Searchable abstracts of presentations at key conferences on calcified tissues
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43rd Annual European Calcified Tissue Society Congress

Rome, Italy
14 May 2016 - 17 May 2016

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Poster Presentations

Cell biology: osteoblasts and bone formation

ba0005p135 | Cell biology: osteoblasts and bone formation | ECTS2016

Up-regulation of inhibitors of DNA binding/differentiation gene during alendronate-induced osteoblast differentiation

Kim Heungyeol

Aim: Alendronate enhances bone morphogenetic proteins (BMP)-mediated osteoblast differentiation. A balanced regulation of inhibitors of DNA binding/differentiation (Ids) plays an important role in BMP-induced osteoblast differentiation. However, there are no studies on the possible roles of Id genes in alendronate-induced osteoblast differentiation. This study investigated the effect of alendronate on the expression of Id genes in osteoblast differentiation.<...

ba0005p137 | Cell biology: osteoblasts and bone formation | ECTS2016

Delta-like 1/fetal antigen 1 (DLK1/FA1) inhibits BMP2-induced osteoblast differentiation by modulating Nfκb signaling pathway: a novel mechanism for regulation of bone formation

Qiu Weimin , Abdallah Basem M. , Kassem Moustapha

Delta-like 1/fetal antigen-1 (DLK1/FA1) is a negative regulator of bone mass in vivo as it inhibits osteoblast (OB) and stimulates osteoclast differentiation. However, the molecular mechanisms underlying these effects are not known. Thus, we studied the effect of DLK1/FA1 on different signaling pathways known to regulate OB differentiation. We identified DLK1/FA1 as an inhibitor of BMP2-induced OB differentiation. Stable overexpression of DLK1/FA1 in C2C12 cells or th...

ba0005p138 | Cell biology: osteoblasts and bone formation | ECTS2016

Osteoblastogenesis is regulated through the interplay between human arrest defective 1 and runt-related transcription factor 2

Park Jong-Wan

Arrest defective 1 was originally identified as an acetyltransferase essential for the life-cycle progression in yeasts. Its human orthologue hARD1 has been known to express the enzymatic activity and to acetylate several targets such as HIF-1alpha, MLCK-1, and beta-catenin. Here, whether hARD1 takes part in pre-osteoblast differentiation toward calcium-deposing osteoblast was explored. ALP staining and alizalin red S staining showed that osteoblast differentiation was negativ...

ba0005p139 | Cell biology: osteoblasts and bone formation | ECTS2016

Human bone marrow-derived mesenchymal stem cells response on calcium- and magnesium-ion-implanted resorbable blast media-treated titanium surface

Lee Heesu , Won Sun , Wook Lee Jae

Purpose: The aims of this study were (1) to assess the topographical and chemical changes of Ca and Mg ion implantation procedures using plasma immersion ion implantation and deposition (PIIID) technique and (2) to evaluate cellular response of human bone marrow-derived mesenchymal stem cells (hBMMSCs) to Ca and Mg ion-implanted titanium surface, compared with resorbable blast media (RBM)-treated titanium surface by observing cell attachment, proliferation and gene expression ...

ba0005p140 | Cell biology: osteoblasts and bone formation | ECTS2016

Matrix metalloproteinases and their inhibitors (TIMPs and RECK): induction of osteoblastic differentiation modulates their protein levels on mineralization onset in human dental pulp stem cells

Paiva Katiucia , Gasparoni Leticia , Bronzeri Cristiane

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

ba0005p141 | Cell biology: osteoblasts and bone formation | ECTS2016

Age-dependent changes in the bone marrow microenvironment

Kusumbe Anjali , Ramasamy Saravana , Itkin Tomer , Lapidot Tsvee , Adams Ralf

Blood vessels define the properties of local microenvironments in the skeletal system, play crucial roles in osteogenesis and provide niches for haematopoietic stem cells. The properties of niche-forming vessels and their changes in the ageing organism remain incompletely understood. We have previously identified a new capillary subtype in the murine skeletal system with distinct morphological, molecular and functional properties. These vessels are CD31hi/Emcnh...

ba0005p142 | Cell biology: osteoblasts and bone formation | ECTS2016

Regulation of genes involved in the integrin signalling pathway induced by titanium with nanotopography

