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8. Effect of mistletoe preparations on cell migration: exploration of 3D experimental models

Author

Scheer, Rainer, Alban, Susanne, Becker, Hans, Beer, André-Michael, Blaschek, Wolfgang, Klein, Reinhild, Kreis, Wolfgang, Leneweit, Gero, Matthes, Harald, Spahn, Günther, Spange, Rainer, Scheer, R ( Rainer ), Alban, S ( Susanne ), Becker, H ( Hans ), Beer, A ( André-Michael ), Blaschek, W ( Wolfgang ), Klein, R ( Reinhild ), Kreis, W ( Wolfgang ), Leneweit, G ( Gero ), Matthes, H ( Harald ), Spahn, G ( Günther ), Spange, R ( Rainer ), Dolder, Alexandra, Avilla-Royo, Eva; https://orcid.org/0000-0002-1002-8491, Weissenstein, U, Baumgartner, Stephan, Simona, Benjamin R, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Simões-Wüst, Ana Paula; https://orcid.org/0000-0002-4489-0952, Scheer, Rainer, Alban, Susanne, Becker, Hans, Beer, André-Michael, Blaschek, Wolfgang, Klein, Reinhild, Kreis, Wolfgang, Leneweit, Gero, Matthes, Harald, Spahn, Günther, Spange, Rainer, Scheer, R ( Rainer ), Alban, S ( Susanne ), Becker, H ( Hans ), Beer, A ( André-Michael ), Blaschek, W ( Wolfgang ), Klein, R ( Reinhild ), Kreis, W ( Wolfgang ), Leneweit, G ( Gero ), Matthes, H ( Harald ), Spahn, G ( Günther ), Spange, R ( Rainer ), Dolder, Alexandra, Avilla-Royo, Eva; https://orcid.org/0000-0002-1002-8491, Weissenstein, U, Baumgartner, Stephan, Simona, Benjamin R, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, and Simões-Wüst, Ana Paula; https://orcid.org/0000-0002-4489-0952

Published
2020

12. Multiaxial mechanical behavior of human fetal membranes and its relationship to microstructure

Author

Buerzle, W., Haller, C., Jabareen, M., Egger, J., Mallik, A., Ochsenbein-Koelble, N., Ehrbar, M., Mazza, E., Buerzle, W., Haller, C., Jabareen, M., Egger, J., Mallik, A., Ochsenbein-Koelble, N., Ehrbar, M., and Mazza, E.

Abstract

This study was directed to the measurement of the mechanical response of fetal membranes to physiologically relevant loading conditions. Characteristic mechanical parameters were determined and their relation to the microstructural constituents collagen and elastin as well as to the pyridinium cross-link concentrations analyzed. 51 samples from twelve fetal membranes were tested on a custom-built inflation device, which allows mechanical characterization within a multiaxial state of stress. Methods of nonlinear continuum mechanics were used to extract representative mechanical parameters. Established biochemical assays were applied for the determination of the collagen and elastin content. Collagen cross-link concentrations were determined by high-performance liquid chromatography measurements. The results indicate a distinct correlation between the mechanical parameters of high stretch stiffness and membrane tension at rupture and the biochemical data of collagen content and pyridinoline as well as deoxypyridinoline concentrations. No correlation was observed between the mechanical parameters and the elastin content. Moreover, the low stretch stiffness is, with a value of 105±31×10−3N/mm much higher for a biaxial state of stress compared to a uniaxial stress configuration. Determination of constitutive model equations leads to better predictive capabilities for a reduced polynomial hyperelastic model with only terms related to the second invariant, I 2, of the right Cauchy-Green deformation tensor. Relevant insights were obtained on the mechanical behavior of fetal membranes. Collagen and its cross-linking were shown to determine membrane's stiffness and strength for multiaxial stress states. Their nonlinear deformation behavior characterizes the fetal membranes as I 2 material

Published
2018

13. 4.31 Cell-Demanded Release of Growth Factors

Author

Ducheyne, Paul, Ducheyne, P ( Paul ), Milleret, V, Lienemann, P S, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Ducheyne, Paul, Ducheyne, P ( Paul ), Milleret, V, Lienemann, P S, and Ehrbar, M; https://orcid.org/0000-0003-2707-4870

Abstract

Stimulation of natural repair processes with growth factors or cytokines has emerged as an alternative to therapies of tissue or organ failure by surgical treatments. To overcome the poor performance of growth factors, biomaterials in which the decision on dose, location and release of growth factors are coupled to cellular activity at the treatment site are being developed. We summarize biomimetic approaches to entrap growth factors within cell-degradable polymer matrices from which the release can be controlled by cell-demanded mechanism. Preclinical results indicating that such biomimetic solutions for localized and prolonged growth factor delivery could improve therapeutic effects are given.

Published
2017

16. pH-controlled recovery of placenta-derived mesenchymal stem cell sheets

Author

Guillaume-Gentil, O, Semenov, O V, Zisch, A H, Zimmermann, R, Vörös, J, Ehrbar, M, University of Zurich, and Ehrbar, M

Subjects
170 Ethics, 2211 Mechanics of Materials, 1502 Bioengineering, 2502 Biomaterials, 610 Medicine & health, 10237 Institute of Biomedical Engineering, 2503 Ceramics and Composites, 10026 Clinic for Obstetrics, 1304 Biophysics
Published
2011

17. Enzyme mediated site-specific surface modification

Author

Sala, A, Ehrbar, M, Trentin, D, Schoenmakers, R G, Vörös, J, Weber, Franz E, University of Zurich, and Ehrbar, M

Subjects
170 Ethics, 10022 Division of Surgical Research, 3104 Condensed Matter Physics, 1607 Spectroscopy, 610 Medicine & health, 10237 Institute of Biomedical Engineering, 1603 Electrochemistry, 3110 Surfaces and Interfaces, 10069 Clinic of Cranio-Maxillofacial Surgery, 10026 Clinic for Obstetrics, 2500 General Materials Science
Published
2010

18. Recent advances in 3D tissue models

Author

Kivelio, A, Ehrbar, M, University of Zurich, Akashi, Misturu, Akagi, Takami, and Matsusaki, MIchiya

Subjects
610 Medicine & health, 10026 Clinic for Obstetrics
Published
2014

19. Locally controlling mesenchymal stem cell morphogenesis by 3D PDGF-BB gradients towards the establishment of an in vitro perivascular niche

Author

Lienemann, P. S., Devaud, Y. R., Reuten, R., Simona, B. R., Karlsson, M., Weber, W., Koch, M., Lutolf, M. P., Milleret, V., Ehrbar, M., Lienemann, P. S., Devaud, Y. R., Reuten, R., Simona, B. R., Karlsson, M., Weber, W., Koch, M., Lutolf, M. P., Milleret, V., and Ehrbar, M.

