des Chondrozyten im Hinblick auf die Arthroseentstehung: Rolle der knorpelspezifischen Wachstumsfaktoren Cartilage-Derived Morphogenetic Proteins

    Directory of Open Access Journals (Sweden)

    Erlacher L

    1999-01-01

    Full Text Available Cartilage-derived morphogenetic proteins-1 and -2 (CDMP-1 and -2 sind Wachstumsfaktoren der Bone morphogenetic protein-Gruppe, die bei der embryonalen Skelettentwicklung maßgeblich beteiligt ist. In vivo führten CDMP-1 und -2 zur Neubildung von Knorpel- und Knochengewebe und mittels in vitro Untersuchungen an Knorpelzellen wurde eine Stimulation der Proteoglykansynthese nachgewiesen. Rezeptorbindungsstudien zeigten eine spezifische Bindung an den BMPR-IB/BMPR-II Rezeptorkomplex. Weiters konnte anhand immunhistochemischer Untersuchungen die Expression von CDMP-1 und -2 in gesundem als auch osteoarthrotischem Gelenksknorpel erbracht werden. In einem in vitro Modell für Osteoarthrose führten beide CDMPs nachhaltig zu einer Steigerung der Proteoglykansynthese und somit zu einer Förderung der Reparaturmechanismen des Knorpelgewebes.

  1. Simultaneous placement of nonvascularized bone graft and dental implant containing recombinant human bone morphogenetic protein-2: the results of ultra-structural examination in dogs%非血管化骨-人重组骨形成蛋白-2复合种植体同期移植的超微结构观察

    Institute of Scientific and Technical Information of China (English)

    李唐新; 郑林卿; 王大章; 陈刚

    2005-01-01

    目的:对于非血管化自体骨移植同期植入种植体,目前仍有争议.近年的研究表明:非血管化自体骨植入后,早期即可有新骨形成.本研究旨在探讨非血管化自体骨-种植体同期植入后种植体的愈合过程,并观察骨形成蛋白对与非血管化骨同期植入的种植体愈合过程的促进作用.方法:健康犬12只,随机分为2组.在犬双侧下颌角区各截取3cm×4cm骨段,实验组骨段内植入含有重组人骨形成蛋白-2的种植体,对照组植入普通纯钛种植体.植入种植体后,将骨块及种植体植回对侧下颌角,并以不锈钢丝固定.术后2、4、6、8及12周各处死2只动物,标本行扫描电子显微镜观察.结果:实验组种植体-骨界面在术后2周即可见明显的新骨形成,术后6~8周,已基本形成骨性结合;术后12周时,可见较为成熟的骨融合.而对照组骨融合在术后6~8周方开始形成,术后12周时仍未完成.实验结果显示,实验组骨融合的时间较对照组至少可提前4周.结论:骨形成蛋白的骨诱导活性可以促使种植体在植入后早期与非血管化骨形成骨融合,从而为提高同期植入种植体的成功率提供了新的途径.%PURPOSE: Simultaneous placement of dental implants with non-vascularized bone graft is still controversial,however, recent researches reveal that new bone formation can be obtained at early stage after bone grafting. The purpose of this study was to investigate the healing process of dental implants simultaneously placed in the nonvascularized grafted bones, and to estimate the effect of bone morphogenetic protein (BMP) on the healing of implants. METHODS: 12mongrel dogs were divided into 2 groups, 3cm×4cm bone segments were harvested from the bilateral mandibular angles.Two types of implants were applied. On the experimental sides, implants containing recombinant human bone morphogenetic protein-2 (rhBMP-2) were implanted, while on the opposite sides

  2. Assessment of bovine biomaterials containing bone morphogenetic proteins bound to absorbable hydroxyapatite in rabbit segmental bone defects Avaliação de biomateriais bovinos contendo proteínas morfogenéticas ósseas absorvidas a hidroxiapatita em defeitos ósseos segmentares em coelhos

    Directory of Open Access Journals (Sweden)

    Evelyn Hasegawa Gonçalves Caporali

    2006-12-01

    Full Text Available PURPOSE: To evaluate the osteo-regenerative capacity of two proprietary bone grafting materials, using a segmental defect model in both radial diaphyses of rabbits. METHODS: The right defect was filled with pooled bone morphogenetic proteins (pBMPs bound to absorbable ultrathin powdered hydroxyapatite (HA mixed with inorganic and demineralized bone matrix and bone-derived collagen, derived from bovine bone (Group A. The left defect was filled with bovine demineralized bone matrix and pBMPs bound to absorbable ultrathin powdered HA (Group B. In both groups, an absorbable membrane of demineralized bovine cortical was used to retain the biomaterials in the bone defects, and to guide the tissue regeneration. The rabbits were euthanized 30, 90 and 150 days after surgery. Radiographic, tomographic and histologic evaluations were carried out on all specimens. RESULTS: At 30 days, the demineralized cortical bone cover was totally resorbed in both groups. HA was totally resorbed from Group A defects, whereas HA persisted in Group B defects. A prominent foreign body reaction was evident with both products, more pronounced in sections from Group B. At 90 days, the defects in Group B exhibited more new bone than Group A. However, at 150 days after surgery, neither treatment had stimulated complete repair of the defect. CONCLUSION: The partial bone healing of the segmental defect occurred with low or none performance of the biomaterials tested.OBJETIVO: Avaliar a capacidade osteo-regenerativa de dois biomateriais utilizando um modelo de defeito segmentar efetuado nas diáfises do rádio de coelhos. MÉTODOS: O defeito direito foi preenchido com pool de proteínas morfogenéticas ósseas (pBMPs e hidroxiapatita em pó ultrafina absorvível (HA combinada com matriz óssea inorgânica desmineralizada e colágeno, derivados do osso bovino (Grupo A. O defeito esquerdo foi preenchido com matriz óssea desmineralizada bovina com pBMPs e hidroxiapatita em p

  3. 基因重叠延伸拼接PCR法钩建骨形态发生蛋白成熟肽真核表达载体的研究%Construction of a eukaryote expression vector containing bone morphogenetic protein-2 mature peptide by SOE-PCR method

    Institute of Scientific and Technical Information of China (English)

    段小红; 陈苏民; 柴玉波; 徐可为

    2002-01-01

    Objective To construct an eukaryote expression vector containing bone morphogenetic protein 2 (BMP 2) mature peptide. Methods Gene splicing by overlapping extension PCR (SOE PCR) method was used to clone BMP2 signal peptide and mature peptide and their fusion fragment.The fusion fragment was cloned into an eukaryote expressing vector pcDNA3.1/myc His(- )A. The sequence of the fusion fragment of BMP2 signal peptide and mature peptide was identified.Results The sequence of the fusion fragment was correct comparing with BMP2 signal peptide and mature peptide published by NCBI.Conclusion The vector pcDNA3.1/myc His(- )A BMP2sm constructed in this experiment was suitable to applying in eukaryotic expression of BMP2.

  4. Bone morphogenetic protein-2-encupsulated PEG-grafted-poly-lactic acid-polycaprolactone nanoparticles promote bone repair%聚乙二醇-骨形态发生蛋白-2-聚乳酸/聚已内酯载基因仿生骨促进骨缺损修复的实验观察

    Institute of Scientific and Technical Information of China (English)

    王湛; 许晓军; 杨军; 丁立峰; 李建军

    2015-01-01

    目的 观察聚乙二醇(PEG)/骨形态发生蛋白(BMP)-2纳米基因复合物及载基因仿生骨与骨髓间充质干细胞(BMSCs)对骨缺损修复的共同作用.方法 通过离子交联法制备PEG/BMP-2纳米基因复合物,通过溶液共混法制备聚乳酸(PLA)/聚已内酯(PCL)载基因仿生骨并接种BMSCs于仿生骨之上,应用免疫组化及免疫印迹检测BMSCs转染后BMP-2蛋白表达;酶联免疫方法检测转染后细胞培养上清中BMP-2分泌情况;实时聚合酶链反应检测转染后细胞BMP-2及骨钙素mRNA表达水平;截除新西兰大耳白兔双侧桡骨中段,植入载基因仿生骨材料,对骨缺损修复部位摄X线片、行HE染色和BMP-2免疫组织化学染色.结果 制备出PEG/BMP-2纳米颗粒及PEG-BMP-2-PLA/PCL载基因仿生骨.PEG/BMP-2纳米颗粒转染BMSCs和载基因仿生骨BMSCs内BMP-2表达量明显上调,骨钙素mRNA表达和碱性磷酸酶活性有所增加;体内实验中,PEG-BMP-2-PLA/PCL载基因仿生骨组与对照组比较BMP-2表达升高,新生骨在骨缺损区域所占面积比也明显增加.结论 PEG-BMP-2-PLA/PCL对骨缺损修复具有良好的效果.%Objective To explore the efficacy of a novel tissue engineered bone in repairing bone defects using poly-lactic acid-polycaprolactone (PLA-PCL) scaffolding seeded with PEG-bone morphogenetic protein-2 (BMP-2) transfected rBMSCs (rabbit bone marrow stromal cells).Methods rBMSCs were harvested,transfected with PEG/BMP-2 or liposome/BMP-2 and then implanted into PLA-PCL tissue engineered bone.The protein level of BMP-2 was assessed by Western blot and immunohistochemistry.Enzyme-linked immunosorbent assay (ELISA) was employed to measure the amount of BMP-2 in culture media.The mRNA levels of BMP-2 and osteocalcin were assayed quantitatively by real-time polymerase chain reaction (PCR).The middle portion of bilateral radius in New Zealand rabbits was excised and implanted with tissue engineered bone.And the modified areas were

  5. Preparation and characterization of recombinant human bone morphogenetic protein-2/poly lactic acid sustained release microspheres%骨形态发生蛋白2/聚乳酸缓释微球的制备及表征

    Institute of Scientific and Technical Information of China (English)

    马立坤; 叶鹏; 黄文良; 田仁元; 邓江

    2014-01-01

    背景:聚乳酸具有良好的生物相容性,是优良的药物缓释载体。  目的:制备重组人骨形态发生蛋白2/聚乳酸缓释微球,考察其理化特性。  方法:采用复乳溶剂挥发法制备重组人骨形态发生蛋白2/聚乳酸缓释微球,进行扫描电镜、激光粒度、Zeta电位、溶胀性能检测及采用ELISA试剂盒检测包封率、载药率及体外释药率。  结果与结论:扫描电镜见重组人骨形态发生蛋白2/聚乳酸缓释微球微球近似圆形,形态较规则,分散性较好,表面光滑。激光粒度分析重组人骨形态发生蛋白2/聚乳酸缓释微球微平均粒径839.6 nm , Zeta 电位(-32.93±3.74)mV,微球溶胀系数1.157±0.059,包封率及载药率分别为(88.943±2.878)%,(0.026±0.001)%;微球在第1天释药约10.199%,随后释药较恒定,至第19天累计释药率为54.643%。说明制备出的重组人骨形态发生蛋白2/聚乳酸缓释微球的粒径达到中华人民共和国药典第10版二部关于亚微球的定义标准及包封率不低于80%的要求,并且在体外具有很好的缓释功能。%BACKGROUND:Poly lactic acid as an excellent delivery has good biocompatibility. OBJECTIVE:To prepare recombinant human bone morphogenetic protein-2 (rhBMP-2)/poly lactic acid (PLA) sustained release microspheres, and to study its physical and chemical properties. METHODS:The rhBMP-2/PLA sustained release microspheres were prepared using w/o/w solvent evaporation method. Scanning electron microscopy, laser particle size, zeta potential, and swel ing properties were detected. ELISA kit was utilized for measurement of encapsulation efficiency, drug-loading rate and in vitro drug release rate. RESULTS AND CONCLUSION:Under the scanning electron microscope, rhBMP-2/PLA sustained release microspheres were approximately circle with excellent dispersion. The uniform spheres were visible with a mean particle size of 839.6 nm. The zeta

  6. Correlation between concentration of serum bone morphogenetic protein-7 and obesity in adult women%血清骨形成蛋白-7浓度与女性肥胖的相关性研究

    Institute of Scientific and Technical Information of China (English)

    曾俊; 姜友昭; 陈兵

    2011-01-01

    Objective To measure serum bone morphogenetic protein-7 (BMP-7) concentration in a dult women and to determine correlation between BMP-7 concentration and obesity, so as to provide a new target for obesity therapy. Methods Sixty-five obese women were enrolled in an obesity group, and one hundred and thirty-four adult women having normal body mass indexes were enrolled in a control group. Clinical data inclu ding age, height, weight, systolic blood pressure, diastolic blood pressure, waist circumference, and hip circ umference were recorded. Then body mass index and waist-hip ratio were calculated. Metabolic parameters such as serum total cholesterol, triglyceride, LDL-cholesterol, HDL-cholesterol, fasting glucose, and uric acid were tested. Serum BMP-7 concentration was detected through enzyme-linked immunosorbent assay (ELISA). The parameters of the two groups were compared. The correlation between serum BMP-7 concentration and met abolic parameters were analyzed. Results Serum BMP-7 concentration in the obesity group was significantly lower than that in the control group ( P < 0. 01 ). Serum BMP-7 concentration was negatively correlated with age, body mass index, systolic blood pressure, waist circumference, total cholesterol, and LDL-cholesterol (r = - 0. 181, - 0. 248, - 0. 143, - 0. 148, - 0. 169, and - 0. 177, respectively, P < 0. 05 ). Multiple stepwise regression analysis results showed that the body mass index was the only independent factor influencing the serum BMP-7 concentration after being adjusted by age, body mass index, systolic blood pressure, waist circumference, total cholesterol, LDL-cholesterol, diastolic blood pressure, mean arterial blood pressure, waist hip ratio, uric acid, triglyceride, HDL-cholesterol, and fasting glucose. Corncltusion In adult women, serum BMP-7 concentration is negatively correlated with body mass index, suggesting that BMP-7 may be a potential target for woman obesity therapy.%目的检

  7. 重组骨形成蛋白-2与珊瑚人工骨复合物应用于拔牙窝修复的动物实验研究%The effects of coral artificial bone composite of recombinant hmnan bone morphogenetic protein-2 on reconstruction of extraction sockets:an experimetal study on dogs

    Institute of Scientific and Technical Information of China (English)

    孔卫东; 林巍; 李小兰; 邓国珍; 沈丽佳

    2001-01-01

    Aim:To evaluate the bone repairing ability of coral artificial bone composite of rhBMP -2(rhBMP-2/CAB) and coral artificial bone(CAB) implanted into immediate extraction sockets. Meth-ods: 12 adult dogs served as the experimental animals. Immediately after extraction of the upper secondand third incisors, the alveolar septum between extraction sockets was resected bilaterally. RhBMP-2/CAB and CAB were implanted respectively into each extraction site. The animals were sacrificed at the 4th, 8th and 12th weeks respectively after implantation. The bone repairing ability of the two grafts wasanalyzed with histologic and image analysis system. Results: RhBMP - 2/CAB has a good effect on therepairing ability of extraction sockets. The implants were absorbed gradually after they were implanted in-to extraction sockets. In the meantime, the new bone was formed in extraction sockets. The implants werereplaced completely by bone at 12 weeks. The ratio of new bone formation of rhBMP-2/CAB was signif-icantly higher than that of CAB at different period( P < 0.05). Conclusion: The repairing ability and ef-fect of rbBMP -2/CAB in extraction sockets are obviously better than those of CAB.%目的:探讨重组骨形成蛋白 - 2(recombinant human bone morphogenetic protein , rhBMP - 2)/ 珊瑚人工骨复合物(复合骨)与珊瑚人工骨(珊瑚骨)在拔牙窝修复中的作用。方法:12只成年狗作为实验动物,拔除两侧上颌第2及第3切牙,并去除牙槽窝之间的牙槽间隔,一侧随即植入复合骨,对侧植人珊瑚骨作为对照。并于植骨后4、8、12周取材,采用组织学观察及计算机图像分析方法,观察比较两种植入材料在拔牙窝内的骨修复能力及修复效果。结果:复合骨具有较强的骨修复作用,植入牙槽窝后,材料被逐渐降解吸收,新骨不断形成,12周后,植入材料完全被成熟的骨组织取代;图像分析结果显示复合骨组新生骨形成的比值明

  8. 骨形态发生蛋白、碱性成纤维细胞生长因子生物材料在关节软骨缺损修复中的生物性能%Biological properties of bone morphogenetic proteins and basic fibroblast growth factor in biological materials for repair of articular cartilage defect

    Institute of Scientific and Technical Information of China (English)

    董君博

    2016-01-01

    BACKGROUND:Articular cartilage regeneration can be regulatedbyautocrineorparacrinesecretionof various cytokines. OBJECTIVE:To analyze biological properties of bone morphogenetic proteins and basic fibroblast growth factor in biological materials for repair of articular cartilage defect. METHODS:Forty New Zealand white rabbits were used and equaly randomized intofourgroups: fibrin, basic fibroblast growth factor, bone morphogenetic protein, and combined treatment (basic fibroblast growth factor combined with bone morphogenetic protein) groups, respectively.Bioactivescaffolds with fibrin, basic fibroblast growth factor,bone morphogenetic protein, and basic fibroblast growth factor combined with bone morphogenetic protein were injected to repair the articular cartilage defect. Therapeutic effect andbiological properties of biological materials were compared. RESULTS AND CONCLUSION:(1) Inthefibrin group,tworabbits appearedto havelimps. Inthebasic fibroblast growth factor group hand functionwaslimited inonerabbit. Inthebone morphogenetic protein group, one had a limpandonewasin a limitation of activity. Inthecombined treatment group,rabbitsrecovered wel andshowedno differencesintheknee joint before and aftersurgery (P  目的:分析骨形态发生蛋白、碱性成纤维细胞生长因子生物材料在关节软骨缺损修复中的生物性能。  方法:选取40只新西兰家兔,随机分为4组,纤维蛋白组、碱性成纤维细胞生长因子组、骨形态发生蛋白组、复合组(骨形态发生蛋白+碱性成纤维细胞生长因子),每组10只。建立兔关节软骨缺损模型,止血彻底后将纤维蛋白、碱性成纤维细胞生长因子、骨形态发生蛋白以及骨形态发生蛋白、碱性成纤维细胞生长因子复合等材料组成的支架分别植入缺损部位。比较不同注射材料在家兔关节软骨缺损中的效果及复合材料的生物性能。  结果与结论:①关节软骨缺损修复情

  9. Changes of Human Recombination Bone Morphogenetic Protein-2 in Bone and Marrow in Tail Suspended%模拟失重下大鼠骨和骨髓中人重组骨形成蛋白-2的变化

    Institute of Scientific and Technical Information of China (English)

    傅崇建; 杨连甲; 曹新生; 陈希哲; 张立藩

    2001-01-01

    目的研究模拟失重骨质和骨髓内人重组骨形成蛋白-2(Human recombination bone morphogeneticprotein-2,rhBMP-2)形态学的变化.方法选用SD大鼠,随机配对分为悬吊组和自由活动组(5只/组),实验期为14、28 d.组织标本行原位杂交.结果尾吊组大鼠骨质和骨髓内rhBMP-2的表达明显弱于对照组(P<0.05);尾吊组14 d rhBMP-2的表达明显强于28 d(P<0.05).结论大鼠后肢去负荷导致骨和骨髓rhBMP-2含量的降低.

  10. BMP-2联合温热化疗对SW480中GDF15和TFF3表达的影响%Effect of bone morphogenetic protein-2 combined with hyperthermic chemotherapy on GDF15 and TFF3 expression in SW480

    Institute of Scientific and Technical Information of China (English)

    毕德利; 王亚旭; 舒宁波; 谢凯

    2012-01-01

    目的:探讨骨形态发生蛋白-2(Bone morphogenetic protein-2,BMP-2)联合温热化疗对SW480中GDF15和TFF3表达的影响.方法:BMP-2作用于大肠癌SW480细胞系,置于43℃温热化疗30 min,培养6h.(1)流式细胞法检测SW480细胞的凋亡;(2) Western blot法检测GDF15和TFF3蛋白表达情况;(3) RT-PCR半定量检测GDF15和TFF3的mRNA表达.结果:BMP-2联合43℃温热化疗后SW480细胞凋亡增加,GDF15和TFF3蛋白表达水平下降,GDF15和TFF3的mRNA表达亦下调.结论:BMP-2联合温热化疗通过抑制大肠癌SW480的GDF15和TFF3蛋白及其相关基因表达,增强抑制SW480的增殖及转移复发的作用;温热疗法联合化疗对GDF15和TFF3表达抑制有协同作用.%Objective: To explore the effect of bone morphogenetic protein-2(BMP-2) combined with hyperthermic chemotherapy on GDF15 and TFF3 expression in SW480. Methods:BMP-2 was acted on SW480 colorectal cancer cell lines,placed in 43 t hyperthermic chemotherapy for 30 min and cultured for 6 h. (1 )Apoptosis of SW480 cells was detected by flow cytometry assay; (2)GDF15 and TFF3 protein expressions were detected by Western blot; (3)GDF15 and TFF3 mRNA expressions were detected by RT-PCR. Results: SW480 cells apoptosis was increased, GDF15 and TFF3 protein levels were decreased after BMP-2 combined with 43 t hyperthermic chemotherapy. GDF 15 and TFF3 mRNA expressions were also reduced. Conclusions: BMP-2 combined hyperthermic chemotherapy can inhibit proliferation and metastasis of SW480 by inhibiting GDF15 and TFF3 protein and mRNA expressions. Hyperthermia combined with chemotherapy has synergistic effect on the inhibition of GDF15 and TFF3 expression.

  11. Experimental Research on Spinal Fusion with Recombinant Human Bone Morphogenetic Protein-2 and Bone Marrow Tromal Cell Composited Tricalcium Phosphate (TCP)%重组人骨形成蛋白2/骨髓间充质干细胞复合磷酸三钙应用于脊柱融合的实验研究

    Institute of Scientific and Technical Information of China (English)

    杨小荣; 吴良绍; 方煌

    2012-01-01

    This paper is aimed to assess the efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2) and bone marrow stromal cell (BMSCs) composited tricalcium phosphate (TCP) in a rat model of posterolateral lumbar intertransverse process fusion. Rat BMSCs were cultured in vitro. Twenty SD rats underwent single-level bilateral intertransverse process spine arthrodesis at L4 and L5. These rats were assigned to two groups according to the graft materials. They received: 10 of the total were treated with the BMSCs with rhBMP-2 and tricalcium phosphate (TCP) as the experimental group,and the other 10 with TCP treatment alone as the control group. All the animals were killed at 4 weeks after surgery and the spine fusion results were assessed by gross inspection, manual palpation, radiography and histology. The fusion rate, the tensile strength and stiffness of the solidly fused levels in the experimental group were statistically higher than that of the controlled group(P<0. 05). These results showed that the spinal fusion could be improved mechanically when rhBMP-2 and BMSCs were added into the TCP.%观察重组人骨形成蛋白2(rhBMP-2)与骨髓间充质干细胞(BMSCs)复合磷酸三钙(TCP)应用于大鼠腰椎横突间融合的愈合情况.体外培养大鼠BMSCs,20只SD大鼠均行单节段双侧L4,5横突间植骨融合术,依植入物的不同分为实验组与对照组,每组10只.实验组植入复合rhBMP-2和BMSCs的TCP,对照组只植入TCP.术后4周处死动物,取出腰椎,通过大体观察、手法检测、影像学、组织学等方法分析融合状况.结果表明应用rhBMP-2和BMSCs的TCP组新骨形成良好,脊柱融合效果好,生物力学强度高,融合率显著高于对照组(P<0.05).结果提示:应用rhBMP-2和BMSCs有利于复合材料成骨和脊柱融合.

