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European Calcified Tissue Society Congress 2013

Lisbon, Portugal
18 May 2013 - 22 May 2013

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European Calcified Tissue Society Congress 2013, 18 - 22 May 2013; Lisbon, Portugal

Poster Presentations

Cell biology: osteoblasts and bone formation

ba0001pp163 | Cell biology: osteoblasts and bone formation | ECTS2013

Hepatic lipase is expressed by osteoblasts and modulates bone remodeling in obesity

Bartelt Alexander , Beil F Timo , Muller Brigitte , Kohne Till , Heine Markus , Yilmaz Tayfun , Heeren Joerg , Schinke Thorsten , Niemeier Andreas

Here we identify the lipolytic enzyme hepatic lipase (HL, encoded by Lipc) as a novel cell-autonomous regulator of osteoblast function. In an unbiased genome-wide expression analysis, we find Lipc – which was formerly thought to be expressed almost exclusively by the liver – to be highly induced upon osteoblast differentiation, as verified by quantitative Taqman analyses of primary osteoblasts in vitro and of bone samples in vivo. ...

ba0001pp164 | Cell biology: osteoblasts and bone formation | ECTS2013

Histologic evaluation of direct pulp capping by the using of calcium hydroxide and octacalcium phosphate in dental pulp of cats

Fereydoon Sargolzaei-aval , Arab Mohammad Reza , Saberi Eshagh Ali

Objective: The aim of this study was to evaluate the pulpal responses to octacalcium phosphate (OCP) and calcium hydroxide (CH) used as direct pulp capping (DPC) materials.Study design: The pulp of 72 premolars teeth of nine cats were selected for this experiment. After the cats had been anesthetized, the teeth were exposed and capped directly with OCP, CH, and no capping materials used as control group. The cavities of all three groups were filled with ...

ba0001pp165 | Cell biology: osteoblasts and bone formation | ECTS2013

Implantation of octacalcium phosphate enhances alveolar ridge in rat mandible

Aval Fereydoon Sargolzaei , Arab Mohammad Reza , Aval Forough Sargolzaei

Background and aim: This study was designed to investigate the process of bone formation caused by implantation of octacalcium phosphate (OCP) at alveolar ridge.Materials and methods: In this descriptive study we used 20 male Sprague–Dawley rats. Synthetic OCP was implanted into the bony defect measuring 3 mm in diameter and 2 mm in depth was surgically created with a bur in the rat mandible. Bone formation at the alveolar ridge was examined histolo...

ba0001pp166 | Cell biology: osteoblasts and bone formation | ECTS2013

Activated protein C increases osteoblast proliferation and BMP2 induced bone formation

Shen Kaitlin , Schindeler Aaron J , Cheng Tegan L , Xue Meilang , Little David G , Jackson Chris J

Introduction: Activated protein C (APC) plays an important role in the cutaneous healing of chronic wounds arising from orthopaedic surgery and has cytoprotective and anti-inflammatory properties which may also assist bone repair. The aim of this study was to examine whether APC could directly influence osteoblasts and increase bone formation in a rodent model.Methods: Proliferation of MG-63 osteoblast-like cells was quantified by MTT assay and direct co...

ba0001pp167 | Cell biology: osteoblasts and bone formation | ECTS2013

Adipogenesis occurs at the expense of osteoblast differentiation in primary osteoblasts deficient in protease-activated receptor 2

Kularathna Pamuditha , Pagel Charles N , Hooper John D , Mackie Eleanor J

The G protein-coupled receptor, protease-activated receptor 2 (PAR2), is expressed by osteoblasts and required for normal skeletal growth and repair. Prostate cancer (PCa) cells commonly secrete proteolytic activators of PAR2 (including matriptase and kallikrein-related peptidase 4) and frequently form osteogenic metastases in bone. This study was undertaken to investigate the hypothesis that PAR2 activators released by PCa cells modulate osteo...

ba0001pp168 | Cell biology: osteoblasts and bone formation | ECTS2013

Distinct potential of osteoblast differentiation of adipose tissue- and bone marrow-derived mesenchymal stem cells

Abuna Rodrigo , de Oliveira Fabiola , Kato Rogerio , Rosa Adalberto , Beloti Marcio

Adult mesenchymal stem cells (MSCs) are of interest in the fields of cell therapy and tissue engineering thanks to their potential of differentiating into distinct cell lineages, e.g. osteoblast, chondrocyte, myoblast, and adipocyte. As the capacity of differentiation may vary according to the cell source, here, we compared the potential of osteoblast differentiation of MSCs derived from either bone marrow or adipose tissue. MSCs from rat bone marrow and adipose tissue were cu...

