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2. Automated Atlas-based Segmentation of Single Coronal Mouse Brain Slices using Linear 2D-2D Registration

Author

Piluso, S��bastien, Souedet, Nicolas, Jan, Caroline, Clouchoux, C��dric, and Delzescaux, Thierry

Subjects
FOS: Computer and information sciences, Mice, Computer Vision and Pattern Recognition (cs.CV), Image and Video Processing (eess.IV), FOS: Electrical engineering, electronic engineering, information engineering, Computer Science - Computer Vision and Pattern Recognition, Animals, Brain, Electrical Engineering and Systems Science - Image and Video Processing
Abstract

A significant challenge for brain histological data analysis is to precisely identify anatomical regions in order to perform accurate local quantifications and evaluate therapeutic solutions. Usually, this task is performed manually, becoming therefore tedious and subjective. Another option is to use automatic or semi-automatic methods, among which segmentation using digital atlases co-registration. However, most available atlases are 3D, whereas digitized histological data are 2D. Methods to perform such 2D-3D segmentation from an atlas are required. This paper proposes a strategy to automatically and accurately segment single 2D coronal slices within a 3D volume of atlas, using linear registration. We validated its robustness and performance using an exploratory approach at whole-brain scale.

Published
2021

5. A modular joint-on-chip approach to study cellular cross-communication in a simulated osteoarthritic micro-environment

Author

Moreira Teixeira, L., Paggi, C., Piluso, S., Leijten, J., Malda, J., van Weeren, R., le Gac, S., Karperien, M., Developmental BioEngineering, TechMed Centre, and Applied Microfluidics for BioEngineering Research

Abstract

INTRODUCTION: Cartilage degeneration and synovitis are key hallmarks of joint degenerative diseases, such as osteoarthritis (OA). The communication between these two tissues is fundamental to maintain both homeostasis and disease onset. However, studying this communication has remained challenging. Herein, we propose a novel modular precision microfluidic platform, that combines a synovial membrane-on-chip and a cartilage-on-chip platform. This seamless strategy enables the facile and in depth study of cross-communication between these two joint components, specificallyvia inflammatory mediators, aiming to replicate the onset of OA. METHODS: Two types of microfluidic PDMS-based chips were designed with actuation chambers to emulate the mechanical forces of the cartilage and synovial membrane. For the cartilage chip, human healthy and OA affected chondrocytes were seeded in an ECM-like hydrogel. The synovial membrane chip was composed of a hydrogel re-enforced by an elastic membrane, which was seeded with synovium fibroblasts. Both chips were connected by a common channel, where synovium mimicking medium (culture medium supplemented with HA) was flowing (60 μl/hr). After both tissues achieved phenotypical maturation, human macrophages were added to the system. The behavior of the immune cells were a key read-out, focusing on their mobility, cytokine and proteinase release profile (ELISA) and polarization ratio between M1 and M2 (qPCR). On-line and end-pointanalysis were conducted after 1, 3 and 7 days. RESULTS & DISCUSSION: The unique chip actuator designs allowed for physiologically relevant stimulation of the cartilage and synovial membrane, by cyclic compression and stretching, respectively. The effect of the mechanical load was determined on the release of inflammatory mediators. The proteinases and inflammatory cytokine profiles are the basis of the fingerprint of the influence of the variables: static vs. mechanically stimulated, healthy vs. OA-affected chondrocytes,and, presence vs. absence of macrophages. CONCLUSIONS: This modular joint-on-chip platform has the potential to provide unprecedented insights in the effects of inflammation of a single joint tissue, the performance of the various joint tissues, and the joint function itself.