Abstract

The perivascular niche is a complex microenvironment containing mesenchymal stem cells (MSCs), among other perivascular cells, as well as temporally organized biochemical and biophysical gradients. Due to a lack of conclusive phenotypic markers, MSCs' identity, heterogeneity and function within their native niche remain poorly understood. The in vitro reconstruction of an artificial three-dimensional (3D) perivascular niche would offer a powerful alternative to study MSC behavior under more defined conditions. To this end, we here present a poly(ethylene glycol)-based in vitro model that begins to mimic the spatiotemporally controlled presentation of biological cues within the in vivo perivascular niche, namely a stably localized platelet-derived growth factor B (PDGF-BB) gradient. We show that 3D-encapsulated MSCs respond to soluble PDGF-BB by proliferation, spreading, and migration in a dose-dependent manner. In contrast, the exposure of MSCs to 3D matrix-tethered PDGF-BB gradients resulted in locally restricted morphogenetic responses, much as would be expected in a native perivascular niche. Thus, the herein presented artificial perivascular niche model provides an important first step towards modeling the role of MSCs during tissue homeostasis and regeneration.

Published
2015

21. Angiogenesis and vascularity for tissue engineering

Author

Largo, R, Ramakrishnan, V, Ehrbar, M, Ziogas, A, Plock, J A, Eberli, D, University of Zurich, and Eberli, D

Subjects
10062 Urological Clinic, 10022 Division of Surgical Research, 610 Medicine & health
Published
2011

22. Cell-demanded release of growth factors

Author

Zisch, A H, Ehrbar, M, University of Zurich, Ducheyne, P, Healy, K E, Hutmacher, D W, Grainger, D W, Kirkpatrick, C J, and Zisch, A H

Subjects
610 Medicine & health, 2700 General Medicine, 10026 Clinic for Obstetrics, 3500 General Dentistry
Published
2011

25. Effects of muCT radiation on tissue engineered bone-like constructs

Author

Kraehenbuehl, T P, Stauber, M, Ehrbar, M, Weber, Franz E, Hall, H, Müller, R, University of Zurich, and Müller, R

Subjects
10022 Division of Surgical Research, 2204 Biomedical Engineering, 610 Medicine & health, 10069 Clinic of Cranio-Maxillofacial Surgery, 10026 Clinic for Obstetrics
Published
2010

26. The effect of matrix characteristics on fibroblast proliferation in 3D gels

Author

Bott, K, Upton, Z, Schrobback, K, Ehrbar, M, Hubbell, J A, Lutolf, M P, Rizzi, S C, University of Zurich, and Rizzi, S C

Subjects
2211 Mechanics of Materials, 1502 Bioengineering, 2502 Biomaterials, 610 Medicine & health, 2503 Ceramics and Composites, 10026 Clinic for Obstetrics, 1304 Biophysics
Published
2010

27. 3D growth factor patterned cell substrates to locally control cellular behavior

Author

Ehrbar, M., Lienemann, P., Metzger, S., Devaud, Y., Kivelio, A-S, Reuten, R., Simona, B., Karlsson, M., Weber, W., Weber, F., Lutolf, M., Milleret, V., Ehrbar, M., Lienemann, P., Metzger, S., Devaud, Y., Kivelio, A-S, Reuten, R., Simona, B., Karlsson, M., Weber, W., Weber, F., Lutolf, M., and Milleret, V.

Published
2014

28. Recent advances in 3D tissue models

Author

Akashi, Misturu, Akagi, Takami, Matsusaki, MIchiya, Akashi, M ( Misturu ), Akagi, T ( Takami ), Matsusaki, M ( MIchiya ), Kivelio, A, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Akashi, Misturu, Akagi, Takami, Matsusaki, MIchiya, Akashi, M ( Misturu ), Akagi, T ( Takami ), Matsusaki, M ( MIchiya ), Kivelio, A, and Ehrbar, M; https://orcid.org/0000-0003-2707-4870

Abstract

Physiologically relevant tissue models that bridge the gap between 2D tissue culture and animal trials would be highly desirable to study the function of tissues in health and disease as well as for the validation of lead compounds during drug development. The field has made impressive advances in 3D culturing cells and organoids in naturally derived materials. Novel, rationally designed, biomimetic materials have been established, which allow the almost individual variation of matrix parameters, such as stiffness, cell adhesion, degradability, or growth factor binding and controlled release. The combination of innovative materials with novel technological platforms such as printing, microfluidics, and additive or preventive manufacturing provides a great potential to build unprecedented, complex tissue models. Here we review recent advances in the design of materials building blocks which allow the formation of 3D structured microenvironments. We will mainly focus on strategies to locally position cell-instructive molecular cues and discuss needs to generate models which would allow the investigator to controllably manipulate cells in their 3D context with the aim to generate complex but yet scalable tissue models.

Published
2014

29. In vivo and in vitro evaluation of flexible, cottonwool-like nanocomposites as bone substitute material for complex defects

Author

Schneider, O D, Weber, Franz, Brunner, T J, Loher, S, Ehrbar, M, Schmidlin, P R, Stark, W J, University of Zurich, and Stark, W J

Subjects
10022 Division of Surgical Research, 1303 Biochemistry, 10066 Clinic of Conservative and Preventive Dentistry, 2502 Biomaterials, 1305 Biotechnology, 1312 Molecular Biology, 2204 Biomedical Engineering, 610 Medicine & health, 10069 Clinic of Cranio-Maxillofacial Surgery, 10026 Clinic for Obstetrics
Published
2009

30. The role of actively released fibrin-conjugated VEGF for VEGF receptor 2 gene activation and the enhancement of angiogenesis

Author

Ehrbar, M, Zeisberger, S M, Raeber, G P, Hubbell, J A, Schnell, C, Zisch, A H, University of Zurich, and Zisch, A H

Subjects
10022 Division of Surgical Research, 2211 Mechanics of Materials, 1502 Bioengineering, 10076 Center for Integrative Human Physiology, 2502 Biomaterials, 570 Life sciences, biology, 610 Medicine & health, 2503 Ceramics and Composites, 10069 Clinic of Cranio-Maxillofacial Surgery, 10026 Clinic for Obstetrics, 1304 Biophysics
Published
2008

32. Enzymatic formation of modular cell-instructive fibrin analogs for tissue engineering

Author

Ehrbar, M, Rizzi, S C, Hlushchuk, R, Djonov, V, Zisch, A H, Hubbell, J A, Weber, F E, Lutolf, M P, University of Zurich, and Hubbell, J A

Subjects
1502 Bioengineering, Fibrin/*chemistry/*pharmacology, 2502 Biomaterials, Cell Culture Techniques/methods, 610 Medicine & health, 2503 Ceramics and Composites, Transglutaminases/*chemistry, Tissue Engineering/*methods, Cell Line, 2211 Mechanics of Materials, 10076 Center for Integrative Human Physiology, Humans, Mesenchymal Stem Cells/*cytology/drug effects/*physiology, 570 Life sciences, biology, Extracellular Matrix/*chemistry, 10026 Clinic for Obstetrics, 1304 Biophysics
Abstract

