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Mukhametov, Ural F., Sergey V. Lyulin, Dmitry Yu Borzunov, and Ilgiz F. Gareev. "Clinical use of bone morphogenetic proteins BMP-2 and BMP-7: analysis of current clinical trials." HERALD of North-Western State Medical University named after I.I. Mechnikov 15, no. 1 (2023): 5–20. http://dx.doi.org/10.17816/mechnikov112617.
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Bone morphogenetic proteins have been used in clinical practice in orthopedics, spine surgery, and maxillofacial surgery for nearly a decade. According to research findings, in most cases the frequency of coalescence when using bone morphogenetic proteins is comparable to or higer that the corresponding indicator when using an autograft. To date, BMP-2 and BMP-7 are commercially available for clinical use and have shown efficacy equal to that of autograft in bone defect replacement.
 This study analyzes existing clinical trials registered on the clinicaltirals.gov website for the therapeutic use of BMP-2 and BMP-7 in pathologies of the musculoskeletal system.
 The search strategy was to use the material from the clinicaltrials.gov website, which focuses on key terms such as bonemorphogenetic protein 2 or BMP-2, bone morphogenetic protein 7 or BMP-7, recombinant bone morphogenetic protein 2 or rhBMP-2, recombinant bone morphogenetic protein 7 or rhBMP-7, InductOs, Op1, bone and diseases of the musculoskeletal system. The inclusion and exclusion criteria were divided into two stages.
 By October 2022, about 85 clinical trials had been registered using BMP-2 and about 12 using BMP-7. Most of the studies are in Phase 2, Phase 23, or Phase 4. Most of them focus on areas such as tibial trauma therapy and spinal surgery. However, only 12 clinical trials using BMP-2 provide meaningful results. All the clinical trials have similar preparation methods, and 12 clinical trials have provided positive results without serious side effects.
 There is a wide potential for clinical use of BMP-2. Many preclinical and clinical studies on the use of BMP-2 and BMP-7 are currently underway; their future results will further explore their therapeutic potential in treating musculoskeletal diseases.
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Herford, Alan S. "Recombinant Human Bone Morphogenetic Protein-2 (Rh BMP-2)." Journal of Oral and Maxillofacial Surgery 63, no. 8 (2005): 14–15. http://dx.doi.org/10.1016/j.joms.2005.05.029.
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Langenfeld, Elaine M., and John Langenfeld. "Bone Morphogenetic Protein-2 Stimulates Angiogenesis in Developing Tumors." Molecular Cancer Research 2, no. 3 (2004): 141–49. http://dx.doi.org/10.1158/1541-7786.141.2.3.
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Abstract Bone Morphogenetic Protein-2 (BMP-2) is highly overexpressed in the majority of patient-derived lung carcinomas. However, a mechanism revealing its role in cancer has not been established. Here we report that BMP-2 enhances the neovascularization of developing tumors. Recombinant BMP-2 stimulated blood vessel formation in tumors formed from A549 cells injected s.c. into thymic nude mice. Recombinant BMP-2 also enhanced angiogenesis in Matrigel plugs containing A549 cells in nude mice. The BMP-2 antagonist noggin abrogated BMP-2-induced angiogenic response. Furthermore, antisense transfection of BMP-2 cDNA resulted in a decrease in blood vessel formation in the Matrigel assays. BMP-2 induced tube formation in both human aortic endothelial cells (HAEC) and umbilical vein endothelial cells. BMP-2 also stimulated proliferation of HAEC. The ability of BMP-2 to activate endothelial cells was further demonstrated by its ability to phosphorylate Smad 1/5/8 and ERK-1/2 and to increase expression of Id1. This study reveals that BMP-2 enhanced the angiogenic response in developing tumors. Furthermore, these data suggest that BMP-2 stimulation of angiogenesis may involve the activation of endothelial cells.
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Fahdia Mazfufah, Nuzli, Retno Wahyu Nurhayati, Ismail Hadisoebroto Dilogo, Dian Anggraini, and Wildan Mubarok. "Signal peptides for optimalization secretion of recombinant protein: An innovation to produce bone morphogenetic protein-2 in mesenchymal stem cells." Bali Medical Journal 13, no. 3 (2024): 1452–61. https://doi.org/10.15562/bmj.v13i2.5020.
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Bone morphogenetic protein 2 (BMP-2) is a protein or growth factor that plays an important role in osteogenic differentiation. Recently, there is an increasing demand for BMP-2, as it is the only osteoinductive growth factor approved by the Food and Drug Administration (FDA) as a substitute for bone grafts. However, recombinant BMP-2 production suffers from low production yields and changes in the biological function of the protein due to the low efficiency of translocation in production hosts. One of the key regulators that influences the efficiency of protein translocation is the signal peptide (SP). SP regulates the process of post-translational protein modification, which determines the fate of the synthesized protein. This literature review aims to explore the role of SP in the process of recombinant protein production in mammalian cells and the utilization of SP in BMP-2 production in the secretome of mesenchymal stem cells (MSCs). The production of recombinant proteins in the MSC secretome is expected to be an innovative therapy because it combines the positive effects of the secretome and BMP-2 together for bone healing.
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Rogers, M. B., V. Rosen, J. M. Wozney, and L. J. Gudas. "Bone morphogenetic proteins-2 and -4 are involved in the retinoic acid-induced differentiation of embryonal carcinoma cells." Molecular Biology of the Cell 3, no. 2 (1992): 189–96. http://dx.doi.org/10.1091/mbc.3.2.189.
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Bone morphogenetic proteins-2 and -4 (BMPs-2 and -4) are transforming growth factor beta-related proteins that can induce bone formation in vivo. We observed that the level of endogenous BMP-2 mRNA increased an average of 11-fold on differentiation of F9 embryonal carcinoma cells into parietal endoderm after treatment with retinoic acid (RA) and cAMP, whereas the message for the closely related BMP-4 decreased 12-fold after this treatment. Therefore, the effects of exogenous recombinant BMP-2 protein on the RA-induced differentiation of F9 embryonal carcinoma cells were investigated. BMP-2 addition altered the growth and morphology of RA-treated but not untreated cells. Moreover, the abundance of several messages was affected by exogenous BMP-2 treatment. Notably, the BMP-2 and -4 messages themselves were reduced by the addition of exogenous BMP-2. The observations suggest that RA, which is known to affect bone morphogenesis, may regulate the osteoinductive proteins, BMP-2 and -4. Furthermore, BMP-2 and -4 may be involved in preimplantation embryogenesis.
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Halloran, Daniel, Hilary W. Durbano, and Anja Nohe. "Bone Morphogenetic Protein-2 in Development and Bone Homeostasis." Journal of Developmental Biology 8, no. 3 (2020): 19. http://dx.doi.org/10.3390/jdb8030019.
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Bone morphogenetic proteins (BMPs) are multi-functional growth factors belonging to the Transforming Growth Factor-Beta (TGF-β) superfamily. These proteins are essential to many developmental processes, including cardiogenesis, neurogenesis, and osteogenesis. Specifically, within the BMP family, Bone Morphogenetic Protein-2 (BMP-2) was the first BMP to be characterized and has been well-studied. BMP-2 has important roles during embryonic development, as well as bone remodeling and homeostasis in adulthood. Some of its specific functions include digit formation and activating osteogenic genes, such as Runt-Related Transcription Factor 2 (RUNX2). Because of its diverse functions and osteogenic potential, the Food and Drug Administration (FDA) approved usage of recombinant human BMP-2 (rhBMP-2) during spinal fusion surgery, tibial shaft repair, and maxillary sinus reconstructive surgery. However, shortly after initial injections of rhBMP-2, several adverse complications were reported, and alternative therapeutics have been developed to limit these side-effects. As the clinical application of BMP-2 is largely implicated in bone, we focus primarily on its role in bone. However, we also describe briefly the role of BMP-2 in development. We then focus on the structure of BMP-2, its activation and regulation signaling pathways, BMP-2 clinical applications, and limitations of using BMP-2 as a therapeutic. Further, this review explores other potential treatments that may be useful in treating bone disorders.
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Hwang, Chang Ju, Alexander R. Vaccaro, James P. Lawrence, et al. "Immunogenicity of bone morphogenetic proteins." Journal of Neurosurgery: Spine 10, no. 5 (2009): 443–51. http://dx.doi.org/10.3171/2009.1.spine08473.
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Object The object of this paper is to review the immunogenicity of bone morphogenetic proteins (BMPs) and to compare the results of the immunogenicity characterization and clinical consequences between recombinant human (rh)BMP-2 and recombinant human osteogenic protein-1 (rhOP-1/BMP-7). Methods The immunogenicity of therapeutic proteins and its clinical effects were reviewed. The characteristics of BMPs were also described in terms of immunogenicity. The methods and results of antibody detection in various clinical trials of rhBMP-2 and rhOP-1 were compared, including the most recent studies using a systematic characterization strategy with both a binding assay and bioassay. Results Similar to all recombinant human proteins, rhBMPs induce immune responses in a select subgroup of patients. Adverse effects from this response in these patients, however, have not been reported with antibody formation to either rhBMP-2 or rhOP-1. Overall, the incidence of antibody formation was slightly higher in rhOP-1 trials than in rhBMP-2 trials. Conclusions Although they occur in a subgroup of patients, the immune responses against rhBMPs have no correlation with any clinical outcome or safety parameter. Clinicians, however, must be aware of the potential complications caused by the immunogenicity of BMPs until more studies clearly elucidate their safety.