Lopes Helena , Freitas Gileade , Florio Pedro , de Oliveira Paulo , Rosa Adalberto , Beloti Marcio

Titanium (Ti) surface with nanotopography induces osteoblast differentiation, at least in part, by upregulating the expression of α1/β1 integrins. Thus, we hypothesized that other potential candidates from the integrin signalling pathway may be involved in the osteogenic potential of nanotopography. In this context, the aim of this study was to compare the expression of integrin family members of cells grown on nanotopography with machined surface. Machined Ti discs ...

ba0005p143 | Cell biology: osteoblasts and bone formation | ECTS2016

RUNX2, osterix and the human sclerostin gene: searching molecular and epidemiological interactions

Perez-Campo Flor , Santurtun Ana , Garcia-Ibarbia Carmen , Pascual Maria A , Valero Carmen , Garces Carlos , Sanudo Carolina , Zarrabeitia Maria T , Riancho Jose A

Sclerostin, encoded by the SOST gene, functions as an inhibitor of the Wnt pathway and thus it is an important regulator of bone homeostasis. The fact that osteoblasts, the only cells expressing SOST, lay buried deeply in the bone matrix, poses intrinsic difficulties to the study of the regulation of this gene. Since RUNX2 and SP7/OSX are two known regulators of the differentiation of cells of the osteoblastic lineage, the aim of this study was to determine t...

ba0005p144 | Cell biology: osteoblasts and bone formation | ECTS2016

Primary osteoblast culture from red fox (Vulpes Vulpes)

Perpetuo Ines Pedro , Felder Alessandro , Pitsillides Andrew , Doube Michael , Orriss Isabel

Effects of whole bone scaling on isolated osteoblast behaviours are unknown. Exhibiting a huge range in size, inbred canines are an ideal species to determine such relationships. We have therefore undertaken initial studies in both male and female red foxes (Vulpes vulpes), the most abundant and accessible wild canid member in the United Kingdom.Femoral heads were removed from five fresh red fox cadavers (see details on table) and bone fragments...

ba0005p145 | Cell biology: osteoblasts and bone formation | ECTS2016

Evaluation of release kinetics and mitogenic capacity of collagen barrier membranes supplemented with the secretome of activated platelets

Mozgan Eva-Maria , Edelmayer Michael , Janjić Klara , Pensch Manuela , Fischer Michael , Agis Hermann

Success in periodontal regeneration through guided tissue regeneration relies on the healing capacity of the host tissue. Platelet preparations are mitogenic and stimulate regeneration through high growth factor levels. Here we assessed collagen barrier membranes (CBM) as carriers for the secretome of activated platelets. We evaluated cell-attachment, the release of the secretome, and cell proliferation.Secretome of washed platelets (washed PSEC) and unw...

ba0005p146 | Cell biology: osteoblasts and bone formation | ECTS2016

Pharmacological activation of the non-canonical TGF-β signaling is a novel strategy to enhance bone formation

Jafari Abbas , Siersbaek Majken , Chen Li , Kassem Moustapha

Identifying novel approaches for enhancing osteoblast (OB) differentiation of human skeletal (mesenchymal) stem cells (hMSC) can lead to development of novel anabolic agents required for efficient bone formation. Transforming growth factor-βs (TGF-β1, 2, 3) are one of the most abundant growth factors in bone and play a key role in regulating bone remodeling. Canonical TGF-β signaling inhibits, whereas components of the non-canonical TGF-β signaling (e.g. Ak...

ba0005p147 | Cell biology: osteoblasts and bone formation | ECTS2016

Skin-derived IL-17A induces bone loss in the absence of adaptive immunity

Uluckan Ozge , Schnabl Jakob , Jimenez Maria , Jeschke Anke , Karbach Susanne , Schinke Thorsten , Waisman Ari , Schett Georg , Wagner Erwin

Inflammatory stimuli can lead to bone loss by mechanisms that are not well understood. We recently showed that skin inflammation induces bone loss in mice and humans. In psoriasis, one of the prototypic IL-17A-mediated inflammatory human skin diseases, low bone formation and bone loss correlates with increased serum IL-17A levels. Similarly, in two mouse models with chronic IL-17A-mediated skin inflammation, K14-IL17Aind and JunB&...