  12. 腺病毒携带骨形态发生蛋白14基因转染脂肪干细胞修复损伤关节软骨%Adipose-derived stem cells transfected with adenovirus carrying bone morphogenetic protein 14 for repair of articular cartilage injury

    Institute of Scientific and Technical Information of China (English)

    马洪斌; 李运祥; 王铭伦

    2015-01-01

    BACKGROUND:The articular cartilage has weak self-repair ability, mainly due to its lack of trophoblast cels in blood vessels and slow cel metabolism. Current treatment methods cannot restore the original function of the cartilage tissue, and cartilage tissue engineering in recent years has garnered increasing attention. OBJECTIVE:To observe the effect of adipose-derived stem cels transfected with bone morphogenetic protein 14 combined with type I colagen sponge scaffold on the repair of articular cartilage injury in the knee of rabbits. METHODS: Adipose-derived stem cels were isolated and cultured from rabbit subcutaneous adipose tissue, and transfected with Ad-CMV-BMP-14-IRES-hrGFP-1. Type I colagen sponge scaffold with the transfected adipose-derived stem cels was used to repair articular cartilage injury in the knee of rabbits. Twelve weeks after operation, the articular tissue was taken for gross assessment and histological evaluation. RESULTS AND CONCLUSION: The expressions of bone morphogenetic protein 14, type II colagen and Sox-9 were higher in cels transfected with bone morphogenetic protein 14 than untransfected ones. At 12 weeks after operation, adipose-derived stem cels transfected with bone morphogenetic protein 14 combined with type I colagen sponge scaffold had good repair effect on articular cartilage injuries, and the injured cartilage tissues were smooth and had good texture, color and integration junction; adipose-derived stem cels combined with type I colagen sponge scaffold could partialy repair the injured cartilage tissues that had similar color and texture to normal tissues, and there was a remarkable boundary between the repaired tissue and normal cartilage tissue;simple type I colagen sponge scaffold was almost colapsed, and no hyaline cartilage tissue formed. These findings indicate that transfection of bone morphogenetic protein 14 can strengthen the ability of adipose-derived stem cels dramaticaly to repair cartilage injuries.%背景

  13. Regulatory Mechanism of Bone Morphogenetic Proteins 6 on Iron Metabolism During Exercise%运动对骨形态发生蛋白6介导的机体铁代谢调节机制的研究进展

    Institute of Scientific and Technical Information of China (English)

    孙娟; 王海涛; 刘玉倩; 王羽; 王金霞

    2012-01-01

    骨形态发生蛋白6(BMP6)为TGF-β超家族中一员,它产生于骨髓源性间质干细胞(BMSC)及造血干细胞,是一种调节成骨细胞和成软骨细胞分化的骨生长因子,在骨缺损的修复中具有重要作用.运动可促进BMP6的表达.研究发现,BMP6也是铁调素的重要的内源调节子,可以上调铁调素的表达,而铁调素是肠铁吸收的负调节子.因此,运动引起的BMP6表达增加,可以促进机体铁的吸收和释放,从而对运动中维持机体铁的动态平衡有重要的调控作用.%Bone morphogenetic protein 6(BMP6) is a member of the transforming growth factor-β family, it is produced by bone marrow-mesenchymal (BMSC) and hematopoietic stem cells. BMP6 is a potent protein for future treatment strategies of bone regeneration as it is a very important regulator of bone ho-meostasis and is a kind of adjusting the osteoblast and cartilage cells of bone growth factors. Moreover,it is also released by osteoclasts as a key bone coupling factor recruiting osteoblasts to the resorption site. So it has a good application potential in all kinds of bone defect repair. Sports can promote expression of BMP6. Recent study shows that BMP6 can up-regulation of hepcidin expression. However,hepcidin is the negative regulators and down-regulation of intestinal iron absorbing. So exercise-induced increased expression of BMP6 and promotes iron absorption and release. Consequently,to maintain iron homeostasis in the sport has an important regulatory role.

  14. Silk fibroin-compound bone cement/recombinant human bone morphogenetic protein-2 to repair sheep vertebral defects%丝素蛋白/双相磷酸钙/半水硫酸钙/重组人骨形态发生蛋白-2骨水泥的制备及修复椎体骨缺损的实验研究

    Institute of Scientific and Technical Information of China (English)

    王根林; 陈广东; 朱雪松; 朱志军; 谢瑞娟; 卢神州; 张波; 夏太宝; 杨惠林

    2015-01-01

    目的 研制丝素蛋白(SF)/双相磷酸钙(BCP)/半水硫酸钙(CSH)/重组人骨形态发生蛋白-2(rhBMP-2)骨水泥,并探讨其在绵羊椎体内的成骨作用. 方法 制备SF/BCP/CSH/rhBMP-2骨水泥,分别在12只绵羊的L2 L3、L4椎体内制作直径为6.0mm、深度为10 mm的圆柱型骨缺损模型,在3个缺损处随机植入SF/BCP/CSH/rhBMP-2骨水泥作为实验组,植入聚甲基丙烯酸甲酯(PMMP)作为对照组,另一椎体缺损处不植入任何材料作为空白对照组.术后3、6个月分别随机处死6只绵羊进行CT、组织学和生物力学检查.结果 CT和组织学检查显示:术后3个月实验组椎体密度与正常椎体相似,骨缺损修复基本完成,术后6个月骨缺损修复完成;对照组术后3、6个月时PMMP无降解,并与骨之间结合疏松,表面无新骨形成;空白对照组术后3、6个月时骨缺损一直存在.生物力学测试显示:术后3、6个月时实验组椎体抗压强度和刚度与正常椎体相比差异无统计学意义(P>0.05). 结论 SF/BCP/CSH/rhBMP-2骨水泥具有良好的成骨作用,在成骨过程中能维持椎体的力学性能,有望成为经皮椎体强化术的一种可降解、具成骨作用的填充剂.%Objective To prepare compound bone cement of silk fibroin/biphasic calcium phosphate/alpha-calcium sulphate hemihydrate/recombinant human bone morphogenetic protein-2 (SF/BCP/CSH/rhBMP-2) and to study its osteogenesis capacity for sheep vertebral defects.Methods Compound bone cement SF/BCP/CSH/rhBMP-2 was prepared and a cylindrical bone defect (6.0 mm in diameter and 10 mm in depth) was created at lumbar vertebrae 2,3 and 4 by open operation in 12 sheep.The injured lumbar vertebrae in each sheep were randomly divided into 3 study groups.The experimental group was implanted with the SF/BCP/CSH/rhBMP-2,the control group with polymethylmethacrylate (PMMA),and the blank control group with nothing.At 3 and 6 months postoperation,6 random sheep were sacrificed for

  15. hsa-miR-654-5p regulates osteogenic differentiation of human bone marrow mesenchymal stem cells by repressing bone morphogenetic protein 2%hsa-miR-654-5p通过抑制骨形态发生蛋白2调控人骨髓基质干细胞成骨分化

    Institute of Scientific and Technical Information of China (English)

    魏均强; 陈华; 郑晓飞; 张伯勋; 王岩; 唐佩福; 余飞; 宋青; 黎檀实

    2012-01-01

    Objective To study the effect of hsa-miR-654-5p in repressing bone morphogenetic protein 2 (BMP2) mRNA and protein in human bone marrow mesenchymal stem cells (hBMSCs), and explore its regulatory role in osteogenic differentiation of hBMSCs. Methods hBMSCs in the 4th passage were cultured for 16 h and transfected with hsa-miR-654-5p followed by further culture for 48 h. qRT-PCR and Western blotting were performed to detect the expressions of BMP2 mRNA and protein. Dual-luciferase reporter gene assay was employed to examine the repression of the BMP2 gene. Results BMP2 mRNA and protein expressions were significantly down-regulated in hBMSCs with hsa-miR-654-5p overexpression. Duallucd-ferase reporter gene assay indicated that the predicted target site of BMP2 was repressed directly by hsa-miR-654-5p, but this repression did not occur at the mutant predicted target site of BMP2. Conclusion hsa-miR-654-5p can directly repress the mRNA and protein expressions of BMP2 by binding to a specific target site. The changes in hsa-miR-654-5p can play an important role in osteogenic differentiation regulation of hBMSCs.%目的 明确萎缩性骨不连组织水平表达上调的hsa-miR-654-5p在人骨髓基质干细胞(hBMSCs)中对其预测靶基因骨形态发生蛋白2(BMP2)mRNA和蛋白的抑制作用,探索其在成骨分化过程中的生物学调控功能.方法 分离培养hBMSCs,将第4代hBMSCs培养16 h后分别按相应体系转染细胞,再培养48 h后取六孔板内细胞提取总RNA和总蛋白,进行实时定量PCR(qRT-PCR)和Western blotting,取24孔板内细胞进行双荧光素酶报告基因检测.结果 当hBMSCs中hsa-miR-654-5p过表达时,BMP2的mRNA和蛋白表达水平均发生明显下调;双荧光素酶报告基因检测提示,BMP2的预测靶位点直接受hsa-miR-654-5p的抑制调控,该靶位点被突变后hsa-miR-654-5p对BMP2的抑制作用消失.结论 hsa-miR-654-5p可通过作用于BMP2的特定靶位点而直接抑制BMP2

  16. Polyphosphate: A Morphogenetically Active Implant Material Serving as Metabolic Fuel for Bone Regeneration.

    Science.gov (United States)

    Müller, Werner E G; Tolba, Emad; Schröder, Heinz C; Wang, Xiaohong

    2015-09-01

    The initial mineralization centers during human bone formation onto osteoblasts are composed of CaCO3 . Those bioseeds are enzymatically formed via carbonic anhydrase(s) in close association with the cell surface of the osteoblasts. Subsequently, the bicarbonate/carbonate anions are exchanged non-enzymatically by inorganic phosphate [Pi ]. One source for the supply of Pi is polyphosphate [polyP] which is a physiological polymer, formed in the osteoblasts as well as in the platelets. The energy-rich acid anhydride bonds within the polyP chain are cleaved by phosphatase(s); during this reaction free-energy might be released that could be re-used, as metabolic fuel, for the maintenance of the steady-state concentrations of the substrates/products during mineralization. Finally it is outlined that polyP, as a morphogenetically active scaffold, is even suitable for 3D cell printing.

  17. 重组人骨形成蛋白2诱导的骨膜细胞构建组织工程骨的实验研究%TISSUE ENGINEERED BONE REGENERATION OF PERIOSTEAL CELLS USING RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN 2 INDUCE

    Institute of Scientific and Technical Information of China (English)

    张超; 胡蕴玉; 熊卓; 张曙明; 颜永年; 崔福斋

    2005-01-01

    目的通过组织工程技术方法,研究细胞-材料复合体体内骨再生的能力. 方法采用材料学自组装技术的原理,以I型胶原蛋白为分子模板,引导钙磷盐在液相中的矿化,制备具有天然骨基质层状结构的羟基磷灰石/胶原复合材料,并以热致分相法制备羟基磷灰石/胶原-聚乳酸[hydroxyapatite/collagen-poly-(L-lactic acid),HAC-PLA]复合三维多孔框架,与重组人骨形成蛋白2(recombinant human bone morphogenetic protein 2,rhBMP-2)诱导分化后的兔骨膜细胞构建组织工程骨,进行体外电镜及组织学观察.将3月龄裸鼠18只,分为3组,每组6只.A组皮下注射经rhBMP-2处理后的骨膜细胞悬液,B组植入未复合细胞的HAC-PLA,C组植入rhBMP-2处理后的细胞-材料复合体.术后8周取材行组织学观察,C组与B组及A组进行比较. 结果三维多孔HAC-PLA框架材料孔隙率大于90%,孔径为50~300 μm.骨膜细胞经500 ng/ml的rhBMP-2处理后,与改善亲水性后的HAC-PLA三维多孔框架复合,构建组织工程骨.电镜显示接种的细胞能在此组织工程骨内部生长增殖,术后8周C组有新骨形成,A组注射部位表面平整,无新生物,B组为纤维组织,无骨及软骨形成. 结论所构建的组织工程骨有望成为骨创伤修复治疗一种新的技术.

  18. 携重组人骨形态发生蛋白2聚乳酸缓释微球的生物支架复合自体松质骨修复骨缺损%Biological scaffold carrying recombinant human bone morphogenetic protein-2/poly bone defects

    Institute of Scientific and Technical Information of China (English)

    马立坤; 邓江; 叶鹏; 佘荣峰; 黄文良; 吕雪峰

    2015-01-01

    目的 探讨含重组人骨形态发生蛋白2(recombinant human Bone Morphogenetic Protein-2,rhBMP-2)聚乳酸(Poly Lactic Acid,PLA)缓释微球的丝素蛋白(Silk Fibroin,SF)/壳聚糖(Chitosan,CS)/纳米羟基磷灰石(nano hydroxyapatite,n-HA)支架复合部分自体松质骨修复骨缺损的疗效.方法 取成年新西兰大白兔60只,随机分4组,制备右侧中段桡骨1.5cm的骨缺损模型.A组:含rhBMP-2/PLA缓释微球的SF/CS/n-HA整合支架复合部分自体松质骨,B组:含rhBMP-2的SF/CS/n-HA支架复合部分自体松质骨,C组:单纯自体松质骨,D组:单纯SF/CS/n-HA支架.术后4、8、12周摄X片、并参照Lane-Sandhu骨缺损修复组织X线评分标准,组织学、并参照Lane-Sandhu组织学评分标准进行评分,骨密度检测评估各组骨缺损修复效果.结果 术后4、8、12周X片和组织学检查的结果显示,A组与C组比较差异无统计学意义(P>0.05),优于其他各组(P<0.05).术后12周骨密度结果示A组与C组比较差异无统计学意义(P>0.05),优于其他各组(P<0.05).结论 含rhBMP-2/PLA缓释微球的SF/CS/n-HA整合支架复合部分自体松质骨与自体松质骨修复兔桡骨缺损的疗效一致,说明可以大大减少自体松质骨的使用,有望成为一种新型、优良的复合骨组织工程支架材料.

  19. 髋关节发育不良髋臼BMP-2和Runx2表达及微结构分析%EXPRESSIONS OF BONE MORPHOGENETIC PROTEIN 2 AND RUNT-RELATED TRANSCRIPTION FACTOR 2 AND MICROARCHITECTURE OF TRABECULAR BONE PERIACETABULA IN ADULT PATIENTS WITH DEVELOPMENTAL DYSPLASIA OF HIP

    Institute of Scientific and Technical Information of China (English)

    刘瑞宇; 王坤正; 李永伟; 柏传毅; 王春生; 党晓谦

    2013-01-01

    目的 探讨髋关节发育不良(developmental dysplasia of the hip,DDH)成人患者髋臼周围骨质微结构变化及与骨质代谢相关生长因子BMP-2和Runx2(runt-related transcription factor 2)表达,分析该类患者人工全髋关节置换术后髋臼假体高松动率的原因. 方法 以2008年3月-9月8例行人工全髋关节置换术的DDH患者作为试验组,男3例,女5例;年龄37~55岁;髋关节脱位程度按照Crowe等评价方法评定为30%~80%.以同期8例行人工髋关节表面置换术的股骨头缺血性坏死(Ficat Ⅱ期)患者作为对照组,男3例,女5例;年龄36~55岁.取两组患者髋臼臼顶内上方松质骨,采用实时定量PCR测量骨组织BMP-2和Runx2表达;Micro-CT扫描观察其微结构,测量骨体积分数(bone volume/total volume,BV/TV),单位体积内骨小梁分支数H(connectivity density,Conn.Dens),骨小梁数目(trabecular number,Tb.N),骨小梁厚度(trabecular thickness,Tb.Th),骨小梁分离度(trabecular separation,Tb.Sp),结构模型指数(structure model index,SMI). 结果 试验组BMP-2及Runx2表达显著低于对照组,差异均有统计学意义(P<0.05).Micro-CT扫描观察示,试验组骨小梁结构稀疏,单一骨小梁直径较粗,对照组骨小梁结构致密,单一骨小梁直径较细.试验组BV/TV、Tb.N显著低于对照组,SMI及Tb.Sp显著高于对照组,差异均有统计学意义(P<0.05);两组Conn.Dens及Tb.Th比较,差异无统计学意义(P>0.05). 结论 DDH患者的髋臼臼顶松质骨处于二低代谢状态,其微结构趋于骨质疏松化,较差的骨质状况可能是DDH患者人工全髋关节置换术后髋臼假体高松动率的原因之一.%Objective To explore the expressions of bone morphogenetic protein 2 (BMP-2) and runt-related transcription facotr 2 (Runx2) and microarchitecture of trabecular bone periacetabula in adult patients with developmental dysplasia of the hip (DDH). Methods Between March and September 2008, the trabecular

  20. Skeletal muscular transformation in osteogenetic process of bone defect repaired with compound vascularized muscle flap with bone morphogenetic protein%骨形态发生蛋白复合带血供肌瓣修复骨缺损成骨过程中骨骼肌的转归

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    taken as the carrier of bone morphogenetic protein (BMP).DESIGN: Observation and comparison were designed in the experiment.SETTING: Experimental Animal Center of Nanfang Hospital affiliated to Southern Medical University.MATERIALS: rhBMP-2, 20 New Zealand healthy white rabbits of clean grade, weighted varied from 1.5 to 2.5 kg, of either gender.METHODS: The experiment was performed in Experimental Animal Center of Nanfang Hospital affiliated to Southern Medical University from January 2000 to August 2002. Ten rabbits were employed and bone defect was modeled 15 mm long on the lower section of radium. After nerve removed,the muscular flap of deep flexor muscle of fingers was implanted into the defect. In addition, the compound of fibrin and rhBMP-2 was grafted into the muscular flap. Four rabbits were sacrificed in 1, 2, 4, 6 and 8 weeks successively under anesthetization for gross observation, histological, TUNEL and ultrastructure examinations of muscular flap on bone defect.MAIN OUTCOME MEASURES: Gross observation, histological, TUNEL and ultrastrncture examinations of muscular flap on bone defect in rabbits tion of muscular flap on radial defect in rabbits: Skeletal muscle was atrophic gradually and muscle fiber almost disappeared in 8 weeks and reamination of muscular flap on radial defect in rabbits: It was displayed in histology that while skeletal muscle was atrophic, a part of cell karyon apmuscular flap of radial defect in rabbits: It was indicated with TUNEL staining that there were large amounts of colored positive karyons or karyon flap in radial defect of rabbits: Skeletal muscular filaments disappeared gradually by time lasting, karyon presented chromatin bordering, pyknosis and fragmentation, indicating typical apoptotic alternations.CONCLUSION: During osteogenetic process of bone defect repaired with compound BMP vascularized muscular flap, muscle fibers were atrophic gradually and apoptotic, terminally, substituted by bone tissues.

  1. 纤维蛋白凝胶复合骨形态发生蛋白和庆大霉素缓释药物对感染性骨缺损的修复%Fibrin glue/bone morphogenetic protein complex plus slow-release gentamicin for repairing infected bone defects in rabbits

    Institute of Scientific and Technical Information of China (English)

    高秋明; 刘兴炎; 董晓萍; 葛宝丰; 白孟海; 陈克明

    2005-01-01

    背景:慢性骨髓炎临床处理较为棘手,手术常需分期进行,目前尚无好的方法予以一期修复.目的:探讨将纤维蛋白凝胶(FG)作为骨形态发生蛋白(BMP)及庆大霉素的共同载体,一期修复感染性骨缺损的可行性.设计:完全随机对照实验研究.单位:解放军兰州军区兰州总医院全军创伤骨科中心.材料:实验在兰州军区兰州总医院骨科研究所完成.对象为体质量1.9~2.4kg的48只成年健康青紫兰兔,雌雄不限,购自甘肃省兰州市生物制品研究所.干预:48只青紫兰兔,制作慢性骨髓炎模型,清创后造成胫骨近侧干骺端内侧1.5 cm长半环形骨缺损,采用3种方法进行处理:A组,植入FG,BMP和庆大霉素复合物;B组,植入FG/BMP复合物,C组,作为空白对照.主要观察指标:术后观察动物一般情况,做骨培养及细菌计数,X射线拍片及组织学检查.结果:A组感染控制及骨修复均良好,感染控制率、再生骨量明显优于B组.B,C两组在感染控制率上无显著差异.C组动物骨修复差.结论:FG,BMP及庆大霉素复合物具有促进成骨及抗感染的双重作用,可用于感染性骨缺损及污染严重的开放性损伤造成的骨缺损的修复.%BACKGROUND: Chronic osteomyelitis is difficult to manage clinically, and two or more operations were commonly needed. No satisfactory method for one-stage repair has been currently available.OBJECTIVE: To examine the possibility of using fibrin glue(FG) as the common carrier for both bone morphogenetic protein(BMP) and gentamicin for one-stage repair of infected bone defects.DESIGN: A completely randomized controlled experiment.SETTING: Center of Orthopaedic Surgery, Lanzhou General Hospital of Lanzhou Area Command of of Chinese PLA.MATERIALS: The experiment was conducted using 48 healthy adult Chinchilla rabbits of either sex on normal diet with body mass of 1.9 to 2.4 kg,provided by the Institute of Biological Products, Lanzhou, Gansu Province

  2. Strontium ranelate promotes osteogenic differentiation of rat bone mesenchymal stem cells through bone morphogenetic protein-2/Smad signaling pathway%雷奈酸锶通过骨形态发生蛋白-2/Smad通路促进骨髓间充质干细胞成骨分化

    Institute of Scientific and Technical Information of China (English)

    吕辉珍; 黄晓丹; 靳思思; 郭润民; 吴文

    2013-01-01

    Objective To explore whether strontium ranelate (Sr) promotes osteoblast lineage differentiation of rat bone mesenchymal stem cells (BMSCs) through the bone morphogenetic protein-2 (BMP-2)/Smad signaling pathway. Methods Cultured rat BMSCs were exposed to different concentrations of Sr, noggin (an inhibitor of BMP-2) or Smadl siRNA. The activity of alkaline phosphatase (ALP) in the exposed cells was detected by colorimetry, and the formation of mineralized nodules was observed with alizarin red staining. The expressions of phosphorylated (p) Smad1/5/8 and Runt-related transcription factor 2 (Runx2) in the cells were detected by Western blotting. Results Exposure to Sr at 0.1 to 10 mmol/L for 1 h markedly increased the expression of p-Smad1/5/8 in the BMSCs, and the increment was the most obvious following 1 mmol/L Sr exposure. Preconditioning with 100 ng/ml noggin for 2 h inhibited Sr-induced up-regulation of p-Smad1/5/8 expressions. Exposure of the cells to 0.1 to 5 mmol/L Sr for 6 h significantly enhanced Runx2 expression, and the peak enhancement occurred following 1 mmol/L Sr exposure. Transfection of the BMSCs with Smadl siRNA decreased the basal level of Smad1/5/ 8 protein expression, and also inhibited Sr-induced up-regulation of p-Smad1/5/8 and Runx2 expressions as well as Sr-induced enhancement of ALP activity and formation of mineralized nodules. Conclusion The BMP-2/Smad pathway is involved in Sr-induced osteoblast differentiation of rat BMSCs.%目的 探讨骨形态发生蛋白-2(BMP-2)/Smad通路在雷奈酸锶(Sr)促进大鼠骨髓间充质干细胞(BMSCs)分化为成骨细胞过程中的作用.方法 体外分离培养大鼠BMSCs,根据实验目的加入不同浓度Sr、BMP-2的拮抗剂noggin及Smad1小干扰RNA(SiRNA).酶标法检测碱性磷酸酶(ALP)活性,茜素红染色检测钙结节,Western blotting法检测磷酸化Smadl/5/8及Runt 相关转录因子-2(Runx2)蛋白的表达.结果 应用0.1~10 mmol/L Sr处理BMSCs细胞1h

  3. Digital subtraction radiographic analysis of the combination of bioabsorbable membrane and bovine morphogenetic protein pool in human periodontal infrabony defects

    Directory of Open Access Journals (Sweden)

    Maria do Carmo Machado Guimarães

    2010-08-01

    Full Text Available OBJECTIVES: This study assessed the bone density gain and its relationship with the periodontal clinical parameters in a case series of a regenerative therapy procedure. MATERIAL AND METHODS: Using a split-mouth study design, 10 pairs of infrabony defects from 15 patients were treated with a pool of bovine bone morphogenetic proteins associated with collagen membrane (test sites or collagen membrane only (control sites. The periodontal healing was clinically and radiographically monitored for six months. Standardized pre-surgical and 6-month postoperative radiographs were digitized for digital subtraction analysis, which showed relative bone density gain in both groups of 0.034 ± 0.423 and 0.105 ± 0.423 in the test and control group, respectively (p>0.05. RESULTS: As regards the area size of bone density change, the influence of the therapy was detected in 2.5 mm² in the test group and 2 mm² in the control group (p>0.05. Additionally, no correlation was observed between the favorable clinical results and the bone density gain measured by digital subtraction radiography (p>0.05. CONCLUSIONS: The findings of this study suggest that the clinical benefit of the regenerative therapy observed did not come with significant bone density gains. Long-term evaluation may lead to a different conclusions.