Published
2019

12. Poster Session 5: Saturday 10 December 2011, 08:30-12:30 * Location: Poster Area

Author

Gong, L., primary, Ye, Z., additional, Zeng, Z., additional, Xia, M., additional, Zhong, Y., additional, Yao, Y., additional, Lee, E., additional, Ionescu, A., additional, Dwivedi, G., additional, Mahadevan, G., additional, Jiminez, D., additional, Frenneaux, M., additional, Steeds, R., additional, Moore, C., additional, Samad, Z., additional, Jackson, K., additional, Castellucci, J., additional, Kisslo, J., additional, Von Ramm, O., additional, D'ascenzi, F., additional, Zaca', V., additional, Cameli, M., additional, Lisi, M., additional, Natali, B., additional, Malandrino, A., additional, Mondillo, S., additional, Barbier, P., additional, Guerrini, U., additional, Franzosi, M., additional, Castiglioni, L., additional, Nobili, E., additional, Colazzo, F., additional, Li Causi, T., additional, Sironi, L., additional, Tremoli, E., additional, Clausen, H., additional, Macdonald, S., additional, Basaggianis, C., additional, Newton, J., additional, Bennati, E., additional, Reccia, R., additional, Bigio, E., additional, Maccherini, M., additional, Chiavarelli, M., additional, Henein, M., additional, Floria, M., additional, Jamart, J., additional, Arsenescu Georgescu, C., additional, Mantovani, F., additional, Barbieri, A., additional, Bursi, F., additional, Valenti, C., additional, Quaglia, M., additional, Modena, M., additional, Kutty, S., additional, Gribben, P., additional, Padiyath, A., additional, Polak, A., additional, Scott, C., additional, Waiss, M., additional, Danford, D., additional, Bech-Hanssen, O., additional, Selimovic, N., additional, Rundqvist, B., additional, Schmiedel, L., additional, Hohmann, C., additional, Katzke, S., additional, Haacke, K., additional, Rauwolf, T., additional, Strasser, R., additional, Tumasyan, L. R., additional, Adamyan, K., additional, Kosmala, W., additional, Derzhko, R., additional, Przewlocka-Kosmala, M., additional, Mysiak, A., additional, Stachowska, B., additional, Jedrzejuk, D., additional, Bednarek-Tupikowska, G., additional, Chrzanowski, L., additional, Kasprzak, J., additional, Wojciechowska, C., additional, Wita, K., additional, Busz-Papiez, B., additional, Gasior, Z., additional, Mizia-Stec, K., additional, Kukulski, T., additional, Gosciniak, P., additional, Sinkiewicz, W., additional, Moelmen, H., additional, Stoylen, A., additional, Thorstensen, A., additional, Torp, H., additional, Dalen, H., additional, Groves, A., additional, Nicholson, G., additional, Lopez, L., additional, Goh, C.-W., additional, Ahn, H., additional, Byun, Y., additional, Kim, J., additional, Park, J., additional, Lee, J., additional, Kim, B., additional, Rhee, K., additional, Kim, K., additional, Yoon, H., additional, Hong, Y., additional, Park, H., additional, Ahn, Y., additional, Jeong, M., additional, Cho, J., additional, Kang, J., additional, Grapsa, J., additional, Dawson, D., additional, Karfopoulos, K., additional, Jakaj, G., additional, Punjabi, P., additional, Nihoyannopoulos, P., additional, Ruisanchez Villar, C., additional, Lerena Saenz, P., additional, Gonzalez Vilchez, F., additional, Gonzalez Fernandez, C., additional, Zurbano Goni, F., additional, Cifrian Martinez, J., additional, Mons Lera, R., additional, Ruano Calvo, J., additional, Martin Duran, R., additional, Vazquez De Prada Tiffe, J., additional, Pietrzak, R., additional, Werner, B., additional, Voillot, D., additional, Huttin, O., additional, Zinzius, P., additional, Schwartz, J., additional, Sellal, J., additional, Lemoine, S., additional, Christophe, C., additional, Popovic, B., additional, Juilliere, Y., additional, Selton-Suty, C., additional, Ishii, K., additional, Furukawa, A., additional, Nagai, T., additional, Kataoka, K., additional, Seino, Y., additional, Shimada, K., additional, Yoshikawa, J., additional, Tekkesin, A., additional, Yildirimturk, O., additional, Tayyareci, Y., additional, Yurdakul, S., additional, Aytekin, S., additional, Jaroch, J., additional, Loboz-Grudzien, K., additional, Bociaga, Z., additional, Kowalska, A., additional, Kruszynska, E., additional, Wilczynska, M., additional, Dudek, K., additional, Kakihara, R., additional, Naruse, C., additional, Hironaka, H., additional, Tsuzuku, T., additional, Cucchini, U., additional, Muraru, D., additional, Badano, L., additional, Solda', E., additional, Tuveri, M., additional, Al Nono, O., additional, Sarais, C., additional, Iliceto, S., additional, Santos, L., additional, Cortez-Dias, N., additional, Ribeiro, S., additional, Goncalves, S., additional, Jorge, C., additional, Carrilho-Ferreira, P., additional, Silva, D., additional, Silva-Marques, J., additional, Lopes, M., additional, Diogo, A., additional, Hristova, K., additional, Vassilev, D., additional, Pavlov, P., additional, Katova, T., additional, Simova, I., additional, Kostova, V., additional, Esposito, R., additional, Santoro, A., additional, Schiano Lomoriello, V., additional, Raia, R., additional, De Palma, D., additional, Dores, E., additional, De Simone, G., additional, Galderisi, M., additional, Zaborska, B., additional, Makowska, E., additional, Pilichowska, E., additional, Maciejewski, P., additional, Bednarz, B., additional, Wasek, W., additional, Stec, S., additional, Budaj, A., additional, Spinelli, L., additional, Morisco, C., additional, Assante Di Panzillo, E., additional, Crispo, S., additional, Di Marino, S., additional, Trimarco, B., additional, Farina, F., additional, Innelli, P., additional, Rapacciuolo, A., additional, Polgar, B., additional, Banyai, F., additional, Rokusz, L., additional, Tomcsanyi, I., additional, Vaszily, M., additional, Nieszner, E., additional, Borsanyi, T., additional, Kerecsen, G., additional, Preda, I., additional, Kiss, R. G., additional, Bull, S., additional, Suttie, J., additional, Augustine, D., additional, Francis, J., additional, Karamitsos, T., additional, Becher, H., additional, Prendergast, B., additional, Neubauer, S., additional, Myerson, S., additional, Lodge, F., additional, Broyd, C., additional, Milton, P., additional, Mikhail, G., additional, Mayet, J., additional, Davies, J., additional, Francis, D., additional, Clavel, M.-A., additional, Ennezat, P.-V., additional, Marechaux, S., additional, Dumesnil, J., additional, Bellouin, A., additional, Bergeron, S., additional, Meimoun, P., additional, Le Tourneau, T., additional, Pasquet, A., additional, Pibarot, P., additional, Herrmann, S., additional, Stoerk, S., additional, Niemann, M., additional, Hu, K., additional, Voelker, W., additional, Ertl, G., additional, Weidemann, F., additional, Aytekin, V., additional, Kogoj, P., additional, Ambrozic, J., additional, Bunc, M., additional, Di Salvo, G., additional, Rea, A., additional, Castaldi, B., additional, Gala, S., additional, D'aiello, A., additional, Mormile, A., additional, Pisacane, F., additional, Pacileo, G., additional, Russo, M., additional, Calabro, R., additional, Nguyen, L., additional, Ricksten, S.-E., additional, Jeppsson, A., additional, Schersten, H., additional, Boerlage-Van Dijk, K., additional, Yong, Z., additional, Bouma, B., additional, Koch, K., additional, Vis, M., additional, Piek, J., additional, Baan, J., additional, Scandura, S., additional, Ussia, G., additional, Caggegi, A., additional, Cammalleri, V., additional, Sarkar, K., additional, Mangiafico, S., additional, Chiaranda', M., additional, Imme', S., additional, Pistritto, A., additional, Tamburino, C., additional, Ring, L., additional, Nair, S., additional, Wells, F., additional, Shapiro, L., additional, Rusk, R., additional, Rana, B., additional, Madrid Marcano, G., additional, Solis Martin, J., additional, Gonzalez Mansilla, A., additional, Bravo, L., additional, Menarguez Palanca, C., additional, Munoz, P., additional, Bouza, E., additional, Yotti, R., additional, Bermejo Thomas, J., additional, Fernandez Aviles, F., additional, Tamayo, T., additional, Denes, M., additional, Balint, O., additional, Csepregi, A., additional, Csillik, A., additional, Erdei, T., additional, Temesvari, A., additional, Fernandez-Pastor, J., additional, Linde-Estrella, A., additional, Cabrera-Bueno, F., additional, Pena-Hernandez, J., additional, Barrera-Cordero, A., additional, Alzueta-Rodriguez, F., additional, De Teresa-Galvan, E., additional, Merlo, M., additional, Pinamonti, M., additional, Finocchiaro, G., additional, Pyxaras, S., additional, Barbati, G., additional, Buiatti, A., additional, Dilenarda, A., additional, Sinagra, G., additional, Kuperstein, R., additional, Freimark, D., additional, Hirsch, S., additional, Feinberg, M., additional, Arad, M., additional, Mitroi, C., additional, Garcia Lunar, I., additional, Monivas Palomero, V., additional, Mingo Santos, S., additional, Beltran Correas, P., additional, Gonzalez Lopez, E., additional, Garcia Pavia, P., additional, Gonzalez Mirelis, J., additional, Cavero Gibanel, M., additional, Alonso