ba0005p148 | Cell biology: osteoblasts and bone formation | ECTS2016

Cationic nacre ethanol soluble matrix has an osteoanabolic effect on human subchondral osteoarthritic osteoblasts and MC3T3-E1 cell line

Zhang Ganggang , Brion Alice , Piet Marie-Helene , Moby Vanessa , Bianchi Arnaud , Mainard Didier , Galois Laurent , Aubriet Frederic , Carre Vincent , Gillet Pierre , Rousseau Marthe

Nacre is able to induce bone-forming cells mineralization, and gains widespread interest in bone regeneration. While, the osteoinductive compounds are not identified yet. The nacre extract, ethanol soluble matrix (ESM), was proven having the capacity to induce the mineralization of mouse MC3T3-E1 cells and to restore mineralization defect in human osteoarthritic osteoblasts. Being tested on the two cells, ESM could induce the formation of mineralization nodules, identified as ...

ba0005p149 | Cell biology: osteoblasts and bone formation | ECTS2016

Enhancement of bone ultra structure preservation using high-pressure freezing and microwave-assisted fixation

Daghma Diaa Eldin S. , Heiss Christian , Khassawna Thaqif El

Despite the improvement in bone’s molecular, metabolic and live-cell imaging, histological investigation remains most crucial in bone biology for diagnostic and research purposes. Therefore, fixation of bone samples – especially for electron microscopy investigation – is critical to the ultrastructural analysis. Up to date, chemical fixation of bone tissue is performed at room temperature resulting in a compromised ultrastructure of bone sample. In this study, w...

ba0005p150 | Cell biology: osteoblasts and bone formation | ECTS2016

GPR39 is negatively regulating osteoblast differentiation and bone formation during aging

Pesic Milena , Ram Gali Guterman , Orenbuch Ayelet , Katchkovsky Svetlana , Levaot Noam

G-protein coupled receptors (GPCRs) are the largest family of cell surface proteins, which are important for bone development, remodeling and diseases. One of the GPCRs, called GPR39, has been found to be expressed in several osteoblastic cell lines. However, its role in bone metabolism has not been investigated yet. In order to elucidate a role for GPR39, we characterized the bone phenotype in GPR39 deficient mice. During aging, at six months old, dynamic histomorphometry dat...

ba0005p151 | Cell biology: osteoblasts and bone formation | ECTS2016

Early deletion of menin in the osteoblast lineage results in decreased bone mass in adult mice

Abi Rafeh Jad , Canaff Lucie , Hendy Geoffrey

Menin, the product of the Men1 tumor suppressor gene, facilitates the cell proliferation control and differentiation induced by the TGF-beta superfamily of ligands critical for bone development and maintenance. Our in vivo studies have shown the importance of menin for proper functioning of the mature osteoblast and maintenance of bone mass in adult mice. In the present study, we examined the in vivo role of menin at earlier stages of the osteoblast lineage through conditional...

ba0005p152 | Cell biology: osteoblasts and bone formation | ECTS2016

Participation of microRNA-34a/RANKL in the osteogenic potential of the Poly(vinylidene-trifluorethylene)/barium titanate membrane

Lopes Helena Bacha , Ferraz Emanuela Prado , de Almeida Adriana Luisa Goncalves , Abuna Rodrigo Paolo Flores , Hassan Mohammad Quamurul , Rosa Adalberto Luiz , Beloti Marcio Mateus

Barrier membranes have been extensively used in dentistry to prevent soft tissue down-growth into the bone defects and to promote alveolar ridge augmentation. Previous studies of our group showed that the Poly(vinylidene-trifluoroethylene)/barium titanate composite (PVDF) enhances both the in vitro osteoblastic differentiation and the in vivo bone repair compared with a commercially available polytetrafluoroethylene (PTFE) membrane. As bone formation may be regulated ...