The molecular engineering of cell-instructive artificial extracellular matrices is a powerful means to control cell behavior and enable complex processes of tissue formation and regeneration. This work reports on a novel method to produce such smart biomaterials by recapitulating the crosslinking chemistry and the biomolecular characteristics of the biopolymer fibrin in a synthetic analog. We use activated coagulation transglutaminase factor XIIIa for site-specific coupling of cell adhesion ligands and engineered growth factor proteins to multiarm poly(ethylene glycol) macromers that simultaneously form proteolytically sensitive hydrogel networks in the same enzyme-catalyzed reaction. Growth factor proteins are quantitatively incorporated and released upon cell-derived proteolytic degradation of the gels. Primary stromal cells can invade and proteolytically remodel these networks both in an in vitro and in vivo setting. The synthetic ease and potential to engineer their physicochemical and bioactive characteristics makes these hybrid networks true alternatives for fibrin as provisional drug delivery platforms in tissue engineering.

Published
2007

33. Engineered fibrin matrices for functional display of cell membrane-bound growth factor-like activities: study of angiogenic signaling by ephrin-B2

Author

Zisch, A H, Zeisberger, S M, Ehrbar, M, Djonov, V, Weber, C C, Ziemiecki, A, Pasquale, E B, Hubbell, J A, University of Zurich, and Zisch, A H

Subjects
10022 Division of Surgical Research, 2211 Mechanics of Materials, 1502 Bioengineering, 2502 Biomaterials, 610 Medicine & health, 2503 Ceramics and Composites, 1304 Biophysics
Published
2004

34. A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells

Author

Muller, K, Engesser, R, Metzger, S, Schulz, S, Kampf, M M, Busacker, M, Steinberg, T, Tomakidi, P, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Nagy, F, Timmer, J, Zubriggen, M D, Weber, W, Muller, K, Engesser, R, Metzger, S, Schulz, S, Kampf, M M, Busacker, M, Steinberg, T, Tomakidi, P, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Nagy, F, Timmer, J, Zubriggen, M D, and Weber, W

Abstract

Growth and differentiation of multicellular systems is orchestrated by spatially restricted gene expression programs in specialized subpopulations. The targeted manipulation of such processes by synthetic tools with high-spatiotemporal resolution could, therefore, enable a deepened understanding of developmental processes and open new opportunities in tissue engineering. Here, we describe the first red/far-red light-triggered gene switch for mammalian cells for achieving gene expression control in time and space. We show that the system can reversibly be toggled between stable on- and off-states using short light pulses at 660 or 740 nm. Red light-induced gene expression was shown to correlate with the applied photon number and was compatible with different mammalian cell lines, including human primary cells. The light-induced expression kinetics were quantitatively analyzed by a mathematical model. We apply the system for the spatially controlled engineering of angiogenesis in chicken embryos. The system’s performance combined with cell- and tissue-compatible regulating red light will enable unprecedented spatiotemporally controlled molecular interventions in mammalian cells, tissues and organisms.

Published
2013

37. Cell-demanded release of growth factors

Author

Ducheyne, P, Healy, K E, Hutmacher, D W, Grainger, D W, Kirkpatrick, C J, Ducheyne, P ( P ), Healy, K E ( K E ), Hutmacher, D W ( D W ), Grainger, D W ( D W ), Kirkpatrick, C J ( C J ), Zisch, A H, Ehrbar, M, Ducheyne, P, Healy, K E, Hutmacher, D W, Grainger, D W, Kirkpatrick, C J, Ducheyne, P ( P ), Healy, K E ( K E ), Hutmacher, D W ( D W ), Grainger, D W ( D W ), Kirkpatrick, C J ( C J ), Zisch, A H, and Ehrbar, M

Published
2011

38. Angiogenesis and vascularity for tissue engineering

Author

Eberli, D, Eberli, D ( D ), Largo, R, Ramakrishnan, V, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Ziogas, A, Plock, J A, Eberli, D, Eberli, D ( D ), Largo, R, Ramakrishnan, V, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Ziogas, A, and Plock, J A

Published
2011

39. Decrease in VEGF expression induces intussusceptive vascular pruning

Author

Hlushchuk, R, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Reichmuth, P, Heinimann, N, Styp-Rekowska, B, Escher, R, Baum, O, Lienemann, P, Makanya, A, Keshet, E, Djonov, V, Hlushchuk, R, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Reichmuth, P, Heinimann, N, Styp-Rekowska, B, Escher, R, Baum, O, Lienemann, P, Makanya, A, Keshet, E, and Djonov, V

Abstract

Diminution of VEGF level induces vascular tree regression by intussusceptive vascular pruning. This observation may allude to the mechanism underlying the "normalization" of tumor vasculature if treated with antiangiogenic drugs. The mechanism described here gives new insights into the understanding of the processes of vasculature regression and hence provides new and potentially viable targets for antiangiogenic and/or angio-modulating therapies during various pathological processes.

Published
2011

40. Elucidating the role of matrix stiffness in 3D cell migration and remodeling

Author

Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Sala, A, Lienemann, P, Ranga, A, Mosiewicz, K, Bittermann, A, Rizzi, S C, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, Lutolf, M P, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Sala, A, Lienemann, P, Ranga, A, Mosiewicz, K, Bittermann, A, Rizzi, S C, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, and Lutolf, M P

Abstract

Reductionist in vitro model systems which mimic specific extracellular matrix functions in a highly controlled manner, termed artificial extracellular matrices (aECM), have increasingly been used to elucidate the role of cell-ECM interactions in regulating cell fate. To better understand the interplay of biophysical and biochemical effectors in controlling three-dimensional cell migration, a poly(ethylene glycol)-based aECM platform was used in this study to explore the influence of matrix cross-linking density, represented here by stiffness, on cell migration in vitro and in vivo. In vitro, the migration behavior of single preosteoblastic cells within hydrogels of varying stiffness and susceptibilities to degradation by matrix metalloproteases was assessed by time-lapse microscopy. Migration behavior was seen to be strongly dependent on matrix stiffness, with two regimes identified: a nonproteolytic migration mode dominating at relatively low matrix stiffness and proteolytic migration at higher stiffness. Subsequent in vivo experiments revealed a similar stiffness dependence of matrix remodeling, albeit less sensitive to the matrix metalloprotease sensitivity. Therefore, our aECM model system is well suited to unveil the role of biophysical and biochemical determinants of physiologically relevant cell migration phenomena.