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Chen, Yuwen, Evelyn Caporali, and Matthew Stewart. "Bone morphogenetic protein 2 stimulates chondrogenesis of equine synovial membrane-derived progenitor cells." Veterinary and Comparative Orthopaedics and Traumatology 29, no. 05 (2016): 378–85. http://dx.doi.org/10.3415/vcot-16-02-0035.
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SummaryObjectives: Bone morphogenetic protein 2 (BMP-2) is critical for skeletal and cartilage development, homeostasis and repair. This study was conducted to clone and characterize equine BMP-2, develop expression constructs for equine BMP-2, and to determine whether BMP-2 can stimulate chondrogenesis of equine synovial membrane-derived progenitor cells (SMPC).Methods: Equine BMP-2 cDNA was amplified from chondrocyte RNA, and then transferred into an expression plasmid and adenoviral vector. Effective expression of equine BMP-2 was confirmed using a BMP reporter cell line. SMPC were isolated from synovium, expanded through two passages and transferred to chondrogenic cultures, with recombinant human (rh) transforming growth factor beta 1 (TGF-[uni03B2]1) or rhBMP-2. Chondro-genesis was assessed by up-regulation of collagen types II and X, and aggrecan mRNA, secretion of collagen type II protein and sulfated glycosaminoglycans (sGAG), and by alkaline phosphatase induction. Chondrogenic stimulation of SMPC by the equine BMP-2 adenovirus was assessed by sGAG secretion and histology.Results: The mature equine BMP-2 peptide is identical to human and murine peptides. Recombinant human BMP-2 and TGF-[uni03B2]1 stimulated equivalent amounts of collagen type II protein in SMPC pellets, but sGAG secretion was doubled by BMP-2. Neither factor stimulated hypertrophic marker expression. The equine BMP-2 adenoviral vector induced chondrogenesis comparably to rhBMP-2 protein, with no indication of hypertrophy.Clinical significance: Bone morphogenetic protein 2 is a potent inducer of SMPC nonhypertrophic chondrogenesis, supporting the use of this combination for articular cartilage repair applications.
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Yao, Shaohua, Ling Li Zhang, Steven Y. Cheng, and Xing Dong Zhang. "Refolding and Purification of rhBMP-2 Expressed as Inclusion Bodies in E.COLI with Hydroxyapatite Chromatography." Key Engineering Materials 288-289 (June 2005): 661–64. http://dx.doi.org/10.4028/www.scientific.net/kem.288-289.661.
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The objective of this study was to develop an efficient method for the production of bioactive bone morphogenetic protein 2 (BMP-2). A recombinant plasmid encoding mature human BMP-2 was transferred and expressed at a high level in E.coli. Most of the aimed proteins existed in inclusion bodies. The non-active recombinant human BMP-2 (rhBMP-2) monomer in inclusion bodies was refolded and simultaneously purified using hydroxyapatite (HA) chromatography. After oxidization of the monomer, the rhBMP-2 dimmer showed biological activity by the induction of alkaline phosphate activity in C2C12 cells. The refolding yield was about 30% and the purity was about 90% just by one chromatography process.
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Paralkar, V. M., B. S. Weeks, Y. M. Yu, H. K. Kleinman, and A. H. Reddi. "Recombinant human bone morphogenetic protein 2B stimulates PC12 cell differentiation: potentiation and binding to type IV collagen." Journal of Cell Biology 119, no. 6 (1992): 1721–28. http://dx.doi.org/10.1083/jcb.119.6.1721.
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Bone morphogenetic protein 2B (BMP 2B, also known as BMP 4) induces cartilage and bone morphogenesis in ectopic extraskeletal sites. BMP 2B is one of several bone morphogenetic proteins which along with activins and inhibins are members of the transforming growth factor-beta (TGF-beta) family. Both BMP 2B and activin A, but not TGF-beta 1, induce rat pheochromocytoma PC12 neuronal cell differentiation and expression of VGF, a nervous system-specific mRNA. PC12 cells exhibited approximately 2,500 receptors per cell for BMP 2B with an apparent dissociation constant of 19 pM. Extracellular matrix components, including fibronectin, laminin, and collagen type IV potentiated the activity of BMP and activin A, with the latter being the most active. Direct experiments demonstrated that radioiodinated BMP 2B bound to collagen type IV better than to either laminin or fibronectin. These data demonstrate a common neurotrophic activity of both BMP 2B and activin A, and suggest that these regulatory molecules alone and in conjunction with extracellular matrix components may play a role in both the development and repair of nervous tissue.
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Subach, Brian R., Regis W. Haid, Gerald E. Rodts, and Michael G. Kaiser. "Bone morphogenetic protein in spinal fusion: overview and clinical update." Neurosurgical Focus 10, no. 4 (2001): 1–6. http://dx.doi.org/10.3171/foc.2001.10.4.4.
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The widespread use of fusion procedures in the management of spinal disorders has led investigators to explore the use of growth and differentiation factors in such procedures. As an adjuvant to allograft bone or as a replacement for harvested autograft, bone morphogenetic proteins (BMPs) appear to improve fusion rates after spinal arthrodesis in both animal models and humans, while reducing the donor-site morbidity previously associated with such procedures. The use of recombinant genetic technology in the production of BMP has improved the efficiency, cost effectiveness, and safety of producing and using such materials. Recombinant human BMP-2 (rhBMP-2), as one of the first factors identified in the process of endochondral bone formation, has been extensively researched over the past decade. The efficacy and dose profile of this differentiation factor in the context of various carrier substrates has been investigated. Based on the encouraging results of preliminary studies, the future role of rhBMP-2 may lie in its replacement of autologous bone grafting and, consequently, the reduced need for instrumented fixation, while concurrently improving overall fusion rates. The authors provide an overview of BMP and review its use in clinical and laboratory settings.
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Jang, Eunhye, Ja-Youn Lee, Eun-Young Lee, and Hyun Seok. "Evaluation of the Bone Regeneration Effect of Recombinant Human Bone Morphogenic Protein-2 on Subperiosteal Bone Graft in the Rat Calvarial Model." Materials 12, no. 10 (2019): 1613. http://dx.doi.org/10.3390/ma12101613.
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The aim of this study was to evaluate the bone regeneration effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on a subperiosteal bone graft in a rat model. A subperiosteal space was made on the rat calvarium, and anorganic bovine bone (ABB), ABB/low bone morphogenetic protein (BMP) (5 µg), and ABB/high BMP (50 µg) were grafted as subperiosteal bone grafts. The new bone formation parameters of bone volume (BV), bone mineral density (BMD), trabecular thickness (TbTh), and trabecular spacing (TbSp) were evaluated by microcomputed tomography (µ-CT), and a histomorphometric analysis was performed to evaluate the new bone formation area. The expression of osteogenic markers, such as bone sialoprotein (BSP) and osteocalcin, were evaluated by immunohistochemistry (IHC). The ABB/high BMP group showed significantly higher BV than the ABB/low BMP (p = 0.004) and control groups (p = 0.000) and higher TbTh than the control group (p = 0.000). The ABB/low BMP group showed significantly higher BV, BMD, and TbTh than the control group (p = 0.002, 0.042, and 0.000, respectively). The histomorphometry showed significantly higher bone formation in the ABB/low and high BMP groups than in the control group (p = 0.000). IHC showed a high expression of BSP and osteocalcin in the ABB/low and high BMP groups. Subperiosteal bone grafts with ABB and rhBMP-2 have not been studied. In our study, we confirmed that rhBMP-2 contributes to new bone formation in a subperiosteal bone graft with ABB.
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T. Ghazi, Ataa, Hayder F. Saloom, and Rana I. Mahmood. "Efficient Delivery of Recombinant Human Bone Morphogenetic Protein (Rhbmp-2) With Cockle Shell Derived Calcium Carbonate Nanoparticles (CaCO3NPs)." Tikrit Journal for Dental Sciences 11, no. 2 (2024): 216–31. http://dx.doi.org/10.25130/tjds.11.2.8.
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Bone morphogenetic protein-2 (BMP-2) has a significant function in the formation of cartilage and bones. Notably, dosing of only BMP-2 protein intravenously is ineffective. Persistent transportation of the stabilized BMP-2 through a carrier has been seen to be essential for enhancing the osteogenesis im pact of BMP-2. The current research built a new system of drug delivery by utilising cockle shell derived calcium carbonate nanoparticles (CaCO3NPs) and studied the efficacy of the delivery system on the recombinant human bone morphogenetic protein (rhBMP-2). rhBMP-2-CaCO3NPs nanoparticles were synthesised by means of a modest precipitation procedure along with mechanical grinding. Fourier-tran sform infrared spectroscopy, UV–Vis spectrophotometer, scanning electron microscope, X-ray powder diffraction, transmission electron microscope, and zeta potential were u tilised for characterising the conjugated rhBMP-2-CaCO3NPs . Cytotoxicity of rhBMP-2, CaCO3NPs and rhBMP-2-CaCO 3NPs was studied by utilising methylthiazol tetrazolium assay against fibroblast (Rat-1) cells in comparison to rhBMP-2 and CaCO3NPs. The outcomes signified bio-stability of CaCO3NPs and lower toxicity for Rat-1 cells. In summary, CaCO3NPs were prepared by a simp le precipitation process. The ensuing nanoparticles could competently entrap rhBMP-2 and generated stable rhBMP-2-CaCO3NPs. A sustained discharge of rhBMP-2 from t he CaCO3NPs was seen. CaCO3NPs loaded with r hBMP-2 demonstrated reasonable bio-compatibility. The outcomes indicated that CaCO3NPs may have significant ability as carrier of therapeutic proteins within bone tissue en gineering.