ba0005p153 | Cell biology: osteoblasts and bone formation | ECTS2016

RhoA and ROCK regulate spreading of osteoblasts

Yang Seungwon , Kim Hyun-Man

When cells detach from or re-attach to the substrates, there is a drastic change in shape. Molecular mechanism by which controls shape upon cell attachment to the substrates was studied in osteoblastic MC3T3-E1 cells in vitro. Most striking feature found in the ovoid cells detached from the substrates was densely packed cortical actin bundles (CAB) encircling cells. Cells lost the CAB upon attachment and spreading on to the substrates. CABs were under control of small...

ba0005p154 | Cell biology: osteoblasts and bone formation | ECTS2016

Low adhesive scaffold collagen promotes the osteogenic differentiation of rat marrow mesenchymal cells

Kunii Saori , Horiuchi Yoshitaka , Morimoto Koichi

Background: Collagen has biocompatibility and biodegradability with tissue or organ, therefore, collagen is the most promising material for tissue engineering. In particular, the binding of collagen to specific cells is considered an essential function to develop scaffolds. However, in some cases the binding inhibits the cell motility. In addition, it is not clear whether collagen is the effective scaffold to promote osteogenic differentiation. These days, we succeeded in deve...

ba0005p155 | Cell biology: osteoblasts and bone formation | ECTS2016

L-Carnitine facilitates mitochondrial activity and osteogenic differentiation in human primary osteoblast culture

Vacante Fernanda , Montesano Anna , Spinello Alice , Bolamperti Simona , Senesi Pamela , Terruzzi Ileana , Luzi Livio , Villa Isabella , Rubinacci Alessandro

Osteoblasts have a high rate of energy consumption during bone formation and bone protein synthesis. Therefore, it is likely that decreased energy production that accompanies aging, could contribute to reduced osteoblast activity, a critical feature of senile osteoporosis, and that this reduction might be counteracted by favoring energy availability. Cells of the osteoblastic lineage generate 40–80% of the requested energy through fatty acid degradation, thus the modulati...

ba0005p156 | Cell biology: osteoblasts and bone formation | ECTS2016

The differential effects of zoledronate and etidronate on the in vivo early osseous healing process of an extracted socket and a tibial defect

Kim In Sook , Lim Shin Saeng , Lee Beomseok , Hwang Soon Jung

Bisphosphonates (BPs) are widely used for treating osteoporosis and preventing osseous metastasis or hypercalcemia in cancers. However, osteonecrosis of the jaw in patients treated with BP after dentoalveolar surgery has been increasing, especially in those treated with strong BPs such as zoledronate (ZA). The pathobiology underlying the occurrence of osteonecrosis only in the jaw bone remains unclear. The objective of this study was to compare the effects of BPs on the in...

ba0005p157 | Cell biology: osteoblasts and bone formation | ECTS2016

Evidence for an osteogenic activity of betaine in human osteoblast in culture

Spinello Alice , Montesano Anna , Senesi Pamela , Villa Isabella , Bolamperti Simona , Vacante Fernanda , Luzi Livio , Rubinacci Alessandro , Terruzzi Ileana

Betaine (BET) is a component of many food. It is an essential osmolyte and a source of methyl groups. BET consumed from food sources and through dietary supplements presents similar bioavailability. BET exerts an antioxidant activity and decreases inflammation states. Dietary supplement with BET are used in many inflammation-connected pathologies, although its mechanism of action is not fully understood. Recent studies have shown that BET stimulates muscle fibers differentiati...

ba0005p158 | Cell biology: osteoblasts and bone formation | ECTS2016

A single 2-day pulse of activin-A leads to a transient change in gene expression eventually followed by reduction in extracellular matrix mineralization

Baroncelli Marta , Drabek Ksenija , Eijken Marco , Peppel Jeroen van de , van Leeuwen Johannes

Activins belong to the transforming growth factor-β superfamily, and they regulate bone formation by controlling both osteoclast and osteoblast behaviour. We have previously shown that activin-A strongly inhibited matrix mineralization in osteoblast cultures, and that activin A-signalling was most effective before the onset of mineralization.The aim of this study was therefore to investigate how an early activin-A pulse affected osteoblast mineraliz...

ba0005p159 | Cell biology: osteoblasts and bone formation | ECTS2016

Osteogenic superiority of bone marrow-derived osteoblastic cells (ALLOB®) over bone marrow-derived mesenchymal stromal cells