Pulpon, L., additional, Pinamonti, B., additional, Zaidi, A., additional, Ghani, S., additional, Sheikh, N., additional, Gati, S., additional, Howes, R., additional, Sharma, R., additional, Sharma, S., additional, Calcagnino, M., additional, O'mahony, C., additional, Coats, C., additional, Cardona, M., additional, Garcia, A., additional, Murphy, E., additional, Lachmann, R., additional, Mehta, A., additional, Hughes, D., additional, Elliott, P., additional, Di Bella, G., additional, Madaffari, A., additional, Donato, R., additional, Mazzeo, A., additional, Casale, M., additional, Zito, C., additional, Vita, G., additional, Carerj, S., additional, Marek, D., additional, Indrakova, J., additional, Rusinakova, Z., additional, Skala, T., additional, Kocianova, E., additional, Taborsky, M., additional, Musca, F., additional, De Chiara, B., additional, Belli, O., additional, Cataldo, S., additional, Brunati, C., additional, Colussi, G., additional, Quattrocchi, G., additional, Santambrogio, G., additional, Spano, F., additional, Moreo, A., additional, Rustad, L., additional, Nytroen, K., additional, Gullestad, L., additional, Amundsen, B., additional, Aakhus, S., additional, Maroz-Vadalazhskaya, N., additional, Shumavetc, V., additional, Kurganovich, S., additional, Seljun, Y., additional, Ostrovskiy, A., additional, Ostrovskiy, Y., additional, Segers, P., additional, Orda, A., additional, Karolko, B., additional, Driessen, M. M. P., additional, Eising, J. B., additional, Uiterwaal, C., additional, Van Der Ent, C. K., additional, Meijboom, F. J., additional, Shang, Q., additional, Tam, L., additional, Sun, J., additional, Sanderson, J., additional, Zhang, Q., additional, Li, E., additional, Yu, C., additional, Arroyo Ucar, E., additional, De La Rosa Hernandez, A., additional, Hernandez Garcia, C., additional, Jorge Perez, P., additional, Lacalzada Almeida, J., additional, Jimenez Rivera, J., additional, Duque Garcia, A., additional, Barragan Acea, A., additional, Laynez Cerdena, I., additional, Kaldararova, M., additional, Simkova, I., additional, Pacak, J., additional, Tittel, P., additional, Masura, J., additional, Tadic, M., additional, Ivanovic, B., additional, Zlatanovic, M., additional, Damjanov, N., additional, Maggiolini, S., additional, Gentile, G., additional, Bozzano, A., additional, Suraci, S., additional, Meles, E., additional, Carbone, C., additional, Tempesta, A., additional, Malafronte, C., additional, Piatti, L., additional, Achilli, F., additional, Luijendijk, P., additional, Stevens, A., additional, De Bruin-Bon, H., additional, Vriend, J., additional, Van Den Brink, R., additional, Vliegen, H., additional, Mulder, B., additional, Chow, V., additional, Ng, A., additional, Chung, T., additional, Kritharides, L., additional, Iancu, M., additional, Serban, M., additional, Craciunescu, I., additional, Hodo, A., additional, Ghiorghiu, I., additional, Popescu, B., additional, Ginghina, C., additional, Styczynski, G., additional, Szmigielski, C. A., additional, Kaczynska, A., additional, Leszczynski, J., additional, Rosinski, G., additional, Kuch-Wocial, A., additional, Slavich, M., additional, Ancona, M., additional, Fisicaro, A., additional, Oppizzi, M., additional, Marone, E., additional, Bertoglio, L., additional, Melissano, G., additional, Margonato, A., additional, Chiesa, R., additional, Agricola, E., additional, Mohammed, M., additional, Cusma-Piccione, M., additional, Piluso, S., additional, Arcidiaco, S., additional, Nava, R., additional, Giuffre, R., additional, Ciraci, L., additional, Ferro, M., additional, Uusitalo, V., additional, Luotolahti, M., additional, Pietila, M., additional, Wendelin-Saarenhovi, M., additional, Hartiala, J., additional, Saraste, M., additional, Knuuti, J., additional, Saraste, A., additional, Kochanowski, J., additional, Scislo, P., additional, Piatkowski, R., additional, Grabowski, M., additional, Marchel, M., additional, Roik, M., additional, Kosior, D., additional, Opolski, G., additional, Bartko, P. E., additional, Graf, S., additional, Khorsand, A., additional, Rosenhek, R., additional, Burwash, I., additional, Beanlands, R., additional, Baumgartner, H., additional, Mundigler, G., additional, Kudrnova, S., additional, Apor, A., additional, Huttl, H., additional, Mori, F., additional, Santoro, G., additional, Oddo, A., additional, Rosso, G., additional, Meucci, F., additional, Pieri, F., additional, Squillantini, G., additional, Gensini, G., additional, Postula, M., additional, Park, D.-G., additional, Hong, J.-Y., additional, Kim, S.-E., additional, Lee, J.-H., additional, Han, K.-R., additional, Oh, D.-J., additional, Dal Bianco, L., additional, Beraldo, M., additional, Peluso, D., additional, Al Mamary, A., additional, Aggeli, C., additional, Felekos, I., additional, Poulidakis, E., additional, Pietri, P., additional, Roussakis, G., additional, Siasos, G., additional, Stefanadis, C., additional, Hoshiba, H., additional, Miyasaka, C., additional, Sato, H., additional, Yamanaka, A., additional, Lilli, A., additional, Baratto, M., additional, Magnacca, M., additional, Comella, A., additional, Poddighe, R., additional, Talini, E., additional, Canale, M., additional, Chioccioli, M., additional, Del Meglio, J., additional, Casolo, G., additional, Kuznetsov, V. A., additional, Melnikov, N. N., additional, Krinochkin, D. V., additional, Calin, A., additional, Enache, R., additional, Beladan, C., additional, Rosca, M., additional, Lupascu, L., additional, Purcarea, F., additional, Calin, C., additional, Gurzun, M., additional, Dulgheru, R., additional, Ciobanu, A., additional, Magda, S., additional, Mihaila, S., additional, Rimbas, R., additional, Margulescu, A., additional, Cinteza, M., additional, Vinereanu, D., additional, Sumin, A. N., additional, Arhipov, O., additional, Yoon, J., additional, Moon, J., additional, Rim, S., additional, Nyktari, E., additional, Patrianakos, A., additional, Solidakis, G., additional, Psathakis, E., additional, Parthenakis, F., additional, Vardas, P., additional, Kordybach, M., additional, Kowalski, M., additional, Kowalik, E., additional, Hoffman, P., additional, Nagy, K. V., additional, Kutyifa, V., additional, Edes, E., additional, Merkely, B., additional, Gerlach, A., additional, Rost, C., additional, Schmid, M., additional, Rost, M., additional, Flachskampf, F., additional, Daniel, W., additional, Breithardt, O., additional, Altekin, E., additional, Karakas, S., additional, Yanikoglu, A., additional, Er, A., additional, Baktir, A., additional, Demir, I., additional, Deger, N., additional, Klitsie, L., additional, Hazekamp, M., additional, Roest, A., additional, Van Der Hulst, A., additional, Gesink- Van Der Veer, B., additional, Kuipers, I., additional, Blom, N., additional, Ten Harkel, A., additional, Farsalinos, K., additional, Tsiapras, D., additional, Kyrzopoulos, S., additional, Avramidou, E., additional, Vasilopoulou, D., additional, Voudris, V., additional, Florianczyk, T., additional, Kalinowski, M., additional, Szulik, M., additional, Streb, W., additional, Rybus-Kalinowska, B., additional, Sliwinska, A., additional, Stabryla, J., additional, Kukla, M., additional, Nowak, J., additional, Kalarus, Z., additional, Florescu, M., additional, Mihalcea, D., additional, Magda, L., additional, Suran, B., additional, Enescu, O., additional, Mincu, R., additional, Salerno, G., additional, Scognamiglio, G., additional, D'andrea, A., additional, Dinardo, G., additional, Gravino, R., additional, Sarubbi, B., additional, Disalvo, G., additional, Liao, J.-N., additional, Sung, S., additional, Chen, C., additional, Park, S., additional, Shin, S., additional, Kim, M., additional, Shim, S., additional, Helvacioglu, F., additional, Ulusoy, O., additional, Duran, C., additional, Kirschner, R., additional, Simor, T., additional, Ambrosio, G., additional, Tran, T., additional, Raman, S., additional, Vidal Perez, R. C., additional, Carreras, F., additional, Leta, R., additional, Pujadas, S., additional, Barros, A., additional, Hidalgo, A., additional, Alomar, X., additional, Pons-Llado, G., additional, Olofsson, M., additional, Boman, K., additional, Ledakowicz-Polak, A., additional, Polak, L., additional, Zielinska, M., additional, Fontana, A., additional, Schirone, V., additional, Mauro, A., additional, Zambon, A., additional, Giannattasio, C., additional, Trocino, G., additional, Dekleva, M., additional, Dungen, H., additional, Inkrot, S., additional, Gelbrich, G., additional, Suzic Lazic, J., additional, Kleut, M., additional, Markovic Nikolic, N., additional, Waagstein, F., additional, Khoor, S., additional, Balogh, N., additional, Simon, I., additional, Fugedi, K., additional, Kovacs, I., additional, Khoor, M., additional, Florian, G., additional, Kocsis, A., additional, Szuszai, T., additional, O'driscoll, J., additional, Saha, A., additional, Smith, R., additional, Gupta, S., additional, Lenkey, Z., additional, Gaszner, B., additional, Illyes, M., additional, Sarszegi, Z., additional, Horvath, I. G., additional, Magyari, B., additional, Molnar, F., additional, Cziraki, A., additional, Elnoamany, M. F., additional, Badran, H., additional, Ebraheem, H., additional, Reda, A., additional, and Elsheekh, N., additional

