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  • 1. Brage, M
    et al.
    Lie, Anita
    Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Ransjö, Maria
    Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Kasprzykowski, F
    Kasprzykowska, R
    Abrahamson, M
    Grubb, A
    Lerner, Ulf
    Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Osteoclastogenesis is decreased by cysteine proteinase inhibitors2004Inngår i: Bone, ISSN 8756-3282, E-ISSN 1873-2763, Vol. 34, nr 3, s. 412-24Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The effects of cystatin C and other cysteine proteinase inhibitors on osteoclast formation and differentiation have been investigated. Cystatin C decreased osteoclast formation stimulated by parathyroid hormone (PTH), 1,25(OH)2-vitamin D3 or interleukin-6 (IL-6) (in the presence of its soluble receptor) as assessed by the number of tartrate-resistant acid phosphatase (TRAP+) multinucleated cells in mouse bone marrow cultures. The inhibitory effect was associated with decreased mRNA expression for the calcitonin receptor as well as decreased number of specific binding sites for 125I-calcitonin, and without any effect on the mRNA expression of receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL). Similarly, the cysteine proteinase inhibitors leupeptin, E-64 and benzyloxycarbonyl-Phe-Ala-diazomethane (Z-FA-CHN2) decreased PTH-stimulated formation of TRAP+ multinucleated cells and binding of 125I-calcitonin. A peptidyl derivative synthesized to mimic part of the proteinase-binding site of cystatin C (benzyloxycarbonyl-Arg-Leu-Val-Gly-diazomethane, or Z-RLVG-CHN2) also decreased PTH-stimulated osteoclast formation. In a 9-day culture, addition of cystatin C during the last 5 days was sufficient to cause substantial inhibition of osteoclast formation. Cystatin C-induced decrease of osteoclast formation was associated with enhanced number of F4/80-positive macrophages and increased mRNA expression of the macrophage receptor c-fms in the bone marrow culture. Osteoclast formation in mouse bone marrow cultures as well as in mouse spleen cell cultures, stimulated by macrophage colony-stimulating factor (M-CSF) and RANKL was also decreased by different cysteine proteinase inhibitors. In addition, cystatin C inhibited M-CSF/RANKL induction of calcitonin receptor mRNA in spleen cell cultures. The inhibitory effect by cystatin C in spleen cells was associated with decreased mRNA expression of RANK and the transcription factor NFAT2. It is concluded that cysteine proteinase inhibitors decrease formation of osteoclasts by interfering at a late stage of pre-osteoclast differentiation.

  • 2.
    Lundgren, Stefan
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Käkkirurgi. Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Sennerby, Lars
    Cricchio, Giovanni
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Käkkirurgi.
    Salata, Luiz
    Palma, Vinnie
    Lundqvist, Carina
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Käkkirurgi.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Rekonstruktiv käkkirurgi: Behandling av den atrofiska posteriora maxillan hos partiellt betandade patienter2008Inngår i: Tandläkartidningen, ISSN 0039-6982, Vol. 100, nr 5, s. 70-71Artikkel i tidsskrift (Annet (populærvitenskap, debatt, mm))
  • 3.
    Matemba, Stephen
    et al.
    Umeå universitet, Medicinsk fakultet, Odontologi. Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Lie, Anita
    Umeå universitet, Medicinsk fakultet, Odontologi.
    Ransjö, Maria
    Umeå universitet, Medicinsk fakultet, Odontologi. Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Regulation of osteoclastogenesis by gap junction communication2006Inngår i: Journal of cellular biochemistry, Vol. 99, nr 2, s. 528-37Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Receptor activator of NF-kappaB ligand (RANKL) is crucial in osteoclastogenesis but signaling events involved in osteoclast differentiation are far from complete and other signals may play a role in osteoclastogenesis. A more direct pathway for cellular crosstalk is provided by gap junction intercellular channel, which allows adjacent cells to exchange second messengers, ions, and cellular metabolites. Here we have investigated the role of gap junction communication in osteoclastogenesis in mouse bone marrow cultures. Immunoreactive sites for the gap junction protein connexin 43 (Cx43) were detected in the marrow stromal cells and in mature osteoclasts. Carbenoxolone (CBX) functionally blocked gap junction communication as demonstrated by a scrape loading Lucifer Yellow dye transfer technique. CBX caused a dose-dependent inhibition (significant > or = 90 microM) of the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells formed in 7- to 8-day marrow cultures stimulated by parathyroid hormone (PTH; 10 nM) or forskolin (FSK; 1 microM). Furthermore, CBX (100 microM) significantly inhibited prostaglandin E2 (PGE2; 10 microM) and 1,25(OH)2-vitamin D3 stimulated osteoclast differentiation in the mouse bone marrow cultures. Consequently, quantitative real-time polymerase chain reaction (PCR) analysis demonstrated that CBX downregulated the expression of osteoclast phenotypic markers, but without having any significant effects on RANK, RANKL, and osteoprotegerin (OPG) mRNA expression. However, the results demonstrated that CBX significantly inhibits RANKL-stimulated (100 ng/ml) osteoclastogenesis in the mouse bone marrow cultures. Taken together, our results suggests that gap junctional diffusion of messenger molecules interacts with signaling pathways downstream RANKL in osteoclast differentiation. Further studies are required to define the precise mechanisms and molecular targets involved. Copyright 2006 Wiley-Liss, Inc.