Published
2011

41. Engineering 3D cell instructive microenvironments by rational assembly of artificial extracellular matrices and cell patterning

Author

Sala, A, Hänseler, P, Ranga, A, Lutolf, M P, Vörös, J, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, Sala, A, Hänseler, P, Ranga, A, Lutolf, M P, Vörös, J, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, and Weber, Franz E; https://orcid.org/0000-0003-1670-2296

Abstract

Engineered artificial microenvironments hold enormous potential as models to study developmental, physiological, pathological, and regenerative processes under highly defined conditions. Such platforms aim at bridging the gap between traditional in vitro 2D culture systems and animal models. By dissecting the biological complexity into an amenable number of parameters, systemic manipulation and study in controllable environments closely resembling the in vivo situation is possible. Novel strategies that address the evaluation of either ECM components, growth factors or cell-cell interactions on cellular behaviour are being developed. However, reliable methods that simultaneously recapitulate the natural instructive microenvironments in terms of cell and matrix composition, biological cues, heterogeneity and geometry are not yet available. Such spatially-defined microenvironments may be necessary to initiate and guide the formation of artificial tissues by morphogenetic processes. In this work, we introduce a flexible strategy that relies on the combination of artificial extracellular matrices with patterning techniques as well as a layer-by-layer approach to mimic rationally-designed instructive milieus. By a rational arrangement of cells and defined biochemical and biophysical extracellular cues, we report control of cell migration and generation of an artificial vascularized bone tissue-like construct.

Published
2011

42. Mechanical loading of mouse caudal vertebrae increases trabecular and cortical bone mass-dependence on dose and genotype

Author

Webster, D, Wasserman, E, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, Bab, I, Müller, R, Webster, D, Wasserman, E, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, Bab, I, and Müller, R

Abstract

Most in vivo studies addressing the skeletal responses of mice to mechanical loading have targeted cortical bone. To investigate trabecular bone responses also we have developed a caudal vertebral axial compression device (CVAD) that transmits mechanical loads to compress the fifth caudal vertebra via stainless steel pins inserted into the forth and sixth caudal vertebral bodies. Here, we used the CVAD in C57BL/6 (B6) and C3H/Hej (C3H) female mice (15 weeks of age) to investigate whether the effect of regular bouts of mechanical stimulation on bone modeling and bone mass was dependent on dose and genotype. A combined micro-computed tomographic and dynamic histomorphometric analysis was carried out at the end of a 4-week loading regimen (3,000 cycles, 10 Hz, 3 x week) for load amplitudes of 0N, 2N, 4N and 8N. Significant increases in trabecular bone mass of 9 and 21% for loads of 4N and 8N, respectively, were observed in B6 mice. A significant increase of 10% in trabecular bone mass occurred for a load of 8N in the C3H strain. For other loads, no significant increases were detected. Both mouse strains exhibited substantial increases in trabecular bone formation rates for all loads, B6: 111% (2N), 86% (4N), 164% (8N), C3H: 41% (2N), 38% (4N), 141% (8N). Significant decreases in osteoclast number of 146 and 93% for a load of 8N were detected in B6 and C3H mice, respectively. These findings demonstrate that the effect of loading on the structural and functional parameters of bone is dose and genotype dependent. The caudal vertebral loading model established here is proposed for further studies addressing the molecular processes involved in the skeletal responses to mechanical stimuli.

Published
2010

43. Enhanced osteoblastic activity and bone regeneration using surface-modified porous bioactive glass scaffolds

Author

San Miguel, B, Kriauciunas, R, Tosatti, S, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Ghayor, C, Textor, M, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, San Miguel, B, Kriauciunas, R, Tosatti, S, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Ghayor, C, Textor, M, and Weber, Franz E; https://orcid.org/0000-0003-1670-2296

Abstract

The potential use as a bone substitute material of a three-dimensional bioactive glass fiber scaffold composed of Na(2)O-K(2)O-MgO-CaO-B(2)O(3)-P(2)O(5)-SiO(2) (BG1) was investigated in this work. Scaffolds were pre-treated with simulated body fluid (SBF) to promote the formation of two different bone-like apatite layers on their surfaces. The topography and roughness of the deposited layers were assessed by scanning electron microscopy (SEM), while the chemical composition and structure using X-ray photoelectron spectroscopy (XPS) and Raman spectroscopy, respectively. Based on surface analysis, the bioactive glass surfaces were ranked from smoothest to roughest: 0 SBF (untreated), 1x SBF and 2x SBF. A calcium-deficient carbonated hydroxyapatite (HCA) layer was present on both SBF-treated scaffolds, with higher number and larger bone-like apatite nodule formation in the 2x SBF case. MC3T3-E1 preosteoblasts showed a more flattened morphology and higher cell proliferation on the nontreated scaffolds; whereas, cells were more elongated and had higher osteoblastic activity on SBF-treated samples. In vivo results in a rabbit calvarial bone defect model showed enhanced bone formation with SBF pretreated scaffolds, compared with untreated ones, commercially available Perioglass particles and empty defects. Our findings demonstrate that the formation of a rough HCA layer on bioactive glass porous scaffolds enhanced preosteoblast maturation in vitro, as well as bone formation in vivo.

Published
2010

44. cAMP enhances BMP2-signaling through PKA and MKP1-dependent mechanisms

Author

Ghayor, C, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, San Miguel, B, Grätz, K W, Weber, Franz E; https://orcid.org/0000-0003-1670-2296, Ghayor, C, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, San Miguel, B, Grätz, K W, and Weber, Franz E; https://orcid.org/0000-0003-1670-2296

Abstract

Recent studies suggest that the elevation of intracellular cyclic adenosine monophosphate (cAMP) and the activation of the protein kinase A regulate BMP-induced osteogenesis. However, the precise mechanisms underlying the enhancing effect of cAMP on BMP2 signaling were not completely revealed. In this study we investigated the effect of elevated cAMP level and PKA activation on the BMP2-induced osteoblastic differentiation in pluripotent C2C12 cells. Alkaline phosphatase activity and its mRNA were consistently induced by BMP2 treatment. The pretreatment of C2C12 cells with Forskolin, a cAMP generating agent, dbcAMP, an analogue of cAMP, or IBMX (3-isobutyl 1-methyl xanthine), and a nonspecific inhibitor of phosphodiesterases elicited further activation of alkaline phosphatase. Furthermore, elevated intracellular cAMP level increased BMP2-induced MKP1. On the other hand, BMP2-induced Erk phosphorylation (p44/p42) and cell proliferation were suppressed in the presence of cAMP. Thus, cAMP might enhance BMP2-induced osteoblastic differentiation by a MKP1-Erk-dependent mechanism.