Published
2011
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13. Poster Session 1: Thursday 8 December 2011, 08:30-12:30 * Location: Poster Area

Author

Vijayan, S., primary, Khanji, M., additional, Ionescu, A., additional, Vijayan, S., additional, Podoleanu, C., additional, Frigy, A., additional, Ugri, A., additional, Varga, A., additional, Podoleanu, D., additional, Incze, A., additional, Carasca, E., additional, Dobreanu, D., additional, Mjolstad, O., additional, Dalen, H., additional, Graven, T., additional, Kleinau, J., additional, Hagen, B., additional, Fu, H., additional, Liu, T., additional, Li, J., additional, Liu, C., additional, Zhou, C., additional, Li, G., additional, Bordese, R., additional, Capriolo, M., additional, Brero, D., additional, Salvetti, I., additional, Cannillo, M., additional, Antolini, M., additional, Grosso Marra, W., additional, Frea, S., additional, Morello, M., additional, Gaita, F., additional, Maffessanti, F., additional, Caiani, E., additional, Muraru, D., additional, Tuveri, F., additional, Dal Bianco, L., additional, Badano, L., additional, Majid, A., additional, Soesanto, A., additional, Ario Suryo Kuncoro, B., additional, Sukmawan, R., additional, Ganesja, M. H., additional, Benedek, T., additional, Chitu, M., additional, Beata, J., additional, Suciu, Z., additional, Kovacs, I., additional, Bucur, O., additional, Benedek, I., additional, Hrynkiewicz-Szymanska, A., additional, Szymanski, F., additional, Karpinski, G., additional, Filipiak, K., additional, Radunovic, Z., additional, Lande Wekre, L., additional, Steine, K., additional, Bech-Hanssen, O., additional, Rundqvist, B., additional, Lindgren, F., additional, Selimovic, N., additional, Jedrzychowska-Baraniak, J., additional, Jozwa, R., additional, Larysz, B., additional, Kasprzak, J., additional, Ripp, T., additional, Mordovin, V., additional, Ripp, E., additional, Ciobanu, A., additional, Dulgheru, R., additional, Dragoi, R., additional, Magda, S., additional, Florescu, M., additional, Mihaila, S., additional, Rimbas, R., additional, Cinteza, M., additional, Vinereanu, D., additional, Benavides-Vallve, C., additional, Pelacho, B., additional, Iglesias, O., additional, Castano, S., additional, Munoz-Barrutia, A., additional, Prosper, F., additional, Ortiz De Solorzano, C., additional, Manouras, A., additional, Sahlen, A., additional, Winter, R., additional, Vardas, P., additional, Brodin, L., additional, Sarvari, S. I., additional, Haugaa, K. H., additional, Zahid, W., additional, Bendz, B., additional, Aaberge, L., additional, Edvardsen, T., additional, Di Bella, G., additional, Pedri, S., additional, Donato, R., additional, Madaffari, A., additional, Zito, C., additional, Stapf, D., additional, Schreckenberg, M., additional, Carerj, S., additional, Yoshikawa, H., additional, Suzuki, M., additional, Kusunose, Y., additional, Hashimoto, G., additional, Otsuka, T., additional, Nakamura, M., additional, Sugi, K., additional, Grapsa, J., additional, Dawson, D., additional, Gin-Sing, W., additional, Howard, L., additional, Gibbs, J., additional, Nihoyannopoulos, P., additional, Smith, B., additional, Coulter, T., additional, Rendon, A., additional, Gorissen, W., additional, Shiran, A., additional, Asmer, I., additional, Adawi, S., additional, Ganaeem, M., additional, Shehadeh, J., additional, Cameli, M., additional, Lisi, M., additional, Righini, F., additional, Maccherini, M., additional, Sani, G., additional, Galderisi, M., additional, Mondillo, S., additional, Kalimanovska-Ostric, D., additional, Nastasovic, T., additional, Jovanovic, I., additional, Milakovic, B., additional, Dostanic, M., additional, Stosic, M., additional, Sasic, I., additional, Sveen, K., additional, Nerdrum, T., additional, Hanssen, K., additional, Dahl-Jorgensen, K., additional, Holte, E., additional, Vegsundvaag, J., additional, Hole, T., additional, Hegbom, K., additional, Wiseth, R., additional, Ikonomidis, I., additional, Lekakis, J., additional, Tritakis, V., additional, Papadakis, I., additional, Kadoglou, N., additional, Tzortzis, S., additional, Trivilou, P., additional, Koukoulis, C., additional, Paraskevaidis, I., additional, Anastasiou-Nana, M., additional, Smedsrud, M. K., additional, Sarvari, S., additional, Gjesdal, O., additional, Beraldo, M., additional, Solda', E., additional, Cucchini, U., additional, Peluso, D., additional, Tuveri, M., additional, Al Mamary, A., additional, Iliceto, S., additional, Dores, H., additional, Abecasis, J., additional, Carvalho, M., additional, Santos, M., additional, Andrade, M., additional, Ribeiras, R., additional, Reis, C., additional, Horta, E., additional, Gouveia, R., additional, Mendes, M., additional, Zaliaduonyte-Peksiene, D., additional, Mizariene, V., additional, Cesnaite, G., additional, Tamuleviciute, E., additional, Jurkevicius, R., additional, Vaskelyte, J., additional, Zaliunas, R., additional, Smarz, K., additional, Zaborska, B., additional, Jaxa-Chamiec, T., additional, Maciejewski, P., additional, Budaj, A., additional, Trifunovic, D., additional, Sobic-Saranovic, D., additional, Stankovic, S., additional, Ostojic, M., additional, Vujisic-Tesic, B., additional, Petrovic, M., additional, Nedeljkovic, I., additional, Banovic, M., additional, Tesic, M., additional, Petrovic, I., additional, Peovska, I., additional, Srbinovska, E., additional, Maksimovic, J., additional, Andova, V., additional, Arnaudova, F., additional, Hristova, E., additional, Otljanska, M., additional, Vavlukis, M., additional, Jovanova, S., additional, Tamborini, G., additional, Fusini, L., additional, Gripari, P., additional, Muratori, M., additional, Pontone, G., additional, Andreini, D., additional, Bertella, E., additional, Ghulam