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Alden, Tord D., Debra D. Pittman, Elisa J. Beres, et al. "Percutaneous spinal fusion using bone morphogenetic protein-2 gene therapy." Journal of Neurosurgery: Spine 90, no. 1 (1999): 109–14. http://dx.doi.org/10.3171/spi.1999.90.1.0109.
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Object. Gene therapy has many potential applications in neurosurgery. One application involves bone morphogenetic protein-2 (BMP-2), a low-molecular-weight glycoprotein that induces bone formation in vivo. Numerous studies have demonstrated that the BMP-2 protein can enhance spinal fusion. This study was undertaken to determine whether direct injection of an adenoviral construct containing the BMP-2 gene can be used for spinal fusion. Methods. Twelve athymic nude rats were used in this study. Recombinant, replication-defective type 5 adenovirus with the cytomegalovirus (CMV) promoter and BMP-2 gene (Ad-BMP-2) was used. A second adenovirus constructed with the CMV promoter and β-galactosidase (β-gal) gene (Ad-β-gal) was used as a control. In three groups (four rats each) 7.5 µl of virus (5 × 108 particles/µl) was injected percutaneously and paraspinally at the lumbosacral junction: Group 1 received Ad-BMP-2 bilaterally; Group 2 received Ad-BMP-2 on the right, Ad-β-gal on the left; and Group 3 received Ad-β-gal bilaterally. Computerized tomography (CT) scans of the lumbosacral spine were obtained at 3, 5, 8, and 12 weeks. At 12 weeks, the animals were killed and underwent histological inspection. Ectopic bone formation was observed both on three-dimensionally reconstructed CT scans and histological examination in all rats at sites treated with Ad-BMP-2. Histological analysis demonstrated bone at different stages of maturity adjacent to the spinous processes, laminae, and transverse processes. Conclusions. Results of this study clearly demonstrated that it is possible to produce in vivo endochondral bone formation by using direct adenoviral construct injection into the paraspinal musculature, which suggests that gene therapy may be useful for spinal fusion in the future.
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Mukhametov, U. F., S. V. Lyulin, D. Yu Borzunov, R. A. Sufianov, and I. F. Gareev. "The risk of tumor with the use of recombinant human bone morphogenetic proteins." Genij Ortopedii 28, no. 4 (2022): 592–98. http://dx.doi.org/10.18019/1028-4427-2022-28-4-592-598.
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Introduction Bone morphogenetic proteins (BMPs) are members of a large family of growth factors known as the transforming growth factor-β (TGF-β) superfamily. BMPs are known for their ability to induce bone formation and successfully used in orthopaedic and neurosurgical applications. Various proteins, such as BMP-2, 4, 7, have been reported to have osteoinductive abilities. Recombinant human bone morphogenetic protein-2 (rhBMP-2) and recombinant human bone morphogenetic protein-7 (rhBMP-7) are widely used for surgical correction of bone defects and spinal fusions. In addition to the effect on bone formation, BMPs also play a role in cell lineage determination, differentiation, proliferation and apoptosis, and BMP receptors are present in many cell types including tumor cells. A large number of studies in vitro and in vivo have examined the role of BMPs as stimulating oncogenesis and metastasis. Therefore, there are some concerns about the use of rhBMPs in clinical practice. Objective In the present study, we aimed to investigate the causal relationship between the use of rhBMPs and oncogenesis by presenting the results of some preclinical and clinical studies. Material and methods For a comprehensive search, we used the following databases: PubMed, Embase, the Cochrane Database and Google Scholar to identifying studies that described a causal relationship between therapeutic use of rhBMPs and oncogenesis. Results The paper represents the findings on the role and identification of molecular mechanisms of BMP involvement in oncogenesis. In addition to that, the studies reporting a risk of oncological diseases with the use of rhBMPs in both preclinical and clinical studies were also analyzed. Conclusion There is a need for further clinical trials in a wide population over a longer timeframe.
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Alden, Tord D., Gerald R. Hankins, Elisa J. Beres, David F. Kallmes, and Gregory A. Helm. "Bone morphogenetic protein gene therapy for the induction of spinal arthrodesis." Neurosurgical Focus 4, no. 2 (1998): E14. http://dx.doi.org/10.3171/foc.1998.4.2.15.
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Gene therapy has many potential applications in neurosurgery. One application involves bone morphogenetic protein-2 (BMP-2), a low-molecular-weight glycoprotein that induces bone formation in vivo. Numerous studies have demonstrated that the BMP-2 protein can enhance spinal fusion. This study was undertaken to determine whether direct injection of an adenoviral construct containing the BMP-2 gene can be used for spinal fusion. Twelve athymic nude rats were used in this study. Recombinant, replication-defective type-5 adenovirus with a universal promoter and BMP-2 gene (Ad-BMP-2) was used. A second adenovirus constructed with a universal promoter and ß-galactosidase (ß-gal) gene (Ad-ß-gal) was used as a control. Seven and one-half microliters of virus was injected percutaneously and paraspinally at the lumbosacral junction in three groups (four animals each): 1) Ad-BMP-2 bilaterally, 2) Ad-BMP-2 on the right, Ad-ß-gal on the left, and 3) Ad-ß-gal bilaterally. Computerized tomography (CT) scans of the lumbosacral spine were obtained at 3, 5, and 12 weeks. At 12 weeks, the animals were killed for histological inspection. Ectopic bone formation was seen both on three-dimensional CT reconstruction and histologically in all rats at sites treated with Ad-BMP-2. Histological analysis revealed bone at different stages of maturity adjacent to the spinous processes, laminae, and transverse processes. This study clearly demonstrated that it is possible to produce in vivo endochondral bone formation by using direct adenoviral construct injection into the paraspinal musculature, which suggests that gene therapy may be useful for spinal fusion in the future.
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Chen, Thomas C. "Recombinant human morphogenetic protein: its future role in spinal fusions." Neurosurgical Focus 4, no. 2 (1998): E13. http://dx.doi.org/10.3171/foc.1998.4.2.14.
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The marriage of basic scientific research and clinical application often leads to profound advances in our understanding of various disease processes and how they may be ameliorated. One such fortuitous combination has been the discovery of bone morphogenetic proteins (BMPs) and their potential application in spinal fusions. The goal of this article is to introduce the neurosurgeon to the basic biology of this protein family, current experimental data (in vitro and in vivo models) demonstrating their effectiveness in enhancing bony fusions, and preliminary clinical trials utilizing BMP in long bone fusions. Using this information, a proposal for the use of BMP in spinal fusions under various clinical scenarios will be discussed.
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Shimaoka, H., Y. Dohi, K. Narikawa, et al. "Comparison of Recombinant Growth/Differentiation Factor-5 (rGDF-5) and Bone Morphogenetic Protein-2 (rBMP-2) in the In Vivo Bone Formation in Porous Hydroxyapatite Ceramic." Key Engineering Materials 284-286 (April 2005): 945–48. http://dx.doi.org/10.4028/www.scientific.net/kem.284-286.945.
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Various recombinant growth factors have been used for promoting osteoblastic differentiation cascade. To compare the growth/differentiation factor-5 (GDF-5) and bone morphogenetic protein-2 (BMP-2) in the in vivo osteogenic potential of bone marrow mesenchymal stem cells (MSCs), the bone formation was assessed by rat subcutaneous implantation of 5 kinds of hydroxyapatite (HA) implants; namely GDF/HA composites, BMP/HA composites, MSCs/HA composites and the MSCs/HA composites supplemented with recombinant mouse GDF-5 (GDF/MSCs/HA) or recombinant human BMP-2 (BMP/MSCs/HA). Neither the GDF/HA nor the BMP/HA composites exhibited any bone formation at any time after implantation. At both 2 and 4 weeks after implantation, obvious de novo bone formation together with active osteoblasts was seen histologically in many pores of the GDF/MSCs/HA and BMP/MSCs/HA composites. The GDF/MSCs/HA and BMP/MSCs/HA composites also showed high alkaline phosphatase (ALP) and osteocalcin expression determined at both the protein and gene levels. Compared with GDF/MSCs/HA, the BMP/MSCs/HA composites exhibited excellent osteogenesis with relatively early osteoblastic phenotype expression. These findings indicate that the two growth factors synergistically enhance de novo bone formation capability of MSCs/HA composites and the importance of ceramic surface to retain and to deliver the molecules of growth factors for the cell differentiation and maturation.
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Hawkins, Bryan J. "Recombinant Bone Morphogenetic Protein-2." Techniques in Foot & Ankle Surgery 6, no. 2 (2007): 80–88. http://dx.doi.org/10.1097/btf.0b013e33180620fc2.
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Shimasaki, Shunichi, R. Kelly Moore, Fumio Otsuka, and Gregory F. Erickson. "The Bone Morphogenetic Protein System In Mammalian Reproduction." Endocrine Reviews 25, no. 1 (2004): 72–101. http://dx.doi.org/10.1210/er.2003-0007.