ba0001pp169 | Cell biology: osteoblasts and bone formation | ECTS2013

Increase of mineral nodules and alkaline phosphatase levels in osteoblasts cultures by using disordered carbon nanotubes and titanium discs

Zancanela Daniela Cervelle , Simao Ana Maria Sper , Matsubara Elaine Yoshiko , Rosolen Jose Mauricio , Ciancaglini Pietro

Biological calcification is a regulated process in which different types of tissues, cells and biomolecules participate in the coordination and regulation of the metabolic events involved in accumulating large amounts of calcium phosphate. This process could be speeded up using carbon nanotubes (CNTs) systems. The objective of our study was to compare cell growth and formation of mineralized matrix nodules by osteoblasts grown in plastic and in titanium (Ti) discs surfaces. Th...

ba0001pp170 | Cell biology: osteoblasts and bone formation | ECTS2013

Calcium transport and phosphomonohydrolase activity by proteoliposomes harboring annexin V and alkaline phosphatase

Bolean Mayte , Simao Ana Maria , Kiffer-Moreira Tina , Hoylaerts Marc , Millan Jose Luis , Ciancaglini Pietro

The biomineralization process is initiated inside matrix vesicles (MVs), with phosphate and calcium ions crystallizing as hydroxyapatite. This process is accomplished by the activities of several proteins, such as annexins (e.g. AnxV) that mediates Ca2+ influx into MVs and tissue-nonspecific alkaline phosphatase (TNAP), a phosphomonohydrolase that uses ATP and PPi as substrates. Dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylserine (DPPS...

ba0001pp171 | Cell biology: osteoblasts and bone formation | ECTS2013

In vitro effect of prolactin on the osteogenic potential of bone marrow mesenchymal stem cells of rats

de Melo Ocarino Natalia , Silvia Silva Santos , Rocha Lorena , Freitas Juneo , Sena Reis Amanda Maria , Serakides Rogeria

The effects of prolactin on bone metabolism have been the subjects of several studies. It is believed that prolactin acts directly influencing the synthesis of bone matrix by stimulating the osteoblastic activity, since receptors for this hormone have been identified in osteoblasts and human mesenchymal stem cells (MSCs). However, no study on the effects of prolactin on the osteogenic differentiation of MSCs was found in the literature. The objective of this study was to verif...

ba0001pp172 | Cell biology: osteoblasts and bone formation | ECTS2013

Differential gene expression of matrix metalloproteinases (MMPs), MMP inhibitors (TIMPs and RECK), and MMP-activator (EMMPRIN/CD147) during osteogenic differentiation from human dental pulp stem cells

Paiva Katiucia , Silva Luiz , Sogayar Mari

Constant remodeling of extracellular matrix (ECM) is a hallmark during physiological conditions, such as stem cell differentiation, embryogenesis and tissue repair. Matrix metalloproteinases (MMP) play a key role in these processes. MMPs, MMP-activator (EMMPRIN/CD147) and MMP-inhibitors (TIMPs and RECK) are responsible for bone matrix remodeling and, probably, determinate the level of its turnover. Mesenchymal stem cells derived from dental pulp are multipotent and have the ca...

ba0001pp173 | Cell biology: osteoblasts and bone formation | ECTS2013

Exogenous polyphosphate is not readily utilized for mineralization in vitro

Ariganello Marianne , Omelon Sidney , Wazen Rima , Variola Fabio , Nanci Antonio

Polyphosphates (polyPs) are inorganic phosphate chains found in many cell types with higher concentrations in bone cells. As a source of inorganic phosphate (Pi) and an effective calcium reservoir due to chelation, PolyPs enable total Ca2+ and PO42− concentrations above those required for apatite saturation. Alkaline phosphatase (ALP) cleaves Pi from polyP, thus polyPs may be involved in apatite mineralization.Aim: ...

ba0001pp174 | Cell biology: osteoblasts and bone formation | ECTS2013

Elevated levels of serotonin decrease bone volume by direct effects on bone turnover in rats

Erjavec Igor , Bordukalo-Niksic Tatjana , Brkljacic Jelena , Pauk Martina , Grgurevic Lovorka , Thompson David D , Paralkar Vishwas M , Cicin-Sain Lipa , Vukicevic Slobodan , Mokrovic Gordana , Kesic Maja , Grcevic Danka

Elevated levels of circulating serotonin have been reported to decrease bone mineral density1. Conversely, reduced serotonin (5HT) in mice lacking TPH1, the rate limiting enzyme for 5HT synthesis, was reported to be anabolic to the skeleton with high osteoblastic activity2. However, in other studies TPH1 deletion led to either an initial increase in BMD due to inhibition of osteoclastic bone resorption3, or had no bone effect4</su...