Ali, S., additional, Bartorelli, A., additional, Pepi, M., additional, Cusma-Piccione, M., additional, Salvia, J., additional, Antonini-Canterin, F., additional, Lentini, S., additional, Donato, D., additional, Miceli, M., additional, Oreto, G., additional, Sachner, R., additional, Rubinshtein, R., additional, Shnapp, M., additional, Gaspar, T., additional, Marchese, A., additional, Deste, W., additional, Sanfilippo, A., additional, Aruta, P., additional, Patane, M., additional, Millan, G., additional, Ussia, G., additional, Tamburino, C., additional, Kujacic, V., additional, Obradovic, S., additional, Crkvenac, Z., additional, Bernard, A., additional, Piquemal, M., additional, Muller, G., additional, Arbeille, P., additional, Charbonnier, B., additional, Broyd, C., additional, Davies, J., additional, Mikhail, G., additional, Mayet, J., additional, Francis, D., additional, Rosca, M., additional, Magne, J., additional, Szymanski, C., additional, Popescu, B., additional, Ginghina, C., additional, Pierard, L., additional, Lancellotti, P., additional, Gonzalez-Mansilla, A., additional, Solis, J., additional, Angulo, R., additional, Perez-David, E., additional, Madrid, G., additional, Garcia-Robles, J., additional, Yotti, R., additional, Prieto, R., additional, Bermejo, J., additional, Fernandez-Aviles, F., additional, Ishikawa, Y., additional, Ishida, T., additional, Osaki, T., additional, Matsuyama, M., additional, Yamashita, H., additional, Ozaki, S., additional, Stevanella, M., additional, Votta, E., additional, Veronesi, F., additional, Alamanni, F., additional, Redaelli, A., additional, Park, S. 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M., additional, Cosin-Sales, J., additional, Dalli, E., additional, Diago, J., additional, Aguilar, J., additional, Ruvira, J., additional, Goncalves, S., additional, Gomes, A., additional, Pinto, F., additional, Tsai, W.-C., additional, Liu, Y.-W., additional, Shih, J.-Y., additional, Huang, Y.-Y., additional, Chen, J.-Y., additional, Tsai, L.-M., additional, Chen, J.-H., additional, Ribeiro, S., additional, Doroteia, D., additional, Santos, L., additional, David, C., additional, Vinhas De Sousa, G., additional, Almeida, A., additional, Iwase, M., additional, Itou, Y., additional, Yasukochi, S., additional, Shiino, K., additional, Inuzuka, H., additional, Sugimoto, K., additional, Ozaki, Y., additional, Gieszczyk-Strozik, K., additional, Sikora-Puz, A., additional, Mizia, M., additional, Lasota, B., additional, Chmiel, A., additional, Lis-Swiety, A., additional, Michna, J., additional, Brzezinska-Wcislo, L., additional, Mizia-Stec, K., additional, Gasior, Z., additional, Luijendijk, P., additional, De Bruin-Bon, H., additional, Zwiers, C., additional, Vriend, J., additional, Van Den Brink, R., additional, Mulder, B., additional, Bouma, B., additional, Brigido, S., additional, Gianfagna, P., additional, Proclemer, A., additional, Plicht, B., additional, Kahlert, P., additional, Kaelsch, H., additional, Buck, T., additional, Erbel, R., additional, Konorza, T., additional, Yoon, H., additional, Kim, K., additional, Ahn, Y., additional, Jeong, M., additional, Cho, J., additional, Park, J., additional, Kang, J., additional, Rha, W., additional, Jansen Klomp, W. W., additional, Brandon Bravo Bruinsma, G., additional, Van 'T Hof, A., additional, Spanjersberg, S., additional, Nierich, A., additional, Bombardini, T., additional, Gherardi, S., additional, Picano, E., additional, Ciarka, A., additional, Herbots, L., additional, Eroglu, E., additional, Van Cleemput, J., additional, Droogne, W., additional, Jasityte, R., additional, Meyns, B., additional, D'hooge, J., additional, Vanhaecke, J., additional, Al Barjas, M., additional, Iskreva, R., additional, Morris, R., additional, Davar, J., additional, Zhao, Y., additional, Holmgren, A., additional, Morner, S., additional, Stepanovic, J., additional, Beleslin, B., additional, Nedeljkovic, M., additional, Mazic, S., additional, Stojanov, V., additional, Piatkowski, R., additional, Kochanowski, J., additional, Scislo, P., additional, Grabowski, M., additional, Marchel, M., additional, Roik, M., additional, Kosior, D., additional, Opolski, G., additional, Tomaszewski, A., additional, Kutarski, A., additional, Tomaszewski, M., additional, Eibel, S., additional, Hasheminejad, E., additional, Mukherjee, C., additional, Tschernich, H., additional, Ender, J., additional, Delithanasis, I., additional, Celutkiene, J., additional, Kenny, C., additional, Monaghan, M., additional, Van Den Oord, S., additional, Ten Kate, G., additional, Akkus, Z., additional, Renaud, G., additional, Sijbrands, E., additional, Ten Cate, F., additional, De Jong, N., additional, Bosch, J., additional, Van Der Steen, A., additional, Schinkel, A., additional, Lisowska, A., additional, Knapp, M., additional, Tycinska, A., additional, Sawicki, R., additional, Kralisz, P., additional, Sobkowicz, B., additional, Chang, S.-A., additional, Lee, S.-C., additional, Kim, E.-Y., additional, Hahm, S.-H., additional, Ahn, G.-T., additional, Sohn, M.-K., additional, Park, S.-J., additional, Choi, J.-O., additional, Park, S.-W., additional, Oh, J.-K., additional, Gursoy, M. O., additional, Gokdeniz, T., additional, Astarcioglu, M., additional, Bayram, Z., additional, Cakal, B., additional, Karakoyun, S., additional, Kalcik, M., additional, Kahveci, G., additional, Yildiz, M., additional, Ozkan, M., additional, Skidan, V., additional, Borowski, A., additional, Park, M., additional, Thomas, J., additional, Ranjbar, S., additional, Hassantash, S., additional, Karvandi, M., additional, Foroughi, M., additional, Davidsen, E. S., additional, Cramariuc, D., additional, Bleie, O., additional, Gerdts, E., additional, Matre, K., additional, Cusma' Piccione, M., additional, Bagnato, G., additional, Mohammed, M., additional, Piluso, S., additional, Oreto, L., additional, Bitter, T., additional, Carvalho, S., additional, Canada, M., additional, Santisteban Sanchez De Puerta, M., additional, Mesa Rubio, M. D., additional, Ruiz Ortiz, M., additional, Delgado Ortega, M., additional, Pena Pena, M. 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J., additional, Vis, J., additional, Van Dijk, A., additional, Van Melle, J., additional, Pieper, P., additional, Vliegen, H., additional, Sieswerda, G., additional, Foukarakis, E., additional, Pitarokilis, A., additional, Kafarakis, P., additional, Kiritsi, A., additional, Klironomos, E., additional, Manousakis, A., additional, Fragiadaki, X., additional, Papadakis, E., additional, and Dermitzakis, A., additional