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Using molecular, cellular, and genetic approaches, recent studies examining the role of the bone morphogenetic protein (BMP) family of growth factors in the reproductive system have led to significant breakthroughs in our understanding of mammalian reproduction and fertility. Gene expression studies have revealed that key components of the BMP system (ligands, receptors, signaling molecules, and binding proteins) exhibit coordinated spatial and temporal expression patterns in fundamental cell types throughout the reproductive system. Availability of recombinant BMPs has enabled functional studies that have demonstrated important biological activities of BMPs in controlling cellular proliferation, differentiation, and apoptosis in reproductive tissues. The physiological importance of the BMP system for mammalian reproduction has been further highlighted by the elucidation of the aberrant reproductive phenotypes of animals with naturally occurring mutations or targeted deletions of certain BMP family genes. Collectively, these studies have established the concept that the BMP system plays a crucial role in fertility in female and male mammals. The purpose of this article is to review the evidence underpinning the importance of the BMP system in mammalian reproduction.
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Villavicencio, Alan T., Sigita Burneikiene, E. Lee Nelson, Ketan R. Bulsara, Mark Favors, and Jeffrey Thramann. "Safety of transforaminal lumbar interbody fusion and intervertebral recombinant human bone morphogenetic protein—2." Journal of Neurosurgery: Spine 3, no. 6 (2005): 436–43. http://dx.doi.org/10.3171/spi.2005.3.6.0436.
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Object. Recombinant human bone morphogenetic protein—2 (rhBMP-2) is being increasingly used for spinal fusion. There are few data regarding its clinical safety, effectiveness, and clinical outcome when applied on an absorbable collagen sponge (ACS) in conjunction with allograft for transforaminal lumbar interbody fusion (TLIF). Methods. Seventy-four consecutive patients undergoing TLIF for degenerative disc disease were divided into five groups depending on whether the patient underwent a minimally invasive or open approach, as well as the number of spinal levels surgically treated. Surgery-related data, fusion results, complications, and clinical outcome were evaluated. The mean follow-up duration was 20.6 months (range 14–28 months). The radiographic fusion rate was 100% at 12 and 24 months after the surgery. No bone overgrowth or other complications related to BMP use were demonstrated. Conclusions. Analysis of the results demonstrated that TLIF combined with a BMP-2—soaked ACS is a feasible, effective, and safe method to promote lumbar fusion. There were no significant intergroup differences in clinical outcome between patients who underwent open compared with minimally invasive procedures. Patient satisfaction rates, however, were higher in the minimally invasive procedure group. The efficacy of BMP-2 was not dependent on which approach was used or the number of spinal levels that were treated.
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Seok, Hyun, Hee-Youl Kim, Dong-Cheol Kang, Jung-Ho Park, and Jong Hoon Park. "Comparison of Bone Regeneration in Different Forms of Bovine Bone Scaffolds with Recombinant Human Bone Morphogenetic Protein-2." International Journal of Molecular Sciences 22, no. 20 (2021): 11121. http://dx.doi.org/10.3390/ijms222011121.
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The aim of this study was to compare the bone regeneration ability of particle and block bones, acting as bone scaffolds, with recombinant human bone morphogenetic protein (rhBMP)-2 and evaluate them as rhBMP-2 carriers. Demineralized bovine bone particles, blocks, and rhBMP-2 were grafted into the subperiosteal space of a rat calvarial bone, and the rats were randomly divided into four groups: particle, block, P (particle)+BMP, and B (block)+BMP groups. The bone volume of the B+BMP group was significantly higher than that of the other groups (p < 0.00), with no significant difference in bone mineral density. The average adipose tissue volume of the B+BMP group was higher than that of the P+BMP group, although the difference was not significant. Adipose tissue formation was observed in the rhBMP-2 application group. Histologically, the particle and B+BMP groups showed higher formation of a new bone. However, adipose tissue and void spaces were also formed, especially in the B+BMP group. Hence, despite the formation of a large central void space, rhBMP-2 could be effectively used with block bone scaffolds and showed excellent new bone formation. Further studies are required to evaluate the changes in adipose tissue.
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Rutherford, Bruce, and Mark Fitzgerald. "A New Biological Approach To Vital Pulp Therapy." Critical Reviews in Oral Biology & Medicine 6, no. 3 (1995): 218–29. http://dx.doi.org/10.1177/10454411950060030401.
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Molecular biology is providing opportunities to develop new strategies or agents for the treatment of a wide variety of diseases. The availability of large amounts of highly purified proteins produced by recombinant DNA techniques is an obvious example. Recent evidence has implicated proteins belonging to the bone morphogenetic protein (BMP) subgroup of the transforming growth factor beta supergene family in tooth formation and dentinogenesis. It has long been known that bone and dentin contain bone morphogenetic protein activity. Recently, recombinant human BMP-2, -4, and -7 (also known as OP-1 ) have been shown to induce reparative dentin formation in experimental models of large direct pulp exposures in permanent teeth. The manner in which these agents act appears unique. New reparative dentin replaces the stimulating agents applied directly to the partially amputated pulp. Hence, the new tissue forms contiguous with, largely superficial to, and not at the expense of the remaining vital pulp tissue. This suggests a therapeutic approach permitting the induction of a predetermined and controlled amount of reparative dentin. Additionally, OP-l has been associated with the formation of reparative dentin after application to a freshly cut but intact layer of dentin. These findings may provide future clinicians with additional options for the treatment of substantially damaged or diseased vital teeth.
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Mitola, Stefania, Cosetta Ravelli, Michela Corsini, et al. "Production and Biochemical Characterization of Dimeric Recombinant Gremlin-1." International Journal of Molecular Sciences 23, no. 3 (2022): 1151. http://dx.doi.org/10.3390/ijms23031151.
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Gremlin-1 is a secreted cystine-knot protein that acts as an antagonist of bone morphogenetic proteins (BMPs), and as a ligand of heparin and the vascular endothelial growth factor receptor 2 (VEGFR2), thus regulating several physiological and pathological processes, including embryonic development, tissue fibrosis and cancer. Gremlin-1 exerts all these biological activities only in its homodimeric form. Here, we propose a multi-step approach for the expression and purification of homodimeric, fully active, histidine-tagged recombinant gremlin-1, using mammalian HEK293T cells. Ion metal affinity chromatography (IMAC) of crude supernatant followed by heparin-affinity chromatography enables obtaining a highly pure recombinant dimeric gremlin-1 protein, exhibiting both BMP antagonist and potent VEGFR2 agonist activities.
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Takami, Akiko, Hirotaka Kato, Ryousuke Takagi, and Tomoyuki Miyashita. "Studies on thePinctada fucataBMP-2 Gene: Structural Similarity and Functional Conservation of Its Osteogenic Potential within the Animal Kingdom." International Journal of Zoology 2013 (2013): 1–9. http://dx.doi.org/10.1155/2013/787323.
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Bone morphogenetic protein (BMP)-2 plays an important role in morphogenesis in both vertebrates and invertebrates. BMP-2 is one of the most powerful bioactive substances known to induce the osteogenic differentiation of mesenchymal cells. We examined the structural and functional conservation ofPinctada fucataBMP-2 in inducing osteogenesis in the murine mesenchymal stem cells, C3H10T1/2. Exposure of C3H10T1/2 cells to the recombinant mature fragment ofPinctada fucataBMP-2 resulted in osteoblastic differentiation. The sequence, SVPKPCCVPTELSSL, within the C-terminal portion ofPinctada fucataBMP-2, is homologous to the knuckle epitope of human BMP-2. This synthetic polypeptide was able to induce differentiation of C3H10T1/2 along the osteoblastic lineage, as confirmed by an increase in alkaline phosphatase activity, and the accumulation of calcium, as determined by von Kossa staining. Furthermore, using immunohistochemical staining, we observed an increased expression of collagen type I, osteopontin, and osteocalcin, which are known markers of osteogenesis. These results show that BMP-2 is conserved, not only in terms of its homology at the amino acid sequence, but also in terms of driving the formation of hard tissues in vertebrates and invertebrates.
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Cyr-Depauw, Chanèle, Jason J. Northey, Sébastien Tabariès, et al. "Chordin-Like 1 Suppresses Bone Morphogenetic Protein 4-Induced Breast Cancer Cell Migration and Invasion." Molecular and Cellular Biology 36, no. 10 (2016): 1509–25. http://dx.doi.org/10.1128/mcb.00600-15.
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ShcA is an important mediator of ErbB2- and transforming growth factor β (TGF-β)-induced breast cancer cell migration, invasion, and metastasis. We show that in the context of reduced ShcA levels, the bone morphogenetic protein (BMP) antagonist chordin-like 1 (Chrdl1) is upregulated in numerous breast cancer cells following TGF-β stimulation. BMPs have emerged as important modulators of breast cancer aggressiveness, and we have investigated the ability of Chrdl1 to block BMP-induced increases in breast cancer cell migration and invasion. Breast cancer-derived conditioned medium containing elevated concentrations of endogenous Chrdl1, as well as medium containing recombinant Chrdl1, suppresses BMP4-induced signaling in multiple breast cancer cell lines. Live-cell migration assays reveal that BMP4 induces breast cancer migration, which is effectively blocked by Chrdl1. We demonstrate that BMP4 also stimulated breast cancer cell invasion and matrix degradation, in part, through enhanced metalloproteinase 2 (MMP2) and MMP9 activity that is antagonized by Chrdl1. Finally, high Chrdl1 expression was associated with better clinical outcomes in patients with breast cancer. Together, our data reveal that Chrdl1 acts as a negative regulator of malignant breast cancer phenotypes through inhibition of BMP signaling.
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Velázquez-González, Cindy Sinaí, Ena Athenea Aguilar-Reyes, Rosa Elvira Núñez-Anita, and Carlos Alberto León-Patiño. "Preparation and Characterization of Chitosan Microspheres for Controlled Release of Rh-BMP-2." MRS Advances 4, no. 59-60 (2019): 3259–67. http://dx.doi.org/10.1557/adv.2019.426.