  • 4.
    Mladenovic, Zivko
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Johansson, Anders
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Willman, Britta
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Shahabi, Kaveh
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Silicon inhibits signaling pathways and cell-cell communication important for osteoclast formation and bone resorption in vitroManuskript (preprint) (Annet vitenskapelig)
    Abstract [en]

    Silicon containing materials are used in bone regeneration, and some of the materials, e.g. Bioactive glass 45S5 (BG), release silicon (Si) ions to the surrounding tissue after implantation. The role of Si in bone biology is debated; nevertheless findings suggest that Si is beneficial for bone formation. A majority of the experimental studies on Si and bone have focused on osteoblasts. The effects of Si on osteoclast formation and function have not been directly addressed. In the present study, we show that ionic dissolution extract from BG inhibit osteoclast bone resorption in an organ culture system as well as osteoclast formation in a mouse bone marrow system and in the RAW264.7 cell line. Si containing cell culture medium was prepared to address the issue whether or not the inhibitory effects with BG dissolution extract were Si ion dependent. The results suggest that the inhibitory effects of Si act directly on osteoclast precursors, by interactions with the RANK/RANKL/OPG signaling pathway as well as with gap junction intercellular communication. However, regulation via osteoblasts cannot be excluded. The inhibitory effect of Si on osteoclasts could be useful for future therapies or treating bone loss in patients, provided that molecular mechanisms are established.

  • 5.
    Mladenovic, Zivko
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Sahlin-Platt, Annika
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Ortodonti.
    Andersson, Britta
    Department of Medicine Solna, Karolinska Institutet, S-171 76 Stockholm,Sweden.
    Johansson, Anders
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Molekylär paradontologi.
    Björn, Erik
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    In vitro study of the biological interface of Bio-Oss: implications of the experimental setup2013Inngår i: Clinical Oral Implants Research, ISSN 0905-7161, E-ISSN 1600-0501, Vol. 24, nr 3, s. 329-335Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Objectives To systematically investigate the biological interface of Bio-Oss by analysing dissolution–precipitation behaviour and osteogenic responses using in vitro experimental systems.

    Material and methods Different concentrations (1–100 mg/ml) of Bio-Oss were incubated in cell culture medium for 24 h before elemental concentrations for calcium, phosphorus and silicon in the medium were analysed with inductive coupled plasma-optical emission spectroscopy. Radioactive calcium-45 isotope labelling technique was used to study possible precipitation of calcium on the Bio-Oss particle. Biological interface of Bio-Oss was studied in osteogenic experiments using mineralization medium and three different sources of cells (primary mouse bone marrow stromal cells, primary rat calvarial cells and MC3T3-E1 mouse pre-osteoblast cell line). Cells were fixed and stained with Toulidine blue, von Kossa or Alizarin Red staining for confirmation of extracellular matrix mineralization.