  4. Role of bone morphogenetic protein 2 in early acetabulum development and dysplastic acetabulum remodeling%BMP-2在髋臼软骨发育早期及发育不良髋臼软骨可逆性恢复过程中的作用研究

    Institute of Scientific and Technical Information of China (English)

    莫越强; 裴新红; 马瑞雪

    2015-01-01

    目的 研究BMP-2在髋臼软骨发育早期及发育不良髋臼软骨可逆性恢复过程中的作用.方法 通过伸髋内收、模拟襁褓体位固定新生大鼠双后肢,建立发育不良髋臼软骨模型.将髋臼标本经HE染色后观察比较正常及发育不良髋臼软骨组织形态学变化特点,同时用ELISA方法和PCR方法分别检测BMP-2、BMP-4、BMP-6、BMP-7的分泌及基因表达情况.将捆绑不同时间后的大鼠松绑,其中部分当场处死,其余大鼠继续喂养,最终至30日龄,建立发育不良髋臼软骨可逆性恢复模型.研究其髋臼软骨组织形态学恢复及BMP-2分泌变化情况.结果 正常大鼠髋臼软骨呈半圆形、容积大、表面光滑.发育不良髋臼软骨髋臼上缘肥厚,软骨发生变性,与周围组织分界不清.髋臼软骨BMP-2的分泌在正常大鼠7日龄和9日龄时出现高峰,分别为(13.7±0.29) ng/ml和(13.9±0.38) ng/ml.而在发育不良髋臼软骨中这一分泌高峰消失.在发育不良髋臼软骨可逆性恢复组,捆绑4d和6d的大鼠,BMP-2的分泌高峰出现延迟,都在15日龄时出现;而在捆绑8d及以上的大鼠,在松绑后继续喂养至30日龄,髋臼软骨组织形态无法恢复正常,并且BMP-2的分泌高峰未出现.结论 BMP-2的分泌可能是髋臼软骨早期发育情况的生物学标记之一.%Objective To explore the early-stage acetabulum development in normal and dysplastic acetabula and elucidate the function of bone morphogenetic protein 2 (BMP-2) in early acetabulum development and dysplastic acetabulum remodeling.Methods The rat model of dysplastic acetabulum was established by maintaining hips in a swaddling position.By analyzing the cartilage histologic characteristics,early-stage acetabulum developments were examined in normal and dysplastic acetabulum animals.Meantime,the mRNA expression and chondrocyte secretion of functional BMP-2,bone morphogenetic protein 4 (BMP-4),bone morphogenetic protein 6 (BMP-6) and bone

  5. 转化生长因子β1和骨形成蛋白2体外诱导成牙本质细胞分化%Transforming growth factor β1 and bone morphogenetic protein 2 induce the differentiation of odontoblasts in vitro

    Institute of Scientific and Technical Information of China (English)

    樊明文; 朱奇; 边专; 张旗

    2002-01-01

    目的观察转化生长因子β1(transforming growth factor β1, TGF-β1)和骨形成蛋白2(bone morphogenetic protein 2,BMP2)对体外培养的鼠牙乳头成牙本质细胞分化的影响. 方法取17 d胎龄小鼠下颌第一磨牙牙胚,胰蛋白酶消化分离牙乳头,置半固态培养基培养6 d,半固态培养基中加入重组TGF-β1或BMP2与肝素, 组织学观察. 结果 TGF-β1或BMP2加肝素可诱导牙乳头周边细胞发生极化,并分泌胞外基质.TGF-β1或BMP2单独加入时未见细胞极化,但基质分泌增加. 结论 TGF-β1和BMP2均能诱导成牙本质细胞的细胞学分化和分泌功能 .

  6. Combined application of vascularized pedicled pectoralis major clavicular periosteal flap and locking plate fixation plus bone morphogenetic protein implantation in the treatment of clavicular defect and nonunion after operation%带血管胸大肌蒂锁骨膜联合锁定钛板固定BMP植入治疗锁骨术后骨不愈合及骨缺损

    Institute of Scientific and Technical Information of China (English)

    孙强; 郑加法

    2009-01-01

    目的 探讨锁骨骨折术后骨缺损、骨折不愈合的临床特点,评价锁定钛板固定骨形态发生蛋白(bone morphogenetic protein,BMP)植入联合带血管胸大肌蒂锁骨膜转位的治疗效果.方法 2004年1月-2008年4月,锁骨骨折不愈合12例,平均年龄42.8岁,均行内固定物取出,清除骨折端纤维瘢痕及硬化骨,锁定钛板同定BMP植入,带血管胸大肌蒂锁骨膜转位覆盖,术后进行功能康复锻炼. 结果 12例患者术后均获得随访,时间8~24个月,平均1.2年,应用Constant-Murley肩关节评分系统进行评价,所有患者均在4~7个月达到临床愈合,部分患者已将内固定物取出. 结论 锁定钛板固定BMP植入联合带血管胸大肌蒂锁骨膜转位治疗锁骨骨缺损、骨不连,可取得良好的临床效果.%Objective To investigate the clinical characteristics of clavicular defect and nonunion after operation and evaluate the clinical effects of vascularized pedicled pectoralis major clavicular periosteal flap and locking plate fixation plus bone morphogenetic protein (BMP) implantation. Methods From January 2004 to April 2008, 12 patients (mean age, 42.8 years old) with clavicular defect and nonunion were treated in Zhongshan Hospital. The internal fixators were removed, and then fibrous scars and sclerotic bones were cleared. The clavicular detects were fixed with locking plate, with implantation of the BMP. The wound was covered with vascularized pedicled pectoralis major clavicular periosteal flap. Postoperative functional rehabilitation exercises were performed. Results All patients were followed up for 8-24 months (mean 1.2 years). The clinical results were evaluated by Constant-Murley scoring system. The clavicular defects were healed in all patients within 4-7 months, and the internal fixators were successfully removed from some of the patients. Conclusions Vascularized pedicled pectoralis major clavicular periosteal flap combined with locking plate fixation

  7. Injectable nano/chitosan/bone morphogenetic protein-2 induces periodontal tissue regeneration%注射型纳米壳聚糖骨形态发生蛋白2复合体诱导牙周组织的再生

    Institute of Scientific and Technical Information of China (English)

    巴格那; 陈华荣; 李婷; 谢富强; 鱼灵会

    2015-01-01

    BACKGROUND:Chitosan hydrogel has good biocompatibility, biodegradability and antibacterial property, which can promote tissue healing and induce bone formation. As a scaffold carrying growth factors, it can ensure the efficient and slow release of exogenous growth factors. OBJECTIVE: To observe the effect of injectable nano/chitosan/bone morphogenetic protein-2 composite to promote periodontal tissue regeneration in rats. METHODS:Fifty-four Wistar rats were randomized into three groups, and then chronic periodontitis model of the second molar was established. After modeling, injectable nano/chitosan/bone morphogenetic protein-2 composite was implanted into the periodontal tissue of the second molar in the experimental group; injectable nano/chitosan hydrogel was implanted in the control grouop; and nothing was implanted in the blank group. At 3, 6, 9 weeks after surgery, gingival bleeding index, probing depth, and tooth mobility were detected. X-ray and histopathological observations were carried out. RESULTS AND CONCLUSION:At 9 weeks after surgery, the probing depth and tooth mobility were both lower in the experimental group than the other two groups (P   目的:观察可注射性纳米壳聚糖骨形态发生蛋白2复合体促进大鼠牙周组织再生的效果。  方法:将54只Wistar大鼠随机均分为3组,建立第二磨牙慢性牙周炎模型,建模成功后,实验组于第二磨牙牙周内植入可注射性纳米壳聚糖骨形态发生蛋白2复合体,对照组于第二磨牙牙周内植入可注射性纳米壳聚糖凝胶,空白组牙周内不植入任何药物。术后3,6,9周,进行牙龈出血指数、探诊深度、松动度、X射线片及组织病理学切片观察。  结果与结论:实验组术后9周的探诊深度、松动度均低于对照组及空白组(P<0.05)。术后9周,实验组牙槽骨高度修复再生至根分叉处,骨小梁致密,分布均匀,可见大量牙骨质样结构、牙周

  8. 骨缝牵引中联合应用骨形态发生蛋白-2和骨保护素的实验研究%The effect of bone morphogenetic protein-2 and osteoprotegerin in trans-sutural distraction osteogenesis

    Institute of Scientific and Technical Information of China (English)

    姚玉胜; 黄华; 常世民; 王程越; 王桂君

    2012-01-01

    目的 探索上颌骨在骨缝牵引时加入缓释骨形态发生蛋白-2 (BMP-2)和骨保护素(OPG)对新骨形成的影响.方法 以24只杂种犬为研究对象,随机分为A、B、C组.通过手术在上颌骨腭横缝植入自制的新型牵引器.A、C组术后5d在牵引区附近注射缓释重组人骨形态发生蛋白-2/聚乳酸-羟基乙酸共聚物/纤维蛋白胶(rhBMP-2/PLGA/FS),B、C组在牵引3周后注射人骨保护素/纤维蛋白胶(rhOPG/FS).牵引1、2、4、6周后处死动物,采集标本进行组织学染色,并通过组织计量学方法检测腭横缝的组织改建情况.结果 A、C组骨缝区成骨细胞功能活跃,透射电镜显示细胞内有大量高尔基复合体、线粒体及粗面内质网.牵引6周时,A、B、C组成骨细胞指数分别为38.5±7.7、35.7±6.5、41.7±11.0,破骨细胞指数分别为5.9±1.0、1.2±0.3、2.8±0.4,骨小梁厚度分别为(38.36±13.28)、(66.20±9.16)、(51.85±9.92) μm;B、C组表现出骨密度增加及破骨细胞指数下降.结论 本实验所用牵引器能促进新骨生成;BMP-2与OPG在骨缝牵引过程中有协同作用,可以促进新骨形成及骨改建.%Objective To determine if locally administered bone morphogenetic protein-2 (BMP-2) and osteoprote-gerin (OPG) improved osteogenesis and new bone formation by trans-sutural distraction osteogenesis. Methods Twenty four dogs were divided into three groups randomly and received new internal trans-sutural distraction osteogenesis treat-ment. Five days after operation, infusion apparatus with double-tube was inserted to submucosa near the distracted zone to deliver controlled release agent of recombinant human bone morphogenetic protein-2/poly Qactic-co-glycolie acid)/ fibrin sealant (rhBMP-2/PLGA/FS) in group A and group C. Recombinant human osteoprotegerin/fibrin sealant (rhOPG/ FS) was injected three weeks later in group B and group C. Histology staining and bone histomorphometry were used to measure the changes of

  9. 人骨形态发生蛋白7基因转染对骨髓间充质干细胞增殖及成骨分化性能的影响%Human bone morphogenetic protein 7 gene transfection for the proliferation and osteogenetic differentiation of the bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    金丹; 裴国献; 王珂; 魏宽海; 陈滨

    2005-01-01

    BACKGROUND: The main aspect of the study in the bone histological engineering is how to maintain and improve theosteogenesis of the osteoblasts in vivo and in vitro. The gene transference may provide a new effective method to deal with theproblem.OBJECTIVE: To discuss the effect of the reverse transcription virus mediated human bone morphogenetic protein7(hBMP-7) gene transfection on the proliferation and osteogenetic differentiation of the bone marrow mesenehymal stemcells (BMSCs) of the rabbits.DESIGN: Cells taken as the study object, grouping control, repeat observation andmeasurement.SETTING: Traumatological and othopaedic lab of a medical university hospital.PARTICIPANTS: The study wascompleted in the Traumatological and Othopaedic Lab in the Affiliated Nanfang Hospital of the Southern Medical University from July 2001 to July 2003. Four New Zealand rabbits,whose weights varied from 1.0 to 1.5 kg, were provided without sexlimit by the Animal Experiment Center of the First Military Medical University of Chinese PLA.METHODS: The reverse transcription virus carriersof the hBMP-7 were constructed, and then the BMSCs were transfected by the virus containing target genes. The expression of the hBMP-7 protein was detected with the immunohistochemical method. The cell proliferation, cycle and ALP synthesis were respectively detected with the MTT method, flow cytometer and NPP method.MAIN OUTCOME MEASURES: Primary results: ① the detection results of the cell proliferation. ② the detection results of the ALP.Secondary results: ① the expression of the hBMP-7 protein in the transfected BMSCs. ② the detection results of the cell cycle.RESULTS: After the BMP-7 gene transfection, there was hBMP-7 positive expression in the BMSCs of the rabbits, using the immunohistochemical detection. There was no significant change in the BMSCs proliferation of the rabbits after the hBMP-7 gene transfection ( P > 0.05). Compared with the ALP synthesis of the transfected BMSCs(294

  10. 丝素蛋白增强型磷酸钙复合rhBMP-2用于绵羊腰椎椎体间融合的实验研究%Experimental study on lumbar interbody fusion with silk fibroin enhanced calcium phosphate cement composite loaded with recombinant human bone morphogenetic protein-2 in sheep

    Institute of Scientific and Technical Information of China (English)

    陈亮; 顾勇; 陈晓庆; 干旻峰; 朱雪松; 杨惠林; 唐天驷

    2010-01-01

    Objective To evaluate the osteogenic characteristics of an injectable silk fibroin (SF) enhanced calcium phosphate cement (CPC) composite loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2) on lumbar interbody fusion in sheep. Methods Twenty-four mature sheep were randomly divided into two groups. Each sheep underwent L1.2, L3.4 and L5.6 lumber interbody fusion, and the three disc spaces were randomly implanted with three of the following materials: SF/CPC, CPC/rhBMP-2, SF/CPC/rhBMP2 and autogenous iliac crest bone. One group was killed at 6 months and the other at 12 months. The fusion segments were observed and analyzed by manual palpation, CT scan, undestructive biomechanical testing, undecalcified histology, and histomorphology. Results The fusion rates of SF/CPC, CPC/rhBMP-2, SF/CPC/rhBMP-2 and autogenous bone assessed by manual palpation were 0, 33.33%, 55.56% and 77.78% respectively at 6 months. At 12 months, the fusion rates improved to 11.11%, 44.44%, 77.78% and 77.78%, respectively.The biomechanical results showed that fusion stiffness was significantly greater in autograft compared with SF/CPC/rhBMP-2, CPC/rhBMP-2, and SF/CPC in 4 degrees of freedom (flexion, extension, right bending, and left bending) at 6 months. The SF/CPC/rhBMP-2 composite showed similar stiffness as autograft, which was significantly greater than CPC/rhBMP-2 and SF/CPC at 12 nonths. Both CPC/rhBMP-2 and SF/CPC/rhBMP-2 showed significantly greater stiffness at 12 months compared with that of at 6 months. The results showed that bone volume was significantly greater in autograft compared with SF/CPC/rhBMP-2, CPC/rhBMP-2, and SF/CPC at 6 months. There was significant difference among ceramic residue among SF/CPC, CPC/rhBMP-2 and SF/CPC/rhBMP-2, with SF/CPC the greatest and SF/CPC/thBMP-2 the least. At 12 months, the bone volume of SF/CPC/rhBMP-2 composite was comparable with autograft, and greater than that of CPC/rhBMP-2 and SF/CPC. The bone volume of SF/CPC, CPC

  11. 骨形成蛋白2基因转染对犬牙髓细胞生物学特性的影响%The effect of gene transfection of bone morphogenetic proteins 2 on the cell biological characteristics of dog dental pulp cells in vitro

    Institute of Scientific and Technical Information of China (English)

    刁志虹; 高毅; 冯艳红; 李威

    2011-01-01

    目的 构建骨形态发生蛋白2 (bone morphogenetic proteins 2,BMP2)绿色荧融合蛋白pEGFP-N1-BMP2真核表达质粒,然后再用其在体外转染犬牙髓细胞(DDPCs)形成BMP2-DDPCs细胞,观察BMP2基因转染对牙髓细胞生物学特性的影响.方法 构建pEGFP-N1 -BMP2真核表达质粒,采用阳离子脂质体转染法将BMP2基因转染体外培养的DDPCs,检测BMP2-DDPCs碱性磷酸酶(ALP)活性,骨钙素(OC)产量和Ⅰ型胶原合成量等指标的测定,研究BMP2基因转染对牙髓细胞生物学特性的影响.结果 成功构建pEGFP-N1 -BMP2真核表达质粒,对构建的BMP2真核重组质粒用XhoⅠ、HindⅢ进行双酶切,其产物进行琼脂糖凝胶电泳后,在1.2、4.7 kb可见两条特异条带;并进行全基因序列测序,报告100%符合,证明pEGFP-N1-BMP2重组质粒构建成功.BMP2-DDPC中ALP活性,OC产量和Ⅰ型胶原合成量增加,与对照组DDPCs、N1 -DDPCs相比差异有统计学意义(P<0.05),通过增强ALP、OC和Ⅰ型胶原等成骨标志物的表达,促进牙髓细胞向成牙本质细胞分化.结论 pEGFP-N1 -BMP2真核表达质粒转染后的牙髓细胞,具有成牙本质细胞的特征.%Objective To construct the eukaryotic expression plasmid of bone morphogenetic proteins 2 (BMP2) and pEGFP-Nl-BMP2, then, using the plasmid to transfect dog dental pulp cells (DDPCs) to form BMP2-DDPCs in vitro so as to observe the effect of BMP2 gene transfection on the cell biological characteristics of dog dental pulp cell. Methods The full length dog BMP2 cDNA was linked into an eukaryotic expression vector pEGFP-Nl. DDPCs were transfected with pEGFP-Nl-BMP2 by lipofectamine transfection. The activity of alkaline phosphatase (ALP) in BMP2-DDPCs was detected,and the contents of type I collagen and osteocalcin (OC) in BMP2-DDPCs were measured. Results The pEGFP-Nl-BMP2 eukaryotic expression plasmid was constructed successfully. The constructed pEGFP-Nl-BMP2 could produce 4. 7kb and 1. 2kb fragments

  12. Development of a morphogenetically active scaffold for three-dimensional growth of bone cells: biosilica-alginate hydrogel for SaOS-2 cell cultivation.

    Science.gov (United States)

    Müller, Werner E G; Schröder, Heinz C; Feng, Qingling; Schlossmacher, Ute; Link, Thorben; Wang, Xiaohong

    2015-11-01

    Polymeric silica is formed from ortho-silicate during a sol-gel formation process, while biosilica is the product of an enzymatically driven bio-polycondensation reaction. Both polymers have recently been described as a template that induces an increased expression of the genes encoding bone morphogenetic protein 2 (BMP-2) and osteoprotegerin in osteoblast-related SaOS-2 cells; simultaneously or subsequently the cells respond with enhanced hydroxyapatite formation. In order to assess whether the biocompatible polymeric silica/biosilica can serve as a morphogenetically active matrix suitable for three-dimensional (3D) cell growth, or even for 3D cell bioprinting, SaOS-2 cells were embedded into a Na-alginate-based hydrogel. Four different gelatinous hydrogel matrices were used for suspending SaOS-2 cells: (a) the hydrogel alone; (b) the hydrogel with 400 μM ortho-silicate; (c) the hydrogel supplemented with 400 μM ortho-silicate and recombinant silicatein to allow biosilica synthesis to occur; and (d) the hydrogel with ortho-silicate and BSA. The SaOS-2 cells showed an increased growth if silica/biosilica components were present in the hydrogel. Likewise intensified was the formation of hydroxyapatite nodules in the silica-containing hydrogels. After an incubation period of 2 weeks, cells present in silica-containing hydrogels showed a significantly higher expression of the genes encoding the cytokine BMP-2, the major fibrillar structural protein collagen 1 and likewise of carbonic anhydrase. It is concluded that silica, and to a larger extent biosilica, retains its morphogenetic/osteogenic potential after addition to Na-alginate-based hydrogels. This property might qualify silica hydrogels to be also used as a matrix for 3D cell printing.

  13. 医用生物蛋白胶对注射型骨形态发生蛋白成骨活性的影响%Influence of fibrin sealant on osteoinductive ability of inject-type bone morphogenetic protein

    Institute of Scientific and Technical Information of China (English)

    王登虎; 刘建; 李丹; 胡蕴玉; 袁志

    2002-01-01

    Objective To observe influence of fibrin sealant(FS) on osteoinductive ability of inject type BMP.Method The inject type BMP power was dissolved in the main glue part or thrombin part of FS, then mixed with the main glue part or thrombin part of FS into gel, observe coagulating time, then implant composite into the thigh muscle pouch of mice to evaluate their capacity to induce new bone formation, and compared to the single BMP implant group.Result There was no difference in the coagulating time between two mixing method, the osteoinductive ability of implants BMP dissolved in the main glue part or thrombin part of FS group was higher than that of simply BMP implant group.Conclusion FS was perfect carrier to inject type BMP.

  14. Combined Effects of Androgen and Growth Hormone on Osteoblast Marker Expression in Mouse C2C12 and MC3T3-E1 Cells Induced by Bone Morphogenetic Protein

    Science.gov (United States)

    Kimura, Kosuke; Terasaka, Tomohiro; Iwata, Nahoko; Katsuyama, Takayuki; Komatsubara, Motoshi; Nagao, Ryota; Inagaki, Kenichi; Otsuka, Fumio

    2017-01-01

    Osteoblasts undergo differentiation in response to various factors, including growth factors and steroids. Bone mass is diminished in androgen- and/or growth hormone (GH)-deficient patients. However the functional relationship between androgen and GH, and their combined effects on bone metabolism, remains unclear. Here we investigated the mutual effects of androgen and GH on osteoblastic marker expression using mouse myoblastic C2C12 and osteoblast-like MC3T3-E1 cells. Combined treatment with dihydrotestosterone (DHT) and GH enhanced BMP-2-induced expression of Runx2, ALP, and osteocalcin mRNA, compared with the individual treatments in C2C12 cells. Co-treatment with DHT and GH activated Smad1/5/8 phosphorylation, Id-1 transcription, and ALP activity induced by BMP-2 in C2C12 cells but not in MC3T3-E1 cells. The insulin-like growth factor (IGF-I) mRNA level was amplified by GH and BMP-2 treatment and was restored by co-treatment with DHT in C2C12 cells. The mRNA level of the IGF-I receptor was not significantly altered by GH or DHT, while it was increased by IGF-I. In addition, IGF-I treatment increased collagen-1 mRNA expression, whereas blockage of endogenous IGF-I activity using an anti-IGF-I antibody failed to suppress the effect of GH and DHT on BMP-2-induced Runx2 expression in C2C12 cells, suggesting that endogenous IGF-I was not substantially involved in the underlying GH actions. On the other hand, androgen receptor and GH receptor mRNA expression was suppressed by BMP-2 in both cell lines, implying the existence of a feedback action. Collectively the results showed that the combined effects of androgen and GH facilitated BMP-2-induced osteoblast differentiation at an early stage by upregulating BMP receptor signaling. PMID:28067796

  15. Bone morphogenetic protein 2 (bmp2) and krüppel-like factor 9 (klf9) cross-regulation in uterine stromal cells promotes timing of uterine endometrial receptivity

    Science.gov (United States)

    Our laboratory has identified a novel progesterone receptor (PGR) co-activator protein, designated Krüppel-like Factor 9 (KLF9), whose absence in mice is associated with subfertility with decreased number of implanting embryos due to altered patterns of proliferation, apoptosis and aberrant P-respon...

  16. Core-shell Hydroxyapatite Combined with Bone Morphogenetic Protein 2 in Periodontal Regeneration Treatmentin Dogs%贝壳多孔羟基磷灰石基骨修复材料和骨形成蛋白-2联合应用引导犬牙周组织再生效果初步评估

    Institute of Scientific and Technical Information of China (English)

    陈斌; 吴文蕾; 孙卫斌; 韩方凯; 张其清

    2011-01-01

    目的 初步评估贝壳多孔羟基磷灰石基骨修复材料及该材料和骨形成蛋白-2联合应用引导比格犬牙周组织再生的效果.方法 选取18月龄比格犬6只,牙周基础治疗后1周,在下颌第二、三、四前磨牙,建立急性牙周骨缺损模型,依照分组情况进行不同治疗.实验组(T组)植入骨修复材料和骨形成蛋白-2;阴性对照组(NC组)植入骨修复材料;空白对照组(BC组)不植入任何材料.实验设计采取同颌同名牙对照,同一只比格犬的3对同颌同名牙分别为:空白对照组和阴性对照组,阴性对照组和实验组,空白对照组和实验组.术后12周,处死动物,Micro-CT检查并对数据进行统计学分析.结果 材料植入后,未见材料溢出,植入局部和全身都未见明显不良反应.3组缺损都有一定程度骨再生,以T组再生组织量最多,BC组最少.Micro-CT结果显示:T组、NC组和BC组的骨再生平均高度为(4.50±0.47) mm、(1.75±0.42) mm和(0.87±0.31) mm.NC组和BC组相比,差异有统计学意义(P<0.05).T组与NC组和BC组相比,差异均有统计学意义(P<0.05),且有临床意义.结论 贝壳多孔羟基磷灰石基骨修复材料和骨形成蛋白-2联合应用于比格犬,可以获得更好的引导组织再生效果.%Objective To evaluate the ability of core-shell hydroxyapatite bone graft material alone and combined with bone morphogenetic protein 2 ( BMP-2) in periodontal regeneration treatment in dogs. Methods Thirty-six defects were created in six 18-months male beagle dogs at the sites of the second, third and fourth mandibular premolars one week later after the dogs were treated with non-surgical periodontal therapy. Different treatments were carried out according to which group the teeth belonged. There are 3 groups. The test group (group T) was treated with core-shell hydroxyapatite bone graft material combined with BMP-2; the negative control group (group NC) was treated with core-shell hydroxyapatite

  17. S-Layer Architectures: Extending the Morphogenetic Potential of S-Layer Protein Self Assembly

    Science.gov (United States)

    2012-07-11

    architectures :  Extending  the  morphogenetic  potential  of   S-­‐layer  protein  self...ray  scattering  and   a   fractal  mean  force  concept:  From  protein  structure  to  nano-­‐disc  assemblies  ...  39...developed   a   broad   range   of   nanometer   scale   architectures   based   on   the   self-­‐ assembly   of  

  18. Dynamic expressions of kielin/chordin-like protein in mouse model of liver fibrosis and changes after bone morphogenetic protein-7 intervention%kielin/chordin样蛋白在肝纤维化模型中的动态表达及骨形成蛋白-7干预后的表达变化

    Institute of Scientific and Technical Information of China (English)

    宋梅; 陈永平; 陈少隆; 陈达之; 阳韬; 林镯

    2012-01-01

    Objective To observe the dynamic expressions of kielin/chordin-like protein (kcp) in mouse model of hepatic fibrosis and the effects of bone morphogenetic protein-7 (BMP-7)intervention on the expressions,and to explore a new target for treatment of fibrosis.Methods A total of 50 healthy male ICR mice were divided into three groups:control group(n=10) ; model group (n=30) and BMP-7 treatment group (n=10).The model group was further divided into three subgroups according to different time points:subgroups of 4,8 and 12 weeks with 10 mice in each subgroup.The mouse model of hepatic fibrosis was established by hypodermic injection of carbon tetrachloride(CCl4 ).The mice in BMP-7 treatment group began to receive human recombmant BMP- 7via intraperitoneal injection after 8 weeks of the first administration of CC14 and lasted for 4 weeks.The serum levels of alanine aminotransferase (ALT),aspartate transaminase (AST) and albumin (Alb) were detected.The pathological changes of liver were observed under optical microscope after HE and Masson staining.The dynamic expressions of kcp mRNA and protein of each group were detected by reverse transcription-polymerase chain reaction,immunohistochemistry and Western blot.The comparison of means among groups was done by univariate ANOVA.Results In model group,ALT and AST levels increased,while Alb level gradually decreased,and peaked at week 12.BMP-7treatment could reduce the changes,and there were significant differences among groups (F=23.501,34.600 and 16.244,respectively; all P<0.05).In normal control group,the expressions of kcp mRNA and protein were low,while those in model group were gradually increased.BMP-7 treatment could achieve remission and the changes were all significantly different among groups (F=30.362 and 10.727,respectively; P<0.01 or 0.05).The expression of kcp mRNA was positively correlated with levels of transforming growth factor (TGF)-β1 mRNA and BMP-7 mRNA (r=0.760 and 0.769,respectively; both P<0

  19. Molecular mechanism of bone formation and regeneration

    Institute of Scientific and Technical Information of China (English)

    Akira Yamaguchi

    2008-01-01

    @@ Bone formation and regeneration are mediated by the coordinate action of various factors. Among these, bone morphogenetic protein (BMP) and runt-related gene 2 (Runx2) play crucial roles in bone formation.