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ABSTRACTThe implantation and controlled release of growth factors can enhance the proliferation and differentiation of cells that promote new bone formation at defect sites. Therefore, chitosan polymer microspheres were prepared by the water-in-oil emulsion (W/O) method and solvent freeze-drying, using glutaraldehyde as an ionic crosslinker, along with the lyophilization of solvents, to microencapsulate growth factors, preventing denaturation. The microspheres were loaded with recombinant bone morphogenetic protein 2 (Rh-BMP-2). They were spherical in shape, with a rough surface ranging in particle size from 0.4 to 1.6 μm. The yield percentage with respect to the polymer was 70% and the BMP-2 load was regulated by the initial protein dose. BMP-2 release experiments were performed for 7 days in PBS solutions at pH 4 and 7.4. The results showed that the protein release rate was only 2% lower at pH 7.4. BMP-2/chitosan microspheres were compatible with the MG-63 cell line (ATCC®CRL-1427™Homo sapiens bone osteosarcoma) and could be considered drug delivery vehicles in bone tissue engineering applications.
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Hu, Ying, Qing-Wei Zhao, Zheng-Cai Wang, et al. "Co-transfection with BMP2 and FGF2 via chitosan nanoparticles potentiates osteogenesis in human adipose-derived stromal cells in vitro." Journal of International Medical Research 49, no. 3 (2021): 030006052199767. http://dx.doi.org/10.1177/0300060521997679.
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Objective To investigate if co-transfection of human bone morphogenetic protein 2 (BMP-2, BMP2) and human fibroblast growth factor 2 (FGF2, FGF2) via chitosan nanoparticles promotes osteogenesis in human adipose tissue-derived stem cells (ADSCs) in vitro. Materials and Methods Recombinant BMP2 and/or FGF2 expression vectors were constructed and packaged into chitosan nanoparticles. The chitosan nanoparticles were characterized by atomic force microscopy. Gene and protein expression levels of BMP-2 and FGF2 in ADSCs in vitro were evaluated by real-time polymerase chain reaction (PCR), western blot, and enzyme-linked immunosorbent assay. Osteocalcin (OCN) and bone sialoprotein (BSP) gene expression were also evaluated by real-time PCR to assess osteogenesis. Results The prepared chitosan nanoparticles were spherical with a relatively homogenous size distribution. The BMP2 and FGF2 vectors were successfully transfected into ADSCs. BMP-2 and FGF2 mRNA and protein levels were significantly up-regulated in the co-transfection group compared with the control group. OCN and BSP mRNA levels were also significantly increased in the co-transfection group compared with cells transfected with BMP2 or FGF2 alone, suggesting that co-transfection significantly enhanced osteogenesis. Conclusions Co-transfection of human ADSCs with BMP2/FGF2 via chitosan nanoparticles efficiently promotes the osteogenic properties of ADSCs in vitro.
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Differ, Christopher, Franka Klatte-Schulz, Nicole Bormann, Susann Minkwitz, Petra Knaus, and Britt Wildemann. "Is NO the Answer? The Nitric Oxide Pathway Can Support Bone Morphogenetic Protein 2 Mediated Signaling." Cells 8, no. 10 (2019): 1273. http://dx.doi.org/10.3390/cells8101273.
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The growth factor bone morphogenetic protein 2 (BMP2) plays an important role in bone development and repair. Despite the positive effects of BMP2 in fracture healing, its use is associated with negative side effects and poor cost effectiveness, partly due to the large amounts of BMP2 applied. Therefore, reduction of BMP2 amounts while maintaining efficacy is of clinical importance. As nitric oxide (NO) signaling plays a role in bone fracture healing and an association with the BMP2 pathway has been indicated, this study aimed to investigate the relationship of BMP2 and NO pathways and whether NO can enhance BMP2-induced signaling and osteogenic abilities in vitro. To achieve this, the stable BMP reporter cell line C2C12BRELuc was used to quantify BMP signaling, and alkaline phosphatase (ALP) activity and gene expression were used to quantify osteogenic potency. C2C12BRELuc cells were treated with recombinant BMP2 in combination with NO donors and substrate (Deta NONOate, SNAP & L-Arginine), NOS inhibitor (LNAME), soluble guanylyl cyclase (sGC) inhibitor (LY83583) and activator (YC-1), BMP type-I receptor inhibitor (LDN-193189), or protein kinase A (PKA) inhibitor (H89). It was found that the NOS enzyme, direct NO application, and sGC enhanced BMP2 signaling and improved BMP2 induced osteogenic activity. The application of a PKA inhibitor demonstrated that BMP2 signaling is enhanced by the NO pathway via PKA, underlining the capability of BMP2 in activating the NO pathway. Collectively, this study proves the ability of the NO pathway to enhance BMP2 signaling.
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SANDHU, HARVINDER S., and SAFDAR N. KHAN. "RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN-2." Journal of Bone and Joint Surgery-American Volume 85 (2003): 89–95. http://dx.doi.org/10.2106/00004623-200300003-00015.
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Park, Shin-Young, Kyoung-Hwa Kim, Sungtae Kim, Yong-Moo Lee, and Yang-Jo Seol. "BMP-2 Gene Delivery-Based Bone Regeneration in Dentistry." Pharmaceutics 11, no. 8 (2019): 393. http://dx.doi.org/10.3390/pharmaceutics11080393.
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Bone morphogenetic protein-2 (BMP-2) is a potent growth factor affecting bone formation. While recombinant human BMP-2 (rhBMP-2) has been commercially available in cases of non-union fracture and spinal fusion in orthopaedics, it has also been applied to improve bone regeneration in challenging cases requiring dental implant treatment. However, complications related to an initially high dosage for maintaining an effective physiological concentration at the defect site have been reported, although an effective and safe rhBMP-2 dosage for bone regeneration has not yet been determined. In contrast to protein delivery, BMP-2 gene transfer into the defect site induces BMP-2 synthesis in vivo and leads to secretion for weeks to months, depending on the vector, at a concentration of nanograms per milliliter. BMP-2 gene delivery is advantageous for bone wound healing process in terms of dosage and duration. However, safety concerns related to viral vectors are one of the hurdles that need to be overcome for gene delivery to be used in clinical practice. Recently, commercially available gene therapy has been introduced in orthopedics, and clinical trials in dentistry have been ongoing. This review examines the application of BMP-2 gene therapy for bone regeneration in the oral and maxillofacial regions and discusses future perspectives of BMP-2 gene therapy in dentistry.
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Deng, Jianye, and Yan Li. "BMP6 Mediates BMP2-Increased Human Trophoblast Invasion." Journal of the Endocrine Society 5, Supplement_1 (2021): A747. http://dx.doi.org/10.1210/jendso/bvab048.1519.
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Abstract TGF-β superfamily proteins play divergent roles in regulating human extravillous trophoblast (EVT) invasion and their coordinated effects are essential for adequate placentation during pregnancy 1. Bone morphogenetic protein 2 (BMP2), which belongs to the BMP subfamily of TGF-β superfamily, has been shown to promote human EVT invasion and the acquisition of endothelial-like phenotype 2,3. It has been reported that BMP2 promotes EVT invasion by up-regulating Activin A, a growth factor which also belongs to TGF-β superfamily. However, whether BMP6 mediates the pro-invasive effect of BMP2 has yet to be determined. Herein, we firstly treated immortalized trophoblast cells (HTR8/SVneo) with recombinant BMP2 protein for 6 and 24 hrs, and our bulk-RNA sequencing results demonstrated significantly increased BMP6 mRNA levels after BMP2 treatment. Furthermore, we confirmed the up-regulatory effects of BMP2 on BMP6 mRNA and protein levels in both HTR8/SVneo and primary EVTs isolated from first-trimester villi. Notably, siRNA-mediated down-regulation of BMP6 significantly attenuated both basal and BMP2-induced cell invasion in HTR8/SVneo cells as measured by Matrigel-coated transwell invasion assay. In summary, our results firstly demonstrated the up-regulatory effect of BMP2 on BMP6 expression in human trophoblasts and identified the mediation role of BMP6 in BMP2-promoted EVT invasion, suggesting the interplay between BMP subfamily members during EVT invasion regulation. Our ongoing research focusing the underlying molecular mechanisms and signaling pathways could further benefit the advancement of diagnostic and therapeutic strategies for EVT invasion dysregulation-related pregnancy disorders, e.g., pre-eclampsia. Reference: (1) Li Yan et al., Trends Endocrinol Metab 2021 18: S1043-2760(20)30266-6. (2) Hong-Jin Zhao et al., FASEB J 2020;34(2):3151-3164. (3) Hong-Jin Zhao et al., Cell Death Dis 2018;9(2):174.
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McMahon, Heather E., Shweta Sharma, and Shunichi Shimasaki. "Phosphorylation of Bone Morphogenetic Protein-15 and Growth and Differentiation Factor-9 Plays a Critical Role in Determining Agonistic or Antagonistic Functions." Endocrinology 149, no. 2 (2007): 812–17. http://dx.doi.org/10.1210/en.2007-1439.