ba0001pp175 | Cell biology: osteoblasts and bone formation | ECTS2013

Identification of a small molecule kinase inhibitor that enhances osteoblast differentiation of human skeletal (mesenchymal) stem cells through regulation of TGFβ signaling

Siersbaek Majken Storm , Jafari Abbas , Zaher Walid , Chen Li , Kassem Moustapha

Identifying novel molecules that enhance human skeletal (mesenchymal) stem cells (hMSC) differentiation into osteoblastic bone forming cells (OB), may lead to development of new bone anabolic drugs. We have identified Kix, a small molecule kinase inhibitor that enhanced ex vivo OB differentiation and reduced apoptosis of hMSC. We found that Kix targeted undifferentiated hMSC populations and not their differentiated progeny. In addition, Kix increased in vivo ...

ba0001pp176 | Cell biology: osteoblasts and bone formation | ECTS2013

The effect of fibroblast growth factor 2 on mesenchymal stromal cell differentiation

Kahkonen Tiina , Ivaska Kaisa K , Harkonen Pirkko

Mesenchymal stromal cells (MSC) have a potential to differentiate to osteoblasts and adipocytes. Differentiation can be stimulated or inhibited by different growth factors, including fibroblast growth factors (FGFs). In this study we evaluated the effect of FGF2 on the osteoblastic and adipocytic differentiation of MSCs in vitro.Mouse MSC-derived cells were cultured in differentiation medium that led to differentiation to osteoblasts in 14 days ...

ba0001pp177 | Cell biology: osteoblasts and bone formation | ECTS2013

Extracellular glucose alters mesenchymal stromal cell growth and differentiation

Virta Anna-Reeta , Ivaska Kaisa K

Disorders of glucose metabolism are associated with adverse skeletal effects. Hyperglycemia impairs the function of osteoblast-like cells but the mechanisms underlying glucose toxicity are poorly understood. In this study we determined the effect of elevated extracellular glucose levels on the proliferation and osteogenic differentiation of mesenchymal stromal cells (MSC).Bone marrow cells were isolated from rat long bones, plastic-adherent MSCs were enr...

ba0001pp178 | Cell biology: osteoblasts and bone formation | ECTS2013

Regulation and function of immunosuppressive molecule human leukocyte antigen G5 in human bone tissue

Deschaseaux Frederic , Gaillard Julien , Langonne Alain , Chauveau Christophe , Naji Abderrahim , Bouacida Amina , Rosset Philippe , Heymann Dominique , de Pinieux Gonzague , Rouas-Freiss Nathalie , Sensebe Luc

Bone-marrow mesenchymal stem cells (MSCs) are the origin of bone-forming cells with immunomodulation potential. Among the generated immunosuppressive molecules there is HLA-G5. HLA-G proteins play a crucial role in promoting the acceptance of allografts. However, the mechanisms regulating the expression of HLA-G5 in human MSCs are unknown. We induced differentiation of human MSCs (harvested from iliac crests of healthy volunteers after their informed consent following approved...

ba0001pp179 | Cell biology: osteoblasts and bone formation | ECTS2013

Mineralizing properties of DMP1 studied in vitro with cellular and acellular 3D collagen model systems mimicking the bone tissue

Silvent Jeremie , Nassif Nadine , Azais Thierry , Helary Christophe , Delgado Sidney , Soncin Fabrice , Giraud-Guille Marie Madeleine , Sire Jean-Yves

Bone is a complex structure associating cells to an extracellular organic phase, including collagen and non collagenous proteins (NCPs), in close association with apatite mineral platelets. Although bone has given rise to extensive studies, the exact part played by NCPs in nucleating or inhibiting the mineral phase remains controversial. The present study aimed to better understand the functions of a major mineralizing protein, dentin matrix phosphoprotein 1 (DMP1), an acidic,...

ba0001pp180 | Cell biology: osteoblasts and bone formation | ECTS2013

N-cadherin governs age-related osteoprogenitor cell determination in mice through modulation of Wnt5a and Wnt10b

Hay Eric , Dieudonne Francois-Xavier , Marty Caroline , Marie Pierre J

Senile osteoporosis and age-related osteopenia are associated with decreased osteoblastogenesis and increased bone marrow adipogenesis. The mechanisms controlling the fate determination of osteoblast to adipocyte differentiation of bone marrow stromal cells (BMSC) during aging are not known. We and others previously showed that the cell-cell adhesion molecule N-cadherin (N-Cadh) expressed in osteoblasts controls bone formation, but little is known about its role in BMSC fate d...