  20. 转化生长因子β1(TGF-β1)和骨形成蛋白2(BMP2)体外联合诱导成牙本质细胞样细胞分化%Transforming Growth Factor β1 (TGFβ1) and Bone Morphogenetic Protein 2 (BMP2) Induce Odontoblast-like Cell Differentiation in vitro

    Institute of Scientific and Technical Information of China (English)

    朱奇; 樊明文; 张旗; 陈智; 边专

    2005-01-01

    目的:观察转化生长因子β1(transforming growth factor β1, TGF-β1)和骨形成蛋白2(bone morphogenetic protein 2,BMP2)联合应用对体外培养的鼠牙乳头成牙本质细胞分化的影响.方法:取17 d胎龄小鼠下颌第一磨牙牙胚,胰蛋白酶消化分离牙乳头,置半固态培养基培养6 d,半固态培养基中单独加入重组TGFβ1、BMP2,或分别与肝素联合,或TGFβ1和BMP2联合,组织学观察牙乳头的形态学变化.结果:TGF-β1或BMP2 单独加入时未见细胞极化, 但基质分泌增加.TGF-β1或BMP2 加肝素可诱导牙乳头周边细胞发生极化,并分泌胞外基质.半固态培养基中同时加入 TGF-β1和BMP2的牙乳头培养6 d后,牙乳头周边细胞出现极化和功能性分化,牙乳头周边胞外基质沉积明显,且可见牙乳头尖形态的维持及从牙乳头尖至牙乳头基底部成牙本质细胞分化梯度的维持.结论:本研究结果证实,在没有内釉上皮和完整的基底膜存在的情况下,TGF-β1和BMP2加肝素可诱导培养的牙乳头出现成牙本质细胞分化,并促进胞外基质的分泌.TGF-β1和BMP2 均能诱导成牙本质细胞的细胞学分化和分泌功能,二者联合应用可协同发挥作用增强诱导效果.

  1. Effects of bone morphogenetic proteins 2 on neural stem cells of 14-day-old fetal rat telencephalon differentiating into cholinergic neurons%骨形态发生蛋白2对14d胎鼠端脑神经干细胞分化为胆碱能神经元的影响

    Institute of Scientific and Technical Information of China (English)

    梁军; 李明秋; 赵富生; 董建将; 李月珍

    2011-01-01

    BACKGROUND: Bone morphogenetic protein-2 (BMP2) may be an extracellular regulatory factor involved in cholinergic differentiation of neuronal precursor cells. OBJECTIVE: To explore the effects of BMP2 on neural stem cells of 14-day-old fetus rat telencephalons induced into cholinergic neurons. METHODS: Fetus telencephalons of 14-day-old SD rats were isolated, and tissues were divisively treated with collagenase typeⅠcontained EDTA following trituration. The cells were raised in polylysine culture plates with serum-free medium. Primary culture medium was changed half after 24 hours, and 10 μg/L BMP2 was added. RESULTS AND CONCLUSION: The adherent monoculture neural stem cells could been obtained using collagenase. The Nestin positve cells were obtained, and the purity was over 99%. After induced by BMP2, the ChAT positve cells were obtained, and the purity was over 97%. BMP2 can induce neural stem cells of 14-day-old fetus rat telencephalons into cholinergic neurons.%背景:骨形态发生蛋白2可能是参与胆碱能神经元前体细胞分化的细胞外调控因子.目的:观察骨形态发生蛋白2在孕14 d胎鼠端脑神经干细胞诱导成胆碱能神经元过程中的作用.方法:取孕14 d胎鼠端脑,用含EDTA的胰酶和Ⅰ型胶原酶消化,无血清培养基培养细胞,种植于涂有多聚赖氨酸的培养板,细胞原代培养24 h后半量换液,加入10 μg/L骨形态发生蛋白2继续培养.结果与结论:胶原酶消化得到的神经干细胞呈单层贴壁生长;Nestin免疫荧光鉴定细胞为阳性,获取的神经干细胞纯度大于99%;ChAT免疫荧光鉴定骨形态发生蛋白2可以将孕14 d胎鼠端脑神经干细胞诱导成胆碱能神经元,细胞纯度大于97%.

  2. 人骨形态发生蛋白2和人成纤维细胞生长因子2双基因共表达腺病毒载体转染人骨髓基质干细胞后的细胞增殖%In vitro proliferation of human bone marrow stromal cells after transfection by denovirus vector co-expressing human bone morphogenetic protein-2 and human fibroblast growth factor 2 genes

    Institute of Scientific and Technical Information of China (English)

    郭伟韬; 王辉; 刘思景; 曾荣; 肖启贤; 陈子秋; 黄云; 王斌; 胡资兵

    2012-01-01

    背景:利用重组腺病毒载体转染外源性基因到组织工程骨的种子细胞是骨缺损基因治疗研究的热点.目的:用人骨形态发生蛋白2和人成纤维细胞生长因子2双基因共表达腺病毒载体转染人骨髓基质干细胞,以探讨基因转染对人骨髓基质干细胞增殖的影响.方法:将Ad-hBMP2-IRES-hFGF2转染至人骨髓基质干细胞中,荧光显微镜观察转染效果,RT-PCR方法观察人骨形态发生蛋白2 cNDA和人成纤维细胞生长因子2 cNDA在人骨髓基质干细胞中的转录情况,Wester blot 方法检测人骨形态发生蛋白2和人成纤维细胞生长因子2蛋白表达情况,锥虫蓝测定细胞活力,流式细胞仪分析其对细胞增殖的影响.结果与结论:转染后人骨形态发生蛋白2和人成纤维细胞生长因子2基因在mRNA水平和蛋白水平均有表达,细胞活力无明显变化,流式细胞仪分析细胞周期中增殖细胞比例明显增加.说明该双基因可高效转染人骨髓基质干细胞,且促进细胞增殖.%BACKGROUND: Transfection of exogenous gene into tissue-engineered seed cells via recombinant adenovirus vector is the key to gene therapy of bone defects.OBJECTIVE: To investigate the effect of gene transfection on the proliferation of human bone marrow stromal stem cellsthat tranfected with adenovirus vectors co-expressing human bone morphogenetic protein-2 (hBMP-2) and humanfibroblast growth factor 2 (hFGF-2).METHODS: The Ad-hBMP2-IRES-hFGF2 plasmids were transfected into human bone marrow stromal stem cells. Theefficiency of transfection was evaluated by fluorescence microscope. Reverse transcriptase polymerase chain reactionwas used to observe the successful transcription of hBMP-2 and hFGF-2 cNDA in bone marrow mesenchymal stem cells.Western blot assay was used to identify the protein expression of hBMP-2 and hFGF-2 genes. The cellular viability wasdetermined by trypan blue staining and the changes of the cell proliferation were

  3. 7-Dehydrocholesterol reductase regulated the palatal development by the sonic hedgehog-bone morphogenetic protein 2 signal pathway%7-脱氢胆固醇还原酶基因沉默对体外培养腭突音猬基因-骨形成蛋白2信号通路的影响

    Institute of Scientific and Technical Information of China (English)

    张岱尊; 许尧祥; 肖文林; 庄翠竹

    2014-01-01

    目的 研究沉默7-脱氢胆固醇还原酶(7-dehydrocholesterol reductase,Dhcr-7)表达对体外培养腭突器官中音猬基因(sonic hedgehog,Shh)-骨形成蛋白2(bone morphogenetic protein 2,BMP-2)信号通路的影响,探讨Dhcr-7参与腭突发育的信号通路.方法 取60只孕期(gestation day,GD) 13.5d小鼠胚胎根据简单随机抽样法平均分为3组:空白对照组(A组):不含胆固醇培养基培养腭突;Dhcr-7基因沉默组(B组):不含胆固醇培养基培养腭突+Dhcr-7-siRNA腺病毒;添加胆固醇组(C组);每组各20只.培养48 h后,A、B组更换不含胆固醇培养基,C组更换含有600 mg/L胆固醇培养基.继续培养72 h后,分别将腭突固定,组织染色和扫描电镜观察其形态变化;分别提取腭突RNA和蛋白质,应用反转录-聚合酶链反应(reverse transcriotion-polymerase chain reaction,RT-PCR)和蛋白质印迹法检测Dhcr-7、Shh和BMP-2表达量的变化.结果 组织染色和扫描电镜显示A组及C组腭突能完全融合,B组腭突未融合.Shh和BMP-2在B组的mRNA和蛋白质的表达量随Dhcr-7表达量降低而降低.B组mRNA和蛋白质的表达量Shh为0.063±0.018和0.092±0.065;BMP-2为0.054±0.018和0.049±0.021;A组mRNA和蛋白质的表达量Shh为0.667±0.093和0.639±0.078;BMP-2为0.591±0.043和0.569±0.081.A、B两组Shh和BMP-2的mRNA和蛋白质的表达量差异分别具有统计学意义(P<0.05);C组Dhcr-7的mRNA表达量(0.074±0.034)和蛋白质表达量(0.075±0.028)基本无变化,与B组(Dhcr-7的mRNA表达量为0.083±0.045;蛋白质表达量为0.067±0.065)相比,差异无统计学意义(P>0.05);RNA和蛋白质的表达量Shh(0.649±0.085和0.608±0.092)和BMP-2(0.578±0.062和0.548±0.065)均明显升高,与B组相比差异有统计学意义(P<0.05).结论 Dhcr-7可影响Shh和BMP-2的表达,Dhcr-7通过Shh-BMP-2信号通路调控腭突发育.%Objective To investigate the effect of 7-dehydrocholesterol reductas(Dhcr-7) gene silencing on the palatal

  4. Effect of bovine bone morphogenetic proteins on radius fracture healing in rabbits Efeito de proteínas morfogenéticas ósseas de origem bovina na consolidação de fraturas induzidas no rádio de coelhos

    Directory of Open Access Journals (Sweden)

    Alfredo Feio da Maia Lima

    2007-08-01

    Full Text Available PURPOSE: To investigate the effect of bovine bone morphogenetic proteins (bBMPs bound to hydroxyapatite plus collagen in the healing of unstable radius fractures. METHODS: A transverse fracture was induced at the mid of the diaphysis in both radii on 15 Norfolk rabbits with average age of 5.5 months and 3.5kg. A mixture of bBMPs bound to thin powdered hydroxyapatite (bBMP-HA and bovine collagen as agglutinant was applied to the right radius fracture site. The left radius fracture was considered control and no treatment was used. After 30, 60 and 90 days (5 rabbits/period the rabbits were euthanized and the radii were collected for histological analysis. RESULTS: The descriptive histological analysis revealed that repair was similar for both forelimbs. The histomorphometric analysis showed that the mean area of newly formed bone was 867442.16 mm², 938743.00 mm² and 779621.06 mm² for the control forelimbs, and 841118.47 mm², 788038.76mm² and 618587.24 mm² for the treated forelimbs at 30, 60 and 90 days, respectively. Thus the newly formed bone area was 12.17% larger in the forelimbs treated with bBMP-HA/collagen than in the control forelimbs (pOBJETIVO: Investigar a influência de Proteínas Morfogenéticas Ósseas de origem bovina (bBMPs ligadas a hidroxiapatita mais colágeno na consolidação de fraturas instáveis do rádio. MÉTODOS: Em 15 coelhos com aproximadamente 5,5 meses de idade e peso médio de 3,5kg foi realizada uma fratura transversa na porção média da diáfise do rádio de ambos os membros. Na fratura do rádio direito foi aplicada mistura de bBMPs ligadas à hidroxiapatita (bBMP-HA e colágeno bovino como aglutinante e na do rádio esquerdo, considerada controle, nenhum tratamento foi usado. Os coelhos (cinco por período foram submetidos à eutanásia aos 30, 60 e 90 dias após a cirurgia para realização do processamento histológico e análise microscópica. RESULTADOS: A análise histológica descritiva revelou que

  5. Bone morphogenetic proteins and the polycystic ovary syndrome

    NARCIS (Netherlands)

    E.L.A.F. van Houten (Leonie); J.S.E. Laven (Joop); Y.V. Louwers (Yvonne); A. McLuskey; A.P.N. Themmen (Axel); J.A. Visser (Jenny)

    2013-01-01

    textabstractBackground: Polycystic Ovary Syndrome (PCOS) is defined by two out of the following three criteria being met: oligo- or anovulation, hyperandrogenism, and polycystic ovaries. Affected women are often obese and insulin resistant. Although the etiology is still unknown, members of the Tran

  6. 重组人骨形态发生蛋白2缓释体对铬磨损颗粒诱导的溶骨效应的影响%Slow-release recombinant human bone morphogenetic protein-2 suppresses chromium wear particle-induced osteolysis in rats

    Institute of Scientific and Technical Information of China (English)

    李干; 李奇; 林荔军; 段鑫; 张西旗

    2012-01-01

    Objective To observe the effect of a slow-release recombinant human bone morphogenetic protein-2 (rhBMP-2) formulation on the expressions of receptor activator of nuclear factor-KB ligand (RANKL) and osteoprotegerin (OPG) in a murine air pouch model of bone implantation. Methods A cranial bone allograft was implanted in the air pouch induced on the back of the recipients. The rat models were then randomized into 5 groups, including a blank control group, chromium particle group, and 3 rhBMP-2 groups receiving 50,100 or 200 μg/L slow-release rhBMP-2 in addition to chromium particles. Three weeks later, the expressions of RANKL and OPG in the air pouch was detected using Western blotting and RT-PCR, and the positively stained area for osteoclasts in the bone graft was determined with TRAP staining for drug effect assessment. Results RANKL and OPG expressions were found in the air pouches in all the 5 groups. RANKL and OPG protein and mRNA expressions, RANKL/OPG ratio and osteoclast staining area in the bone graft were the highest in chromium particle group (P0.05). Conclusion Chromium particles can cause osteolysis by increasing the RANKL/OPG ratio in rats, and intervention with slow-release rhBMP-2 can significantly promote bone formation and suppress bone resorption by decreasing RANKL/OPG ratio.%目的 建立大鼠植骨气囊模型,观察在不同浓度重组人骨形态发生蛋白2(rhBMP-2)缓释体干预下气囊内组织的破骨细胞分化因子(RANKL)、骨破坏素(OPG)表达情况.方法 在大鼠背部注入空气形成气囊,取同源大鼠的颅骨植入气囊内.将已制成的植骨气囊模型大鼠分成5组:空白组、铬颗粒组、50 μg/L rhBMP-2缓释体+铬颗粒组、100 μg/L rhBMP-2缓释体+铬颗粒组和200 μg/L rhBMP-2缓释体+铬颗粒组.各组分别用药处理,2周后取出囊腔内组织进行RANKL、OPG的Wester-blot、Rt-PCR检测,并对囊腔内骨片行TRAP染色,用计算机图像分析技术测定骨片破骨细胞染

  7. The expression of uncoupling protein 2 and bone morphogenetic protein 2 in the kidney of iodine deficiency rats%碘缺乏大鼠肾脏解耦联蛋白2与骨形态发生蛋白2的表达

    Institute of Scientific and Technical Information of China (English)

    田秀标; 韩颖; 刘艳; 房辉; 阎玉琴; 林来祥

    2013-01-01

    Objective To detect the expression of uncoupling protein 2 (UCP2) and bone morphogenetic protein 2 (BMP2) in the kidney of Wistar rats,and explore the changes and distribution of them.Methods Female Wistar rats were divided into mild iodine deficiency group (MIDG group),severe iodine deficiency group(SIDG group) and normal iodine group as control (CL group) with radom number table method,and fed adaptively for one week before the experiment with ten rats in each group (everyday total iodine intaking were 5.00,1.24,10.00 μg/d,respectively).Observed the expression of UCP2 and B MP2 in isolated kidney of the experimental rats through immunohistochemistry.And the results were analyzed with related statistic methods.Results Compared with CL group,the level of free T3,total T3,total T4 in MIDG group declined (t =2.54,1.81,4.41,P < 0.05),while the level of free T4 did not decrease obviously,and the difference had no statistical significance.And the level of free T3,free T4,total T3,total T4 in SIDG group declined evidently (t =6.68,20.25,5.38,21.56,P < 0.01).Compared with MIDG group,the level ofblood hormone in SIDG group declined more obviously (t =3.19,15.45,6.93,13.48,P < 0.05).Immunohistochemistry showed that UCP2 and BMP2 expressed mainly in the cortical distal renal tubular and less in glomeruli.Compared with CL group,the expression of UCP2 and BMP2 in SIDG group and MIDG group were both decreased significantly (t =5.72,8.79,8.74,18.63,P < 0.01).Compared MIDG group,the expression in SIDG group declined more obviously (t =7.43,11.77,P < 0.01).Meanwhile,the relevant analysis revealed that the tendencies of decline of UCP2 and BMP2 were consistent with the level of free T4 (r =0.81,0.82).Conclusion The deficiency of iodine can cause hypothyroidism which can lead to the decline of UCP2 and BMP2 in kidney of rats.%目的 通过测定碘缺乏大鼠肾脏解耦联蛋白(UCP)2、骨形态发生蛋白(BMP)2的表达,探讨其表达的改变与分布.方法

  8. BMP 活性多肽复合羟基磷灰石聚乳酸在大鼠体内异位成骨的实验研究%A study on the ectopic osteogenesis experiment of bone morphogenetic protein active polypeptide combined with hydroxyapatite and poly lactic acid in rats

    Institute of Scientific and Technical Information of China (English)

    王硕; 谢惠敏; 甘少磊; 任卫卫; 陈秉耀; 张增亮; 韦兴

    2015-01-01

    Objective To evaluate osteoinductive potential of bone morphogenetic protein ( BMP ) III and BMPIV active polypeptides in animals by detecting ectopic bone formation ability of the 2 active polypeptides in different concentrations in rats, and to elect the best polypeptide in the best concentration by comparing with the osteoinductive activity of 0.4 g / L BMPI.Methods A total of 48 Sprague-Dawley ( SD ) rats were randomly divided into 8 groups ( groups A, B, C, D, E, F, G, H ), with 6 rats in each group. The composite materials of 0.2 g / L, 0.4 g / L and 0.8 g / L BMPIII and 0.2 g / L, 0.4 g / L and 0.8 g / L BMPIV with hydroxyapatite and poly lactic acid ( HA / PLA ) were implanted into superifcial muscles of the hips of the rats in group A, B, C, D, E and F respectively. The rats in the experimental control group ( group G ) received the composite materials of 0.4 g / L BMPI with HA / PLA and the rats in the blank control group ( group H ) only received HA / PLA. At 3 weeks after the operation, the anteroposterior X-ray films of the back were taken. The X-ray and CT images were observed at 5 weeks after the operation. The samples were taken out, stained by HE and Masson and observed under the light microscope at 3 and 5 weeks after the operation respectively. Using the method of nonparametric statistics, the osteogenic properties of the rats were analyzed at 5 weeks after the operation.Results All the rats underwent operation successfully, and their diets, activities and wound healing were ifne. All the rats were alive. The hardness of implantation materials was increased in each group. ( 1 ) According to the X-ray iflms, the following results were found. At 3 weeks after the operation, there were slightly fuzzy shadows in the implantation area, which were more obvious in group C and F. At 5 weeks after the operation, there were slight shadows in group A and D, which were more obvious in group C and F. There were no shadows in group H. ( 2 ) The CT images

  9. 15-zinc finger protein Bloody Fingers is required for zebrafish morphogenetic movements during neurulation.

    Science.gov (United States)

    Sumanas, Saulius; Zhang, Bo; Dai, Rujuan; Lin, Shuo

    2005-07-01

    A novel zebrafish gene bloody fingers (blf) encoding a 478 amino acid protein containing fifteen C(2)H(2) type zinc fingers was identified by expression screening. As determined by in situ hybridization, blf RNA displays strong ubiquitous early zygotic expression, while during late gastrulation and early somitogenesis, blf expression becomes transiently restricted to the posterior dorsal and lateral mesoderm. During later somitogenesis, blf expression appears only in hematopoietic cells. It is completely eliminated in cloche, moonshine but not in vlad tepes (gata1) mutant embryos. Morpholino (MO) knockdown of the Blf protein results in the defects of morphogenetic movements. Blf-MO-injected embryos (morphants) display shortened and widened axial tissues due to defective convergent extension. Unlike other convergent extension mutants, blf morphants display a split neural tube, resulting in a phenotype similar to the human open neural tube defect spina bifida. In addition, dorsal ectodermal cells delaminate in blf morphants during late somitogenesis. We propose a model explaining the role of blf in convergent extension and neurulation. We conclude that blf plays an important role in regulating morphogenetic movements during gastrulation and neurulation while its role in hematopoiesis may be redundant.