Pietri Sandra , Dubout Helene , Ena Sabrina , Hoste Candice , Bastianelli Enrico

Bone therapeutics is a bone cell therapy company addressing high unmet medical needs in the field of bone fracture repair, more specifically in non-union and delayed-union fractures where the bone repair process is impaired. The company has developed a unique allogeneic osteoblastic cell product (ALLOB®) derived from bone marrow which is currently tested in humans in the indication of delayed-union fractures. The purpose of the study was to directly compare ALL...

ba0005p160 | Cell biology: osteoblasts and bone formation | ECTS2016

Effect of melatonin on in vitro bone remodelling and blood supply

Gohin Stephanie , Pereira Marie , Belgrave Naomi Weekes , Lee Ji Sun , Tompkins Hayley , Orriss Isabel , Chenu Chantal

Melatonin is a neuro-hormone released primarily from the pineal gland, which has been shown to have bone anabolic effect, although it is still unclear whether its skeletal action is directly mediated by receptors expressed on bone cells or is indirect. In this study, we examined melatonin’s effects on bone cellular activities in vitro and tested whether it modifies angiogenesis and blood flow to bone, which are both essential for bone formation.<p class="abst...

ba0005p161 | Cell biology: osteoblasts and bone formation | ECTS2016

Vaspin regulates the osteogenic differentiation of MC3T3-E1 through the PI3K-Akt/miR-34c loop

Liu Yuan , Xu Feng , Pei Hong-Xia , Yuan Ling-Qing

Vaspin (visceral adipose tissue-derived serine protease inhibitor) is a newly discovered adipokine that widely participates in diabetes mellitus, polycystic ovarian syndrome and other disorders of metabolism. However, the effect of vaspin on the regulation of osteogenesis and the mechanism responsible are still unclear. Here, we found that vaspin could attenuate the ALP activities, osteocalcin secretion and Runx2 expression in the preosteoblast cell line MC3T3-E1 in a dose-dep...

ba0005p162 | Cell biology: osteoblasts and bone formation | ECTS2016

RCOR2 is a novel regulator of osteoblast differentiation

Tarkkonen Kati , Al Majidi Rana , Valensisi Cristina , Saastamoinen Lauri , Hawkins David , Kiviranta Riku

Epigenetic mechanisms regulating osteoblast differentiation are still inadequately described. In a genome wide transcriptional profiling of MC3T3-E1 osteoblastic cell line, we identified RCOR2 as a significantly upregulated gene during a differentiation time-course from proliferative to mature osteoblasts. Similar expression profile of RCOR2 was found in mouse calvarial osteoblasts. RCOR2 belongs to CoREST/RCOR family of proteins that regulate action of lysine-specific histone...

ba0005p163 | Cell biology: osteoblasts and bone formation | ECTS2016

Inhibition of histone demethylase LSD1 suppresses osteoblast differentiation

Al Majidi Rana , Tarkkonen Kati , Kiviranta Riku

Post-translational modifications of histone N-terminal tail domains affect the local chromatin conformation and serve as a dynamic regulatory layer for controlling gene transcription during cell differentiation. However, the role of epigenetic modifications in the regulation of osteoblast differentiation is still not fully understood. Recently, we identified RCOR2 as a novel regulator of osteoblast differentiation in a genome-wide transcription profiling of mouse MC3T3-E1 cell...

ba0005p164 | Cell biology: osteoblasts and bone formation | ECTS2016

Natural uranium triggers autophagy in osteoblasts

Pierrefite-Carle Valerie , Santucci-Darmanin Sabine , Breuil Veronique , Vidaud Claude , Creff Gaelle , Den Auwer Christophe , Carle Georges

Bone is a complex organ constituted of a mineralized matrix generated by osteoblasts (OB). Bone matrix is a major storage site for minerals but also for toxicants from the environment. Among them, uranium, a natural element of the earth crust, has a dual toxicity due to its radiological effects as an alpha emitter and its chemical effects due to its metal properties. In the case of natural uranium, the chemical toxicity is predominant. Uranium level in drinking water is usuall...