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Two highly homologous oocyte-secreted growth factors, bone morphogenetic protein (BMP)-15 and growth and differentiation factor (GDF)-9, are known to control folliculogenesis and ovulation through direct effects on granulosa cells in the developing follicles. Although much is known about the expression and biology of these proteins, the impact of posttranslational modifications of BMP-15 and GDF-9 is unknown. Here, we report that: 1) recombinant human (rh) BMP-15 and rhGDF-9 are phosphorylated; 2) the phosphorylation is essential for bioactivity; and 3) the dephosphorylated forms of rhBMP-15 and rhGDF-9 can abolish the bioactivity of rhBMP-15, rhGDF-9, and rhBMP-7, but not rh activin A. These results indicate that the phosphorylation state of rhBMP-15 and rhGDF-9 is a determinant of their agonistic and antagonistic activities.
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Chen, Wei, Caiqian Zhang, Yeqing Wu, and Xiuping Su. "Soluble expression and purification of high-bioactivity recombinant human bone morphogenetic protein-2 by codon optimisation in Escherichia coli." Protein Engineering, Design and Selection 32, no. 3 (2019): 153–57. http://dx.doi.org/10.1093/protein/gzz028.
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Abstract We developed a simple method of preparing recombinant human bone morphogenetic protein-2 (rhBMP-2) with high biological activity. This rhBMP-2 was overproduced in Escherichia coli as a fusion protein with thioredoxin 6xHis-tag at its amino terminus. The cDNA fragment of human bone morphogenetic protein-2 (hBMP-2) fused to the secretion signal of alkaline phosphatase (PhoA) was expressed under T7 promoter in E. coli. After DNA sequence confirmation, the recombinant vector pETpho-bmp2 was transformed into E. coli BL21 (DE3). rhBMP-2 was produced by the recombinant strain pETpho-bmp2/BL21 (DE3) in a soluble form with an yield of 6.2 mg/L culture. Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) results showed that the molecular weight of the product was approximately 28 kD. Moreover, rhBMP-2 was secreted as a dimer with a natural structure. rhBMP-2, purified by Ni Nitrilotriacetic acid Agarose (Ni-NTA) affinity chromatography, was used to examine osteosarcoma MG-63 cells and assay the alkaline phosphatase (ALP) activity. Results showed that rhBMP-2 induced MG-63 cell differentiation. When the final concentration was 500 ng/mL, the effect was more remarkable and ALP activity reached 525% compared with that of the control group.
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Rocque, Brandon G., Mick P. Kelly, Joseph H. Miller, Yiping Li, and Paul A. Anderson. "Bone morphogenetic protein–associated complications in pediatric spinal fusion in the early postoperative period: an analysis of 4658 patients and review of the literature." Journal of Neurosurgery: Pediatrics 14, no. 6 (2014): 635–43. http://dx.doi.org/10.3171/2014.8.peds13665.
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Object Use of recombinant human bone morphogenetic protein–2 has risen steadily since its approval by the FDA for use in anterior lumbar interbody fusion in 2002. The FDA has not approved the use of bone morphogenetic protein (BMP) in children. Age less than 18 years or lack of evidence of epiphyseal closure are considered by the manufacturer to be contraindications to BMP use. In light of this, the authors performed a query of the database of one of the nation's largest health insurance companies to determine the rate of BMP use and complications in pediatric patients undergoing spinal fusion. Methods The authors used the PearlDiver Technologies private payer database containing all records from United Health-Care from 2005 to 2011 to query all cases of pediatric spinal fusion with or without BMP use. A review of the literature was also performed to examine the complications associated with BMP use in pediatric spinal fusion. Results A total of 4658 patients underwent spinal fusion. The majority was female (65.4%), and the vast majority was age 10–19 years (94.98%) and underwent thoracolumbar fusion (93.13%). Bone morphogenetic protein was used in 1752 spinal fusions (37.61%). There was no difference in the rate of BMP use when comparing male and female patients or age 10 years or older versus less than 10 years. Anterior cervical fusions were significantly less likely to use BMP (7.3%). Complications occurred in 9.82% of patients treated with versus 9.88% of patients treated without BMP. The complication rate was nearly identical in male versus female patients and in patients older versus younger than 10 years. Comparison of systemic, wound-related, CNS, and other complications showed no difference between groups treated with and without BMP. The reoperation rate was also nearly identical. Conclusions Bone morphogenetic protein is used in a higher than expected percentage of pediatric spinal fusions. The rate of acute complications in these operations does not appear to be different in patients treated with versus those treated without BMP. Caution must be exercised in interpreting these data due to the many limitations of the administrative database as a data source, including the short length of follow-up.
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Suzuki, A., S. Nishimatsu, A. Shoda, K. Takebayashi, K. Murakami, and N. Ueno. "Biochemical properties of amphibian bone morphogenetic protein-4 expressed in CHO cells." Biochemical Journal 291, no. 2 (1993): 413–17. http://dx.doi.org/10.1042/bj2910413.
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The biochemical properties of recombinant amphibian bone morphogenetic protein-4 (BMP-4), the cDNA of which has been cloned recently by screening of a Xenopus cDNA library, was characterized. The protein was expressed by the transfection of Chinese hamster ovary (CHO) cells with the cDNA cloned into expression vectors bearing a cytomegalovirus promoter or a simian virus 40 promoter. Northern-blot analysis showed that the latter vector was more efficient for Xenopus BMP-4 expression. Specific antiserum against Xenopus BMP-4 peptide demonstrated that the protein is synthesized as a large precursor, processed to the mature form and then secreted from the cells as a homodimer. Analysis of the biological activity in the conditioned medium revealed that Xenopus BMP-4 has a potent alkaline phosphatase-inducing activity on mouse osteoblastic cells.
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Zhang, Heng, Wen Zhang, Guangchao Bai, Lei Gao, and Kuanxin Li. "Bone Morphogenetic Protein-7 (BMP-7) Promotes Neuronal Differentiation of Bone Marrow Mesenchymal Stem Cells (BMSCs) In Vitro." BioMed Research International 2021 (January 27, 2021): 1–9. http://dx.doi.org/10.1155/2021/7239783.
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This study is aimed at investigating the effects of bone morphogenetic protein-7 (BMP-7) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells in vitro. The rat BMSCs were isolated and identified, which were divided into the control, empty, recombinant rhBMP-7 transfection, and Lv-BMP-7 transfection groups. BMSCs were induced under different conditions. CCK-8 assay was performed to detect cell proliferation. ALP was used to detect cell activity. Cellular morphology after induction was observed. Immunofluorescence was conducted to detect the expression and location of nerve cell markers. Quantitative real-time PCR and Western blot analysis were performed to detect the mRNA and protein expression levels, respectively. The rhBMP-7 and Lv-BMP-7 promoted the proliferation of BMSCs, accompanied with increased ALP activities. Morphological observations revealed that rhBMP-7 and Lv-BMP-7 induced BMSCs to differentiate into neuron-like cells. Immunofluorescence revealed that the rhBMP-7 and Lv-BMP-7 groups showed positive expression of MAP-2 and Nfh in BMSCs. MAP-2 was mainly distributed in the cell body and cellular protrusion, while Nfh was mainly distributed in the cytoplasm and cell protrusion. Positive mRNA and protein expressions of MAP-2 and Nfh were observed in the cells of the rhBMP-7 and Lv-BMP-7 groups, and the expression levels were significantly higher than the control and empty groups. Both exogenous BMP-7 (rhBMP-7) and endogenous BMP-7 (Lv-BMP-7) can induce BMSCs to differentiate into neuron-like cells highly expressing the neuronal markers MAP-2 and Nfh.
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Walker, David H., and Neill M. Wright. "Bone morphogenetic proteins and spinal fusion." Neurosurgical Focus 13, no. 6 (2002): 1–13. http://dx.doi.org/10.3171/foc.2002.13.6.4.
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Bone morphogenetic proteins (BMPs) have increasingly become a focus of research in the laboratory, with animal models, and in human clinical trials for the treatment of spinal disorders. Basic science research has elucidated the putative mechanism of action of BMPs, and the efficacy of BMPs in inducing bone formation has been evaluated in multiple animal models of anterior and posterior spinal fusion. Not only has BMP been shown to improve the quality and amount of bone formation when used as a supplement to autograft, it has also been shown to promote superior fusion in the absence of autograft, even in high-risk fusion models involving the use of nicotine or nonsteroidal antiinflam-matory agents. Both completed and ongoing clinical trials have demonstrated the efficacy of recombinant BMP, leading to the first BMP product being approved for clinical use earlier this year. Animal models and clinical trials have also been used to evaluate the safety of BMPs. Although few complications have been reported, BMPs can induce heterotopic bone formation, especially when placed adjacent to exposed neural elements. Potentially more serious, antibody formation has been seen in up to 38% of patients in some clinical trials. No clinical sequelae have been reported despite the development of antibodies against BMP, a naturally occurring human protein implicated in processes other than osteoinduction. The future directions of biological manipulation of the osteoinduction process include further understanding of the interactions of the BMP subtypes, the interactions of BMP with its receptors, and exploring other molecules capable of osteoinduction.
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Halloran, Daniel, Venu Pandit, Kelechi Chukwuocha, and Anja Nohe. "Methyl-Beta-Cyclodextrin Restores Aberrant Bone Morphogenetic Protein 2-Signaling in Bone Marrow Stromal Cells Obtained from Aged C57BL/6 Mice." Journal of Developmental Biology 12, no. 4 (2024): 30. http://dx.doi.org/10.3390/jdb12040030.