ba0001pp181 | Cell biology: osteoblasts and bone formation | ECTS2013

The impairement of bone formation and mineralization in BSP−/− mouse calvaria cell cultures is partly rescued by increasing cell density

Bouet Guenaelle , Bouleftour Wafa , David Marchat , Marie-Therese Linossier , Mireille Thomas , Jane Aubin E , Laurence Vico , Luc Malaval

Bone sialoprotein regulates osteoblast activity and bone formation. In knockout (BSP−/−) mouse bone marrow (BM) stromal cell cultures, the pool of osteoprogenitor (OP) cells (CFU-F number) is not different from wild-type (+/+), nor is their early differentiation (same numbers of alkaline phosphatase positive colonies=CFU-ALP, although these are smaller), while the number of osteoblast, mineralized colonies (CFU-OB) is dramatically reduced. Because ossifi...

ba0001pp182 | Cell biology: osteoblasts and bone formation | ECTS2013

Evaluation of bone formation capacities of human adipose-derived stromal cells cultured in platelet growth factor-enriched plasma medium.

Guilloton Fabien , Rabani Vahideh , Brennan Meadhbh , Bassi Giulio , Krampera Mauro , Layrolle Pierre , Sensebe Luc , Deschaseaux Frederic

Human adipose-derived stromal cells (ASCs) exhibit strong plasticity and proliferation potentials. In addition, ASCs are easy to harvest and are found at high frequency in adipose tissue samples. This gives us opportunities for their use in bone regeneration therapy. We thus evaluated the bone formation potential of ASCs in vitro and in vivo.ASCs were isolated from subcutaneous adipose tissue (following Local Ethical Guideline and after...

ba0001pp183 | Cell biology: osteoblasts and bone formation | ECTS2013

Development of mice models to study implant osseointegration and failure in alveolar bone

Mouraret Sylvain , Bardet Claire , Hunter Dan J , Popelut Antoine , Brunski John B , Chaussain Catherine , Bouchard Philippe , Helms Jill A

Many of our assumptions concerning oral implant osseointegration are extrapolated from experimental models studying skeletal tissue repair in long bones rather than in oral bones. This discrepancy between clinical practice and experimental research hampers our understanding on how alveolar bone forms or resorbs around implants and how osseointegration of oral implants can be improved. To overcome this disconnect, we have developed a mouse model which mimics oral implant placem...

ba0001pp184 | Cell biology: osteoblasts and bone formation | ECTS2013

Activation of β-catenin signalling enhances the osteogenic gene response to mechanical loading in mesenchymal stem cells

Nemitz Claudia , Jakob Franz , Ignatius Anita , Liedert Astrid

Introduction: Wnt/β-catenin signalling and mechanical loading are able to inhibit adipogenesis and to stimulate osteoblastogenesis of mesenchymal stem cells1,2. The involvement of β-catenin signalling in mechanically induced bone formation has already been shown in vivo using a tibia loading model3. The aim of this study was to investigate the influence of the activation of β-catenin on the osteogenic and adip...

ba0001pp185 | Cell biology: osteoblasts and bone formation | ECTS2013

DNA-damage, survival, differentiation, and matrix mineralization in vitro of a murine multipotent mesenchymal precursor cell line

Lau Patrick , Hu Yueyuan , Hellweg Christine E , Baumstark-Khan Christa , Reitz Gunther

Radiation therapy is one of the most effective and indispensable treatment modalities for cancer patients. Known tissue complications caused by radiation-induced stem cell depletion, may result in structural and functional alterations of the surrounding matrix. Although, studies have demonstrated that ionizing radiation can induce apoptosis and senescence, little is known about the effects of therapeutic irradiation concerning the commitment of mesenchymal stem cells to the os...

ba0001pp186 | Cell biology: osteoblasts and bone formation | ECTS2013

Involvement of Runx2 in the differentiation process of osteoblastic precursor cells after radiation exposure

Hu Yueyuan , Lau Patrick , Hellweg Christine , Baumstark-Khan Christa , Reitz Guenther

Astronauts on exploratory space missions will experience a complex environment that includes microgravity and radiation. While the deleterious effects of unloading on bone are well established, fewer studies have focused on the effects of radiation. Space radiation produces distinct biological damages which, up to now, little is known about the correlation between radiation exposure and bone tissue. In our study we used osteoblastic precursor cells to investigate the radiation...

ba0001pp187 | Cell biology: osteoblasts and bone formation | ECTS2013

Implantation of octacalcium phosphate enhances long bone's repair in rats

Arab Mohammad Reza , Aval Fereydoon Sargolzaei , Aval Forugh Sargolzaei

Background: This study was designed to investigate the process of bone formation caused by implantation of octacalcium phosphate (OCP) in rat tibiae.Methods: We used 25 young male Sprague–Dawley rats. A full thickness standardized trephine defect, 3-mm in diameter, was surgically created on the superior end of right and left tibia. Amount of 6-μg synthetic octacalcium phosphate was implanted into a bony defect on the right tibia as an experimen...