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2011
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14. Poster session V * Saturday 11 December 2010, 08:30-12:30

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Pham, Q. H., primary, Von Lueder, T. G., additional, Namtvedt, S. K., additional, Rosjo, H., additional, Omland, T., additional, Steine, K., additional, Timoteo, A. T., additional, Mota Carmo, M., additional, Simoes, M., additional, Branco, L. M., additional, Ferreira, R. C., additional, Kato, R., additional, Ito, J., additional, Tahara, T., additional, Yokoyama, Y., additional, Ashikaga, T., additional, Satoh, Y., additional, Na, J. O., additional, Hong, H. E., additional, Kim, M. N., additional, Shin, S. Y., additional, Choi, C. U., additional, Kim, E. J., additional, Rha, S. W., additional, Park, C. G., additional, Seo, H. S., additional, Oh, D. J., additional, Ticulescu, R., additional, Brigido, S., additional, Vriz, O., additional, Sparacino, L., additional, Popescu, B. A., additional, Ginghina, C., additional, Carerj, S., additional, Nicolosi, G. L., additional, Antonini-Canterin, F., additional, Onaindia Gandarias, J. J., additional, Romero, A., additional, Laraudogoitia, E., additional, Velasco, S., additional, Quintana, O., additional, Cacicedo, A., additional, Rodriguez, I., additional, Alarcon, J. A., additional, Gonzalez, J., additional, Lekuona, I., additional, Subinas, A., additional, Abdula, G., additional, Lund, L. H., additional, Winter, R., additional, Brodin, L., additional, Sahlen, A., additional, Masaki, M., additional, Cha, Y. M., additional, Yuasa, T., additional, Dong, K., additional, Dong, Y. X., additional, Mankad, S. V., additional, Oh, J. K., additional, Vallet, F., additional, Lequeux, B., additional, Diakov, C., additional, Sosner, P., additional, Christiaens, L., additional, Coisne, D., additional, Kihara, C., additional, Murata, K., additional, Wada, Y., additional, Uchida, K., additional, Ueyama, T., additional, Okuda, S., additional, Susa, T., additional, Matsuzaki, M., additional, Cho, E. J., additional, Choi, K. Y., additional, Kwon, B. J., additional, Kim, D. B., additional, Jang, S. W., additional, Cho, J. S., additional, Jung, H. O., additional, Jeon, H. K., additional, Youn, H. J., additional, Kim, J. H., additional, Cikes, M., additional, Bijnens, B., additional, Velagic, V., additional, Kopjar, T., additional, Milicic, D., additional, Biocina, B., additional, Gasparovic, H., additional, Almuntaser, I., additional, Brown, A., additional, Foley, B., additional, Mulvihill, N., additional, Crean, P., additional, King, G., additional, Murphy, R., additional, Takata, Y., additional, Taniguchi, M., additional, Nobusada, S., additional, Sugawara, M., additional, Toh, N., additional, Kusano, K., additional, Itoh, H., additional, Wellnhofer, E., additional, Kriatselis, C., additional, Nedios, S., additional, Gerds-Li, J. H., additional, Fleck, E., additional, Poulsen, M. K., additional, Henriksen, J. E., additional, Dahl, J., additional, Johansen, A., additional, Haghfelt, T., additional, Hoilund-Carlsen, P. F., additional, Beck-Nielsen, H., additional, Moller, J. E., additional, Dankowski, R., additional, Wierzchowiecki, M., additional, Michalski, M., additional, Nowicka, A., additional, Szymanowska, K., additional, Pajak, A., additional, Poprawski, K., additional, Szyszka, A., additional, Kasner, M., additional, Westermann, D., additional, Schultheiss, H. P., additional, Tschoepe, C., additional, Watanabe, T., additional, Iwai-Takano, M., additional, Kobayashi, A., additional, Machii, H., additional, Takeishi, Y., additional, Paelinck, B. P., additional, Van Herck, P. L., additional, Bosmans, J. M., additional, Vrints, C. J., additional, Lamb, H. J., additional, Doltra, A., additional, Vidal, B., additional, Silva, E., additional, Poyatos, S., additional, Mont, L., additional, Berruezo, A., additional, Castel, A., additional, Tolosana, J. 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V., additional, Tagliamonte, E., additional, Cirillo, T., additional, Coppola, A., additional, Marinelli, U., additional, Romano, C., additional, Riccio, G., additional, Citro, R., additional, Astarita, C., additional, Capuano, N., additional, Quaranta, G., additional, Desiderio, A., additional, Frattini, S., additional, Faggiano, P., additional, Zilioli, V., additional, Locantore, E., additional, Longhi, S., additional, Bellandi, F., additional, Faden, G., additional, Triggiani, M., additional, Dei Cas, L., additional, Dalsgaard, M., additional, Kjaergaard, J., additional, Iversen, K., additional, Hassager, C., additional, Dinh, W., additional, Nickl, W. N., additional, Smettan, J. S., additional, Koehler, T. K., additional, Scheffold, T. D., additional, Coll Barroso, M. C. B., additional, Guelker, J. G., additional, Fueth, R. F., additional, Kamperidis, V., additional, Hadjimiltiades, S., additional, Sianos, G., additional, Efthimiadis, G., additional, Karvounis, H., additional, Parcharidis, G., additional, Styliadis, I. H., additional, Velasco Del Castillo, M. S., additional, Onaindia, J. J., additional, Telleria, M., additional, Carstensen, H. G., additional, Nordenberg, C., additional, Sogaard, P., additional, Fritz-Hansen, T., additional, Bech, J., additional, Galatius, S., additional, Jensen, J. S., additional, Mogelvang, R., additional, Bartko, P. E., additional, Graf, S., additional, Rosenhek, R., additional, Burwash, I. G., additional, Bergler-Klein, J., additional, Clavel, M.-A., additional, Baumgartner, H., additional, Pibarot, P., additional, Mundigler, G., additional, Kirilmaz, B., additional, Eser, I., additional, Tuzun, N., additional, Komur, B., additional, Dogan, H., additional, Taskiran Comez, A., additional, Ercan, E., additional, Cusma-Piccione, M., additional, Zito, C., additional, Oreto, G., additional, Piluso, S., additional, Tripepi, S., additional, Oreto, L., additional, Longordo, C., additional, Ciraci, L., additional, Di Bella, G., additional, Piatkowski, R., additional, Kochanowski, J., additional, Scislo, P., additional, Grabowski, M., additional, Marchel, M., additional, Roik, M., additional, Kosior, D., additional, Opolski, G., additional, Sknouril, L., additional, Dorda, M., additional, Holek, B., additional, Gajdusek, L., additional, Chovancik, J., additional, Branny, M., additional, Fiala, M., additional, Szymanski, P., additional, Lipczynska, M., additional, Klisiewicz, A., additional, Hoffman, P., additional, Jander, N., additional, Minners, J., additional, Martin, G., additional, Zeh, W., additional, Allgeier, M., additional, Gohlke-Baewolf, C., additional, Gohlke, H., additional, Nistri, S., additional, Porciani, M. C., additional, Attanasio, M., additional, Abbate, R., additional, Gensini, G. F., additional, Pepe, G., additional, Duncan, R. F., additional, Piantadosi, C., additional, Nelson, A. J., additional, Wittert, G., additional, Dundon, B., additional, Worthley, M. I., additional, Worthley, S. G., additional, Jung, P., additional, Berlinger, K., additional, Rieber, J., additional, Sohn, H. Z., additional, Schneider, P., additional, Leibig, M., additional, Koenig, A., additional, Klauss, V., additional, Tomkiewicz-Pajak, L., additional, Kolcz, J., additional, Olszowska, M., additional, Pieculewicz, M., additional, Podolec, P., additional, Przewlocki, T., additional, Suchon, E., additional, Sobien, B., additional, Wilkolek, P., additional, Ziembicka, A., additional, Hlawaty, M., additional, Van De Bruaene, A., additional, Hermans, H., additional, Buys, R., additional, Vanhees, L., additional, Delcroix, M., additional, Voigt, J.