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During aging, disruptions in various signaling pathways become more common. Some older patients will exhibit irregular bone morphogenetic protein (BMP) signaling, which can lead to osteoporosis (OP)—a debilitating bone disease resulting from an imbalance between osteoblasts and osteoclasts. In 2002, the Food and Drug Administration (FDA) approved recombinant human BMP-2 (rhBMP-2) for use in spinal fusion surgeries as it is required for bone formation. However, complications with rhBMP-2 arose and primary osteoblasts from OP patients often fail to respond to BMP-2. Although patient samples are available for study, previous medical histories can impact results. Consequently, the C57BL/6 mouse line serves as a valuable model for studying OP and aging. We find that BMP receptor type Ia (BMPRIa) is upregulated in the bone marrow stromal cells (BMSCs) of 15-month-old mice, consistent with prior data. Furthermore, conjugating BMP-2 with Quantum Dots (QDot®s) allows effective binding to BMPRIa, creating a fluorescent tag for BMP-2. Furthermore, after treating BMSCs with methyl-β-cyclodextrin (MβCD), a disruptor of cellular endocytosis, BMP signaling is restored in 15-month-old mice, as shown by von Kossa assays. MβCD has the potential to restore BMPRIa function, and the BMP signaling pathway offers a promising avenue for future OP therapies.
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Peng, Wu-Xun, and Lei Wang. "Adenovirus-Mediated Expression of BMP-2 and BFGF in Bone Marrow Mesenchymal Stem Cells Combined with Demineralized Bone Matrix For Repair of Femoral Head Osteonecrosis in Beagle Dogs." Cellular Physiology and Biochemistry 43, no. 4 (2017): 1648–62. http://dx.doi.org/10.1159/000484026.
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Background: This study investigated the effect of using adenovirus-mediated expression of bone morphogenetic protein 2 (Ad-BMP-2) and basic fibroblast growth factor (bFGF) in bone marrow mesenchymal stem cells (BMSCs) in combination with a demineralized bone matrix (DBM) to repair osteonecrosis of the femoral head (ONFH) in Beagle dogs. Methods: A total of 30 Beagle dogs were selected for the isolation of BMSCs, which were cultured and transfected with the recombinant adenovirus vector Ad-BMP2-bFGF-GFP (carrying BMP-2 and bFGF) or a control adenovirus plasmid (encoding green fluorescent protein (Ad-GFP)). The expression of the transfected BMP-2 and bFGF proteins was detected by Western blotting. After transfection, the BMSCs were induced to undergo osteoblastic differentiation. The DBM was prepared to construct a DBM/BMSC complex. Beagle models of canine femoral head defects and necrosis were established and divided into control, DBM, DBM/BMSC, vector Ad-BMP2-bFGF-GFP and Ad-GFP groups. The composite graft was then implanted, and new bone morphology was visualized via X-ray at 3, 6 and 12 weeks after the operation. Hematoxylin and eosin (HE) staining and Masson’s trichrome staining were used to identify new bone formation. Immunohistochemistry was performed to calculate the density of new blood vessels. The compressive and bending strength of the BMSCs was evaluated at 12 weeks after the operation. Results: BMSCs were successfully isolated. The protein expression of BMP-2 and bFGF was significantly higher in the Ad-BMP-2/bFGF group than the normal and Ad-GFP groups. Compared with the control group, at 12 weeks after the operation, the DBM, DBM/BMSC, vector Ad-BMP2-bFGF-GFP and Ad-GFP groups showed a larger area of new bone, higher X-ray scores, greater neovascularization density, and increased compressive and bending strength. The most significant modifications occurred in thevector Ad-BMP2-bFGF-GFP group. Conclusion: The results indicate that the use of Ad-BMP-2/bFGF-modified BMSCs in conjunction with DBM could successfully repair ONFH in a dog model by promoting bone formation and angiogenesis.
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Haugen, Morgan J., and A. L. Johnson. "Bone morphogenetic protein 2 inhibits FSH responsiveness in hen granulosa cells." REPRODUCTION 140, no. 4 (2010): 551–58. http://dx.doi.org/10.1530/rep-10-0211.
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Prior to follicle selection into the preovulatory hierarchy, hen granulosa cells from prehierarchal follicles remain undifferentiated, as defined in part by the virtual absence of LHR mRNA expression and inability to produce progesterone. It has previously been proposed that prior to follicle selection, granulosa cells are actively maintained in an undifferentiated state by epidermal growth factor receptor ligands (EGFRL) signaling via the MAP kinase/extracellular regulated kinase pathway. Moreover, there is recent evidence that EGFRL/MAP kinase signaling modulates FSH receptor (FSHR) transcription, in part, via inhibitor of differentiation/DNA-binding (ID) proteins. In the present studies with undifferentiated granulosa, recombinant human (rh) bone morphogenetic protein 2 (BMP2) induced the phosphorylation of SMAD1/5/8, and blocked transforming growth factor β and FSH-induced FSHR expression and progesterone production. Significantly, BMP2 rapidly induced mRNAs encoding betacellulin and EGF, plus ID proteins (ID1, ID3, and ID4). Alternatively, the bioactivity of BMPs can be modulated by one or more BMP antagonists, including noggin (NOG). NOG mRNA is expressed by both hen granulosa and theca tissues from prehierarchal follicles. Pretreatment of cultured granulosa with rh NOG reversed both the stimulatory effects of BMP2 on ID1, ID3, and ID4 expression and the inhibitory effects of BMP2 on FSHR mRNA levels and progesterone production. Collectively, these data provide evidence that prior to follicle selection, BMP2 signaling contributes toward maintaining granulosa cells in an undifferentiated state. The actions of BMP2 are, at least in part, mediated indirectly via enhanced EGFRL expression and ERBB receptor-mediated MAP kinase signaling, and can be modulated by the autocrine/paracrine production of NOG.
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Shaffrey, Christopher I., and Justin S. Smith. "Editorial: Recombinant human bone morphogenetic protein–2." Journal of Neurosurgery: Spine 18, no. 2 (2013): 109–11. http://dx.doi.org/10.3171/2012.9.spine12476.
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Mukherjee, Aditi, Elizabeth M. Wilson, and Peter Rotwein. "Selective Signaling by Akt2 Promotes Bone Morphogenetic Protein 2-Mediated Osteoblast Differentiation." Molecular and Cellular Biology 30, no. 4 (2009): 1018–27. http://dx.doi.org/10.1128/mcb.01401-09.
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ABSTRACT Mesenchymal stem cells are essential for repair of bone and other supporting tissues. Bone morphogenetic proteins (BMPs) promote commitment of these progenitors toward an osteoblast fate via functional interactions with osteogenic transcription factors, including Dlx3, Dlx5, and Runx2, and also can direct their differentiation into bone-forming cells. BMP-2-stimulated osteoblast differentiation additionally requires continual signaling from insulin-like growth factor (IGF)-activated pathways. Here we identify Akt2 as a critical mediator of IGF-regulated osteogenesis. Targeted knockdown of Akt2 in mouse primary bone marrow stromal cells or in a mesenchymal stem cell line, or genetic knockout of Akt2, did not interfere with BMP-2-mediated signaling but resulted in inhibition of osteoblast differentiation at an early step that preceded production of Runx2. In contrast, Akt1-deficient cells differentiated normally. Complete biochemical and morphological osteoblast differentiation was restored in cells lacking Akt2 by adenoviral delivery of Runx2 or by a recombinant lentivirus encoding wild-type Akt2. In contrast, lentiviral Akt1 was ineffective. Taken together, these observations define a specific role for Akt2 as a gatekeeper of osteogenic differentiation through regulation of Runx2 gene expression and indicate that the closely related Akt1 and Akt2 exert distinct effects on the differentiation of mesenchymal precursors.
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Li, Wei, Richard M. Salmon, He Jiang, and Nicholas W. Morrell. "Regulation of the ALK1 ligands, BMP9 and BMP10." Biochemical Society Transactions 44, no. 4 (2016): 1135–41. http://dx.doi.org/10.1042/bst20160083.
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Bone morphogenetic protein (BMP)9 and BMP10 are high affinity ligands for activin receptor-like kinase 1 (ALK1), a type I BMP receptor mainly expressed on vascular endothelial cells (ECs). ALK1-mediated BMP9/BMP10 signalling pathways have emerged as essential in EC biology and in angiogenesis. Several genetic mutations in the genes encoding the ligands and receptors of this pathway have been reported in two cardiovascular diseases, pulmonary arterial hypertension (PAH) and hereditary haemorrhagic telangiectasia (HHT). Administration of recombinant BMP9 reverses experimental PAH in preclinical rodent models. Dalantercept, an Fc-fusion protein of the extracellular domain of ALK1 and a ligand trap for BMP9 and BMP10, is in phase II clinical trials for anti-tumour angiogenesis. Understanding the regulation of BMP9 and BMP10, at both gene and protein levels, under physiological and pathological conditions, will reveal essential information and potential novel prognostic markers for the BMP9/BMP10-targeted therapies.
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Zhang, Yan Hong, Liang Jun Zhu, and Ju Ming Yao. "Studies on Recombinant Human Bone Morphogenetic Protein 2 Loaded Nano-Hydroxyapatite/Silk Fibroin Scaffolds." Advanced Materials Research 175-176 (January 2011): 253–57. http://dx.doi.org/10.4028/www.scientific.net/amr.175-176.253.