  10. 骨形态发生蛋白2聚乳酸缓释微球对兔骨髓间充质干细胞相容性研究%Bone morphogenetic protein 2 poly lactic acid sustained release microspheres for compatibility between rabbit bone marrow mesenchymal stem cell research

    Institute of Scientific and Technical Information of China (English)

    马立坤; 邓江; 黄文良; 叶鹏; 田仁元; 吕雪峰

    2015-01-01

    Objective To investigate the influence of recombinant human bone morphogenetic protein‐2(rhBMP‐2)of poly lactic acid(PLA) release microspheres for compatibility of rabbit bone marrow mesenchymal stem cells(BMSCs) .Methods The rh‐BMP‐2‐PLA release microspheres were prepared by w/o/w multiple emulsion volatilizing method and then cocultured BMSCs .The effects of rhBM P‐2‐PLA release microspheres on the cytotoxicity and relative proliferation rate by MTTassay .Evaluation of mate‐rials biocompatibility by inverted microscope and scanning electron microscope(SEM) .Results The rhBMP‐2‐PLA release micro‐spheres in various concentration of leaching solution and BMSCs training of uninfected cells .Experimental group and control group in 4 different time cell proliferation OD values by analysis of repeated measurement variance between time OD values were statisti‐cally significant(P=0 .000) ,the experimental group and control group OD values are statistically significant(P=0 .025) ,the exper‐imental group higher than the control group ,experimental group OD value time there was a significant interaction effect and the group number ,the change trend are obviously different(P=0 .006) .Inverted microscope to observe materials normal cell prolifera‐tion ,SEM found that vaccination cells surrounding rhBMP‐2‐PLA release microspheres of 7 days later ,the cells grew well and split proliferation activity .Conclusion rhBMP‐2‐PLA release microspheres of BMSCs has non‐toxic and has compatibility .%目的:观察重组人骨形态发生蛋白2(rhBMP‐2)聚乳酸(PLA)缓释微球对兔骨髓间充质干细胞(BMSCs)相容性。方法通过复乳干燥法制备rhBMP‐2‐PLA缓释微球,并与兔BMSCs共培养,采用MTT法检测rhBMP‐2‐PLA缓释微球对细胞的毒性及增殖,并通过倒置显微镜及扫描电镜等评价材料的细胞相容性。结果 rhBM P‐2‐PLA缓释微球各浓度浸提液与BM‐SCs培养得出

  11. BMP15外显子1 SNPs检测及其与邵伯鸡母系产蛋性状的关联性分析%Detection of SNPs of Bone Morphogenetic Protein 15 Gene Exon1 and Its Association with Egg Production Traits in Female Line of Shaobo Chicken

    Institute of Scientific and Technical Information of China (English)

    李春苗; 黎寿丰; 赵振华; 黄华云; 薛龙岗

    2012-01-01

    本研究旨在阐明骨形态发生蛋自15(Bone morphogenetic protein l5,BMP15)基因多态性与邵伯鸡母系产蛋性状之间的关系,为鸡繁殖性状的标记辅助选择提供科学依据.采用PCR-RFLP技术检测261只邵伯鸡母系BMP15的基因多态性,用最小二乘法分析该基因多态性与邵伯鸡母系产蛋性状的关系.发现BMP15基因外显子1序列中存在3个多态位点C397T、A474G和C594T,其中C397T位点C→T的突变使亮氨酸变为苯丙氨酸,经RFLP检测,3个多态位点均发现3种基因型.x2检验表明,邵伯鸡母系在这3个位点均处于Hardy-Weinberg平衡.用最小二乘法分析这3个位点的多态性与邵伯鸡母系产蛋性状之间的关系,结果发现,C397T位点TT基因型个体的开产日龄显著早于CT型个体(P<0.05),TT型个体的300日龄产蛋数显著高于CT型个体(P<0.05) ;A474G位点AA、AG和GG型个体间的各性状差异均不显著(P>0.05) ;C594T位点CC型个体的开产日龄显著早于CT与TT型个体.3个位点的合并基因型TTAATT对开产日龄、开产体质量、开产蛋质量、300日龄平均蛋质量、300日龄产蛋数均有显著影响(P<0.05).对于邵伯鸡母系而言,TTAATT是最有利基因型,本研究结果初步表明,BMP 15基因合并基因型TTAATT可以作为邵伯鸡母系产蛋性状潜在的DNA分子标记.%The objective of the present study were to elucidate the relationship between polymorphisms of bone morphogenetic protein 15 (BMP15) gene and egg production traits in female line of Shaobo chicken and to provide a scientific basis for marker-assisted selection for reproductive traits of chicken. Polymorphisms of exonl of BMP15 gene were detected in 261 female line of Shaobo chicken by PCR-RFLP. The relationship between polymorphisms of BMP15 gene and egg production traits in female line of Shaobo chicken was analyzed by least squares linear model. Three polymorphic sites C397T, A474G and C594T were found in exonl of BMP15, and C

  12. 重组人类骨形态蛋白2复合自固化磷酸钙材料在体内的血管化%Vascularization of autosetting calcium phosphate cultivated with recombinant human bone morphogenetic protein 2

    Institute of Scientific and Technical Information of China (English)

    王炜; 玛依拉·吾甫尔; 阿不都赛米·艾买提; 艾合买提江·玉素甫

    2011-01-01

    BACKGROUND: Deep fascia promotes vascularization of tissue-engineered bone, which is a mature technology, but different animal species and different implant materials can result in a great difference in vascularization.OBJECTIVE: To compare the vascularization of autosetting calcium phosphate cement cultivated with recombinant human bone morphogenetic protein 2 (rhBMP-2) and a single autosetting calcium phosphate cement implant in beagle dogs with deep fasica flaps. METHODS: Twelve healthy adult beagle dogs were respectively implanted with autosetting calcium phosphate cement cultivated with rhBMP-2 in the left thigh (experimental sides) and a single autosetting calcium phosphate cement in right thigh (control sides). The vascularization in each condition was assessed by experiment study (Physical, Massion stain, the hematoxylin-eosin stain, ⅧRAg marked) at time intervals of 2, 4, 8 and 16 weeks after operation.RESULTS AND CONCLUSION: In experimental groups and in control groups, vascularization was found. New vessels invaded in scaffold with time. In experimental groups, the amount of vessels and the expression of ⅧRAg were stronger than those in control groups at 2, 4, 8, and 16 weeks. The deep fasica flaps have great effect on the vascularization. The deep fasica flap binding with rhBMP-2 is proved to be the better in vascularization of autosetting calcium phosphate cement.%背景:深筋膜瓣促进组织工程骨血管化是一种成熟的技术,但动物种属不同、植入材料不同均会使血管化的结果产生较大的差异.目的:比较复合重组人类骨形态蛋白2的自固化磷酸钙人工骨和单纯的自固化磷酸钙人工骨在比格犬带蒂筋膜瓣内的血管再生能力.方法:分别将复合重组人骨形态蛋白2的自固化磷酸钙人工骨和单纯的自固化磷酸钙人工骨包裹于12只成年比格犬腰背部两侧带蒂深筋膜瓣中,于术后第2,4,8,16周各随机选取3只动物摘取血管化标本,进行大

  13. Effect of overexpression of vascular endothelial growth factor 165 on the osteogenic potential of bone morphogenetic protein%VEGF165过表达对BMP2成骨细胞分化影响的实验研究

    Institute of Scientific and Technical Information of China (English)

    张聪; 刘洪美; 李庆伟; 陈国武; 梁啸; 孟纯阳

    2015-01-01

    [目的]探讨腺病毒转染血管内皮生长因子165(vascular endothelial growth factor 165,VEGF165)对骨形态发生蛋白(bone morphogenetic protein 2,BMP2)促成骨细胞分化的抑制性作用研究.[方法]采用密度梯度离心法分离兔骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs),取第3代BMSCs进行细胞表型鉴定并作为细胞实验对象,使用Ad-BMP2和Ad-BMP2-VEGF165载体体外转染BMSCs,倒置荧光显微镜下观察GFP表达变化;同时ELISA和Westem blot检测BMP2和VEGF165蛋白的表达.然后,应用成骨细胞诱导培养液定向诱导BMSCs向成骨细胞分化.实验分3组:Ad-BMP2-VEGF165转染BMSCs组(Ad-BMP2-VEGF165组),Ad-BMP2转染BMSCs组(Ad-BMP2组),BMSCs组(对照组).分别在成骨细胞诱导培养后7、14、21 d,通过Real-time PCR分析ALP和OC mRNA相对表达水平和碱性磷酸酶(alkaline phosphatase,ALP)活性测定、骨钙素(Osteocalcin,OC)免疫组化染色来评价各组BMSCs成骨分化潜能的影响. [结果]第3代细胞表面高表达CD29(99.82%)、CD44 (94.14%),低表达CD14 (3.11%)、CD34 (0.34%);腺病毒转染后第5 h BMP2和hVEGF165蛋白水平表达最高;成骨细胞诱导培养14和21 d后,Ad-BMP2组成骨相关基因表达、OC免疫组化和ALP活性表达最高,结果与其他两组相比较差异具有统计学意义(P<0.05);而对照组表达最弱,与Ad-BMP2-VEGF165组比较差异具有统计学意义(P<0.05).[结论]腺病毒载体Ad-BMP2与Ad-BMP2-VEGF165转染BMSCs后,均具有明显促进BMSCs体外诱导成骨细胞分化潜能,但Ad-BMP2诱导作用更为显著,同时说明VEGF165可能对BMSCs成骨细胞分化起抑制作用.

  14. Poly(DL-lactic-glycolic acid)/tricalcium phosphate scaffolds loaded with recombinant human bone morphogenetic protein-2 chitosan microspheres for treatment of bone defects in rabbits%骨形态发生蛋白-2壳聚糖微球复合组织工程支架修复兔股骨髁部骨缺损的实验研究

    Institute of Scientific and Technical Information of China (English)

    黄鑫; 吕荣; 王军; 郝赋; 孟国林; 刘建; 袁志; 熊卓; 李国臣; 白峰; 禚文昆; 马煜

    2010-01-01

    Objective To investigate the feasibility and efficacy of using the poly(DL-lactic-glycolic acid)/tricalcium phosphate(PLGA/TCP)scaffolds loaded with recombinant human bone morphogenetic protein-2(rhBMP-2)chitosan microspheres to treat bone defects in rabbits.Methods The rhBMP-2 chitosan microspheres were incorporated into the PLGA/TCP scaffold.The composites of rhBMP-2 microspheres and the PLGA/TCP were then used to bridge the bone defects(diameter: 0.6 cm,depth: 1.0 cm)at the condyle of the left femur in rabbits.Altogether 45 rabbits were randomized into 5 groups in the present study.In group A(n = 10)the defects were not treated.The PLGA scaffold was used in group B(n = 10),the rhBMP-2/PLGA scaffold in group C(n = 10),and the rhBMP-2 micorsphere/PLGA scaffold in group D(n = 10).Group E(n = 5)was the normal control.Ⅹ-ray,Micro-CT and histological studies were conducted to identify new bone formation in groups A to D at 4 and 12 weeks after surgery.Results At 4 weeks postoperatively,plenty new bone and mature trabecular bone were observed,and most of the PLGA/TCP scaffolds were degraded in group D.At 12 weeks postoperatively,the PLGA/TCP scaffolds and chitosan microspheres were completely degraded,and the bone defects were filled with mature bone in group D.In addition,the ratio of new bone formation was significantly higher in group D(74.25%±8.91%)than in group B(5.78% ±1.21%)and group C(37.26% ±6.45%)(P < 0.05).Conclusion The PLGA/TCP composites loaded with rhBMP-2 microspheres are effective in stimulating new bone formation when used to bridge bone defects in rabbits,highlighting their potential application in clinical settings.%目的 探讨重组人骨形态发生蛋白-2(rhBMP-2)壳聚糖缓释微球复合聚乳酸-聚羟乙酸/磷酸三钙(PLGA/TCP)支架修复骨缺损的可行性和有效性.方法 取健康成年新西兰大白兔45只,在40只实验动物股骨髁部制备0.6 cm×1.0 cm骨缺损.实验分4组:A组:缺损组,B组:用PLGA/TCP空白

  15. Molecular Cloning and Sequence Analysis of Full-Length cDNA Encoding Human Bone Morphogenetic Protein-7%人骨形态发生蛋白-7全长基因cDNA的克隆和序列分析

    Institute of Scientific and Technical Information of China (English)

    李新友; 刘淼; 李曙明; 姚煜; 王全颖; 杨广笑

    2003-01-01

    Objective: To clone the full-length human bone morphogenetic protein-7 ( BMP-7) gene and analyse its sequence, to aid in investigation of its function and structure. Methods: TotalRNA was isolated from Chinese fetal kidney by the acid guanidinium thiocyanate phenol-chloroformmethod. Two overlapping segments of human BMP-7 cDNA were obtained by reverse transcription(RT)-PCR. Fallowing application, the two segments were ligated to each other and subcloned intoPGEM-7 easy vector to form PEGM-T easy/hBMP-7 recombinant plasmid. Sanger dideoxy chain-ter-mination method was used to sequence the cDNA. Results: There was 750 bp fragment obtained RT-PCR using # 2 primer from 5' end of BMP-7 gene (PCR by using # 2 and # 1) ,and 540 bp frag-merit from 3' end was generated by RT-PCR using # 4 primer (PCR using # 3 and # 4). Full-lengthcDNA encoding BMP-7 was obtained by religation of two segments. When compared with hBMP-7 se-quence in Gene bank (XM30619) ,our full-length BMP- 7 cDNA has a G instead of a T at nucleotide862. This change results in valine substituting for phenylalanine in the protein. Conclusion: This isthe first time that BMP-7 cDNA was successfully cloned from Chinese fetal kidney. BMP-7 cDNAplays an important role in healing injuries of the osteo-articular system. This makes BMP-7 is an at-tractive target for various clinical applications.%目的:克隆人骨形态发生蛋白-7(BMP-7)全长基因,并进行测序及序列分析,研究其结构和功能.方法:采用异硫氰酸胍一步法从人胎儿肾中提取总RNA,利用逆转录-聚合酶链反应(RT-PCR)的方法,分段扩增出hBMP-7全长基因cDNA,与PGEM-T easy连接成PGEM-T easy/hBMP-7重组质粒,采用Sanger双脱氧链终止法测定基因序列.结果:以4号特异引物引导的RT-PCR扩增出BMP-7基因3'端540 bp片段,以2号特异引物引导的RT-PCR扩增出BMP-7基因5'端750 bp片段,重组连接两片段得到BMP-7全长基因cDNA.与Genebank上发表的hBMP-7序列(XM30619)

  16. TiO2 nanotubes functionalized with recombinant human bone morphogenetic protein-2 enhance biological activity in vitro%二氧化钛纳米管阵列加载重组人骨形成蛋白2的体外生物活性研究

    Institute of Scientific and Technical Information of China (English)

    孙子环; 夏荣; 孙磊; 胡小晔; 闵曦; 徐基亮

    2015-01-01

    目的 探讨二氧化钛纳米管阵列加载重组人骨形成蛋白2 (recombinant human bone morphogenetic protein-2,rhBMP-2)对小鼠骨髓间充质干细胞(bone mesenchymal stem cell,BMSC)早期活性的影响,为钛种植体表面生物化学改性提供实验依据.方法 利用阳极氧化技术在纯钛片表面制备双层二氧化钛纳米管阵列,化学接枝rhBMP-2(实验组),以机械抛光的纯钛为空白对照组,阴性对照A组为二氧化钛纳米管组、阴性对照B组为二氧化钛纳米管+羰基二咪唑组.用场发射扫描电镜观察各组形貌并用X射线光电子能谱仪检测各组元素.各组试件与BMSC共培养,检测第1天各组细胞黏附铺展情况(每组样本量为3),第1、3、5天各组细胞增殖A值及第5、7、11天各组碱性磷酸酶活性(每组每个时间点样本量为3).结果 场发射扫描电镜示实验组表面可见粟粒状颗粒物,X射线光电子能谱仪示实验组氮峰明显增高.场发射扫描电镜示第1天实验组细胞黏附铺展良好,细胞间连接广泛而紧密,均优于其他3组.第1天各组细胞增殖效果不明显;第3、5天实验组A值(3.295±0.153、3.823±0.059)均显著高于空白对照组(2.479±0.064、3.131±0.096)、阴性对照A组(2.715±0.075、3.371±0.047)及阴性对照B组(2.756±0.132、3.637±0.047) (P<0.05);第5、7、11天实验组碱性磷酸酶活性(0.0477±0.0287、0.0615±0.0016、0.0667±0.0018)均显著高于其他3组(P<0.05).结论 二氧化钛纳米管阵列可通过生物化学法加载rhBMP-2并具备良好的生物相容性.%Objective To investigate the effect of TiO2 nanotube arrays covalently modified by recombinant human bone morphogenetic protein-2(rhBMP-2) on the early bioactivity of mesenchymal stem cells(MSC) in vitro and to provide experimental evidence for the biochemical modification of titanium implants.Methods In the experiment group,double titanium nanotube arrays were prepared by anodization,and were chemically

  17. 高糖对骨形态发生蛋白-2、胰岛素样生长因子-1基因转染大鼠骨髓基质干细胞增殖的影响%Effects of transfection of bone morphogenetic protein-2 and insulin-like growth factor-1 gene on the proliferation of bone marrow mesenchymal stem cells with high glucose condition

    Institute of Scientific and Technical Information of China (English)

    吴建军; 邢德国; 王亮; 田克立; 刘中浩; 宫明智

    2011-01-01

    目的 观察高糖环境下骨形态发生蛋白-2(BMP-2)和胰岛素样生长因子-1(IGF-1)基因转染大鼠骨髓基质干细胞(BMSC)后BMSC的增殖.方法 用Ad-BMP-2和Ad-IGF-1转染大鼠BMSC,Wester blot检测蛋白表达.噻唑蓝(MTT)比色法及流式细胞术检测细胞增殖.结果 Western blot检测到转染组细胞中有目的 蛋白表达.MTT结果显示第5天细胞增殖能力达到高峰,5 d光吸收值:A~E组分别为0.324±0.027、0.319±0.017、0.622±0.028、0.626±0.020、0.778±0.031.流式细胞术结果显示A~E组细胞处于增殖期的比重分别为23.92±3.07、23.51±2.11、34.37±6.85、35.04±1.45、42.56±1.15.结论 高糖环境下BMP-2和IGF-1基因转染均能促进BMSC增殖,联合转染对BMSC增殖有协同效应.%Objective To observe the proliferation of bone marrow stromal cell (BMSC) transfected by bone morphogenetic protein-2 (BMP-2) and insulin-like growth factor-1 ( IGF-1 ) gene under high glucose condition.Methods Ad-BMP-2 and Ad-IGF-1 transfected rat BMSC,protein expression of BMSC were detected by Western blotting analysis.Methyl thiazol tetrazolium (MTT) assay and flow cytometry to observe the proliferation potential of BMSC.Results In the Western blotting analysis,positive signal lane of protein was observed in transfected group.MTT assay show that proliferation reached the peak in all groups on the fifth day,and the absorbency values of A to E group were 0.324 ± 0.027,0.319 ± 0.017,0.622 ±0.028,0.626 ± 0.020,0.778 ± 0.031.Flow cytometry show that the proliferative percentage from A to E group were 23.92 ±3.07,23.51 ±2.11,34.37 ±6.85,35.04 ± 1.45,42.56 ± 1.15.Conclusion BMP-2 or IGF-1 can promote the proliferation of BMSC under high glucose condition,but the combined has the synergy effect.

  18. The morphogenetic MreBCD proteins of Escherichia coli form an essential membrane-bound complex

    DEFF Research Database (Denmark)

    Kruse, Thomas; Bork-Jensen, Jette; Gerdes, Kenn

    2005-01-01

    spherical, enlarged and finally lysed. Depletion of each mre gene separately conferred similar gross changes in cell morphology and viability. Thus, the three proteins encoded by mreBCD are all essential and function in the same morphogenetic pathway. Interestingly, the presence of a multicopy plasmid......D. In contrast, MreB and MreD did not interact in this assay. Thus, we conclude that the E. coli MreBCD form an essential membrane-bound complex. Curiously, MreB did not form cables in cell depleted for MreC, MreD or RodA, indicating a mutual interdependency between MreB filament morphology and cell shape. Based...

  19. 重组人胶原绑定骨形态发生蛋白2在大肠杆菌中的表达、纯化与复性%Expression,purification and renaturation of recombinant human collagen-binding bone morphogenetic protein-2 from Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    吴乃蓬; 王宇; 宋佳; 武振旭; 高田林; 冯祥汝; 付川; 王宗良; 王春艳

    2016-01-01

    目的:利用大肠杆菌表达体系制备带有胶原结合结构域(CBD)的骨形态发生蛋白2(BMP2),研究CBD-BMP2表达、纯化及复性的条件和方法。方法:将具有胶原结合能力的CBD基因序列克隆入 BMP2基因序列的 N端,构建重组蛋白表达质粒 pet21b/CBD-BMP2,转化入工程性大肠杆菌 BL21菌株内;37℃条件下添加诱导剂异丙基硫代半乳糖苷(IPTG)持续诱导表达;采用 Ni-NTA亲和层析柱进行纯化;运用超纯水稀释复性法对纯化后的CBD-BMP2进行复性;0.22μm微孔滤膜对复性后蛋白除菌,通过除菌前后蛋白浓度比值计算回收率;聚丙烯酰胺凝胶电泳(SDS-PAGE)检测重组蛋白表达、纯化以及复性;BCA蛋白定量法测定蛋白浓度。结果:重组质粒 pet21 b/CBD-BMP2在工程性大肠杆菌中得到充分表达;CBD-BMP2以包涵体形式表达;SDS-PAGE分析,8 mol·L-1尿素存在条件下目的蛋白溶解于裂解液上清中,经纯化后目的蛋白单体存在于洗脱液B中,单体相对分子质量约为14000;稀释复性后SDS-PAGE分析,相对分子质量14000及28000处可见2条清晰条带,重组蛋白单链成功复性为二聚体结构,相对分子质量约为28000;过滤除菌前后目的蛋白浓度分别为110和80 mg·L-1,回收率约为73%。结论:重组CBD-BMP2载体成功转化至大肠杆菌内,CBD-BMP2蛋白得到了高效的表达和复性。建立了利用原核表达体系制备重组CBD-BMP2蛋白的实验方法。%Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of

  20. 血管内皮生长因子和骨形成蛋白2诱导犬恒牙原位牙髓再生%Regeneration of dental pulp tissue in mature dog teeth with apical periodontitis using vascular endothelial growth factor and bone morphogenetic protein 2

    Institute of Scientific and Technical Information of China (English)

    周俊; 王兆晶; 陈文瑨; 陈文霞

    2016-01-01

    目的 通过选择犬根尖孔发育完成的恒牙建立根尖周炎模型,探索血管内皮生长因子(VEGF)和骨形态生成蛋白2(BMP2)诱导原位牙髓再生的可能性.方法 2只10~12月龄的杂种犬,选择根尖孔发育完成的14颗恒前牙建立根尖周炎模型,分别将VEGF(VEGF组)、BMP2(BMP2组)单独和VEGF+BMP2联合(VEGF+BMP2组)与水凝胶复合植入感染控制后的根管腔内,对照组仅植入水凝胶.8周后组织学观察根管内组织再生情况.结果 植入8周后,VEGF组和VEGF+BMP2组根管腔内可见含有大量成纤维样细胞和血管的新生组织形成;而BMP2组和对照组根管腔内见均质状物质,未见细胞、血管形成.结论 VEGF或VEGF+BMP2复合水凝胶支架可以诱导犬根尖发育成熟的根尖周炎患牙在根管腔内生成含有血管的疏松结缔组织.%Objective To investigate the feasibility of dental pulp regeneration in mature teeth with apical periodontitis on situ using vascular endothelial growth factor(VEGF)and bone morphogenetic protein 2(BMP2). Methods Apical periodontitis model was established in 14 mature anterior teeth in 2 dogs(10-12 months). The disinfected root canals were filled with peptide hydrogel scaffold composited with different cytokines:VEGF group,BMP2 group,VEGF+BMP2 group and a control group(without cytokines). Eight weeks after the operation,a histological observation was undertaken to evaluate the regeneration tissue in the root canals. Results Eight weeks after the operation,newly formed vascularized connective tissue were found in the root canals which filled with VEGF and VEGF+BMP2. No cells and vessels were observed in the root canals in BMP2 group and control group. Conclusion VEGF alone or combinated with BMP2 can induce pulp-like tissue regeneration within the root canals of the mature teeth with apical periodontitis.