ba0001pp188 | Cell biology: osteoblasts and bone formation | ECTS2013

Intracellular calcium fluxes in human bone cells in osteoporotic and osteoarthritic patients

celi Monica , Gasbarra Elena , Frank Claudio , Cutarelli Alessandro , Cinci Giulio Fioravanti , Tarantino Umberto

We studied changes in intracellular Ca2+ concentration in bone cell cultures obtained from human subjects with osteoporosis and osteoarthritis, to evaluate differences between these patients and healthy subjects. We enrolled 36 patients: 12 undergoing primary total hip arthroplasty for osteoporotic femoral fractures (group A, mean age range 57–80), 12 for hip osteoarthritis (group B, mean age range 57–80), and 12 healthy subjects who suffered a high-energy...

ba0001pp189 | Cell biology: osteoblasts and bone formation | ECTS2013

Integrins and cadherins in mesenchymal stem cells from dental tissues: possible implication in the osteogenic differentiation process

Di Benedetto Adriana , Carbone Claudia , Oranger Angela , Brunetti Giacomina , Lorenzo Lo Muzio , Colucci Silvia , Grano Maria , Mori Giorgio

Numerous studies have reported beneficial effects of multipotent mesenchymal stem cells (MSCs) in tissue repair and regeneration. These multipotent cells can be isolated from many different adult tissues and give rise to different cell lineages. The most well-characterized source for adult stem cells is still adult bone marrow, however in the past decade, subpopulations of stem cells have been isolated from dental tissues. Dental pulp has been identified as a promising source ...

ba0001pp190 | Cell biology: osteoblasts and bone formation | ECTS2013

Role of vitamin D and K on human osteoblasts in vitro on primary cultures derived from osteoporotic and normal patients

Roscetti Gianna , Marini Mario , Arango Emiliano , Tarantino Umberto

This study is focused on the effects of the synergic use of vitamins D and K on human osteoblasts primary cultures derived from osteoporotic and normal patients. The aim of this work is the evaluation of the different cellular behaviour in response to the lipophilic vitamins stimulation. We included 20 osteoporotic and 20 control patients in age between 35 and 50 and in age between 55 and 85. All patients gave informed consent for using bone samples as a source of bone cells. ...

ba0001pp191 | Cell biology: osteoblasts and bone formation | ECTS2013

Nuclear translocation of oxytocin receptor mediates increased gene expression in osteoblasts

Di Benedetto Adriana , Cuscito Concetta , Colaianni Graziana , Tamma Roberto , Nico Beatrice , Calvano Damiana , Zambonin Carlo , Corcelli Michelangelo , Zallone Alberta

The neuro-hypophiseal hormone oxytocin (OT) is a novel anabolic regulator of bone mass (Tamma et al. PNAS, 2009), upregulating expression of critical osteoblast transcription factors. These effects are mediated by oxytocin receptor, a GPCR expressed by osteoblasts. Recently an increasing number of reports indicates that GPCRs could be targeted to the nuclear membrane; prostaglandin receptors, endothelin receptors and β-adrenergic receptors among others (...

ba0001pp192 | Cell biology: osteoblasts and bone formation | ECTS2013

Moderate hypothermia induces growth arrest in normal human osteoblast cells but retained mitochondrial metabolism in vitro

Aisha Mohd Din , Nor-Ashikin Mohamed Noor Khan , Sharanizax Ab. Rahim , Nawawi Hapizah , Kapitonova Marina , Froemming Gabriele Ruth Anisah

Ablation of osteosarcoma cells by sublethal hypothermia before radiation may increase sarcoma tissue sensitivity by inducing growth arrest. Normal cells that are not lethally damaged by hypothermia and radiation can undergo DNA repair thus promoting cell survival. Nevertheless, understanding of the response of normal bone forming osteoblast cells towards hypothermia is necessary before administering on osteosarcoma cells. In this study we evaluated the response of short-term m...

ba0001pp193 | Cell biology: osteoblasts and bone formation | ECTS2013

Normal human osteoblast cells exerts an adaptive effect towards moderate hypothermia by retaining bone metabolism and cellular function in vitro

Aisha Mohd Din , Nor-Ashikin Mohamed Noor Khan , Sharaniza Ab. Rahim , Nawawi Hapizah , Kapitonova Marina , Froemming Gabriele Ruth Anisah