    Results Elemental analysis of the cell culture medium demonstrated a significant decrease of calcium and phosphorus and a dose-dependent release of silicon to the medium after incubation with Bio-Oss. A significant decrease of calcium and phosphorus in the medium occurred even at low concentrations of Bio-Oss. Uptake of calcium on the Bio-Oss particle was confirmed with radioactive calcium-45 isotope labelling technique. In osteogenic experiments with Bio-Oss (<1 mg/ml), matrix mineralization around the Bio-Oss particles were demonstrated in all three cell types with von Kossa and Alizarin Red staining.

    Conclusion Dissolution–precipitation reactions occur at the surface of Bio-Oss, and osteogenic responses are seen at the biological interface. The concentration of Bio-Oss is a key factor for the experimental in vitro results, and may also have implications for the clinic.

  • 6.
    Mladenovic, Zivko
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Sahlin-Platt, Annika
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Andersson,, Martin
    Department of Chemical and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden.
    Shchukarev, Andrey
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Investigation of surface reactions and solid-solution interfaces of three bonegraft substitute materials incubated in cell culture mediumArtikkel i tidsskrift (Annet vitenskapelig)
  • 7.
    Mladenovic, Zivko
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Sahlin-Platt, Annika
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Bengtsson, Åsa
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Shchukarev, Andrey
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Surface characterization of bone graft substitute materials conditioned in cell culture medium2010Inngår i: Surface and Interface Analysis, ISSN 0142-2421, E-ISSN 1096-9918, Vol. 42, nr 6-7, s. 452-456Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Biomaterials are widely used in clinical practice as bone graft substitutes for treating patients with bone defects. A molecular level understanding of the chemical processes at the interface between the biomaterial and the biological environment is crucial to succeed in tissue regeneration and to predict the treatment outcome. In this study, we used three different bone graft substitute materials (BioGlass 45S5—synthetic, Bio-Oss—bovine derived and Algipore—derived from algae) which were incubated in an α-minimum essential medium (α-MEM) during 1, 3 and 7 days. Initial surface composition of the biomaterials and the chemistry of their solid–solution interface were monitored by XPS with a fast-frozen samples technique. The XPS analysis showed that the equilibrium at the solid-solution interface is reached within 24 h. The Na/Cl atomic ratio at equilibrium indicates a negatively charged surface for Bio-Oss. In contrast, the other two materials gained a positive surface charge, which resulted in pronounced adsorption of amino acids at the interface from the medium. The surface chemical reconstruction and charge generation mechanism responsible for this effect are discussed with regard to bulk composition of the materials and possible proliferation and differentiation cell patterns that could be expected at the interface. Copyright © 2010 John Wiley & Sons, Ltd.

  • 8.
    Ransjö, Maria
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Oral cellbiologi.
    Lundgren, Stefan
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Käkkirurgi.
    Det växer fast - om implantat och benbildning.: På bettet hela livet, om  odontologisk vetenskap i Umeå. pp2006Inngår i: På bettet hela livet, om  odontologisk vetenskap i Umeå.: En bok från Forskningens dag 2006, Medicinska fakulteten, Umeå universitet. Författare Svante Twetman / [ed] Medicinska fakulteten vid Umeå universitet, Umeå: Umeå universitet , 2006, s. 67-82Kapittel i bok, del av antologi (Annet (populærvitenskap, debatt, mm))
  • 9.
    Ransjö, Maria
    et al.
    Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Sahli, J
    Lie, Anita
    Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Expression of connexin 43 mRNA in microisolated murine osteoclasts and regulation of bone resorption in vitro by gap junction inhibitors.2003Inngår i: Biochemical and Biophysical Research Communications - BBRC, ISSN 0006-291X, E-ISSN 1090-2104, Vol. 18;303, nr 4, s. 1179-1185Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Several studies have demonstrated that connexin 43 (Cx43) mediates signals important for osteoblast function and osteogenesis. The role of gap junctional communication in bone resorption is less clear. We have investigated the expression of Cx43 mRNA in osteoclasts and bone resorption cultures and furthermore, the functional importance of gap junctional communication in bone resorption. RT-PCR analysis demonstrated Cx43 mRNA expression in mouse bone marrow cultures and in osteoclasts microisolated from the marrow cultures. Cx43 mRNA was also expressed in bone resorption cultures with osteoclasts and osteoblasts/stromal cells incubated for 48h on devitalized bone slices. An up-regulation of Cx43 mRNA was detected in parathyroid (PTH)-stimulated (0.1 nM) bone resorption. Two inhibitors of gap junction communication, 18alpha-glycyrrhetinic acid (30 microM) and oleamide (100 microM), significantly inhibited PTH- and 1,25-(OH)(2)D(3)-stimulated osteoclastic pit formation. In conclusion, our data indicate a functional role for gap junction communication in bone resorption.