Published
2009

45. The role of actively released fibrin-conjugated VEGF for VEGF receptor 2 gene activation and the enhancement of angiogenesis

Author

Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Zeisberger, S M, Raeber, G P, Hubbell, J A, Schnell, C, Zisch, A H, Ehrbar, M; https://orcid.org/0000-0003-2707-4870, Zeisberger, S M, Raeber, G P, Hubbell, J A, Schnell, C, and Zisch, A H

Abstract

A major challenge for therapeutic delivery of angiogenic agents such as vascular endothelial growth factor (VEGF) is to achieve sustained, low dose signaling leading to durable neovessel formation. To this end, we recently created a variant of VEGF(121), TG-VEGF(121) that directly binds to fibrin and gets released locally in proteolysis-triggered manner. Here we combined noninvasive biophotonic monitoring of VEGF receptor 2 gene activation in transgenic VEGFR2-luc mice and histomorphometry to compare endothelial activation and long-term neovascularization by actively released TG-VEGF(121)versus passively released, diffusible wild-type VEGF(121) in subcutaneous fibrin implants. Monitoring in real-time over 3 weeks of luciferase signal driven by the VEGFR2 promoter revealed endothelial activation in skin exposed to wild-type VEGF(121), but no detectable elevation over fibrin alone by TG-VEGF(121). Histology at 3 weeks, however, demonstrated that TG-VEGF(121) promoted vessel growth significantly more effectively and reliably than wild-type VEGF(121). The majority of vessels surviving to 3 weeks contained stabilizing smooth muscle cells. Yet, by 6 weeks, no extra vessels induced by exogenous VEGF were left. In conclusion, release of fibrin-conjugated variant TG-VEGF(121) elicited lower VEGFR2-luc activation than wild-type VEGF(121) yet significantly more vascularization. In the absence of true physiological demand, even stabilized vessels are ultimately regressed.