  1. The correlation analysis between environmental factors, bone morphogenetic protein-4 and transforming growth factor beta-3 polymorphisms in nonsyndromic cleft lip with or without cleft palate%BMP4、TGF-β3基因多态性与环境暴露在非综合征性唇腭裂发生中的交互作用

    Institute of Scientific and Technical Information of China (English)

    林健燕; 栾荣生; 郭泽强; 林新勤; 汤洪洋; 陈苑萍

    2010-01-01

    目的 探讨环境暴露因素、骨形态生成蛋白4(BMP4)基因、转化生长因子β3(transforming growth factor beta-3,TGF-β3)基因之间的交互作用在非综合征性唇腭裂(nonsyndromic cleft lip and cleft palate,NSCLP)发生中的可能作用.方法 通过问卷调查获取环境暴露资料.用聚合酶链反应(PCR)-限制性片段长度多态性(restriction fragment length polymorphism,RFLP)技术对对照组(200例)和NSCLP组(200例)各基因位点的多态性进行检测.采用多因子降维法(multifactor djmensionality reduction,MDR)分析基因之间、基因与环境之间的交互作用关系,并对筛选的交互作用关系用Logistic回归进行验证.结果 BMP4 T538C、TGF-β3 C641A和TGF-β3G15572-三个单核苷酸多态性(single nucleotide polymorphism,SNP)位点间的交互作用与NSCLP的发生无关联.基因与环境交互作用分析发现,BMP4 T538C与母亲妊娠早期被动吸烟、母亲妊娠早期感染史对NSCLP的发生具有交互作用;TGF-β3G15572-与母亲妊娠早期被动吸烟、母亲妊娠早期感染史、父亲知晓妊娠前吸烟、父亲知晓妊娠前饮酒、母亲妊娠早期补充维生素对NSCLP的发生具有交互作用.经Logistic回归验证,结果一致.结论 NSCLP是基因与环境因素共同作用的结果,易感基因多态性影响着个体对环境因素的反应,研究它们之间的相互关系对阐明NSCLP的病因及发病机制具有重要意义.%Objective To investigate the effects of interactions among environmental factors, bone morphogenetic protein-4 ( BMP4 ) and transforming growth factor beta-3 ( TGF-β3 ) polymorphisms on nonsyndromic cleft lip and cleft palate (NSCLP) . MethodsThe data of environmental exposures were collected with questionnaires.Genotypes were determined with techniques of polymerase chain reactionrestriction fragment length polymorphism. Interactions between genes, environmental factors and NSCLP were analyzed using multifactor dimensionality

  2. 骨形态发生蛋白与碱性成纤维细胞生长因子联合修复软骨缺损的效果评价%Effects of recombinant human bone morphogenetic protein combined with basic fibroblast growth factor on the repair of articular cartilage defects

    Institute of Scientific and Technical Information of China (English)

    朱国华; 蔡建平; 郭翠玲; 廖家新; 刘勇; 罗洪涛; 许国华; 胡红涛

    2012-01-01

    背景:多种细胞生长因子在骨软骨代谢过程中的协同作用越来越受到重视,但目前复合细胞生长因子修复软骨缺损报道较少,且修复效果尚无定论.目的:探讨骨形态发生蛋白和碱性成纤维细胞生长因子联合应用修复关节软骨缺损的效果.方法:24 只日本大耳白兔建立骨软骨缺损模型后随机等分为4 组,对照组缺损处仅填塞明胶海绵,其他3 组在对照组基础上,缺损处分别注射骨形态发生蛋白和碱性成纤维细胞生长因子、骨形态发生蛋白、碱性成纤维细胞生长因子.结果与结论:大体观察显示联合应用2 种细胞因子后,软骨缺损面基本修复但稍不平整,单独使用其中1 种细胞因子缺损面未完全修复,对照组无明显修复.联合应用2 种细胞因子缺损部位软骨细胞数多于其他3 组(P < 0.05),且Ⅱ型胶原免疫组化染色深于其他组.提示联合应用骨形态发生蛋白和碱性成纤维细胞生长因子可以促进关节软骨损伤的修复,疗效优于单独应用骨形态发生蛋白或碱性成纤维细胞生长因子.%BACKGROUND: The synergy of various cell growth factors attracts more and more attention in the course of cartilage metabolism.However, there are few reports of repairing cartilage defects with combined cell growth factors, and the effect remains unknown atpresent.OBJECTIVE: To study the repairing effect of recombinant human bone morphogenetic protein (rhBMP) combined with basicfibroblast growth factor (bFGF) on articular cartilage defects.METHODS: After the model of articular cartilage defects was made, 24 Japan big-eared white rabbits were randomly divided intofour groupsforintervention: rhBMP combined with bFGF (group A), single rhBMP (group B), single bFGF (group C), the fourthgroup was without injection and just filled with gelatin sponge (group D).RESULTS AND CONCLUSION: In general observation, articular cartilage defects were basically repaired but slightly

  3. Preliminary research on the expression of sclerostin mediated by bone morphogenetic protein 2 in cementoblast%骨形态发生蛋白2对成牙骨质细胞中硬化蛋白表达调控机理的研究

    Institute of Scientific and Technical Information of China (English)

    陈悦; 李书琴; 黄兰; 戴红卫

    2016-01-01

    目的:探索成牙骨质细胞OCCM-30中骨形态发生蛋白2(BMP2)对硬化蛋白(SOST)表达的调控机制。方法用2种质量浓度的BMP2(50、100 ng·mL-1)处理成牙骨质OCCM-30细胞3、5、7 d,相同体积的PBS液为对照组,采用实时荧光定量聚合酶链反应(RT-PCR)、免疫印迹法检测SOST mRNA和蛋白的表达情况。将OCCM-30细胞分为5组:空白对照组、BMP2组、BMP2+dorsomorphin组、BMP2+SB202190组、BMP2+PD98059组,根据分组分别加入100 ng·mL-1的BMP2和相应的试剂共培养,于3、5 d时检测SOST mRNA和蛋白的表达情况。结果100 ng·mL-1 BMP2对SOST表达的上调作用强于50 ng·mL-1 BMP2,且有时间依赖性(P<0.05)。BMP2+dorsomorphin组、BMP2+SB202190组、BMP2+ PD98059组的SOST mRNA水平和蛋白质水平均降低,其中BMP2+dorsomorphin组降低最明显(P<0.05)。结论成牙骨质细胞中BMP2主要是通过Smad信号通路介导上调SOST的表达。%Objective This research explores the regulatory role of bone morphogenetic protein 2 (BMP2) in the expression of sclerostin in OCCM-30 cementoblast. Methods OCCM-30 cementoblasts were treated with 50 and 100 ng·mL−1 BMP2 for 3, 5, and 7 days. SOST mRNA was detected by real-time quantitative polymerase chain reaction (RT-PCR). Western blot analysis was employed to detect the sclerostin levels in the nucleus. Five groups were prepared for the experiments: control, BMP2, BMP2+dorsomorphin, BMP2+SB202190, and BMP2+PD98059. OCCM-30 was pretreated with BMP2 for 3 and 5 days, and then the sclerostin and SOST mRNA levels were measured. Results RT-PCR and Western blot analyses showed that BMP2 upregulated the expression of SOST in a concentration-dependent manner. SOST expression increased with time (P<0.05). Moreover, sclerostin levels of BMP2+dorsomorphin, BMP2+SB202190, and BMP2+PD98059 groups were lower than that of the BMP2 group, and the sclerostin level in BMP2+dorsomorphin

  4. Bone morphogenetic proteins-ERK5 signaling pathway mediates osteogenic differentiation of human periodontal ligament stem cells%BMPs-ERK5信号通路调控人牙周膜干细胞成骨分化的研究

    Institute of Scientific and Technical Information of China (English)

    蒋琳; 周鹏飞; 王佳; 张燕

    2016-01-01

    目的 比较骨形态发生蛋白(bone morphogenetic proteins,BMP)2、7、9对人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)成骨分化的诱导作用,探讨BMPs-ERK5信号通路在成骨分化中的作用.方法 首先采用组织块酶消化法分离培养原代hPDLSCs并鉴定,利用腺病毒载体介导的GFP、BMP2、BMP7和BMP9(Ad-GFP、Ad-BMP2、Ad-BMP7、Ad-BMP9)分别感染hPDLSCs,通过早期成骨分化指标碱性磷酸酶活性定性和定量检测、茜素红染色检测晚期成骨分化指标钙盐结节沉积情况、qRT-PCR检测细胞成骨分化相关基因骨钙蛋白和骨桥蛋白mRNA表达,比较发现BMP9对细胞成骨分化的作用最强;采用Ad-BMP9感染hPDLSCs 24 h后,检测BMP9作用PDLSCs后对ERK5磷酸化水平的影响和ERK5信号通路特异性抑制剂BIX02189预处理后BMP9-ERK5通路对hPDLSCs成骨分化的影响.结果 ①分离培养的hPDLSCs为成纤维细胞样,呈漩涡状生长,抗波形蛋白表达阳性,抗角蛋白表达阴性,具有间充质属性和克隆形成能力.②BMP2、BMP7和BMP9均有促进细胞成骨分化作用,其中3组的碱性磷酸酶定量结果和成骨分化相关基因相对表达量与GFP组比较,差异具有统计学意义(P<0.05),且BMP9效力最强.③Western blot检测显示BMP9增强p-ERK5的表达.而抑制剂BIX02189呈剂量依赖性地抑制p-ERK5的表达.BIX02189预处理细胞后、感染Ad-BMP9的结果显示,BMP9对细胞的促成骨作用较未处理组明显降低,其中BMP9组的碱性磷酸酶定量分析结果与成骨分化相关基因相对表达量与BIX02189处理组比较,差异具有统计学意义(P<0.05).结论 BMP9具有较强的促hPDLSCs成骨分化作用,ERK5信号转导途径在BMP9诱导hPDLSCs成骨分化过程中发挥了正向调节作用.

  5. 骨形态发生蛋白2/7异二聚体对人脂肪间充质干细胞成骨分化的促进作用%Promoted role of bone morphogenetic protein 2/7 heterodimer in the osteogenic differentiation of human adipose-derived stem cells

    Institute of Scientific and Technical Information of China (English)

    张晓; 刘云松; 吕珑薇; 陈彤; 吴刚; 周永胜

    2016-01-01

    Objective:To investigate the role of bone morphogenetic protein 2/7 heterodimer (BMP-2/7)in the osteogenesis of human adipose-derived stem cells (hASCs).Methods:hASCs were exposed to three different treatments in vitro:osteogenic medium with 1 50 μg/L BMP-2/7 (experimental group), osteogenic medium alone (OM group)and proliferation medium (PM group).After 1 ,4 and 7 days of osteogenic induction,the amount of cellular DNA was measured to investigate the cytotoxicity.After 7 and 1 4 days,alkaline phosphatase (ALP)staining and quantification were performed to test the activity of ALP.After 21 and 28 days,the calcification deposition was determined by Alizarin Red S (ARS)stai-ning and quantification.The expressions of the osteoblast-related genes were tested on days 1 ,4,7 and 1 4.In the in vivo study,6 nude mice were used and 4 groups were set and implanted subcutaneously into the back of nude mice:(1 )β-TCP scaffold only (scaffold control group );(2 )β-TCP scaffold with hASCs cultured by PMin vitro for 1 week (PMcontrol group);(3)β-TCP scaffold with hASCs cultured by OM in vitro for 1 week (OM control group);(4)β-TCP scaffold with hASCs cultured by OM with 1 50 μg/L BMP-2/7 in vitro for 1 week (test group).After 4 weeks of implantation,histological staining was performed to evaluate the in vivo osteogenesis of hASCs.Results:After induction for 1 day,there was no significant difference between the experimental group and the PM group on the cellular DNA con-tent (P>0.05 ).After 4 days,the cellular DNA content increased under the stimulation of BMP-2/7 (P0.05).ALP ac-tivity was higher by the induction of BMP-2/7 than in OMalone and PM(P<0.05).More mineraliza-tion deposition and more expressions of osteoblast-related genes such as Runx2,ALP,COL-1 A1 and OC were determined in the experimental group at different time points (P<0.05).HE staining showed that, in the test group and OM control group,the extracellular matrix (ECM)with eosinophilic staining were observed

  6. Enzymatically synthesized inorganic polymers as morphogenetically active bone scaffolds: application in regenerative medicine.

    Science.gov (United States)

    Wang, Xiaohong; Schröder, Heinz C; Müller, Werner E G

    2014-01-01

    In recent years a paradigm shift in understanding of human bone formation has occurred that starts to change current concepts in tissue engineering of bone and cartilage. New discoveries revealed that fundamental steps in biomineralization are enzyme driven, not only during hydroxyapatite deposition, but also during initial bioseed formation, involving the transient deposition and subsequent transformation of calcium carbonate to calcium phosphate mineral. The principal enzymes mediating these reactions, carbonic anhydrase and alkaline phosphatase, open novel targets for pharmacological intervention of bone diseases like osteoporosis, by applying compounds acting as potential activators of these enzymes. It is expected that these new findings will give an innovation boost for the development of scaffolds for bone repair and reconstruction, which began with the use of bioinert materials, followed by bioactive materials and now leading to functional regenerative tissue units. These new developments have become possible with the discovery of the morphogenic activity of bioinorganic polymers, biocalcit, bio-polyphosphate and biosilica that are formed by a biogenic, enzymatic mechanism, a driving force along with the development of novel rapid-prototyping three-dimensional (3D) printing methods and bioprinting (3D cell printing) techniques that may allow a fabrication of customized implants for patients suffering in bone diseases in the future.

  7. 骨形成蛋白家族成员2、7对小鼠胚胎肝干细胞分化的体外研究%The effecf of bone morphogenetic proteins 2, 7 in inducing murine embryonic stem cells into hepatic cells in vitro

    Institute of Scientific and Technical Information of China (English)

    陈聪; 康权; 罗庆; 迭小红; 田雯

    2013-01-01

    目的 探讨骨形成蛋白2、7(BMP2、BMP7)对小鼠胚胎肝干细胞(HP14.5)定向诱导分化为肝细胞样细胞的影响.方法 将表达BMP2、BMP7、肝细胞生长因子(HGF)和绿色荧光蛋白(GFP)的重组腺病毒作为4个组分别感染HP14.5,诱导其分化,在病毒感染后的第1、4、7天通过检测荧光素酶报告基因读数,在第7天通过细胞免疫荧光染色,观察肝细胞标志物白蛋白(ALB)的表达情况;并在第4、7、10天通过PAS染色、尿素氮合成功能检测,观察诱导后HP14.5向肝细胞方向的分化成熟度.结果 BMP2组、HGF组荧光素酶读数较GFP对照组明显上升;免疫荧光染色显示,诱导7d后BMP2组、HGF组细胞质内表达肝细胞特有的ALB,而GFP对照组几乎无表达;糖原染色可见BMP2组、HGF组胞质存在紫红色颗粒,呈阳性反应;尿素合成功能检测显示BMP2组、HGF组培养液中尿素氮随时间而升高.BMP7组诱导后,细胞免疫荧光染色、荧光素酶活性、PAS染色和尿素合成检测均呈阴性或弱阳性反应.结论 BMP2具有一定的诱导HP14.5向成熟肝细胞分化的作用,并初步具备肝细胞的合成分泌功能,而BMP7对其无诱导作用.%Objective To explore the effect of recombinant adenovirus-mediated bone morphogenetic proteins 2, 7 (Adv-BMP2, Adv-BMP7) in inducing transformation of murine embryonic hepatic progenitor cells to mature hepatic-like cells. Methods HP14.5 cells were divided into 4 groups, and then infected by recombinant adenovirus expressing BMP2, BMP7, hepatocyte growth factor (HGF), and green fluorescent protein (GFP), respectively. For investigating the differential regulation of HP14.S cells, the luciferase report gene was detected at the 1st, 4th and 7th day post infection, the expression of hepatocyte marker albumin (ALB) was detected at the 7th day after infection by cellular immunofluorescence assay. The maturation and differentiation of HP14.S cells were examined by PAS staining

  8. Association of serum bone morphogenetic protein-6 and hepcidin levels with anemia in patients with rheumatoid arthritis%类风湿关节炎患者血清骨形态发生蛋白6和铁调素与贫血的关系

    Institute of Scientific and Technical Information of China (English)

    邬秀娣; 张振; 陈勇; 谢斌华; 干敏芝; 黄娴倩; 刘磊

    2013-01-01

    Objective To investigate the association of serum bone morphogenetic protein- 6 (BMP- 6) and hepcidin (Hepc) levels with anemia in patients with rheumatoid arthritis(RA). Methods Blood samples were col ected from 99 patients with RA,19 patients with systemic lupus erythematosus (SLE) and 40 healthy controls. Serum BMP- 6, Hepc and IL- 6 were measured by ELISA, and serum iron(SI), ferritin(SF), soluble transferrin receptor(sTfR) levels were also determined. The correlation of BMP- 6 and Hepc levels with anemia and disease activity score for 28 joints(DAS28) was analyzed by SPSS software. Results In 99 RA patients, 59 (59.6%) were complicated with anemia, including 30 patients (52.5%) with anemia of chronic disease (ACD) having sTfR/logSF index value5.1) were lower than those with low DAS28 scores (≤5.1), while serum BMP- 6 and Hepc levels were higher in RA patients with high DAS28 scores than those with low DAS28 scores(P<0.05). In RA patients, the level of serum BMP- 6 was positively correlated with Hepc, IL- 6, SI and DAS28 scores (P<0.01). And the level of serum Hepc was positively correlated with IL- 6 and DAS28 scores(P<0.05). Conclusion The serum levels of BMP- 6 and Hepc are high in RA patients, es-pecial y in patients with ACD or high DAS28 scores. The correlation between BMP- 6, Hepc and IL- 6 suggests that BMP- 6 and Hepc may induce ACD in RA and might be used as therapeutic target for ACD in RA.%目的探讨类风湿关节炎(RA)患者骨形态发生蛋白6(BMP-6)和铁调素(Hepc)与贫血的关系。方法采用酶联免疫吸附法测定99例RA患者、19例系统性红斑狼疮(SLE)患者、40例健康志愿者的血清BMP-6、Hepc水平,同时检测RA患者的红细胞计数(RBC)、血红蛋白水平(HGB)、白细胞介素6(IL-6)、血清铁(SI)、血清可溶性转铁蛋白受体(sTfR)、铁蛋白(SF),分析比较RA组与SLE组和健康对照组、RA患者贫血组与非贫血组、慢性病性贫血(ACD)与慢

  9. Inhibitory effect of meloxicam on osteogenic differentiation in mesenchymal stem cells induced by bone morphogenetic protein 9%美洛昔康对骨形态发生蛋白9诱导间充质干细胞骨向分化的抑制作用

    Institute of Scientific and Technical Information of China (English)

    周龙洋; 杨秋珺; 牟钰钦; 刘映孜; 周岐新; 蒋青松; 何百成

    2012-01-01

    OBJECTIVE To investigate the effect of meloxicam on bone morphogenetic protein 9 (BMP9) induced osteogenic differentiation in mesenchymal stem cells. METHODS The infected C3H10T1/2 cells with BMP9 recombinant adenovirus were interfered with meloxicam 5, 10 and 20 μmol·L-1. The alkaline phosphatase activity (ALP) on day 5, day 7 and day 9 after treatment was determined. The mRNA expression of osteocalcin (OCN) was detected on day 9 and day 11 by RT-PCR, and so did the Dlx-5 mRNA on day 1, day 3 and day 5. The matrix mineralization with alizarin red S staining on day 14 and day 20 was assayed. The transcriptional activity of BMP-Smad signaling with Smad binding site luciferase reporter was measured. RESULTS ALP in BMP9 group was much higher than in normal control group ( P < 0.01) on day 5, day 7 and day 9, but the ALP activities were reduced when treated with BMP9 combined with meloxicam (P<0.05). Compared with normal control group, the mRNA expression of osteocalcin and Dlx-5 apparently increased in BMP9 group. However, the function of BMP9 was attenuated by combination with meloxicam (P<0.05). Compared with normal control group, the matrix mineralization in BMP9 group was increased, but decreased when BMP9 combined with meloxicam. Compared with normal control group, BMP9 promoted the firefly luciferase activities of BMPR-Smad reporter plasmids in C3H10T1/2 cells (P <0. 01), but decreased when BMP9 and meloxicam were combined (P < 0.05). CONCLUSION Meloxicam can inhibit osteogenic differentiation induced by BMP9 in mesenchymal stem cells, which may be mediated by inhibiting the BMP-Smad signaling activation.%目的 探讨美洛昔康对骨形态发生蛋白9(BMP 9)诱导间充质干细胞成骨分化的影响.方法 用BMP 9编码序列的重组腺病毒感染C3H10T1/2细胞,并同时加入美洛昔康5,10和20 μmol·L-1对BMP9的作用进行干预.采用化学发光法检测第5天、第7天和第9天的碱性磷酸酶(ALP)活性,RT-PCR方法分别于第9

  10. 骨形态发生蛋白7基因变异与新疆维吾尔族肥胖人群的相关性%Genetic variations of bone morphogenetic protein 7 associated with obesity in Xinjiang Uygur populations

    Institute of Scientific and Technical Information of China (English)

    严治涛; 李南方; 郭艳英; 姚晓光; 王红梅; 胡君丽

    2011-01-01

    BACKGROUND: Studies show that bone morphogenetic protein 7 (BMP-7) gene maps to chromosome 20q13, a susceptibility locus for obesity. BMP-7 represents a strong biological and positional candidate for a susceptibility factor for obesity.OBJECTIVE: To investigate the association between the genetic variations of functional region in BMP-7 gene and obesity in Xinjiang Uygur populations.METHODS: A case-control study was conducted based on epidemiological investigation. 1063 Uygur populations were selected.The exons, segmental introns and the promoter regions of BMP-7 gene were sequenced in 48 Uygur obese patients (male/female: 24/24). Representative variations were selected according to the minor allele frequency and linkage disequilibrium and genotyped using the TaqMan polymerase chain reaction method in 1063 Uygur individuals, a relatively isolated general population in a relatively homogeneous environment and a case-control study was conducted to analyze the association between genetic variations of BMP-7 gene and obesity.RSSULTS AND CONCLUSION: Five novel and eight known variations in the BMP-7 gene were identified. Two genotypedistributions, rs60254222 and rs17480735, followed Hardy-Weinberg equilibrium. There were significant differences in AA, AG, GG genotype distribution frequency of rs6025422 variation between obese and control groups in old adults (32.2%, 55.7%,12.2% vs. 23.8%, 48.3%, 27.9%; P < 0.01).There were significant differences in means of body mass index, waist circumference and waist-to-hip ratio were significantly decreased among individuals with AA, AG and GG genotypes of rs60254222 in old adults (P < 0.05). Logistic regression analysis showed that GG genotype of rs60254222 variation might be a obesity protective factor in old adults (OR = 0.578, 95% confidence interval 0.401-0.833, P = 0.003). Results showed that rs60254222 polymorphism in BMP-7 gene may be associated with obesity in Uygur populations.%背景:研

  11. Induction of Bone Matrix Protein Expression by Native Bone Matrix Proteins in C2C12 Culture

    Institute of Scientific and Technical Information of China (English)

    ZHEN-MING HU; SEAN A. F. PEEL; STEPHEN K. C. HO; GEORGE K. B. SANDOR; CAMERON M. L. CLOKIE

    2009-01-01

    Objective To study the expression of bone matrix protein (BMP) induced by bovine bone morphogenetic proteins (BMPs) in vitro. Methods Type I collagen, osteopontin (OPN), osteonectin (ON), osteocalcin (OC), and bone sialoprotein (BSP) were detected by immunohistochemistry in C2C12 cultured from day 1 to day 28. Results The signaling of bone matrix protein expression became weaker except for type I collagen, OC and BSP after 5 days. Fourteen days after culture, the positive signaling of type I collagen, OPN, ON, OC, and BSP was gradually declined, and could be detected significantly as compared with that of the negative control on day 28. BMP assay showed that the Ikaline phosphatase (ALP) activity was higher in C2C12 culture than in the control during the 14-day culture. Also, total protein and DNA significantly increased during the 14-day culture. High levels of ALP were seen in preosteoblasts and osteoblsts in vivo and in differentiating ostcoblasts in vitro. ALP was well recognized as a marker reflecting osteoblastic activity. Conclusion Native bovine BMP induces conversion of myoblasts into osteoblasts, produces type 1 collagen, and plays significantly role in osteoinduction and bone matrix mineralization of C2C12 in vitro.