  • 10.
    Sahlin-Platt, Annika
    et al.
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Oral cellbiologi.
    Örtengren, Ulf
    Mladenovic, Zivko
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Oral cellbiologi.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Oral cellbiologi.
    Effects of Dyract AP and released ionic products on periodontal ligament cells and bone marrow cultures2008Inngår i: Dental Materials, ISSN 0109-5641, E-ISSN 1879-0097, Vol. 24, nr 12, s. 1623-1630Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    OBJECTIVES: The aim of this work was to investigate the release of inorganic ionic products from specimens of the polyacid-modified composite resin Dyract AP (DAP) and furthermore, to analyze the biological effect of DAP and the medium extract in human periodontal ligament (PDL) cells and mouse bone marrow cell (BMC) cultures.

    METHODS: Ion release from DAP specimens immersed in cell culture medium was analyzed with inductively coupled plasma optical emission spectroscopy (ICP-OES). Cells were cultured with either DAP specimens or with DAP media extract and effects on cell proliferation, osteoblastic gene expression and mineralization capacity were analyzed with direct-contact tests, neutral red (NR) uptake, quantitative real-time PCR and a bone nodule formation assay.

    RESULTS: ICP-OES analysis of DAP extract demonstrated a significant increase in fluoride, strontium and silica. PDL cells demonstrated normal growth pattern in the direct-contact tests with the material. DAP extracts produced a dose-dependent stimulation of cell proliferation and concomitant inhibition of osteoblast specific markers and nodule formation.

    SIGNIFICANCE: The compomer may have possible bioactive properties due to ions leaching out from the filler component.

  • 11.
    Shchukarev, Andrey
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Mladenovic, Zivko
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Ortodonti.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi, Ortodonti.
    Surface characterization of bone graft substitute materials conditioned in cell culture medium. 2. Protein adsorption2012Inngår i: Surface and Interface Analysis, ISSN 0142-2421, E-ISSN 1096-9918, Vol. 44, nr 8, s. 919-923Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Three bone graft substitute materials (Bioglass 45S5, Bio-Oss (R) and Algipore (R)) were conditioned in a-minimum essential medium (alpha-MEM), with the addition of 10% fetal bovine serum (FBS), for 1 and 7?days. The chemistry of their solid-solution interface was monitored by X-ray photoelectron spectroscopy, using fast-frozen sample technique, and compared to that reported for original alpha-MEM. FBS added to the biological medium causes significant changes in the interface after only 1day of conditioning. Interfacial chemical composition and N 1s spectra show immediate adsorption of proteins at the surface of all three biomaterials, independent of their surface charge and chemical composition. However, the atomic ratio C/N and the C 1s spectra indicate a different orientation of adsorbed serum proteins, which is dependent on the particle's surface charge. Moreover, the adsorption of serum proteins at the surface of Bio-Oss causes a charge reversal at the interface, as evidenced by the change in the atomic ratio of Na/Cl. In addition to the particle's surface charge, the formation of the protein interfacial layer at the surface of the biomaterial seems to be the second major phenomenon important for subsequent cell recognition and the initiation of biomineralization. Copyright (c) 2012 John Wiley & Sons, Ltd.