Published
2008

48. Poster session 2

Author

Perez-Pomares, J. M., primary, Ruiz-Villalba, A., additional, Ziogas, A., additional, Segovia, J. C., additional, Ehrbar, M., additional, Munoz-Chapuli, R., additional, De La Rosa, A., additional, Dominguez, J. N., additional, Hove-Madsen, L., additional, Sankova, B., additional, Sedmera, D., additional, Franco, D., additional, Aranega Jimenez, A., additional, Babaeva, G., additional, Chizh, N., additional, Galchenko, S., additional, Sandomirsky, B., additional, Schwarzl, M., additional, Seiler, S., additional, Steendijk, P., additional, Huber, S., additional, Maechler, H., additional, Truschnig-Wilders, M., additional, Pieske, B., additional, Post, H., additional, Simrick, S., additional, Kreutzer, R., additional, Rao, C., additional, Terracciano, C. M., additional, Kirchhof, P., additional, Fabritz, L., additional, Brand, T., additional, Theveniau-Ruissy, M., additional, Parisot, P., additional, Francou, A., additional, Saint-Michel, E., additional, Mesbah, K., additional, Kelly, R. G., additional, Wu, H.-T., additional, Sie, S.-S., additional, Chen, C.-Y., additional, Kuan, T.-C., additional, Lin, C. S., additional, Ismailoglu, Z., additional, Guven, M., additional, Yakici, A., additional, Ata, Y., additional, Ozcan, S., additional, Yildirim, E., additional, Ongen, Z., additional, Miroshnikova, V., additional, Demina, E., additional, Rodygina, T., additional, Kurjanov, P., additional, Denisenko, A., additional, Schwarzman, A., additional, Rubanenko, A., additional, Shchukin, Y., additional, Germanov, A., additional, Goldbergova, M., additional, Parenica, J., additional, Lipkova, J., additional, Pavek, N., additional, Kala, P., additional, Poloczek, M., additional, Vasku, A., additional, Parenicova, I., additional, Spinar, J., additional, Gambacciani, C., additional, Chiavacci, E., additional, Evangelista, M., additional, Vesentini, N., additional, Kusmic, C., additional, Pitto, L., additional, Chernova, A., additional, Nikulina, S. U. Y., additional, Arvanitis, D. A., additional, Mourouzis, I., additional, Pantos, C., additional, Kranias, E. G., additional, Cokkinos, D. V., additional, Sanoudou, D., additional, Vladimirskaya, T. E., additional, Shved, I. A., additional, Kryvorot, S. G., additional, Schirmer, I. M., additional, Appukuttan, A., additional, Pott, L., additional, Jaquet, K., additional, Ladilov, Y., additional, Archer, C. R., additional, Bootman, M. D., additional, Roderick, H. L., additional, Fusco, A., additional, Sorriento, D., additional, Santulli, G., additional, Trimarco, B., additional, Iaccarino, G., additional, Hagenmueller, M., additional, Riffel, J., additional, Bernhold, E., additional, Katus, H. A., additional, Hardt, S. E., additional, Maqsood, A., additional, Zi, M., additional, Prehar, S., additional, Neyses, L., additional, Ray, S., additional, Oceandy, D., additional, Khatami, N., additional, Wadowski, P., additional, Wagh, V., additional, Hescheler, J., additional, Sachinidis, A., additional, Mohl, W., additional, Chaudhry, B., additional, Burns, D., additional, Henderson, D. J., additional, Bax, N. A. M., additional, Van Marion, M. H., additional, Shah, B., additional, Goumans, M. J., additional, Bouten, C. V. C., additional, Van Der Schaft, D. W. J., additional, Van Oorschot, A. A. M., additional, Maas, S., additional, Braun, J., additional, Van Tuyn, J., additional, De Vries, A. A. F., additional, Gittenberger-De Groot, A. C., additional, Bageghni, S., additional, Drinkhill, M. J., additional, Batten, T. F. C., additional, Ainscough, J. F. X., additional, Onate, B., additional, Vilahur, G., additional, Ferrer-Lorente, R., additional, Ybarra, J., additional, Diez-Caballero, A., additional, Ballesta-Lopez, C., additional, Moscatiello, F., additional, Herrero, J., additional, Badimon, L., additional, Martin-Rendon, E., additional, Clifford, D. M., additional, Fisher, S. A., additional, Brusnkill, S. J., additional, Doree, C., additional, Mathur, A., additional, Clarke, M., additional, Watt, S. M., additional, Hernandez-Vera, R., additional, Kavanagh, D., additional, Yemm, A. I., additional, Frampton, J., additional, Kalia, N., additional, Terajima, Y., additional, Shimizu, T., additional, Tsuruyama, S., additional, Ishii, H., additional, Sekine, H., additional, Hagiwara, N., additional, Okano, T., additional, Vrijsen, K. R., additional, Chamuleau, S. A. J., additional, Sluijter, J. P. G., additional, Doevendans, P. F. M., additional, Madonna, R., additional, Delli Pizzi, S., additional, Di Donato, L., additional, Mariotti, A., additional, Di Carlo, L., additional, D'ugo, E., additional, Teberino, M. A., additional, Merla, A., additional, T, A., additional, De Caterina, R., additional, Kolker, L., additional, Ali, N. N., additional, Maclellan, K., additional, Moore, M., additional, Wheeler, J., additional, Harding, S. E., additional, Fleck, R. A., additional, Rowlinson, J. M., additional, Kraenkel, N., additional, Ascione, R., additional, Madeddu, P., additional, O'sullivan, J. F., additional, Leblond, A. L., additional, Kelly, G., additional, Kumar, A. H. S., additional, Metharom, P., additional, Buneker, C. K., additional, Alizadeh-Vikali, N., additional, Hynes, B. G., additional, O'connor, R., additional, Caplice, N. M., additional, Noseda, M., additional, De Smith, A. J., additional, Leja, T., additional, Rao, P. H., additional, Al-Beidh, F., additional, Abreu Pavia, M. S., additional, Blakemore, A. I., additional, Schneider, M. D., additional, Stathopoulou, K., additional, Cuello, F., additional, Ehler, E., additional, Haworth, R. S., additional, Avkiran, M., additional, Morawietz, H., additional, Eickholt, C., additional, Langbein, H., additional, Brux, M., additional, Goettsch, C., additional, Goettsch, W., additional, Arsov, A., additional, Brunssen, C., additional, Mazilu, L., additional, Parepa, I. R., additional, Suceveanu, A. I., additional, Suceveanu, A. P., additional, De Man, F. S., additional, Guignabert, C., additional, Tu, L., additional, Handoko, M. L., additional, Schalij, I., additional, Fadel, E., additional, Postmus, P. E., additional, Vonk-Noordegraaf, A., additional, Humbert, M., additional, Eddahibi, S., additional, Del Giudice, C., additional, Anastasio, A., additional, Fazal, L., additional, Azibani, F., additional, Bihry, N., additional, Merval, R., additional, Polidano, E., additional, Samuel, J.-L., additional, Delcayre, C., additional, Zhang, Y., additional, Mi, Y. M., additional, Ren, L. L., additional, Cheng, Y. P., additional, Guo, R., additional, Liu, Y., additional, Jiang, Y. N., additional, Kokkinos, A. D., additional, Tretjakovs, P., additional, Jurka, A., additional, Bormane, I., additional, Mikelsone, I., additional, Reihmane, D., additional, Elksne, K., additional, Krievina, G., additional, Verbovenko, J., additional, Bahs, G., additional, Lopez-Andres, N., additional, Rousseau, A., additional, Calvier, L., additional, Akhtar, R., additional, Labat, C., additional, Cruickshank, K., additional, Diez, J., additional, Zannad, F., additional, Lacolley, P., additional, Rossignol, P., additional, Hamesch, K., additional, Subramanian, P., additional, Li, X., additional, Thiemann, A., additional, Heyll, K., additional, Dembowsky, K., additional, Chevalier, E., additional, Weber, C., additional, Schober, A., additional, Yang, L., additional, Kim, G., additional, Gardner, B., additional, Earley, J., additional, Hofmann-Bowman, M., additional, Cheng, C.-F., additional, Lian, W.