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Bone Morphogenetic Protein 2 (BMP-2) is a member of the transforming growth factor superfamily. It plays an important role in stimulating osteoblast differentiation and bone formation, and has been widely utilized in clinical bone repairing by implantation. In this study, the nano-hydroxyapatite (nHA)/silk fibroin (SF) porous scaffolds were fabricated for the sustained delivery of recombinant human bone morphogenetic protein-2 (rhBMP-2), and then used to address the hypothesis that rhBMP-2 delivered from the scaffolds could enhance the bone formation in vitro. We optimized an effective method using a prokaryotic expression system to produce rhBMP-2. The rhBMP-2 was expressed, purified and renatured in vitro. And then the rhBMP-2 was loaded onto the nHA/SF scaffolds. The bioactivities of rhBMP-2-loaded nHA/SF scaffolds were assessed in vitro. The results showed that the rhBMP-2 promoted the osteoblasts adhesion and proliferation on the nHA/SF scaffolds. Also, the rhBMP-2 released from the nHA/SF scaffold stimulated a significant increase in alkaline phosphatase (ALP) activity of osteoblasts in vitro. These results demonstrated that the rhBMP-2-loaded nHA/SF scaffolds could promote the bone regeneration and showed potential applications in the bone tissue engineering.
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Erickson, D. M., S. E. Harris, D. D. Dean, et al. "Recombinant bone morphogenetic protein (BMP)-2 regulates costochondral growth plate chondrocytes and induces expression of BMP-2 and BMP-4 in a cell maturation-dependent manner." Journal of Orthopaedic Research 15, no. 3 (1997): 371–80. http://dx.doi.org/10.1002/jor.1100150309.
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Fuchs, Thomas, Josef Stolberg-Stolberg, Philipp A. Michel, et al. "Effect of Bone Morphogenetic Protein-2 in the Treatment of Long Bone Non-Unions." Journal of Clinical Medicine 10, no. 19 (2021): 4597. http://dx.doi.org/10.3390/jcm10194597.
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Background: Delayed fracture healing continues to cause significant patient morbidity and an economic burden to society. Biological stimulation of non-unions includes application of recombinant bone morphogenetic protein-2 (rhBMP-2). However, rhBMP-2 use continues to be a matter of controversy as literature shows scarce evidence for treatment effectiveness. Questions: The objective of this study was to evaluate the effectiveness of rhBMP-2 treatment on long bone non-unions measuring union rate and time to union. Furthermore, we assess risk factors for treatment failure. Methods and patients: A total of 91 patients with non-unions of long bones were treated with rhBMP-2 (n = 72) or standard care without BMP (n = 19) at our institution. Patient characteristics, comorbidities, nicotine consumption, and complications were recorded. Bone healing was assessed by plane X-rays and clinical examination. Patients were followed up with for 24 months. Results: Overall, there was significantly faster bone healing after rhBMP-2 application compared to the no-BMP group (p < 0.001; HR = 2.78; 95% CI 1.4–5.6). Union rates differed significantly between rhBMP-2 compared to the no-BMP group (89% vs. 47%; p < 0.001). At the humerus, there was neither a significantly higher union rate in the rhBMP-2 (83%) compared to the no-BMP group (50%) (p = 0.26; n = 12) nor a faster bone healing with a median time of 9 months in both groups (HR = 2.01; 95% CI 0.49–8.61; p = 0.315). The 33 femora treated using rhBMP-2 healed significantly faster than 9 femora in the no-BMP group (HR = 2.93; 95% CI 1.00–8.4; p = 0.023) with significant differences in union rate with 85% and 44%, respectively (p = 0.022). Regarding tibia non-unions, 25 out of 27 (93%) healed with a median of 9 months after rhBMP-2 application with no significant difference in the no-BMP group (33%) in time to union (p = 0.097) but a significantly higher union rate (p = 0.039). There was no effect of comorbidities, age, sex, soft tissue damage, or nicotine use on time to union, union rate, or secondary interventions. Conclusion: Consistent with the literature, overall, significantly higher union rates with reduced time to union were achieved after rhBMP-2 application. Femoral and tibial non-unions in particular seem to profit from rhBMP-2 application.
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Jepsen, S., J. E. Lüttges, H. K. Albers, B. Fleiner, and D. C. Rueger. "Repair processes of the pulp following implantation of recombinant human hone morphogenetic protein (BMP)." British Journal of Oral and Maxillofacial Surgery 33, no. 5 (1995): 331. http://dx.doi.org/10.1016/0266-4356(95)90063-2.
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Ren, Bin, Oliver B. Betz, Daniel Seitz, et al. "Osteogenic Differentiation of Human Adipose-Derived Stem Cells Seeded on a Biomimetic Spongiosa-like Scaffold: Bone Morphogenetic Protein-2 Delivery by Overexpressing Fascia." International Journal of Molecular Sciences 23, no. 5 (2022): 2712. http://dx.doi.org/10.3390/ijms23052712.
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Human adipose-derived stem cells (hADSCs) have the capacity for osteogenic differentiation and, in combination with suitable biomaterials and growth factors, the regeneration of bone defects. In order to differentiate hADSCs into the osteogenic lineage, bone morphogenetic proteins (BMPs) have been proven to be highly effective, especially when expressed locally by route of gene transfer, providing a constant stimulus over an extended period of time. However, the creation of genetically modified hADSCs is laborious and time-consuming, which hinders clinical translation of the approach. Instead, expedited single-surgery gene therapy strategies must be developed. Therefore, in an in vitro experiment, we evaluated a novel growth factor delivery system, comprising adenoviral BMP-2 transduced fascia tissue in terms of BMP-2 release kinetics and osteogenic effects, on hADSCs seeded on an innovative biomimetic spongiosa-like scaffold. As compared to direct BMP-2 transduction of hADSCs or addition of recombinant BMP-2, overexpressing fascia provided a more uniform, constant level of BMP-2 over 30 days. Despite considerably higher BMP-2 peak levels in the comparison groups, delivery by overexpressing fascia led to a strong osteogenic response of hADSCs. The use of BMP-2 transduced fascia in combination with hADSCs may evolve into an expedited single-surgery gene transfer approach to bone repair.
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Tzachanis, Dimitrios, Esther M. Lafuente, Lequn Li, Alla Berezovskaya, Gordon J. Freeman, and Vassiliki A. Boussiotis. "Twisted Gastrulation (Tsg), an Evolutionarily Conserved, Secreted, Morphogenetic Protein Enhances TGF-b Signaling in T Lymphocytes." Blood 106, no. 11 (2005): 341. http://dx.doi.org/10.1182/blood.v106.11.341.341.
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Abstract We have previously determined that quiescent T cells express Tob, a member of the novel APRO gene family, which functions to repress T cell activation. Tob is as a transcriptional regulator and may regulate mRNA transcription. To identify genes that are regulated by Tob, Jurkat T cells were transfected with Tob cDNA or empty vector. Differential gene expression was determined by suppression subtractive hybridization. Twisted Gastrulation (Tsg) was one of the genes suppressed by Tob. Tsg is an evolutionarily conserved secreted protein that interacts with Drosophila Decapentaplegic (Dpp) and its vertebrate orthologs BMP2/4, and also the extracellular Dpp/BMP inhibitors short gastrulation (sog) and chordin. As a result, Tsg affects the binding of Dpp/BMP2/4 to their receptors and subsequent downstream signaling. BMPs belong to a family of secreted signaling molecules the founder of which, TGF-b, is essential for immune homeostasis and maintenance of T cell quiescence. In the thymus, Tsg functions as a regulator of thymocyte development by inhibiting BMP2/4 signaling which arrests thymocyte differentiation. Here, we examined Tsg expression and function in mature CD4+ human T cells. Tsg mRNA was expressed at very low levels in unstimulated T cells and was highly upregulated after activation by TCR/CD3 and either CD28, IL-2 or PMA. To determine the function of Tsg, we generated recombinant human Tsg tagged with V5 epitope. Based on its effects on thymocytes, we hypothesized that Tsg may regulate BMP2/4-mediated effects on mature T cells. Culture of CD4+ T cells with various concentrations of BMP2 or BMP4 in the presence or absence of anti-CD3 mAb did not affect T cell proliferation or cytokine production. Addition of Tsg to cultures containing BMP2/4 did not alter T cell responses. Because our results showed that Tsg mRNA was induced after activation, we tested pre-activated, polyclonal CD4+ alloreactive T cell lines. Neither BMP2 nor BMP4 had any effect on proliferation of alloreactive T cell lines in the presence or absence of anti-CD3 mAb. Strikingly, Tsg induced a significant inhibitory effect on CD3-mediated proliferation and cytokine production, including IL-2, IL-4, IFN-g and IL-10. This effect was not altered by the presence of BMP2 or BMP4. In contrast, Tsg enhanced the inhibitory effect of TGF-b1 suggesting that Tsg regulates TGF-b and not BMP downstream signaling in pre-activated, mature CD4+ T cells. Consistent with this hypothesis, Tsg did not affect phosphorylation of the BMP-specific Smad1, but induced phosphorylation of the TGF-b-specific Smad2 and mediated DNA binding on Smad3/4 consensus sites. In vitro association assays using recombinant Tsg-V5 and TGF-b revealed a direct interaction of these proteins as determined by co-immunoprecipitation followed by western blot with V5-specific and TGF-b-specific antibodies. Moreover, soluble anti-TGF-b receptor reversed the inhibitory effect of Tsg on pre-activated T cells either in the presence or in the absence of TGF-b, providing functional evidence for the biological significance of the Tsg/TGF-b interaction. Our results show that Tsg is a potent agonist of TGF-b downstream signaling in human CD4+ T cells and suggest that enhancement of TGF-b mediating signaling by Tsg may represent a novel target for molecular intervention for induction of transplantation tolerance.