-U., additional, Budts, W., additional, De Cillis, E., additional, Acquaviva, T., additional, Basile, D., additional, Bortone, A. S., additional, Kalimanovska-Ostric, D., additional, Nastasovic, T., additional, Vujisic-Tesic, B., additional, Jovanovic, I., additional, Milakovic, B., additional, Dostanic, M., additional, Stosic, M., additional, Frogoudaki, A., additional, Andreou, K., additional, Parisis, J., additional, Triantafyllidi, E., additional, Gaitani, S., additional, Paraskevaidis, J., additional, Anastasiou-Nana, M., additional, De Pasquale, G., additional, Kuehn, A., additional, Petzuch, K., additional, Mueller, J., additional, Meierhofer, C., additional, Fratz, S., additional, Hager, A., additional, Hess, J., additional, Vogt, M., additional, Attenhofer Jost, C. H., additional, Dearani, J. A., additional, Scott, C. G., additional, Burkhart, H. M., additional, Connolly, H. M., additional, Vitarelli, A., additional, Battaglia, D., additional, Caranci, F., additional, Padella, V., additional, Continanza, G., additional, Dettori, O., additional, Capotosto, L., additional, Vitarelli, M., additional, De Cicco, V., additional, Cortez Morichetti, M., additional, Mohanan Nair, K. K., additional, Sasidaharan, B., additional, Thajudeen, A., additional, Tharakan, J. M., additional, Mertens, L., additional, Ahmad, N., additional, Kantor, P. K., additional, Grosse-Wortmann, L., additional, Friedberg, M. K., additional, Bernard, Y. F., additional, Morel, M. A., additional, Descotes-Genon, V., additional, Jehl, J., additional, Meneveau, N., additional, Schiele, F., additional, Kaldararova, M., additional, Simkova, I., additional, Tittel, P., additional, Masura, J., additional, Trojnarska, O., additional, Szczepaniak, L., additional, Mizia -Stec, K., additional, Cieplucha, A., additional, Bartczak, A., additional, Grajek, S., additional, Tykarski, A., additional, Gasior, Z., additional, Babovicvuksanovic, D., additional, Bonnichsen, C. R., additional, Morgan, G. J., additional, Slorach, C., additional, Hui, W., additional, Sarkola, T., additional, Lee, K. J., additional, Chaturvedi, R., additional, Benson, L., additional, Bradley, T., additional, Iancu, M. E., additional, Ghiorghiu, I., additional, Serban, M., additional, Craciunescu, I., additional, Hodo, A., additional, Morgan, J., additional, Roche, L., additional, Lee, K., additional, Milanesi, O., additional, Favero, V., additional, Padalino, M., additional, Biffanti, R., additional, Cerutti, A., additional, Maschietto, N., additional, Reffo, E., additional, Vida, V., additional, Stellin, G., additional, Irtyuga, O., additional, Gamazin, D., additional, Voronkina, I., additional, Tsoyi, N., additional, Gudkova, E., additional, Moiseeva, O., additional, Aggeli, C., additional, Kazazaki, C., additional, Felekos, I., additional, Lagoudakou, S., additional, Roussakis, G., additional, Skoumas, J., additional, Pitsavos, C., additional, Stefanadis, C., additional, Cueff, C., additional, Keenan, N., additional, Steg, P. G., additional, Cimadevilla, C., additional, Ducrocq, G., additional, Vahanian, A., additional, Messika-Zeitoun, D., additional, Petrella, L., additional, Mazzola, A. M., additional, Villani, C. V., additional, Giancola, R. G., additional, Ciocca, M. C., additional, Di Eusanio, D. E. M., additional, Nolan, S., additional, Ionescu, A., additional, Skaug, T. R., additional, Amundsen, B. H., additional, Hergum, T., additional, Torp, H., additional, Haugen, B. O., additional, Lopez Aguilera, J., additional, Mesa Rubio, D., additional, Ruiz Ortiz, M., additional, Delgado Ortega, M., additional, Villanueva Fernandez, E., additional, Cejudo Diaz Del Campo, L., additional, Toledano Delgado, F., additional, Leon Del Pino, M., additional, Romo Pena, E., additional, Suarez De Lezo Cruz-Conde, J., additional, De Marco, E., additional, Colucci, A., additional, Comerci, G., additional, Gabrielli, F. A., additional, Natali, R., additional, Garramone, B., additional, Savino, M., additional, Lotrionte, M., additional, Sonaglioni, A., additional, Loperfido, F., additional, Zdravkovic, M., additional, Perunicic, J., additional, Krotin, M., additional, Ristic, M., additional, Vukomanovic, V., additional, Zaja, M., additional, Radovanovic, S., additional, Saric, J., additional, Zdravkovic, D., additional, Cotrim, C., additional, Almeida, A. R., additional, Miranda, R., additional, Almeida, A. G., additional, Picano, E., additional, Carrageta, M., additional, D'andrea, A., additional, Cocchia, R., additional, Riegler, L., additional, Golia, E., additional, Scarafile, R., additional, Caso, P., additional, Russo, M. G., additional, Bossone, E., additional, Calabro', R., additional, Noman, H., additional, Adel, A., additional, Elfaramawy, A. M. R., additional, Abdelraouf, M., additional, Elnaggar, W. A. E. L., additional, Baligh, E., additional, Sargento, L., additional, Silva, D., additional, Goncalves, S., additional, Ribeiro, S., additional, Vinhas Sousa, G., additional, Almeida, A., additional, Lopes, M., additional, Rodriguez-Manero, M., additional, Aguado Gil, L., additional, Azcarate, P., additional, Lloret Luna, P., additional, Macias Gallego, A., additional, Castano, S. A. R. A., additional, Garcia, M., additional, Pujol Salvador, C., additional, Barba, J., additional, Redondo, P., additional, Tomasoni, L., additional, Sitia, S., additional, Atzeni, F., additional, Gianturco, L., additional, Ricci, C., additional, Sarzi-Puttini, P., additional, Turiel, M., additional, De Gennaro Colonna, V., additional, Uejima, T., additional, Jaroch, J., additional, Polombo, C., additional, Hughes, A., additional, Vinereanu, D., additional, Evanvelista, A., additional, Leftheriotis, G., additional, Fraser, A. G., additional, Lewczuk, A., additional, Sobkowicz, B., additional, Tomaszuk-Kazberuk, A., additional, Sawicki, R., additional, Hirnle, T., additional, Michalski, B. W., additional, Filipiak, D., additional, Kasprzak, J. D., additional, Lipiec, P., additional, Dalen, H., additional, Mjolstad, O. C., additional, Klykken, B. E., additional, Graven, T., additional, Martensson, M., additional, Olsson, M., additional, Brodin, L.-A., additional, Enache, R., additional, Leiballi, E., additional, Penhall, A., additional, Perry, R., additional, Altman, M., additional, Sinhal, A., additional, Bennetts, J., additional, Chew, D. P., additional, Joseph, M. X., additional, Larsen, L. H., additional, Kristensen, T., additional, Kober, L. V., additional, Kofoed, K. F., additional, Moscoso Costa, F., additional, Ribeiras, R., additional, Brito, J., additional, Boshoff, S., additional, Neves, J., additional, Teles, R., additional, Canada, M., additional, Andrade, M. J., additional, Gouveia, R., additional, Silva, A., additional, Miskovic, A., additional, Poerner, T. P., additional, Stiller, C. S., additional, Goebel, B. G., additional, Moritz, A. M., additional, Stefani, L., additional, Galanti, G. G., additional, Moraldo, M., additional, Bergamini, C., additional, Pabari, P. A., additional, Dhutia, N. M., additional, Malaweera, A. S. N., additional, Willson, K., additional, Davies, J., additional, Hughes, A. D., additional, Xu, X. Y., additional, Francis, D. P., additional, Jasaityte, R., additional, Amundsen, B., additional, Barbosa, D., additional, Loeckx, D., additional, Kiss, G., additional, Orderud, F., additional, Robesyn, V., additional, Claus, P., additional, D'hooge, J., additional, Nao, T., additional, Miura, T., additional, Shams, K., additional, Samir, S., additional, Samir, R., additional, El-Sayed, M., additional, Anwar, A. M., additional, Nosir, Y., additional, Galal, A., additional, Chamsi-Pasha, H., additional, Ciobanu, A., additional, Dulgheru, R., additional, Bennett, S., additional, De Luca, A., additional, Toncelli, L., additional, Cappelli, F., additional, Cappelli, B., additional, Vono, M. C. R., additional, Galanti, G., additional, Zorman, Y., additional, Yilmazer, M. S., additional, Akyildiz, M., additional, Gurol, T., additional, Aydin, A., additional, Dagdeviren, B., additional, and Kalangos, A., additional