-S., additional, Lin, H., additional, Jinjolia, N. J., additional, Abuladze, G. A., additional, Tvalchrelidze, S. H. T., additional, Khamnagadaev, I., additional, Shkolnikova, M., additional, Kokov, L., additional, Miklashevich, I., additional, Drozdov, I., additional, Ilyich, I., additional, Bingen, B. O., additional, Askar, S. F. A., additional, Ypey, D. L., additional, Van Der Laarse, A., additional, Schalij, M. J., additional, Pijnappels, D. A., additional, Roney, C. H., additional, Ng, F. S., additional, Chowdhury, R. A., additional, Chang, E. T. Y., additional, Patel, P. M., additional, Lyon, A. R., additional, Siggers, J. H., additional, Peters, N. S., additional, Obergrussberger, A., additional, Stoelzle, S., additional, Bruggemann, A., additional, Haarmann, C., additional, George, M., additional, Fertig, N., additional, Moreira, D., additional, Souza, A., additional, Valente, P., additional, Kornej, J., additional, Reihardt, C., additional, Kosiuk, J., additional, Arya, A., additional, Hindricks, G., additional, Adams, V., additional, Husser, D., additional, Bollmann, A., additional, Camelliti, P., additional, Dudhia, J., additional, Dias, P., additional, Cartledge, J., additional, Connolly, D. J., additional, Nobles, M., additional, Sebastian, S., additional, Tinker, A., additional, Opel, A., additional, Daimi, H., additional, Haj Khelil, A., additional, Be Chibani, J., additional, Barana, A., additional, Amoros, I., additional, Gonzalez De La Fuente, M., additional, Caballero, R., additional, Aranega, A., additional, Kelly, A., additional, Bernus, O., additional, Kemi, O. J., additional, Myles, R. C., additional, Ghouri, I. A., additional, Burton, F. L., additional, Smith, G. L., additional, Del Lungo, M., additional, Sartiani, L., additional, Spinelli, V., additional, Baruscotti, M., additional, Difrancesco, D., additional, Mugelli, A., additional, Cerbai, E., additional, Thomas, A. M., additional, Aziz, Q., additional, Khambra, T., additional, Addlestone, J. M. A., additional, Cartwright, E. J., additional, Wilkinson, R., additional, Song, W., additional, Marston, S., additional, Jacquet, A., additional, Mougenot, N. M., additional, Lipskaia, A. J., additional, Paalberends, E. R., additional, Stam, K., additional, Van Dijk, S. J., additional, Van Slegtenhorst, M., additional, Dos Remedios, C., additional, Ten Cate, F. J., additional, Michels, M., additional, Niessen, H. W. M., additional, Stienen, G. J. M., additional, Van Der Velden, J., additional, Read, M. I., additional, Andreianova, A. A., additional, Harrison, J. C., additional, Goulton, C. S., additional, Kerr, D. S., additional, Sammut, I. A., additional, Wallner, M., additional, Von Lewinski, D., additional, Kindsvater, D., additional, Saes, M., additional, Morano, I., additional, Muegge, A., additional, Buyandelger, B., additional, Kostin, S., additional, Gunkel, S., additional, Vouffo, J., additional, Ng, K., additional, Chen, J., additional, Eilers, M., additional, Isaacson, R., additional, Milting, H., additional, Knoell, R., additional, Cattin, M.-E., additional, Crocini, C., additional, Schlossarek, S., additional, Maron, S., additional, Hansen, A., additional, Eschenhagen, T., additional, Carrier, L., additional, Bonne, G., additional, Coppini, R., additional, Ferrantini, C., additional, Olivotto, I., additional, Belardinelli, L., additional, Poggesi, C., additional, Leung, M. C., additional, Messer, A. E., additional, Copeland, O., additional, Marston, S. B., additional, Mills, A. M., additional, Collins, T., additional, O'gara, P., additional, Thum, T., additional, Regalla, K., additional, Macleod, K. T., additional, Prodromakis, T., additional, Chaudhry, U., additional, Darzi, A., additional, Yacoub, M. H., additional, Athanasiou, T., additional, Bogdanova, A., additional, Makhro, A., additional, Hoydal, M., additional, Stolen, T. O., additional, Johnssen, A. B., additional, Alves, M., additional, Catalucci, D., additional, Condorelli, G., additional, Koch, L. G., additional, Britton, S. L., additional, Wisloff, U., additional, Bito, V., additional, Claus, P., additional, Vermeulen, K., additional, Huysmans, C., additional, Ventura-Clapier, R., additional, Sipido, K. R., additional, Seliuk, M. N., additional, Burlaka, A. P., additional, Sidorik, E. P., additional, Khaitovych, N. V., additional, Kozachok, M. M., additional, Potaskalova, V. S., additional, Driesen, R. B., additional, Galan, D. T., additional, De Paulis, D., additional, Arnoux, T., additional, Schaller, S., additional, Pruss, R. M., additional, Poitz, D. M., additional, Augstein, A., additional, Braun-Dullaeus, R. C., additional, Schmeisser, A., additional, Strasser, R. H., additional, Micova, P., additional, Balkova, P., additional, Hlavackova, M., additional, Zurmanova, J., additional, Kasparova, D., additional, Kolar, F., additional, Neckar, J., additional, Novak, F., additional, Novakova, O., additional, Pollard, S., additional, Babba, M., additional, Hussain, A., additional, James, R., additional, Maddock, H., additional, Alshehri, A. S., additional, Baxter, G. F., additional, Dietel, B., additional, Altendorf, R., additional, Daniel, W. G., additional, Kollmar, R., additional, Garlichs, C. D., additional, Sirohi, R., additional, Roberts, N., additional, Lawrence, D., additional, Sheikh, A., additional, Kolvekar, S., additional, Yap, J., additional, Arend, M., additional, Walkinshaw, G., additional, Hausenloy, D. J., additional, Yellon, D. M., additional, Posa, A., additional, Szabo, R., additional, Szalai, Z., additional, Szablics, P., additional, Berko, M. A., additional, Orban, K., additional, Murlasits, Z. S., additional, Balogh, L., additional, Varga, C., additional, Ku, H. C., additional, Su, M. J., additional, Chreih, R.-M., additional, Ginghina, C., additional, Deleanu, D., additional, Ferreira, A. L. B. J., additional, Belal, A., additional, Ali, M. A., additional, Fan, X., additional, Holt, A., additional, Campbell, R., additional, Schulz, R., additional, Bonanad, C., additional, Bodi, V., additional, Sanchis, J., additional, Morales, J. M., additional, Marrachelli, V., additional, Nunez, J., additional, Forteza, M. J., additional, Chaustre, F., additional, Gomez, C., additional, Chorro, F. J., additional, Csont, T., additional, Fekete, V., additional, Murlasits, Z., additional, Aypar, E., additional, Bencsik, P., additional, Sarkozy, M., additional, Varga, Z. V., additional, Ferdinandy, P., additional, Duerr, G. D., additional, Zoerlein, M., additional, Dewald, D., additional, Mesenholl, B., additional, Schneider, P., additional, Ghanem, A., additional, Rittling, S., additional, Welz, A., additional, Dewald, O., additional, Becker, E., additional, Peigney, C., additional, Bouleti, C., additional, Galaup, A., additional, Monnot, C., additional, Ghaleh, B., additional, Germain, S., additional, Timmermans, A., additional, Ginion, A., additional, De Meester, C., additional, Sakamoto, K., additional, Vanoverschelde, J.-L., additional, Horman, S., additional, Beauloye, C., additional, Bertrand, L., additional, Maroz-Vadalazhskaya, N., additional, Drozd, E., additional, Kukharenko, L., additional, Russkich, I., additional, Krachak, D., additional, Seljun, Y., additional, Ostrovski, Y., additional, Martin, A.-C., additional, Le Bonniec, B., additional, Lecompte, T., additional, Dizier, B., additional, Emmerich, J., additional, Fischer, A.-M., additional, Samama, C.