ba0005p165 | Cell biology: osteoblasts and bone formation | ECTS2016

Canine osteoblasts from trabecular, cortical and subchondral bone present differences in alkaline phosphatase activity

Perpetuo Ines Pedro , Shah Mittal , Parsons Kevin , Orriss Isabel , Doube Michael , Pitsillides Andrew , Meeson Richard

Hip osteoarthritis is a cause of significant morbidity to people and their canine companions. Medical management is frequently insufficient, leading to surgery to relieve pain and regain mobility. Hip replacements are not without potential complications, including loosening and infection. Currently, there is a focus on uncemented implants to decrease these problems, however these rely on the biology of the femur for osseointegration and long-term stability. It has been previou...

ba0005p166 | Cell biology: osteoblasts and bone formation | ECTS2016

Primary osteoblast culture from domestic dog (Canis lupus familiaris)

Perpetuo Ines Pedro , Meeson Richard , Pitsillides Andrew , Doube Michael , Orriss Isabel

Effects of whole bone scaling on isolated osteoblast behaviour are unknown. With two orders of magnitude range in body mass, dog breeds are well-suited to determine such relationships.Femoral heads from three canine hip replacement surgeries were collected. Bone fragments were washed in PBS+AB/AM, trypsin-digested and incubated in 0.2% collagenase. Cells from resultant supernatant were seeded in DMEM+10% FCS+AB/AM at 37 °C, 5% CO2, g...

ba0005p167 | Cell biology: osteoblasts and bone formation | ECTS2016

Implication of autophagy in a preclinical mouse model of bone ageing and of osteoporosis

Camuzard Olivier , Breuil Veronique , Santucci-Darmanin Sabine , Battaglia Severine , Heymann Dominique , Carle Georges , Pierrefite-Carle Valerie

Although estrogen deficiency has been considered for a long time as the main factor leading to osteoporosis (OP), several lines of evidence highlight the role of oxidative stress increase with age as a key factor in this pathology. Autophagy acts as a central mechanism allowing damaged macromolecules and organelles to be degraded and recycled, such as mitochondria, the main source of reactive oxygen species (ROS). Recent publications have shown that autophagy is a new actor in...

ba0005p168 | Cell biology: osteoblasts and bone formation | ECTS2016

Two different domains of fibronectin stimulate osteoblast differentiation by activating distinct integrins

Sens Carla , Rau Katrin , Nakchbandi Inaam

Fibronectin is produced in different isoforms by osteoblasts during differentiation. In contrast to the circulating Isoform, which lacks extra domains, the fibronectin laid down by the osteoblasts in the matrix contains one or both extra-domains called EDA and EDB. We had found that deletion of fibronectin in osteoblasts decreased BMD. This is mediated by diminished osteoblast differentiation and cannot be corrected by the circulating isoform of fibronectin, suggesting a role ...

ba0005p169 | Cell biology: osteoblasts and bone formation | ECTS2016

Iron deficiency increases osteoblast function via Wnt5a

Baschant Ulrike , Platzbecker Uwe , Rauner Martina , Hofbauer Lorenz

Iron overload due to hemochromatosis or chronic blood transfusions has been implicated in the development of osteoporosis. However, the impact of iron overload or iron deficiency on stromal cell functions and the underlying mechanisms are poorly defined. Since the Wnt signaling pathway is a critical regulator of bone remodelling, we aimed to analyse the effects of iron overload and iron deficiency on osteoblast function and further define the role of Wnt signaling in these pro...

ba0005p170 | Cell biology: osteoblasts and bone formation | ECTS2016

Matrix vesicle-mediated mineralization depends on the balance between annexins and fetuin-A which may be modulated by TNAP and calcium channel inhibitors

Strzelecka-Kiliszek Agnieszka , Bozycki Lukasz , Roszkowska Monika , Pikula Slawomir

Bone mineralization is initiated by matrix vesicles (MVs), cell-derived structures released into the extracellular matrix which are nucleation sites for hydroxyapatite (HA) formation. It is suggested that annexins are mineralization-stimulating membrane proteins that exhibit ion channel activity and facilitate the influx of Ca2+ into MVs. The process is also regulated via enzymatic degradation of inhibitory pyrophosphate by tissue-nonspecific alkaline phosphatase (T...