ATP-mediated mineralization of MC3T3-E1 osteoblast cultures

Nakano, Yukiko; Addison, William N.; Kaartinen, Mari T.

doi:10.1016/j.bone.2007.06.011

Cited by 74 publications

(63 citation statements)

References 89 publications

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“…Moreover, pyrophosphate and phosphate produced by ATP hydrolysis are important for mineralization in osteoblasts. 11,38) We found that ALP activity in TBR31-2 cells is increased by BMP-2, indicating that ATP hydrolysis leads to decreased amounts of ATP. Thus, our results indicate that the difference in the ATP-induced increase in [Ca 2ϩ ] i might be related to the stage of differentiation, and it may be caused by a variety of factors as described above.…”

Section: Discussionmentioning

confidence: 79%

Effects of ATP on the Intracellular Calcium Level in the Osteoblastic TBR31-2 Cell Line

Nishii

Nejime

Yamauchi

et al. 2009

Biological & Pharmaceutical Bulletin

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The development of osteoblasts is regulated by various hormones including parathyroid hormone (PTH), 1) 1a,25-dihydroxyvitamin D 3 (1a,25(OH)D 3 ), 2) estrogen 3) and glucocorticoids 4) as well as other local factors, such as bone morphogenetic proteins (BMPs), hedgehogs and the transcription factor, core-binding factor a-1 (Cbfa1). 5) In recent years, extracellular nucleotides have also been considered to be regulating factors in bone cells; for example, the release of ATP from osteoblasts into the extracellular space after stimulation by mechanical strain [6][7][8][9] and stimulating factors, 6) the presence of functional P2 receptors in osteoblasts, 6,10) or the role of extracellular nucleotides in autocrine/paracrine signaling in the cell. 6,[10][11][12] It is generally accepted that immortalization of various cell types by simian virus (SV) 40 T-antigen could at least lead to some stabilization of cell-type specific functions and that the growth and differentiation of cell lines established by the temperature-sensitive (ts) mutant of SV 40 T-antigen gene could be controlled by temperature shifts. [13][14][15][16] Yanai et al. tried to establish many stromal cell lines from the bone marrow of transgenic mice harboring ts SV 40 T-antigen gene. [17][18][19][20] TBR31-2 is one of those stromal cell lines, and Negishi et al. reported that TBR31-2 cells have characteristics of undifferentiated cells that have the potential to express the multipotential cell lineages, including osteoblasts during longterm culture. 21) In addition, as their differentiation is strictly controlled by the temperature shift and culture medium, they are advantageous when examining the molecular mechanism separating the proliferative phase from differentiation phase.The role of the increase in intracellular calcium ion level ([Ca 2ϩ ] i ) induced by ATP in regulating TBR31-2 cells has not yet been investigated. Therefore, we investigated the effects of extracellular ATP on [Ca 2ϩ ] i in TBR31-2 cells induced to differentiate toward osteoblasts. MATERIALS AND METHODSMaterials RITC80-7 medium and plastic tissue culture flasks were obtained from Asahi Techno Glass (Tokyo, Japan), and other materials were obtained from the following sources: fetal bovine serum (FBS) and alpha-modified minimum essential medium (a-MEM) from Gibco-BRL Laboratories (Great Island, NY, U.S.A.); human recombinant epidermal growth factor (EGF) from Wakunaga Pharmaceutical (Tokyo, Japan); human recombinant BMP-2 and L-ascorbic acid from Wako Pure Chemicals (Osaka, Japan); bicinchoninic acid (BCA) protein assay reagent from Pierce (Rockford, IL, U.S.A.); ATP from Oriental Yeast Co., Ltd. (Tokyo, Japan); uridine 5Ј-triphosphate (UTP), ADP, a,bmethylene ATP (a,b-mATP), MRS2179 (a selective P2Y 1 receptor antagonist), U73122 (a phospholipase C inhibitor), and thapsigargin (TSG, a calcium pump inhibitor) from Sigma Chemical Co. (St. Louis, MO, U.S.A.); pyridoxalphosphate-6-azophenyl-2Ј,4Ј-disulphonic acid (PPADS, a non-specific P2 receptor antagonist) from Tocri...

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Section: Discussionmentioning

confidence: 79%

Effects of ATP on the Intracellular Calcium Level in the Osteoblastic TBR31-2 Cell Line

Nishii

Nejime

Yamauchi

et al. 2009

Biological & Pharmaceutical Bulletin

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“…Confluent MC3T3-E1 cells form a more continuous layer of cells and mineralized extracellular matrix (20)(21)(22). We suppose that HA scaffold fabricated by SFF makes bone mineralization accelerate as the amount of osteoid on the preosteoblast surface is increased.…”

Section: Resultsmentioning

confidence: 99%

Biological Advantages of Porous Hydroxyapatite Scaffold Made by Solid Freeform Fabrication for Bone Tissue Regeneration