-M., additional, Godier, A., additional, Mogensen, S., additional, Furchtbauer, E. M., additional, Aalkjaer, C., additional, Choong, W. L., additional, Jovanovic, A., additional, Khan, F., additional, Daniel, J. M., additional, Dutzmann, J. M., additional, Widmer-Teske, R., additional, Guenduez, D., additional, Sedding, D., additional, Castro, M. M., additional, Cena, J. J. C., additional, Cho, W. J. C., additional, Goobie, G. G., additional, Walsh, M. P. W., additional, Schulz, R. S., additional, Dutzmann, J., additional, Preissner, K. T., additional, Sones, W., additional, Kotlikoff, M., additional, Serizawa, K., additional, Yogo, K., additional, Aizawa, K., additional, Hirata, M., additional, Tashiro, Y., additional, Ishizuka, N., additional, Varela, A., additional, Katsiboulas, M., additional, Tousoulis, D., additional, Papaioannou, T. G., additional, Vaina, S., additional, Davos, C. H., additional, Piperi, C., additional, Stefanadis, C., additional, Basdra, E. K., additional, Papavassiliou, A. G., additional, Hermenegildo, C., additional, Lazaro-Franco, M., additional, Sobrino, A., additional, Bueno-Beti, C., additional, Martinez-Gil, N., additional, Walther, T., additional, Peiro, C., additional, Sanchez-Ferrer, C. F., additional, Novella, S., additional, Ciccarelli, M., additional, Franco, A., additional, Dorn, G. W., additional, Cseplo, P., additional, Torok, O., additional, Springo, Z. S., additional, Vamos, Z., additional, Kosa, D., additional, Hamar, J., additional, Koller, A., additional, Bubb, K. J., additional, Ahluwalia, A., additional, Stepien, E. L., additional, Gruca, A., additional, Grzybowska, J., additional, Goralska, J., additional, Dembinska-Kiec, A., additional, Stolinski, J., additional, Partyka, L., additional, Zhang, H., additional, Sweeney, D., additional, Thomas, G. N., additional, Fish, P. V., additional, Taggart, D. P., additional, Cioffi, S., additional, Bilio, M., additional, Martucciello, S., additional, Illingworth, E., additional, Caporali, A., additional, Shantikumar, S., additional, Marchetti, M., additional, Martelli, F., additional, Emanueli, C., additional, Meloni, M., additional, Al Haj Zen, A., additional, Sala-Newby, G., additional, Del Turco, S., additional, Saponaro, C., additional, Dario, B., additional, Sartini, S., additional, Menciassi, A., additional, Dario, P., additional, La Motta, C., additional, Basta, G., additional, Santiemma, V., additional, Bertone, C., additional, Rossi, F., additional, Michelon, E., additional, Bianco, M. J., additional, Castelli, A., additional, Shin, D. I., additional, Seung, K. B., additional, Seo, S. M., additional, Park, H. J., additional, Kim, P. J., additional, Baek, S. H., additional, Choi, Y. S., additional, Her, S. H., additional, Kim, D. B., additional, Lee, J. M., additional, Park, C. S., additional, Rocchiccioli, S., additional, Cecchettini, A., additional, Pelosi, G., additional, Citti, L., additional, Parodi, O., additional, Trivella, M. G., additional, Michel-Monigadon, D., additional, Burger, F., additional, Dunoyer-Geindre, S., additional, Pelli, G., additional, Cravatt, B., additional, Steffens, S., additional, Didangelos, A., additional, Mayr, U., additional, Yin, X., additional, Stegemann, C., additional, Shalhoub, J., additional, Davies, A. H., additional, Monaco, C., additional, Mayr, M., additional, Lypovetska, S., additional, Grytsenko, S., additional, Njerve, I. U., additional, Pettersen, A. A., additional, Opstad, T. B., additional, Bratseth, V., additional, Arnesen, H., additional, Seljeflot, I., additional, Dumitriu, I. E., additional, Baruah, P., additional, Antunes, R. F., additional, Kaski, J. C., additional, Trapero, I., additional, Benet, I., additional, Alguero, C., additional, Chaustre, F. J., additional, Mangold, A., additional, Puthenkalam, S., additional, Distelmaier, K., additional, Adlbrecht, C., additional, Lang, I. M., additional, Koizumi, T., additional, Inoue, I., additional, Komiyama, N., additional, Nishimura, S., additional, Korneeva, O. N., additional, Drapkina, O. M., additional, Fornai, L., additional, Angelini, A., additional, Kiss, A., additional, Giskes, F., additional, Eijkel, G., additional, Fedrigo, M., additional, Valente, M. L., additional, Thiene, G., additional, Heeren, R. M. A., additional, Padro, T., additional, Casani, L., additional, Suades, R., additional, Bertoni, B., additional, Carminati, R., additional, Carlini, V., additional, Pettinari, L., additional, Martinelli, C., additional, Gagliano, N., additional, Noppe, G., additional, Buchlin, P., additional, Marquet, N., additional, Baeyens, N., additional, Morel, N., additional, Baysa, A., additional, Sagave, J., additional, Dahl, C. P., additional, Gullestad, L., additional, Carpi, A., additional, Di Lisa, F., additional, Giorgio, M., additional, Vaage, J., additional, Valen, G., additional, Vafiadaki, E., additional, Papalouka, V., additional, Terzis, G., additional, Spengos, K., additional, Manta, P., additional, Gales, C., additional, Genet, G., additional, Dague, E., additional, Cazorla, O., additional, Payre, B., additional, Mias, C., additional, Ouille, A., additional, Lacampagne, A., additional, Pathak, A., additional, Senard, J. M., additional, Abonnenc, M., additional, Da Costa Martins, P., additional, Srivastava, S., additional, Gautel, M., additional, De Windt, L., additional, Comelli, L., additional, Lande, C., additional, Ucciferri, N., additional, Ikonen, L., additional, Vuorenpaa, H., additional, Kujala, K., additional, Sarkanen, J.-R., additional, Heinonen, T., additional, Ylikomi, T., additional, Aalto-Setala, K., additional, Capros, H., additional, Sprincean, N., additional, Usurelu, N., additional, Egorov, V., additional, Stratu, N., additional, Matchkov, V., additional, Bouzinova, E., additional, Moeller-Nielsen, N., additional, Wiborg, O., additional, Gutierrez, P. S., additional, Aparecida-Silva, R., additional, Borges, L. F., additional, Moreira, L. F. P., additional, Dias, R. R., additional, Kalil, J., additional, Stolf, N. A. G., additional, Zhou, W., additional, Suntharalingam, K., additional, Brand, N., additional, Vilar Compte, R., additional, Ying, L., additional, Bicknell, K., additional, Dannoura, A., additional, Dash, P., additional, Brooks, G., additional, Tsimafeyeu, I., additional, Tishova, Y., additional, Wynn, N., additional, Oyeyipo, I. P., additional, Olatunji, L. A., additional, Maegdefessel, L., additional, Azuma, J., additional, Toh, R., additional, Raaz, U., additional, Merk, D. R., additional, Deng, A., additional, Spin, J. M., additional, Tsao, P. S., additional, Tedeschi, L., additional, Taranta, M., additional, Naldi, I., additional, Grimaldi, S., additional, Cinti, C., additional, Bousquenaud, M., additional, Maskali, F., additional, Poussier, S., additional, Marie, P. Y., additional, Boutley, H., additional, Karcher, G., additional, Wagner, D. R., additional, Devaux, Y., additional, Torre, I., additional, Psilodimitrakopoulos, S., additional, Iruretagoiena, I., additional, Gonzalez-Tendero, A., additional, Artigas, D., additional, Loza-Alvarez, P., additional, Gratacos, E., additional, Amat-Roldan, I., additional, Murray, L., additional, Carberry, D. M., additional, Dunton, P., additional, Miles, M. J., additional, Suleiman, M.-S., additional, Kanesalingam, K., additional, Taylor, R., additional, Mc Collum, C. N., additional, Parniczky, A., additional, Solymar, M., additional, Porpaczy, A., additional, Miseta, A., additional, Lenkey, Z. S., additional, Szabados, S., additional, Cziraki, A., additional, Garai, J., additional, Myloslavska, I., additional, Menazza, S. M., additional, Canton, M. C., additional, Di Lisa, F. D. L., additional, Oliveira, S. H. V., additional, Morais, C. A. S., additional, Miranda, M. R., additional, Oliveira, T. T., additional, Lamego, M. R. A., additional, Lima, L. M., additional, Goncharova, N. S., additional, Naymushin, A. V., additional, Kazimli, A. V., additional, Moiseeva, O. M., additional, Carvalho, M. G., additional, Sabino, A. P., additional, Mota, A. P. L., additional, Sousa, M. O., additional, Niessner, A., additional, Richter, B., additional, Hohensinner, P. J., additional, Rychli, K., additional, Zorn, G., additional, Berger, R., additional, Moertl, D., additional, Pacher, R., additional, Wojta, J., additional, Huelsmann, M., additional, Kukharchik, G., additional, Nesterova, N., additional, Pavlova, A., additional, Gaykovaya, L., additional, Krapivka, N., additional, Konstantinova, I., additional, Sichinava, L., additional, Prapa, S., additional, Mccarthy, K. P., additional, Kilner, P. J., additional, Xu, X. Y., additional, Johnson, M. R., additional, Ho, S. Y., additional, Gatzoulis, M. A., additional, Stoupel, E. G., additional, Garcia, R., additional, Merino, D., additional, Montalvo, C., additional, Hurle, M. A., additional, Nistal, J. F., additional, Villar, A. V., additional, Perez-Moreno, A., additional, Gilabert, R., additional, and Ros, E., additional

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2012
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