Over the years, it has been demonstrated that the ability to maintain body core temperature in older adult’s declines with age. Temperature is a vital physical factor for cell growth and a downshift in core body temperature (<37 °C) might have a direct affect on maintaining bone density or repair fractures. Disruption in any of the cellular processes involved in bone remodelling leads to a net loss of bone mineral density and bone loss. Therefore our study looked...

ba0001pp194 | Cell biology: osteoblasts and bone formation | ECTS2013

Black tea polyphenols suppress adverse effects of TNFα-induced inflammation in osteoblast cells

Zulkipli Husna , Din Aisha Mohd , Salim Norita , Froemming Gabriele Anisah , Ismail Aletza Mohd , Nawawi Hapizah

Introduction: Most chronic inflammatory bone diseases are characterized by loss of bone density due to an increase of osteoclastic activity without equally increased osteoblast activity which in turn is leading to an imbalance in bone repair and remodelling. Several studies have reported that green tea rich in polyphenols especially catechins could improve bone mass and structure and eventually increase bone formation. Data on black tea, also rich in polyphenols especially the...

ba0001pp195 | Cell biology: osteoblasts and bone formation | ECTS2013

Microarray reveals positive effects of green and black tea polyphenols on TNFα-induced changes of gene expression

Zulkipli Husna , Salim Norita , Froemming Gabriele Anisah , Ismail Aletza Mohd , Nawawi Hapizah

Introduction: Recent studies have found anti-inflammatory, antioxidant and bone forming properties of green (GTP) and black tea (BTP) polyphenols. However most of these studies are focussed on specific genes or pathways. We wanted to know if GTP and BTP could help to reduce symptoms of chronic inflammation especially bone loss and what are the possible genes and pathways involved. We were especially interested in unexplored pathways which may play a role in regaining bone heal...

ba0001pp196 | Cell biology: osteoblasts and bone formation | ECTS2013

The effect of enamel matrix derivative on human gingival fibroblasts cultured on zirconium disc surfaces

Lee Heesu , Pae Ahran , Kwon Yong-Dae , Ko Seonghee

Purpose: To investigate the effect of enamel matrix derivative (Emdogain) on the attachment, growth behavior and the genetic effect of human gingival fibroblasts (HGF) cultured on zirconium disc surfaces.Materials and methods: HGF cells were cultured on i) zirconium discs without enamel matrix derivative (EMD), ii) zirconium discs with EMD 25 μg/ml, and iii) zirconium discs with EMD 100 μg/ml. The cell proliferation activity was evaluated throu...

ba0001pp197 | Cell biology: osteoblasts and bone formation | ECTS2013

EGF suppresses BMP2-induced osteogenic differentiation through the up-regulation of Smurf1 expression

Lee Hye-Lim , Woo Kyung Mi , Ryoo Hyun-Mo , Kim Gwan-Shik , Baek Jeong-Hwa

Although EGF has been known to inhibit osteoblast differentiation, its molecular mechanism has not been clearly elucidated. Smurf1 acts as a negative regulator of BMP signaling by inducing ubiquitination and proteasomal degradation of BMP type I receptor and R-Smads. In this study, we investigated the effect of EGF on the expression of Smurf1 and the role of Smurf1 in EGF-induced inhibition of BMP2-induced osteogenesis. EGF increased Smurf1 expression which was blocked by trea...

ba0001pp198 | Cell biology: osteoblasts and bone formation | ECTS2013

Estrogen effect on the sclerostin induction by BMP-2 in human mesenchymal stromal cells

Kim In Sook , Yang Hoon Joo , Song Yun Mi , Ryu Soo Jin , Kim Ri Youn , Hwang Soon Jung

Introduction: Estrogen therapy decreases circulating levels of sclerostin, a protein product of SOST which increase in postmenopausal women. However, the mechanisms of estrogen on the expression of SOST remain unclear. This study was hypothesized that estrogen modulates SOST expression by interfering bone morphogenic protein (BMP) signaling on the basis that BMP is an inducer of SOST in osteoblasts.Description of methods: We investigated the expression o...

ba0001pp199 | Cell biology: osteoblasts and bone formation | ECTS2013

Effect of β-cryptoxanthin on the differentiation of human bone-marrow stromal stem-cells treated with pioglitazone

Casado-Diaz Antonio , Santiago-Mora Raquel , Dorado Gabriel , Quesada-Gomez Jose Manuel

Pioglitazone is a drug of the thiazolidinedione (TZD) class used to treat type 2 diabetes mellitus. TZD is an agonist of peroxisome proliferator-activated receptor γ (PPAR-γ) that improves insulin sensitivity, glucose and lipid metabolism and inflammation. However, TZD induces bone marrow adiposity with suppression of osteogenesis, that could contribute to bone loss and osteoporotic fractures. β-Cryptoxanthin is a carotenoid with antioxidant properties abundant ...