  • 12.
    Shchukarev, Andrey
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Ransjö, Maria
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    Mladenović, Živko
    Umeå universitet, Medicinska fakulteten, Institutionen för odontologi.
    To build or not to build: the interface of bone graft substitute materials in biological media from the view point of the cells2011Inngår i: Biomaterials science and engineering / [ed] Rosario Pignatello, Published by InTech , 2011, s. 287-308Kapittel i bok, del av antologi (Fagfellevurdert)
    Abstract [en]

    These contribution books collect reviews and original articles from eminent experts working in the interdisciplinary arena of biomaterial development and use. From their direct and recent experience, the readers can achieve a wide vision on the new and ongoing potentials of different synthetic and engineered biomaterials. Contributions were not selected based on a direct market or clinical interest, than on results coming from very fundamental studies which have been mainly gathered for this book. This fact will also allow to gain a more general view of what and how the various biomaterials can do and work for, along with the methodologies necessary to design, develop and characterize them, without the restrictions necessarily imposed by industrial or profit concerns. The book collects 22 chapters related to recent researches on new materials, particularly dealing with their potential and different applications in biomedicine and clinics: from tissue engineering to polymeric scaffolds, from bone mimetic products to prostheses, up to strategies to manage their interaction with living cells.

  • 13. Smith, R
    et al.
    Ransjö, Maria
    Umeå universitet, Medicinsk fakultet, Odontologi, Oral cellbiologi.
    Tatarczuch, L
    Song, SJ
    Pagel, C
    Morrison, JR
    Pike, RN
    Mackie, EJ
    Activation of protease-activated receptor-2 leads to inhibition of osteoclast differentiation.2004Inngår i: Journal of Bone and Mineral Research, ISSN 0884-0431, E-ISSN 1523-4681, Vol. 19, nr 3, s. 507-516Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    PAR-2 is expressed by osteoblasts and activated by proteases present during inflammation. PAR-2 activation inhibited osteoclast differentiation induced by hormones and cytokines in mouse bone marrow cultures and may protect bone from uncontrolled resorption. INTRODUCTION: Protease-activated receptor-2 (PAR-2), which is expressed by osteoblasts, is activated specifically by a small number of proteases, including mast cell tryptase and factor Xa. PAR-2 is also activated by a peptide (RAP) that corresponds to the "tethered ligand" created by cleavage of the receptor's extracellular domain. The effect of activating PAR-2 on osteoclast differentiation was investigated. MATERIALS AND METHODS: Mouse bone marrow cultures have been used to investigate the effect of PAR-2 activation on osteoclast differentiation induced by parathyroid hormone (PTH), 1,25 dihydroxyvitamin D3 [1,25(OH)2D3], and interleukin-11 (IL-11). Expression of PAR-2 by mouse bone marrow, mouse bone marrow stromal cell-enriched cultures, and the RAW264.7 osteoclastogenic cell line was demonstrated by RT-PCR. RESULTS: RAP was shown to inhibit osteoclast differentiation induced by PTH, 1,25(OH)2D3, or IL-11. Semiquantitative RT-PCR was used to investigate expression of mediators of osteoclast differentiation induced by PTH, 1,25(OH)2D3, or IL-11 in mouse bone marrow cultures and primary calvarial osteoblast cultures treated simultaneously with RAP. In bone marrow and osteoblast cultures treated with PTH, 1,25(OH)2D3, or IL-11, RAP inhibited expression of RANKL and significantly suppressed the ratio of RANKL:osteoprotegerin expression. Activation of PAR-2 led to reduced expression of prostaglandin G/H synthase-2 in bone marrow cultures treated with PTH, 1,25(OH)2D3, or IL-11. RAP inhibited PTH- or 1,25(OH)2D3-induced expression of IL-6 in bone marrow cultures. RAP had no effect on osteoclast differentiation in RANKL-treated RAW264.7 cells. CONCLUSION: These observations indicate that PAR-2 activation inhibits osteoclast differentiation by acting on cells of the osteoblast lineage to modulate multiple mediators of the effects of PTH, 1,25(OH)2D3, and IL-11. Therefore, the role of PAR-2 in bone may be to protect it from uncontrolled resorption by limiting levels of osteoclast differentiation.

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