  12. 骨形成蛋白7干预肝纤维化小鼠肝组织E3泛素连接酶Arkadia表达的实验研究%Effects of bone morphogenetic protein-7 therapy on E3 ubiquitin ligase expression in mouse liver with experimentally induced fibrosis

    Institute of Scientific and Technical Information of China (English)

    申春燕; 陈永平; 阳韬; 陆小蒟; 李春艳; 林镯; 宋梅

    2012-01-01

    均<0.01);与12周模型组相比,BMP-7干预组Arkadia、Smad7、TGF β1的蛋白质表达量明显降低(t值分别为23.438、11.667和42.889,P值均<0.01).结论 Arkadia在肝纤维化进展过程中呈上升趋势,BMP-7具有抗肝纤维化作用并可以抑制纤维化过程中Arkadia的表达.%Objective This study explored the dynamic expression of the E3 ubiquitin-protein ligase gene,Arkadia,in response to carbon tetrachloride (CCl4)-induced liver fibrosis in a mouse model and investigated the differential expression that occurs following treatment with the anti-fibrotic bone morphogenetic protein-7 (BMP-7).Methods Thirty healthy male imprinting control region (ICR) mice were randomly assigned to three groups:normal (control; n =6),CCl4-induced model group (model; n =18),and CCl4-induced model with BMP-7 treatment group (treatment; n =6).The model group was further divided into three subgroups (n =6 each) for analysis at 4,8 and 12 weeks after fibrosis induction.Liver fibrosis was induced by hypodermic injections of 60% CCl4/peanut oil (5 mL/kg)to the hind legs of mice two-times per week in alternating legs for a period of 12 weeks.At week 9,the treatment group of CC14-induced mice were given an intraperitoneal injection of BMP-7 (300 pg/g) simultaneously with that day's hypodermic injection of 60% CCl4/peanut oil,and then every other day for a period of four weeks.The pathological changes in liver tissues were observed after staining with hematoxylin-eosin (HE) and Masson's trichrome.Messenger RNA (mRNA) and protein expression of Arkadia in liver were evaluated using reverse transcription-polymerase chain reaction and immunohistochemistry and Western blotting,respectively.Results Mouse models of liver fibrosis were successfully established by CCl4 exposure.Arkadia,Stmad7 and TGF-β1 mRNA levels were up-regulated in the model group in a time-dependent manner (vs.control group),and BMP-7 treatment led to significant down-regulation of the CCl4-induced

  13. 重组人BMP-2修饰的β磷酸三钙/胶原材料制备及其诱导成牙性能的初步研究%PREPARATION OF RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN 2 DECORATED β TRICALCIUM PHOSPHATE/COLLAGEN AND PRELIMINARY STUDIES ON ITS PROPERTIES OF INDUCING TOOTH FORMATION

    Institute of Scientific and Technical Information of China (English)

    张文涛; 刘建华; 王慧明; 李志勇

    2011-01-01

    性良好,可作为牙组织工程支架材料的良好选择.%Objective To explore a novel nanometer biomaterial which could induce the regeneration of tooth tissues intelligently, and to evaluate the feasibility of using this kind of biomaterial as the scaffold for tooth tissue engineering by investigating the role it plays in tooth tissue engineering.Methods The scaffold for tooth tissue engineering containing recombinant human bone morphogenetic protein 2 (rhBMP-2) was prepared by mixing nanoscale β tricalcium phosphate (β-TCP)/collagen particles.Forty-six 8-10 weeks old specific pathogen free Sprague Dawley (SD) rats, including 34 females and 12 males, weighing 250-300 g, were involved in this study.Tooth germs were removed under a stereomicroscope from the mandible of newborn SD rat, then digested and suspended.Scanning electronic microscope (SEM), adhesion rate of cells, and MTT assay were used to evaluate the effects of the scaffold on the tooth germ cells cultured in vitro.The tissue engineered tooth germ which was constructed by tooth germ cells and scaffold was transplanted under SD rat's kidney capsule as the experimental group (n=12); the tooth germ cells (cell-control group, n=12) or scaffold without cells (material-control group, n=4) were transplanted separately as control groups.Specimens were harvested to perform general and histological observations at 4 and 8 weeks after transplantation.Results β-TCP/collagen showed a loose and porous appearance with soft texture and excellent hydrophilicity.Tooth germ cells grew well and could attach to the scaffold tightly 3 days after coculture.The adhesion rates of tooth germ cells were 27.20% ± 2.37%,44.52% ± 1.87%, and 73.81% ± 4.15% when cocultured with scaffold for 4, 8, and 12 hours, respectively.MTT assay showed that the cell proliferation status of experimental group was similar to that of the control group, showing no significant difference (P >0.05).Some white calcified specimens

  14. 大鼠脊髓损伤后骨形态发生蛋白2表达变化的规律及意义%Dynamic expression of bone morphogenetic protein 2 and its clinical significance after spinal cord injury in rat

    Institute of Scientific and Technical Information of China (English)

    马敏杰; 贺西京; 王国毓; 臧全金; 李锋涛; 刘宇

    2011-01-01

    脊髓灰质区实施.%Objective:To observe the dynamic expression of bone morphogenetic protein 2 (BMP2) after spinal cord injury in rat,especially the changes on different time point and in different site,and to investigate the optimal time point and site for inhibiting BMP2 expression.Method:A total of 40 Sprague Dawley rats were randomly assigned into 5 groups: 1-day-after-impact group,3-day-after-impact group,7-day-after-impact group,14-day-after-impact group and sham operation group with 8 rats in each group.After making skin incision centered as T10,the whole T10 and T9 and T11 partial laminar were removed to expose the spinal cord. Spinal cord injury models were created by NYU impactor. After that,behavior testing of all rats was assessed by using the Basso-Beattie-Bresnahan (BBB) locomotor rating scale before and after the perfusion. Each group of rats were perfused with 4% paraformaldehyde in heart on the 1st,3th,7th,14th day-afterimpact,and then HE staining was used to observe the difference between the sham operation group and test groups. Meanwhile,immunohistochemistry was used to evaluate the level of BMP2 in the injuried region. Result: BBB score showed no significant difference among the spinal cord injury groups at the same time point (P> 0.05).By HE staining, the tissue structure of sham operation group showed clear and orderly,on the contrary, it was disorderly in rest groups. According to the result of immunohistochemistry,significant difference was detected between the model group and sham operation group (P<0.05).Compared with the sham operation group,the mean number of BMP2 positive cells in 1-day-after-impact group was 13.70±4.81 and 7.23±3.14 for the gray and white matter respectively,9.37±3.61 and 5.63±2.23 in 3-day-after-impact group,6.17±1.81 and 4.17±1.24 in 7-day-after-impact group,and 4.36±1.60 and 1.87±1.13 in 14-day-after-impact group.The BMP2-positive cells in gray matter increased more significantly than white matter (P<0.01).Conclusion:The number of BMP2

  15. Mistura de proteínas morfogenéticas ósseas, hidroxiapatita, osso inorgânico e colágeno envolta por membrana de pericárdio no preenchimento de defeito ósseo segmentar em coelhos Mixture of bone morphogenetic protein, hydroxyapatite, inorganic bone and collagen interposed by pericardium barrier membrane in the filling of the segmental bone defect in rabbits

    Directory of Open Access Journals (Sweden)

    R.B. Ciani

    2006-02-01

    Full Text Available Avaliou-se o uso de biomaterial de origem bovina na regeneração de defeitos ósseos segmentares empregando-se 12 coelhos, fêmeas, da raça Norfolk, com idade de seis meses e pesos entre 3 e 4,5kg. Realizou-se falha segmentar bilateral de um centímetro de comprimento na diáfise do rádio, com inclusão do periósteo. No membro direito, o defeito foi delimitado por membrana de pericárdio liofilizada, contendo em seu interior mistura de proteínas morfogenéticas ósseas adsorvidas a hidroxiapatita, colágeno liofilizado e osso inorgânico. No membro esquerdo, o defeito não recebeu tratamento. Radiografias foram obtidas ao término do procedimento cirúrgico e aos sete, 30, 60, 90, 120 e 150 dias de pós-operatório. Após eutanásia de seis coelhos aos 60 dias e seis aos 150 dias de pós-cirúrgico, os resultados radiográficos e histológicos mostraram que a regeneração óssea foi inibida nos defeitos segmentares tratados com o biomaterial.Biomaterials of bovine origin in regenerating segmental bone defects were evaluated. Twelve six-month old Norfolk rabbits, weighting 3 to 4.5kg were used. A 1cm long segmental defect was created in the radial diaphysis, including the periosteum, of both forelimbs. In the right forelimb, the defect was filled using a mixture of bone morphogenic proteins adsorbed to hydroxyapatite, agglutinant of lyophilized collagen in granules and anorganic cortical bone in granules delimited by a pericardial membrane. In the left forelimb, the defect did not receive treatment and served as a control. Radiographies were taken immediately after surgery and at seven, 30, 60, 90, 120 and 150 days post-operatively. Six rabbits were euthanized at 60 days and the other six at 150 days post-surgery for histological evaluation. Radiographic and histological results revealed that bone regeneration was inhibited in the segmental defects receiving biomaterials.

  16. Multi-protein delivery by nanodiamonds promotes bone formation.

    Science.gov (United States)

    Moore, L; Gatica, M; Kim, H; Osawa, E; Ho, D

    2013-11-01

    Bone morphogenetic proteins (BMPs) are well-studied regulators of cartilage and bone development that have been Food and Drug Administration (FDA)-approved for the promotion of bone formation in certain procedures. BMPs are seeing more use in oral and maxillofacial surgeries because of recent FDA approval of InFUSE(®) for sinus augmentation and localized alveolar ridge augmentation. However, the utility of BMPs in medical and dental applications is limited by the delivery method. Currently, BMPs are delivered to the surgical site by the implantation of bulky collagen sponges. Here we evaluate the potential of detonation nanodiamonds (NDs) as a delivery vehicle for BMP-2 and basic fibroblast growth factor (bFGF). Nanodiamonds are biocompatible, 4- to 5-nm carbon nanoparticles that have previously been used to deliver a wide variety of molecules, including proteins and peptides. We find that both BMP-2 and bFGF are readily loaded onto NDs by physisorption, forming a stable colloidal solution, and are triggered to release in slightly acidic conditions. Simultaneous delivery of BMP-2 and bFGF by ND induces differentiation and proliferation in osteoblast progenitor cells. Overall, we find that NDs provide an effective injectable alternative for the delivery of BMP-2 and bFGF to promote bone formation.

  17. The composite of bone morphogenetic protein-2 and poly(lactic-co-glycolic acid)-tricalcium phosphate plus different vascularized tissues to repair large segmental radial defects in sheep%复合骨形态发生蛋白-2的人工骨结合带血供组织修复羊桡骨大段骨缺损的实验研究

    Institute of Scientific and Technical Information of China (English)

    徐建强; 周密; 张树明; 李长庚; 杨飞; 孙锁柱; 王长江; 王克利

    2012-01-01

    目的 探讨复合骨形态发生蛋白-2的聚乳酸-乙醇酸共聚物/磷酸三钙(PLGA-TCP-BMP-2)人工骨结合不同自体带血供组织移植修复羊桡骨大段骨缺损的效果. 方法 将30只绵羊制作成30 mm桡骨骨缺损模型,随机分为5组:PLGA-TCP-BMP-2人工骨及带血供的长段尺骨组(A组)、PLGA-TCP-BMP-2人工骨及带血供的屈指长肌肌腹组(B组)、PLGA-TCP-BMP-2人工骨及带血供的尺骨骨膜组(C组)、PLGA-TCP-BMP-2人工骨组(D组)及不植人任何材料组(E组),每组6只.各组均以钢板固定桡骨缺损区.术后当天及4、12、24周行手术部位X线摄片,术后24周处死动物行CT及组织学检查. 结果 X线片及CT检查显示:术后24周A、B、C组桡骨缺损处完全成骨修复,皮质骨与髓腔的轮廓清晰;D组亦能完全修复,但新生骨密度及髓腔轮廓清晰度均不如前3组;E组无有效骨痂形成.Samantha放射学评分和Lane-Sandhu放射学评分显示:A组评分高于其他4组,差异均有统计学意义(P<0.05).组织学检查结果显示:A组新生骨完全修复骨缺损区;B、C组新生骨与断端皮质骨融合,新生骨痂较为纤细;D组新生板层骨及骨陷窝排列较为紊乱;E组无骨连接表现.A、B、C、D、E组Lane-Sandhu组织学评分平均分别为(11.3±0.6)、(10.8±0.8)、(10.7±0.9)、(10.2±1.1)、(4.5±1.2)分,5组比较差异有统计学意义(F=5.891,P=0.026),其中A组评分高于其他4组,差异均有统计学意义(P<0.05).结论 PLGA-TCP-BMP-2人工骨结合不同自体带血供组织移植能满意修复羊桡骨30 mm的骨缺损.%Objective To study effects of the artificial composite bone of bone morphogenetic protein (BMP-2) and poly ( lactic-co-glycolic acid) -tricalcium phosphate(PLGA-TCP) with different vascularized tissues on repairing the large segmental radial defects in sheep. Methods Defects of 30 mm were made in the middle radial segments in 30 sheep which were randomized into 5 groups.In group A

  18. Power-frequency electromagnetic field promotes mRNA expressions of bone morphogenetic protein-2 and transforming growth factor-beta 1 in bone marrow mesenchymal stem cells%工频电磁场对小鼠骨髓间充质干细胞骨形态发生蛋白2和转化生长因子β1mRNA表达的刺激

    Institute of Scientific and Technical Information of China (English)

    陈继革; 吴华; 葛保健; 方真华

    2007-01-01

    BACKGROUND:Studies confirm that electromagnetic field (EMF) can promote the synthesis and secretion of many bone growth factors,and some growth factors can induce the osteoblastic directional differentiation of bone marrow mesenchymal stem cells (MSCs).OBJECTIVE: To investigate the effect of power-frequency EMF on mRNA expressions of bone morphogenetic protein-2 (BMP-2) and transforming growth factor-beta 1 (TGF-β1) in mouse bone marrow MSCs cultured in vitro.DESTGN: Single sample, block design, observation and controlled animal trial.SETTING: Department of Traumatic Surgery, Tongji Hospital Affiliated to Tongji Medical .College, Huazhong University of Science and Technology.MATERIALS: This trial was carried out in the laboratory of Department of Traumatic Surgery, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology during February to December 2005. ①Twenty Kunming mice of clean grade were selected for harvest of bone marrow MSCs. ②Magnetic field generator,which could generate EMF with 0 to 100 mT field strength and successive adjustable 50 Hz sinusoidal wave, was developed by Wuhan Naval University of Engineering. ③ Primer was all synthesized by Saibaisheng Bioengineering Co.,Ltd., Beijing.NETHODS: ① The involved mice were sacrificed by cervical dislocation. Bilateral femora and tibia were harvested. Bone marrow MSCs were isolated and cultured in vitro, and the second generation of cells were used for the trial. ②Different intensities of EMF stimulation tests: Negative control group, positive control group, EMF 0.4, 0.8 and 1.6 mT stimulation groups were set. Five bottles of cells of the second generation were chosen from each group for test. The cells in the negative control group and positive control group were not exposed to EMF. But medium containing osteogenic inductor(10-8 mol/L dexamethasone, 10 mmol/L β-sodium glycerophosphate and 50 mg/L Vitamin C included) was added in the positive control group

  19. 3D生物打印构建聚乳酸羟基乙酸/纳米羟基磷灰石支架骨形态发生蛋白2缓释复合体的实验研究%3D-bioprinting manufacturing polylactic-co-glycolic acid/nano-hydroxyapatite scaffold/bone morphogenetic protein-2 sustained release composite

    Institute of Scientific and Technical Information of China (English)

    臧晓龙; 孙健; 李亚莉; 陈立强; 杨学财; 梁立卿; 杜国庆

    2016-01-01

    BACKGROUND:Tissue-engineered bone scaffold fabricated by 3D-bioprinting technique has good controlability in morphology and structure. However, construction of tissue-engineered bone/cel growth factor complex and time-dose effect of sustained-release factors are needed to be further researched. OBJECTIVE:To fabricate a sustained-release composite of polylactic-co-glycolic acid (PLGA)/nano-hydroxyapatite (n-HA) scaffold carrying bone morphogenetic protein-2 (BMP-2) using 3D-bioprinting technique, and test the biological properties of the PLGA/n-HA scaffold carrying BMP-2 and the sustained-release properties, thereby to discuss its feasibility as the tissue-engineered bone scaffold composite. METHODS:Temperature-sensitive chitosan hydrogel was prepared using chitosan andβ-glycerophosphate to construct a sustained-release composite, chitosan nanoparticles carrying BMP-2 . 3D-bioprinting technique was utilized to fabricate the PLGA/n-HA scaffold carrying BMP-2. Biological features of the scaffold composite were tested, and time-dose effect of BMP-2 sustained-release was observed. RESULTS AND CONCLUSION:The average pore size of the scaffold-cytokine composite was (431.31±18.40)μm, and the porosity was (73.64±1.82)%. The cumulative release rate of BMP-2 from the scaffold-cytokine composite that effectively controled the burst release during 48 hours and 30 days were suitable for the physiological needs. In conclusion, the porosity, pore size, release property, degradation rate, and mechanical strength of the scaffold-cytokine composite al meet the biological requirements of tissue-engineered bone construction.%背景:3D生物打印技术制备的工程骨支架,其形态、结构可控性好,但对组织工程骨细胞生长因子复合体的构建及缓释细胞因子的时效、量效特点有待进一步研究。  目的:应用3D生物打印技术制备聚乳酸羟基乙酸/纳米羟基磷灰石支架骨形态发生蛋白2缓释复合体,检测聚

  20. Positive modulator of bone morphogenic protein-2

    Science.gov (United States)

    Zamora, Paul O.; Pena, Louis A.; Lin, Xinhua; Takahashi, Kazuyuki

    2009-01-27

    Compounds of the present invention of formula I and formula II are disclosed in the specification and wherein the compounds are modulators of Bone Morphogenic Protein activity. Compounds are synthetic peptides having a non-growth factor heparin binding region, a linker, and sequences that bind specifically to a receptor for Bone Morphogenic Protein. Uses of compounds of the present invention in the treatment of bone lesions, degenerative joint disease and to enhance bone formation are disclosed.

  1. Evaluation of bone morphogenic proteins in periodontal practice.

    Science.gov (United States)

    Kaur, Supreet; Grover, Vishakha; Kaur, Harkiran; Malhotra, Ranjan

    2016-01-01

    Forty years ago Marshal R. Urist discovered a substance in bone matrix that had inductive properties for the development of bone and cartilage, until date, at least 20 bone morphogenetic proteins (BMPs) have been identified, some of which have been shown in vitro to stimulate the process of stem cell differentiation into osteoblasts in human and animal models. The purpose of this paper is to give a brief overview of BMPs and to review critically the clinical data currently available on the use of BMPs in various periodontal applications. The literature on BMPs was reviewed. A comprehensive search was designed. The articles were independently screened for eligibility. Articles with authentic controls and proper randomization and pertaining specifically to their role in periodontal applications were included. The available literature was analyzed and compiled. The analysis indicates BMPs to be a promising, as well as an effective novel approach to reconstruct and engineer the periodontal apparatus. Here, we represent several articles, as well as recent texts that make up a special and an in-depth review on the subject. On the basis of the data provided in the studies that were reviewed BMPs provide revolutionary therapies in periodontal practice.

  2. 肾血流彩色多普勒检测与骨形态发生蛋白-7检测在2型糖尿病肾病早期诊断中的价值%Evaluation of color Doppler ultrasound of renal blood flow combined with the detection of bone morphogenetic protein-7 in early diagnosis of type 2 diabetic nephropathy

    Institute of Scientific and Technical Information of China (English)

    张之杰; 于宁; 王正滨; 闫志梅; 孟冬梅; 刘荣桂; 丁兆艳; 史锋锋

    2012-01-01

    目的 探讨彩色多普勒超声检查肾血流与骨形态发生蛋白-7(BMP-7)水平检测在2型糖尿病肾病中的诊断价值.方法 检测90例糖尿病肾病患者与30例健康志愿者血BMP-7水平,并经彩色多普勒超声测量肾血流动力学参数,比较各组血BMP-7水平、肾段动脉及肾叶间动脉阻力指数(RI).结果 与对照组比较,随着糖尿病肾损害程度加重,BMP-7水平逐渐降低(P<0.01),肾段动脉、肾叶间动脉收缩期最大血流速度及舒张末期最低血流速度逐渐减慢,RI增高(P<0.01).BMP-7水平与肾段动脉、叶间动脉RI间存在显著负相关(r=-0.603,P<0.01;r=-0.652,P<0.01).结论 彩色多普勒超声测量肾血流动力学参数联合检测血BMP-7水平对早期预测糖尿病肾病的肾损害程度有重要价值.%Objective To evaluate the significance of color Doppler ultrasound examination of renal blood flow combined with the detection of bone morphogenetic protein-7(BMP-7)in early diagnosis of type 2 diabetic nephropathy.Methods Blood BMP-7 level was tested in 90 patients with type 2 diabetic nephropathy and 30 controls,and parameters of renal blood flow were measured by color Doppler ultrasound examination.Blood BMP-7 level as well as resistant index(RI)of segmental renal artery(SRA)and interlobar renal artery(IRA),were compared between these two groups.Results Compared with controls,blood BMP-7 level gradually decreased with the aggravation of diabetic kidney damage(P<0.01).The peak systolic velocity(Vmax)and the end diastolic velocity(Vmin)of SRA and TRA were slowed gradually,while RI increased(P<0.01).Blood BMP-7 level was negatively correlated with IRA's and SRA's RI of IRA and SRA(r =-0.603,P<0.01;r =-0.652,P<0.01).Conclusions Color Doppler ultrasound examination of renal blood flow combined with detection of BMP-7 might play an important role in early diagnosis of type 2 diabetic nephropathy.

  3. 以钙磷骨水泥为载体重组人血管内皮细胞生长因子与重组人骨形态发生蛋白-2复合体对异体脱蛋白骨修复骨关节缺损的促进效应%Calcium phosphate cement attaching with recombinant human vascular endothelial growth factor and recombinant human bone morphogenetic protein-2 promotes deproteinized osteoarticular allograft to repair osteoarticular defect

    Institute of Scientific and Technical Information of China (English)

    瞿向阳; 蒋电明; 安洪

    2007-01-01

    8、12及16周A组血管内皮细胞生长因子和BMP-2的面积积分吸光度值明显高于B、C组,差异有统计学意义(P<0.01).③墨汁灌注微血管分析结果显示A组术后各期移植物内新生血管较多,部分趋向成熟,与B、C组比较均有统计学意义(P<0.01).④术后4周时各组手术侧膝关节较对侧血流量均增高,但A组显著增高,8周时进一步增高,12周时达到顶峰,16周时略有下降,但仍较B、C组高,差异有显著性意义(P<0.01).⑤A组三点抗弯曲应力,强于B、C组(P<0.01).结论:重组脱蛋白骨关节材料具有较强的再血管化和成骨能力,可早期与受体达到骨愈合修复骨关节缺损并最终成为自身组织.%BACKGROUND: Deproteinized bone has three diamensions frame profitting bone cells to adhere to, new capillaries and interstitial cells to enter, osteoblasts to differentiate and extracellular matrix to synthesize. Calcium phosphate cement (CPC) is a new in-ceramic bone cement that can degradate, which has plasticity, no heat production, invariably mechanics intension and porosity.OBJECTIVE: To explore the ability of CPC attaching with human vascular endothelial growth factor (rhVEGF) and recombinant human bone morphogenetic protein-2 (rhBMP-2) in promoting deproteinized osteoarticular allograft to repair osteoarticular defect.DESIGN: Randomly control observation.SETTING: Department of Orthopaedics, the First Affiliated Hospital, Chongqing Medical University.MATERIALS: A total of 42 adult hybridization dogs of both genders and weighing (10±0.5) kg were provided by Animal Center of Chongqing Medical University. RhVEGF was provided by Beijing Boaosen Biotechnology Co., Ltd.; rhBMP-2 by Guangzhou Dahui Biotechnology Co., Ltd.; CPC by Shanghai Ruibang Biotechnology Co., Ltd.METHODS: The experiment was carried out in the Laboratory of Orthopaedics (Municipal Laboratory), Clinical College,Chongqing Medical University from March to September 2006. A total of 36

  4. The Deep-Sea Natural Products, Biogenic Polyphosphate (Bio-PolyP and Biogenic Silica (Bio-Silica, as Biomimetic Scaffolds for Bone Tissue Engineering: Fabrication of a Morphogenetically-Active Polymer

    Directory of Open Access Journals (Sweden)

    Florian Draenert

    2013-03-01

    Full Text Available Bone defects in human, caused by fractures/nonunions or trauma, gain increasing impact and have become a medical challenge in the present-day aging population. Frequently, those fractures require surgical intervention which ideally relies on autografts or suboptimally on allografts. Therefore, it is pressing and likewise challenging to develop bone substitution materials to heal bone defects. During the differentiation of osteoblasts from their mesenchymal progenitor/stem cells and of osteoclasts from their hemopoietic precursor cells, a lineage-specific release of growth factors and a trans-lineage homeostatic cross-talk via signaling molecules take place. Hence, the major hurdle is to fabricate a template that is functioning in a way mimicking the morphogenetic, inductive role(s of the native extracellular matrix. In the last few years, two naturally occurring polymers that are produced by deep-sea sponges, the biogenic polyphosphate (bio-polyP and biogenic silica (bio-silica have also been identified as promoting morphogenetic on both osteoblasts and osteoclasts. These polymers elicit cytokines that affect bone mineralization (hydroxyapatite formation. In this manner, bio-silica and bio-polyP cause an increased release of BMP-2, the key mediator activating the anabolic arm of the hydroxyapatite forming cells, and of RANKL. In addition, bio-polyP inhibits the progression of the pre-osteoclasts to functionally active osteoclasts. Based on these findings, new bioinspired strategies for the fabrication of bone biomimetic templates have been developed applying 3D-printing techniques. Finally, a strategy is outlined by which these two morphogenetically active polymers might be used to develop a novel functionally active polymer.

  5. Microspectroscopic evidence of cretaceous bone proteins.

    Directory of Open Access Journals (Sweden)

    Johan Lindgren

    Full Text Available Low concentrations of the structural protein collagen have recently been reported in dinosaur fossils based primarily on mass spectrometric analyses of whole bone extracts. However, direct spectroscopic characterization of isolated fibrous bone tissues, a crucial test of hypotheses of biomolecular preservation over deep time, has not been performed. Here, we demonstrate that endogenous proteinaceous molecules are retained in a humerus from a Late Cretaceous mosasaur (an extinct giant marine lizard. In situ immunofluorescence of demineralized bone extracts shows reactivity to antibodies raised against type I collagen, and amino acid analyses of soluble proteins extracted from the bone exhibit a composition indicative of structural proteins or their breakdown products. These data are corroborated by synchrotron radiation-based infrared microspectroscopic studies demonstrating that amino acid containing matter is located in bone matrix fibrils that express imprints of the characteristic 67 nm D-periodicity typical of collagen. Moreover, the fibrils differ significantly in spectral signature from those of potential modern bacterial contaminants, such as biofilms and collagen-like proteins. Thus, the preservation of primary soft tissues and biomolecules is not limited to large-sized bones buried in fluvial sandstone environments, but also occurs in relatively small-sized skeletal elements deposited in marine sediments.