ba0001pp200 | Cell biology: osteoblasts and bone formation | ECTS2013

Premixed acidic calcium phosphate cement as a local delivery system for simvastatin

Montazerolghaem Maryam , Engqvist Hakan , Ott Marjam

In 1999 Mundy et al. showed that simvastatin, a drug administered for high cholesterol levels, had a profound stimulatory effect on osteoblasts. Since then other studies have also confirmed that simvastatin enhances bone formation; however, the lack of a local delivery system have restricted its clinical use. We have used premixed acidic calcium phosphate cement as a local delivery system for simvastatin. To confirm that the simvastatin released retained its activity,...

ba0001pp201 | Cell biology: osteoblasts and bone formation | ECTS2013

Primary Human Bone Cells treated with Parathyroid Hormone or Dexamethasone show Effects on micro-RNA Expression Patterns Assessed by Second Generation Sequencing

Laxman Navya , Rubin Carl-Johan , Mallmin Hans , Nilsson Olle , Tellgren-Roth Christian , Kindmark Andreas

Introduction: Micro-RNAs (miRNAs) are important post-transcriptional regulators. By binding to complementary RNA strands, they affect mRNA levels and/or mRNA translation. We have previously identified ~90 miRNAs with significant expression levels, with a subset of miRNAs exhibiting interindividual and/or gender differences in expression. In the present project, we have investigated the impact of treatment of parathyroid hormone (PTH) and dexamethasone (DEXA) on global miRNA ex...

ba0001pp202 | Cell biology: osteoblasts and bone formation | ECTS2013

Expression and function of glutamate transporters in mouse primary osteoblasts

Xie Wenjie , Dolder Silvia , Siegrist Mark , Wetterwald Antoinette , Hofstetter Willy

Introduction: Osteoblast lineage cells express glutamate receptors and secrete glutamate, which acts as an autocrine factor to promote cellular differentiation and activation. However, the mechanisms by which glutamate regulates these functions, remain unclear.Methods: Primary osteoblasts were isolated from calvaria of 2–3 days old mice. The cells were treated with inhibitors of glutamate transporters, namely the Scl1a1 and Scl1a3 inhibitor L...

ba0001pp203 | Cell biology: osteoblasts and bone formation | ECTS2013

Connectivity Map-based discovery of novel compounds that induce osteoblast differentiation

Brum A M , van de Peppel J , van Kerkwijk A , Janssen M , Schreuders-Koedam M , Strini T , Eijken M , van Leeuwen J P T M , van der Eerden B C J

Osteoporosis is a common skeletal disorder characterized by low bone mass leading to increased bone fragility and fracture susceptibility. Little is currently known about what specific factors stimulate osteoblast differentiation from human mesenchymal stem cells (hMSCs). Therefore, the aim for this project is to determine novel factors and mechanisms involved in human bone production which can be targeted to treat osteoporosis, using gene expression profiling and bioinformati...

ba0001pp204 | Cell biology: osteoblasts and bone formation | ECTS2013

Non-canonical BMP signaling in bone healing

Sanchez-Duffhues Gonzalo , de Vinuesa Amaya Garcia , Kloen Peter , Goumans Marie-Jose , ten Dijke Peter

The healing of bone fractures is a tightly regulated process where released growth factors and cytokines interplay within an inflammatory environment in order to reestablish the functional bone. Recent studies have suggested that endothelial cells may dedifferentiate into mesenchymal multipotent cells via a mechanism called endothelial-to-mesenchymal-transition (EndoMT). Transforming growth factor-β (TGF-β) plays a critical role inducing EndoMT. Subsequent differenti...

ba0001pp205 | Cell biology: osteoblasts and bone formation | ECTS2013

Thrombin receptor deficiency leads to osteopetrosis by decreasing the RANKL/OPG ratio

van der Eerden BCJ , Tudpor K , Jongwattapapisan P , Woudenberg-Vrenken TE , Bindels RJM , Hoenderop JGJ , van Leeuwen JPTM

Communication between osteoblasts and osteoclasts is crucial for bone remodeling. Thrombin and its thrombin receptor (TR; PAR-1) are expressed in osteoclasts and osteoblasts, respectively. To date, the physiological roles of thrombin and TR in bone metabolism have not been elucidated. Therefore, we fully characterized the bone phenotype of mice lacking the thrombin receptor.We performed bone microarchitectural analyses of the femurs of 10–12 week ol...

ba0001pp206 | Cell biology: osteoblasts and bone formation | ECTS2013

Serum sclerostin does not reflect its expression in bone, but is related to bone mineral density