Kwon

Kim²,

Kim³

et al. 2013

Artificial Organs

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Presently, commercially available porous bone substitutes are manufactured by the sacrificial template method, direct foaming method, and polymer replication method (PRM). However, current manufacturing methods provide only the simplest form of the bone scaffold and cannot easily control pore size. Recent developments in medical imaging technology, computer-aided design, and solid freeform fabrication (SFF), have made it possible to accurately produce porous synthetic bone scaffolds to fit the defected bone shape. Porous scaffolds were fabricated by SFF and PRM for a comparison of physical and mechanical properties of scaffold. The suggested threedimensional model has interconnected cubic pores of 500 mm and its calculated porosity is 25%. Whereas hydroxyapatite scaffolds fabricated by SFF had connective macropores, those by PRM formed a closed pore external surface with internally interconnected pores. SFF was supposed to be a proper method for fabricating an interconnected macroporous network. Biocompatibility was confirmed by testing the cytotoxicity, hemolysis, irritation, sensitization, and implantation. In summary, the aim was to verify the safety and efficacy of the scaffolds by biomechanical and biological tests with the hope that this research could promote the feasibility of using the scaffolds as a bone substitute.

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“…Furthermore, ALP degrades the ubiquitous mineralization inhibitor, pyrophosphate (PPi), resulting in the formation of hydroxyapatite crystals within the collagen fibrils. [42] BMP2 significantly increases the ALP activity via the formation of heteromeric complexes of type I and type II serine-threonine kinase receptors, which are transmitted downstream by the canonical S mad 1/5/8 pathway. [43] Additionally, the Arg-DG conjugate was hypothesized to facilitate the formation of these complexes.…”

Section: Effect Of Arg-dg Conjugate On Alp Activitymentioning

confidence: 99%

Synthesis and biological evaluation of arginyl–diosgenin conjugate as a potential bone tissue engineering agent

Liao

Jung

et al. 2017

Chem Biol Drug Des

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Water-soluble arginyl-diosgenin (Arg-DG) conjugate was designed, synthesized, and evaluated for a biological activity. The Arg-DG conjugate was characterized using FT-IR, 1 H NMR, 13 C NMR, and HPLC-MS analyses, followed by a biological activity evaluation. Compared with DG, the Arg-DG conjugate showed a decreased cytotoxicity against L929 cells and an increased antiproliferative activity against hepatocellular cells. The safety of the Arg-DG conjugate was confirmed using the highly sensitive Alamar Blue assay, which indicated that it increased the cellular metabolic activity at suitable concentrations. The Arg-DG conjugate promoted an endothelial tube formation as well. Furthermore, the Arg-DG conjugate improved the bone morphogenetic protein 2 (BMP2)-induced osteoblastic differentiation with synergistic effects on alkaline phosphatase (ALP) activity and mineralization. These results suggest that the Arg-DG conjugate developed in this study has great potentials for biomedical applications such as bone tissue engineering. K E Y W O R D SALP assay, arginyl-diosgenin conjugate, bone tissue engineering, cytotoxicity, tube formation assay | INTRODUCTIONDiosgenin [(25R)-spirost-5-en-3ß-ol] (DG) is a major naturally occurring steroid saponin, which is abundantly distributed in the rootstock of yams (Dioscorea sp.) and other plants such as Smilax bockii warb and Trigonella foenum graecum. [1,2] DG has exhibited biological effects in the treatment of various diseases such as hypercholesterolemia, climacteric syndromes including diabetic neuropathy and even cancer. [3][4][5][6] It has also been claimed to have an osteogenic property. DG was reported to inhibit osteoclastogenesis and enhance osteoblastic cell differentiation and the synthesis of cellular bone matrix protein. [7,8] It has been hypothesized that DG could be a potential activator of bone formation and potential therapeutic agent for bone-related diseases including osteoporosis.[8]However, the medical application of DG is considerably limited because of its solubility problem and associated toxicity. The aqueous solubility of DG was determined to be 0.7 ng/ml (1.69 nm).[9] It was also revealed that the low solubility of DG led to poor bioavailability and gastrointestinal mucosal toxicity. [10] As poor solubility has been recognized as a frequently encountered challenge in the field of pharmaceutics, different approaches have been employed to improve the solubility and bioavailability of poorly watersoluble compounds including DG. [11] For instance, an inclusion of complexation of DG and β-cyclodextrin was formed. [12] Iron oxide nanoparticles loaded with DG were prepared. [13] Improving the solubility of DG and decreasing its intrinsic cytotoxicity or at least avoiding any additional toxicity against normal cells are the major challenges to its biomedical application. Numerous efforts have been made to chemically modify this compound to improve its applicable biological activities. Diosgenyl saponin, dioscin analogues were designed to improve its anticancer a...

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ATP-mediated mineralization of MC3T3-E1 osteoblast cultures

Cited by 74 publications

References 89 publications

Effects of ATP on the Intracellular Calcium Level in the Osteoblastic TBR31-2 Cell Line

Effects of ATP on the Intracellular Calcium Level in the Osteoblastic TBR31-2 Cell Line

Biological Advantages of Porous Hydroxyapatite Scaffold Made by Solid Freeform Fabrication for Bone Tissue Regeneration

Synthesis and biological evaluation of arginyl–diosgenin conjugate as a potential bone tissue engineering agent

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