  6. 骨形成蛋白2基因转染的人牙周膜成纤维细胞的生物学特性%The Biological Characterization of Bone Morphogenetic Protein 2 Gene Transfected Human Periodontal Ligament Fibroblasts

    Institute of Scientific and Technical Information of China (English)

    司晓辉; 刘正

    2001-01-01

    Objective To establish the human periodontal ligament fibroblasts(HPDLFs)that express BMP2 and observe their biologicl characterization. Methods A phagemid expression vector for BMP2 (pBK-B2) was transfected into HPDLFs by using LipofectAMINE. The BMP2 expression was determined by the immunohistochemical ABC method. The alkaline phosphatase (ALP) activity, osteocalcin (OC) production and capacity of mineralization were measured in the transfected cells. Results BMP2 protein was expressed in HPDLFs after gene transfection. The BMP2 gene transfected cells showed prominently elevated ALP activity, OC production and increase in mineralized nodules. Conclusion The results indicate that BMP2 is expressed in HPDLFs and is involved in inducing differ- entiation of HPDLFs into osteoblast-like cells.%目的建立表达骨形成蛋白2(BMPS)的人牙周膜成纤维细胞(HPDLFS)并观察其生物学特性。方法利用脂质体将BMP2噬菌粒表达载体pBK-B2转染至HPDLFs,免疫组化ABC法检测BMP2基因的表达,并检测转染细胞的碱性磷酸酶(ALP)活力、骨钙素(OC)含量和矿化能力。结果转染BMP2基因后,HPDLFs内有BMP2蛋白的表达,ALP活力、OC含量和矿化结节数量均显著增加。结论BMP2基因在HPDLFs中得到表达并促进其向成骨样细胞分化。

  7. Ablation of the Sam68 RNA binding protein protects mice from age-related bone loss.

    Directory of Open Access Journals (Sweden)

    Stéphane Richard

    2005-12-01

    Full Text Available The Src substrate associated in mitosis of 68 kDa (Sam68 is a KH-type RNA binding protein that has been shown to regulate several aspects of RNA metabolism; however, its physiologic role has remained elusive. Herein we report the generation of Sam68-null mice by homologous recombination. Aged Sam68-/- mice preserved their bone mass, in sharp contrast with 12-month-old wild-type littermates in which bone mass was decreased up to approximately 75%. In fact, the bone volume of the 12-month-old Sam68-/- mice was virtually indistinguishable from that of 4-month-old wild-type or Sam68-/- mice. Sam68-/- bone marrow stromal cells had a differentiation advantage for the osteogenic pathway. Moreover, the knockdown of Sam68 using short hairpin RNA in the embryonic mesenchymal multipotential progenitor C3H10T1/2 cells resulted in more pronounced expression of the mature osteoblast marker osteocalcin when differentiation was induced with bone morphogenetic protein-2. Cultures of mouse embryo fibroblasts generated from Sam68+/+ and Sam68-/- littermates were induced to differentiate into adipocytes with culture medium containing pioglitazone and the Sam68-/- mouse embryo fibroblasts shown to have impaired adipocyte differentiation. Furthermore, in vivo it was shown that sections of bone from 12-month-old Sam68-/- mice had few marrow adipocytes compared with their age-matched wild-type littermate controls, which exhibited fatty bone marrow. Our findings identify endogenous Sam68 as a positive regulator of adipocyte differentiation and a negative regulator of osteoblast differentiation, which is consistent with Sam68 being a modulator of bone marrow mesenchymal cell differentiation, and hence bone metabolism, in aged mice.

  8. Role of Soy Protein on Bone Turnover

    Directory of Open Access Journals (Sweden)

    A Haghighian roudsari

    2004-03-01

    Full Text Available Bone mass loss is one of the commonest menopause symptoms, resulting from cessation of estrogen production. Compounds which have estrogen – like biological activity similar to “Isoflavones” present in plants especially soy, may reduce bone loss in postmenopausal women, because as they are similar in structure to estrogens. This study, therefore, was undertaken to assess the effect of soy protein on bone metabolism biomarkers in postmenopausal women with osteopenia. This “before and after” clinical trial was carried out, on 15 postmenopausal women with osteopenia, between 45 to 64 years of age. The subjects were asked to consume 35 gram/day of soy protein for 12 weeks. Blood and urine samples, were taken at 0, 6 and 12 weeks of the study. Anthropometric measurements and a 2-day dietary recall were done at the beginning of the study, and at the 6 and 12 weeks. The food consumption data were analyzed by “Food Proccessor” software. Repeated measurement analysis was utilized to determine the changes in biochemical indices, anthropometric and dietary data. P-values less than 0.05 were considered as significant. Comparison of weight, BMI, physical activity and dietary intake of subjects during the study did not show any significant differences. Soy protein consumption, showed significant reductions in deoxypyridinoline (biochemical marker of bone resorption and significant increase in total alkaline phosphatase ( biochemical marker of bone formation.There were no significant differences in serum osteocalcin, C- telopeptide, insulin- like growth factor binding protein 3 (IGFBP3, and type-I- collagen telopeptides. Considering the beneficial effects of soy protein consumption on bone metabolism biomarkers, inclusion of this inexpensive and available food item in postmenopausal women diet, may reduce bone loss and could be recommended for the prevention of osteoporosis.

  9. Role of Soy Protein on Bone Turnover

    Directory of Open Access Journals (Sweden)

    A Haghighian roudsari

    2004-11-01

    Full Text Available Bone mass loss is one of the commonest menopause symptoms, resulting from cessation of estrogen production. Compounds which have estrogen – like biological activity similar to “Isoflavones” present in plants especially soy, may reduce bone loss in postmenopausal women, because as they are similar in structure to estrogens. This study, therefore, was undertaken to assess the effect of soy protein on bone metabolism biomarkers in postmenopausal women with osteopenia. This “before and after” clinical trial was carried out, on 15 postmenopausal women with osteopenia, between 45 to 64 years of age. The subjects were asked to consume 35 gram/day of soy protein for 12 weeks. Blood and urine samples, were taken at 0, 6 and 12 weeks of the study. Anthropometric measurements and a 2-day dietary recall were done at the beginning of the study, and at the 6 and 12 weeks. The food consumption data were analyzed by “Food Proccessor” software. Repeated measurement analysis was utilized to determine the changes in biochemical indices, anthropometric and dietary data. P-values less than 0.05 were considered as significant. Comparison of weight, BMI, physical activity and dietary intake of subjects during the study did not show any significant differences. Soy protein consumption, showed significant reductions in deoxypyridinoline (biochemical marker of bone resorption and significant increase in total alkaline phosphatase ( biochemical marker of bone formation.There were no significant differences in serum osteocalcin, C- telopeptide, insulin- like growth factor binding protein 3 (IGFBP3, and type-I- collagen telopeptides. Considering the beneficial effects of soy protein consumption on bone metabolism biomarkers, inclusion of this inexpensive and available food item in postmenopausal women diet, may reduce bone loss and could be recommended for the prevention of osteoporosis.

  10. Signaling by bone morphogenetic proteins directs formation of an ectodermal signaling center that regulates craniofacial development.

    Science.gov (United States)

    Foppiano, Silvia; Hu, Diane; Marcucio, Ralph S

    2007-12-01

    We previously described a signaling center, the Frontonasal Ectodermal Zone (FEZ) that regulates growth and patterning of the frontonasal process (FNP). The FEZ is comprised of FNP ectoderm flanking a boundary between Sonic hedgehog (Shh) and Fibroblast growth factor 8 (Fgf8) expression domains. Our objective was to examine BMP signaling during formation of the FEZ. We blocked BMP signaling throughout the FNP prior to FEZ formation by infecting chick embryos at stage 10 (HH10) with a replication-competent avian retrovirus encoding the BMP antagonist Noggin. We assessed gene expression patterns in the FNP 72 h after infection (approximately HH22) and observed that Shh expression was reduced or absent. In the mesenchyme, we observed that Bmp2 transcripts were absent while the Bmp4 expression domain was expanded proximally. In addition to the molecular changes, infected embryos also exhibited facial malformations at 72 and 96 h after infection suggesting that the FEZ did not form. Our data indicate that reduced cell proliferation, but not apoptosis, in the mesenchyme contributed to the phenotype that we observed. Additionally, adding exogenous SHH into the mesenchyme of RCAS-Noggin-infected embryos did not restore Bmp2 and Bmp4 to a normal pattern of expression. These data indicate that BMP signaling mediates interactions between tissues in the FNP that regulate FEZ formation; and that the correct pattern of Bmp2 and Bmp4, but not Bmp7, expression in the FNP mesenchyme requires signaling by the BMP pathway.

  11. In anemia of multiple myeloma hepcidin is induced by increased bone-morphogenetic protein-2

    Science.gov (United States)

    Hepcidin is the principal iron-regulatory hormone and pathogenic factor in anemia of inflammation. Patients with multiple myeloma (MM) frequently present with anemia. We showed that MM patients had increased serum hepcidin, which inversely correlated with hemoglobin, suggesting that hepcidin contrib...

  12. Connective tissue growth factor and bone morphogenetic proteins in diabetic nephropathy

    NARCIS (Netherlands)

    Nguyen, T.Q.

    2008-01-01

    Diabetes mellitus is a severe and rapidly growing problem in health care, accounting for approximately 150 million patients worldwide. Patients with diabetes are at increased risk to develop diabetic nephropathy, which is currently the most important cause of end-stage renal disease in large parts o

  13. Neuroendocrine Differentiation in Prostate Cancer: Role of Bone Morphogenetic Protein-6 and Macrophages

    Science.gov (United States)

    2010-07-01

    7-10-2010 11 Fig 6. When RAW 264.7 cells were treated with SB203580 and BMP-6, ChIP assay using GATA4 antibody no longer amplified...neuroendocrine differentiation was no longer observed. Mechanistically, series of studies including shRNA knockdowns and immunoprecipitation assays have...et al., 2005). These mice express the human diphtheria toxin receptor (DTR) under the control of cd11b, a macrophage-specific promoter. Because the

  14. Bone morphogenetic protein 2 signaling negatively modulates lymphatic development in vertebrate embryos

    DEFF Research Database (Denmark)

    Dunworth, William P; Cardona-Costa, Jose; Bozkulak, Esra Cagavi

    2014-01-01

    signaling in zebrafish embryos and mouse embryonic stem cell-derived embryoid bodies substantially decrease the emergence of LECs. Mechanistically, BMP2 signaling induces expression of miR-31 and miR-181a in a SMAD-dependent mechanism, which in turn results in attenuated expression of prospero homeobox...

  15. Bone morphogenetic proteins regulate osteoprotegerin and its ligands in human vascular smooth muscle cells

    DEFF Research Database (Denmark)

    Knudsen, Kirsten Quyen Nguyen; Olesen, Ping; Ledet, Thomas

    2007-01-01

    in the transformation of human vascular smooth muscle cells (HVSMC) to osteoblast-like cells. In this study, we evaluated the effect of BMP-2, BMP-7 and transforming growth factor beta (TGF-beta1) on the secretion and mRNA expression of OPG and its ligands receptor activator of nuclear factor-kappabeta ligand (RANKL...

  16. Bone morphogenetic protein activity and connective tissue growth factor in renal and vascular disease

    NARCIS (Netherlands)

    Leeuwis, J.W.

    2011-01-01

    Response to renal injury is dependent on growth factors that determine how resident cells act, which cells are attracted to the site of injury, and how these resident cells, together with infiltrating cells and their surrounding matrix act together. These mechanisms are not confined to the kidney an

  17. Functional assay, expression of growth factors and proteins modulating bone-arrangement in human osteoblasts seeded on an anorganic bovine bone biomaterial

    Directory of Open Access Journals (Sweden)

    O Trubiani

    2010-07-01

    Full Text Available The basic aspects of bone tissue engineering include chemical composition and geometry of the scaffold design, because it is very important to improve not only cell attachment and growth but especially osteodifferentiation, bone tissue formation, and vascularization. Geistlich Bio-Oss® (GBO is a xenograft consisting of deproteinized, sterilized bovine bone, chemically and physically identical to the mineral phase of human bone.In this study, we investigated the growth behaviour and the ability to form focal adhesions on the substrate, using vinculin, a cytoskeletal protein, as a marker. Moreover, the expression of bone specific proteins and growth factors such as type I collagen, osteopontin, bone sialoprotein, bone morphogenetic protein-2 (BMP-2, BMP-7 and de novo synthesis of osteocalcin in normal human osteoblasts (NHOst seeded on xenogenic GBO were evaluated. Our observations suggest that after four weeks of culture in differentiation medium, the NHOst showed a high affinity for the three dimensional biomaterial; in fact, cellular proliferation, migration and colonization were clearly evident. The osteogenic differentiation process, as demonstrated by morphological, histochemical, energy dispersive X-ray microanalysis and biochemical analysis was mostly obvious in the NHOst grown on three-dimensional inorganic bovine bone biomaterial. Functional studies displayed a clear and significant response to calcitonin when the cells were differentiated. In addition, the presence of the biomaterial improved the response, suggesting that it could drive the differentiation of these cells towards a more differentiated osteogenic phenotype. These results encourage us to consider GBO an adequate biocompatible three-dimensional biomaterial, indicating its potential use for the development of tissue-engineering techniques.

  18. Low-dose rhBMP2/7 heterodimer to reconstruct peri-implant bone defects: a micro-CT evaluation

    NARCIS (Netherlands)

    Wang, J.; Zheng, Y.; Zhao, J.; Liu, T.; Gao, L.; Gu, Z.; Wu, G.

    2012-01-01

    Objectives To delineate the dynamic micro-architectures of bone induced by low-dose bone morphogenetic protein (BMP)-2/7 heterodimer in peri-implant bone defects compared to BMP2 and BMP7 homodimer. Material and Methods Peri-implant bone defects (8 mm in diameter, 4 mm in depth) were created surroun

  19. Regulation of Bone Metabolism.

    Science.gov (United States)

    Shahi, Maryam; Peymani, Amir; Sahmani, Mehdi

    2017-04-01

    Bone is formed through the processes of endochondral and intramembranous ossification. In endochondral ossification primary mesenchymal cells differentiate to chondrocytes and then are progressively substituted by bone, while in intramembranous ossification mesenchymal stem cells (MSCs) differentiate directly into osteoblasts to form bone. The steps of osteogenic proliferation, differentiation, and bone homeostasis are controlled by various markers and signaling pathways. Bone needs to be remodeled to maintain integrity with osteoblasts, which are bone-forming cells, and osteoclasts, which are bone-degrading cells.In this review we considered the major factors and signaling pathways in bone formation; these include fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), wingless-type (Wnt) genes, runt-related transcription factor 2 (RUNX2) and osteoblast-specific transcription factor (osterix or OSX).

  20. Regulation of Bone Metabolism

    Science.gov (United States)

    Shahi, Maryam; Peymani, Amir; Sahmani, Mehdi

    2017-01-01

    Bone is formed through the processes of endochondral and intramembranous ossification. In endochondral ossification primary mesenchymal cells differentiate to chondrocytes and then are progressively substituted by bone, while in intramembranous ossification mesenchymal stem cells (MSCs) differentiate directly into osteoblasts to form bone. The steps of osteogenic proliferation, differentiation, and bone homeostasis are controlled by various markers and signaling pathways. Bone needs to be remodeled to maintain integrity with osteoblasts, which are bone-forming cells, and osteoclasts, which are bone-degrading cells.In this review we considered the major factors and signaling pathways in bone formation; these include fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), wingless-type (Wnt) genes, runt-related transcription factor 2 (RUNX2) and osteoblast-specific transcription factor (osterix or OSX). PMID:28367467

  1. Protein growth factors loaded highly porous chitosan scaffold: A comparison of bone healing properties

    Energy Technology Data Exchange (ETDEWEB)

    Nandi, Samit K., E-mail: samitnandi1967@gmail.com [Department of Veterinary Surgery and Radiology, West Bengal University of Animal and Fishery Sciences, Kolkata (India); Kundu, Biswanath, E-mail: biswa_kundu@rediffmail.com [Bioceramics and Coating Division, CSIR—Central Glass and Ceramic Research Institute, Kolkata (India); Basu, Debabrata [Bioceramics and Coating Division, CSIR—Central Glass and Ceramic Research Institute, Kolkata (India)

    2013-04-01

    Present study aimed to investigate and compare effectiveness of porous chitosan alone and in combination with insulin like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) in bone healing. Highly porous (85 ± 2%) with wide distribution of macroporous (70–900 μm) chitosan scaffolds were fabricated as bone substitutes by employing a simple liquid hardening method using 2% (w/v) chitosan suspension. IGF-1 and BMP-2 were infiltrated using vacuum infiltration with freeze drying method. Adsorption efficiency was found to be 87 ± 2 and 90 ± 2% for BMP-2 and IGF-1 respectively. After thorough material characterization (pore details, FTIR and SEM), samples were used for subsequent in vivo animal trial. Eighteen rabbit models were used to evaluate and compare control (chitosan) (group A), chitosan with IGF-1 (group B) and chitosan with BMP-2 (group C) in the repair of critical size bone defect in tibia. Radiologically, there was evidence of radiodensity in defect area from 60th day (initiated on 30th day) in groups B and C as compared to group A and attaining nearly bony density in most of the part at day 90. Histological results depicted well developed osteoblastic proliferation around haversian canal along with proliferating fibroblast, vascularization and reticular network which was more pronounced in group B followed by groups C and A. Fluorochrome labeling and SEM studies in all groups showed similar outcome. Hence, porous chitosan alone and in combination with growth factors (GFs) can be successfully used for bone defect healing with slight advantage of IGF-1 in chitosan samples. - Highlights: ► Fabrication and characterization of porous chitosan with or without IGF-1 and BMP-2 ► Highly porous growth factor loaded chitosan studied in animal subjects for 3 months ► Parameters studied: histopathology, radiology and fluorochrome labeling ► IGF-1 loaded porous chitosan found to be very effective for bone defect healing.

  2. A strategy to quantitate global phosphorylation of bone matrix proteins.

    Science.gov (United States)

    Sroga, Grażyna E; Vashishth, Deepak

    2016-04-15

    Current studies of protein phosphorylation focus primarily on the importance of specific phosphoproteins and their landscapes of phosphorylation in the regulation of different cellular functions. However, global changes in phosphorylation of extracellular matrix phosphoproteins measured "in bulk" are equally important. For example, correct global phosphorylation of different bone matrix proteins is critical to healthy tissue biomineralization. To study changes of bone matrix global phosphorylation, we developed a strategy that combines a procedure for in vitro phosphorylation/dephosphorylation of fully mineralized bone in addition to quantitation of the global phosphorylation levels of bone matrix proteins. For the first time, we show that it is possible to enzymatically phosphorylate/dephosphorylate fully mineralized bone originating from either cadaveric human donors or laboratory animals (mice). Using our strategy, we detected the difference in the global phosphorylation levels of matrix proteins isolated from wild-type and osteopontin knockout mice. We also observed that the global phosphorylation levels of matrix proteins isolated from human cortical bone were lower than those isolated from trabecular bone. The developed strategy has the potential to open new avenues for studies on the global phosphorylation of bone matrix proteins and their role in biomineralization as well for other tissues/cells and protein-based materials.

  3. A novel insertion mutation in the cartilage-derived morphogenetic protein-1 (CDMP1 gene underlies Grebe-type chondrodysplasia in a consanguineous Pakistani family

    Directory of Open Access Journals (Sweden)

    Ansar Muhammad

    2008-11-01

    Full Text Available Abstract Background Grebe-type chondrodysplasia (GCD is a rare autosomal recessive syndrome characterized by severe acromesomelic limb shortness with non-functional knob like fingers resembling toes. Mutations in the cartilage-derived morphogenetic protein 1 (CDMP1 gene cause Grebe-type chondrodysplasia. Methods Genotyping of six members of a Pakistani family with Grebe-type chondrodysplasia, including two affected and four unaffected individuals, was carried out by using polymorphic microsatellite markers, which are closely linked to CDMP1 locus on chromosome 20q11.22. To screen for a mutation in CDMP1 gene, all of its coding exons and splice junction sites were PCR amplified from genomic DNA of affected and unaffected individuals of the family and sequenced directly in an ABI Prism 310 automated DNA sequencer. Results Genotyping results showed linkage of the family to CDMP1 locus. Sequence analysis of the CDMP1 gene identified a novel four bases insertion mutation (1114insGAGT in exon 2 of the gene causing frameshift and premature termination of the polypeptide. Conclusion We describe a 4 bp novel insertion mutation in CDMP1 gene in a Pakistani family with Grebe-type chondrodysplasia. Our findings extend the body of evidence that supports the importance of CDMP1 in the development of limbs.

  4. The Structure and Function of Non-Collagenous Bone Proteins

    Science.gov (United States)

    Hook, Magnus; McQuillan, David J.

    1997-01-01

    The research done under the cooperative research agreement for the project titled 'The structure and function of non-collagenous bone proteins' represented the first phase of an ongoing program to define the structural and functional relationships of the principal noncollagenous proteins in bone. An ultimate goal of this research is to enable design and execution of useful pharmacological compounds that will have a beneficial effect in treatment of osteoporosis, both land-based and induced by long-duration space travel. The goals of the now complete first phase were as follows: 1. Establish and/or develop powerful recombinant protein expression systems; 2. Develop and refine isolation and purification of recombinant proteins; 3. Express wild-type non-collagenous bone proteins; 4. Express site-specific mutant proteins and domains of wild-type proteins to enhance likelihood of crystal formation for subsequent solution of structure.

  5. Novel silk protein barrier membranes for guided bone regeneration.

    Science.gov (United States)

    Smeets, Ralf; Knabe, Christine; Kolk, Andreas; Rheinnecker, Michael; Gröbe, Alexander; Heiland, Max; Zehbe, Rolf; Sachse, Manuela; Große-Siestrup, Christian; Wöltje, Michael; Hanken, Henning

    2016-10-12

    This study assesses the biocompatibility of novel silk protein membranes with and without modification, and evaluates their effect on facilitating bone formation and defect repair in guided bone regeneration. Two calvarian bone defects 12 mm in diameter were created in each of a total of 38 rabbits. Four different types of membranes, (silk-, hydroxyapatite-modified silk-, β-TCP-modified silk- and commonly clinically used collagen-membranes) were implanted to cover one of the two defects in each animal. Histologic analysis did not show any adverse tissue reactions in any of the defect sites indicating good biocompatibility of all silk protein membranes. Histomorphometric and histologic evaluation revealed that collagen and β-TCP modified silk membranes supported bone formation (collagen: bone area fraction p = 0.025; significant; β-TCP modified silk membranes bone area fraction: p = 0.24, not significant), guided bone regeneration and defect bridging. The bone, which had formed in defects covered by β-TCP modified silk membranes, displayed a more advanced stage of bone tissue maturation with restoration of the original calvarial bone microarchitecture when compared to the bone which had formed in defects, for which any of the other test membranes were used. Micro-CT analysis did not reveal any differences in the amount of bone formation between defects with and without membranes. In contrast to the collagen membranes, β-TCP modified silk membranes were visible in all cases and may therefore be advantageous for further supporting bone formation beyond 10 weeks and preventing soft tissue ingrowth from the periphery. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2016.

  6. Dietary protein: an essential nutrient for bone health.

    Science.gov (United States)

    Bonjour, Jean-Philippe

    2005-12-01

    Nutrition plays a major role in the development and maintenance of bone structures resistant to usual mechanical loadings. In addition to calcium in the presence of an adequate vitamin D supply, proteins represent a key nutrient for bone health, and thereby in the prevention of osteoporosis. In sharp opposition to experimental and clinical evidence, it has been alleged that proteins, particularly those from animal sources, might be deleterious for bone health by inducing chronic metabolic acidosis which in turn would be responsible for increased calciuria and accelerated mineral dissolution. This claim is based on an hypothesis that artificially assembles various notions, including in vitro observations on the physical-chemical property of apatite crystal, short term human studies on the calciuric response to increased protein intakes, as well as retrospective inter-ethnic comparisons on the prevalence of hip fractures. The main purpose of this review is to analyze the evidence that refutes a relation of causality between the elements of this putative patho-physiological "cascade" that purports that animal proteins are causally associated with an increased incidence of osteoporotic fractures. In contrast, many experimental and clinical published data concur to indicate that low protein intake negatively affects bone health. Thus, selective deficiency in dietary proteins causes marked deterioration in bone mass, micro architecture and strength, the hallmark of osteoporosis. In the elderly, low protein intakes are often observed in patients with hip fracture. In these patients intervention study after orthopedic management demonstrates that protein supplementation as given in the form of casein, attenuates post-fracture bone loss, increases muscles strength, reduces medical complications and hospital stay. In agreement with both experimental and clinical intervention studies, large prospective epidemiologic observations indicate that relatively high protein intakes

  7. Retention of in vitro and in vivo BMP-2 bioactivities in sustained delivery vehicles for bone tissue engineering

    NARCIS (Netherlands)

    Kempen, Diederik H. R.; Lu, Lichun; Hefferan, Teresa E.; Creemers, Laura B.; Maran, Avudaiappan; Classic, Kelly L.; Dhert, Wouter J. A.; Yaszemski, Michael J.

    2008-01-01

    In this study, we investigated the in vitro and in vivo biological activities of bone morphogenetic protein 2 (BMP-2) released from four sustained delivery vehicles for bone regeneration. BMP-2 was incorporated into (1) a gelatin hydr