Bravenboer Nathalie , Visser Ruben , den Heijer Martin , Heijboer Annemieke

Sclerostin is a major negative regulator of osteoblastic activity. Serum sclerostin has a weak positive association with BMD but contradictory results have been described concerning associations with fractures. These contradictions could be explained by the fact that serum sclerostin does not reflect its action in bone. In this study we question whether serum sclerostin is associated with its expression in bone. In addition we aimed to detect associations between sclerostin in...

ba0001pp207 | Cell biology: osteoblasts and bone formation | ECTS2013

In vitro 3D osteoblast-osteocyte co-culture mechanical loading model

Vazquez Marisol , Evans Bronwen , Evans Sam , Ralphs Jim , Riccardi Daniela , Mason Deborah

Introduction: Normal mechanical loading potently induces bone formation via effects on osteocytes. Current investigations of mechanical loading of bone do not reflect the interactions of the cells within it, mostly focusing on mechanical loading of osteoblasts in monolayers. Existing 3D models do not elucidate the osteoblast-osteocyte interactions that regulate mechanically-induced bone formation. We developed a novel in vitro 3D co-culture model of bone1 t...

ba0001pp208 | Cell biology: osteoblasts and bone formation | ECTS2013

The positional origins of human osteoblasts dictate growth and differentiation potential and capacity for paracrine vascular cell cross-talk via VEGF

Shah Mittal , Gburcik Valentina , Sankey Andrew , Reilly Peter , Emery Roger , Clarkin Claire , Pitsillides Andrew

Successful long-term, cementless fixation of human shoulder components in osteoporotic (OP) and osteoarthritic (OA) patients poses major challenges. The possibility that enhanced osseointegration may rely on both the region of bone targeted and its relationship with the vasculature remains unexplored. We hypothesise that bone cells derived from subchondral (SC), cortical (C) and trabecular (Trb) bone regions exhibit differing osteogenic potential, which will be diminished in b...

ba0001pp209 | Cell biology: osteoblasts and bone formation | ECTS2013

Bone marrow stromal cells of female BAG-1 heterozygous mice exhibit reduced osteogenic potential

Greenhough Joanna , Townsend Paul , Oreffo Richard , Tare Rahul

The co-chaperone protein, Bcl-2-associated athanogene 1 (BAG-1) is expressed ubiquitously in bone including cells of the osteoblast-lineage and, plays an important role in cell proliferation, apoptosis and differentiation by regulating signalling and transcription. The study aims to elucidate the function of BAG-1 in osteoblast development by examining differences in osteogenic differentiation of bone marrow stromal cells (BMSCs) from Bag-1+/−</sup...

ba0001pp210 | Cell biology: osteoblasts and bone formation | ECTS2013

Open source software for semi-automated histomorphometry of bone resorption and formation parameters

van 't Hof Rob , Rose Lorraine , Daroszewska Anna

Histomorphometric analysis is an essential technique to measure bone formation and resorption parameters. Here we present three novel open source image analysis packages that allow the rapid semi-automated analysis of histomorphometric bone resorption, osteoid, and calcein double labelling parameters.Mice were injected with calcein 5 and 2 days before killing. Tibia and vertebrae were fixed in formalin, embedded in methylmethacrylate and the blocks secti...

ba0001pp211 | Cell biology: osteoblasts and bone formation | ECTS2013

Rab27a is involved in bone formation by osteoblasts

Coxon Fraser , Douglass Angela , Hughes Alun , Helfrich Miep , Seabra Miguel , Tolmachova Tanya

The Rab family GTPases Rab27a and Rab27b play an important role in the trafficking of lysosome-related organelles in specialised cells, such as melanocytes. Since secretory lysosomes, also considered a lysosome-related organelle, are important for osteoclast and osteoblast function, we hypothesised that Rab27 plays a role in bone physiology. In support of this, a recent study demonstrated impaired transport of RANK ligand to the plasma membrane in osteoblasts from mice lacking...

ba0001pp212 | Cell biology: osteoblasts and bone formation | ECTS2013

Bone-forming cultures of rat and mouse calvarial osteoblasts: key differences in protocols

Orriss Isabel , Hajjawi Mark , Huesa Carmen , MacRae Vicky , Arnett Timothy

The in vitro culture of calvarial osteoblasts from neonatal rodents remains an important method for studying the regulation of osteoblast function. Widespread use of transgenics has created a particular need for a reliable, simple method that allows the differentiation and bone-forming activity of mouse osteoblasts to be studied directly. We have established such a method and have identified key differences in optimal culture conditions between mouse and rat osteoblas...