Published
2010
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16. Cause of oral ulcers in HIV-infected patients

Author

Piluso, S., Ficarra, G., Lucatorto, F.M., Orsi, A., Dionisio, D., Stendardi, L., and Eversole, L.R.

Abstract

Objectives: To study the cause and clinical aspects of oral ulcers in HIV-infected patients. Study design: Forty-one consecutive HIV-positive patients with long-standing oral ulcers were examined; 19 were evaluated by biopsy. From these 19 cases, viral, bacterial, and fungal cultures and biopsies were taken in each patient. When indicated, special microbial stains were undertaken to identify bacteria or fungi. Ten cases without granulomatous bacterial fungal or lymphomatous features were available for in situ hybridization to detect viral DNA of herpes simplex virus 1 and 2, cytomegalovirus, varicella-zoster virus, and Epstein-Barr virus. Results: Most of the oral ulcers occurred in patients with severe immunodepression. Median CD4 T-lymphocyte count was 60 cell/mm^3 (range, 3 to 335). It was ascertained that nine (47%) patients had nonspecific aphthous-like ulcers, and ulcers caused by herpes group viruses were identified in six (31,5%) patients. One (5%) person was diagnosed with non-Hodgkin's lymphoma; and in one (5%) patient, multiple ulcers were an expression of lues maligna. Two ulcers (10.5%) in the palate harbored mycotic granulomatous foci (cryptococcosis, histoplasmosis). In this population, almost all of these ulcers were found to be large, persistent, and painful. Conclusions: Nontumefactive oral ulcers in HIV-positive patients may be a source of diagnostic difficulties because of the diverse array of underlying pathologic entities and multiplicity of etiologic agents. Biopsy should always be performed on long-standing ulcers because either infection or a neoplastic process may be extant. In the absence of infection or neoplasm, such lesions are then designated as ulcers not otherwise specified.

Published
1996
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17. Postoperative skeletal pain: Development of non-opioid treatment strategies

Author

Steverink, Jasper Gerard, Malda, J., Verlaan, J.J., and Piluso, S.

Subjects
bone pain, local anesthetic, bupivacaine, hydrogel, opioid-free, spine surgery, postoperative pain
Abstract

This thesis attempts to better understand postoperative skeletal pain, quantify its severity and provide directions for its improved treatment. Chapter 2 described the current clinical practice in patients undergoing spine surgery regarding postoperative pain and its treatment. Most patients developed moderate opioid-related adverse drug events (ORADEs) while still experiencing pain. To understand the severity of postoperative skeletal pain, an immunohistochemical study quantifying bone sensory innervation was performed on human bone samples from various anatomical locations in Chapter 3. The highest sensory innervation density was found in the periosteum. The anatomical location receiving most sensory innervation was the thoracic spine. Chapter 4 consists of a literature review on bone pain. This chapter identified four pathways involved in generation and maintenance of bone pain. These pathways were the activation, sensitization and sprouting of sensory nerve fibers as well as central sensitization. The high incidence of postoperative ORADEs illustrated the delicate balance between analgesic and side effects of current postoperative pain treatment. Its dense innervation identified the periosteum as a source of pain and potential target for postoperative analgesia. The second aim was to assess the safety and efficacy of a non-opioid anesthetic (bupivacaine) when applied in high concentrations in musculoskeletal tissues. The toxic effects of bupivacaine in bone, muscle and neural tissue, articular cartilage and the intervertebral disc in vitro were reviewed in Chapter 5. All reported effects in vitro were shown to be reversible in animal and clinical studies, with the exception of chondrotoxicity. The absence of regenerative capacity in vitro likely limits the translation of laboratory findings. Chapter 6 assesses the local toxic effects of high bupivacaine concentrations in a rat model for skeletal surgery. Rats underwent skeletal implantation of a cannula and screw, to allow bolus administration or sustained infusion of bupivacaine in three concentrations at the surgical site. Histological analysis of implant sites after 28 days did not reveal differences between bupivacaine concentrations. The third aim concerns the development of a clinically relevant hydrogel-based bupivacaine sustained release formulation for use in musculoskeletal surgery. Chapter 7 described the development and in vitro optimization of a ring-shaped hydrogel. The hydrogel displayed tunable mechanical properties and drug-releasing behavior. Drug release was sustained for 72 hours in vitro. The material was implantable, biodegradable and cytocompatible. Chapter 8 revealed that implantation of up to 8 hydrogel rings yielded sustained but low bupivacaine plasma levels, with correspondingly high surgical site levels for 72 hours. Drug release followed a first-order profile. Hydrogels elicited a mild tissue response and degraded in situ. In Chapter 9, a bridging study was performed in sheep according to Good Laboratory Practice (GLP). Sheep received either 8 pedicle screws, 8 pedicle screws combined with unloaded hydrogels, or 8 pedicle screws with bupivacaine-loaded hydrogels. Plasma bupivacaine levels remained tenfold below toxic thresholds, and were sustained for longer compared to subcutaneous infiltration of bupivacaine. Local tissue reaction and screw osseointegration were similar between groups. Taken together, the results indicate biocompatibility, biodegradability, large systemic safety margins and extended drug release of the bupivacaine-loaded hydrogels in vivo. The final aim of this thesis was to provide directions for a clinical trial, assessing safety and feasibility of the bupivacaine-loaded hydrogel for use in spine surgery. Chapter 10 presents a protocol for a phase 1B clinical trial, in which female and male adult patients with degenerative spinal disease and scheduled spine surgery are included. The patients will receive 4 or 6 hydrogels, to be co-implanted with pedicle screws. The primary outcome of the study is systemic safety of the bupivacaine hydrogel formulation as defined by the Cmax.

Published
2023

18. Noninvasive Imaging of Transgene Expression in Neurons Using Chemical Exchange Saturation Transfer MRI.

Author

Flament J, Pépin J, Maugard M, Gaudin M, Cohen L, Jan C, Valette J, Piluso S, Delzescaux T, and Bonvento G

Abstract

Advances in gene therapy, especially for brain diseases, have created new imaging demands for noninvasive monitoring of gene expression. While reporter gene imaging using co-expression of fluorescent protein-encoding gene has been widely developed, these conventional methods face significant limitations in longitudinal in vivo applications. Magnetic resonance imaging (MRI), specifically chemical exchange saturation transfer (CEST) MRI, provides a robust noninvasive alternative that offers unlimited depth penetration, reliable spatial resolution, and specificity toward particular molecules. In this study, we explore the potential of CEST-MRI for monitoring gene expression in neurons. We designed a CEST polypeptide reporter expressing 150 arginine residues and evaluated its expression in the living brain after viral vector delivery. A longitudinal study performed at one and 2 months postinjection showed that specific CEST signal was observable. In particular, the CEST contrast exhibited distinct peaks at 0.75 and 1.75 ppm, consistent with the expected hydroxyl and guanidyl protons resonance frequencies. Histological study confirmed the specific neuronal expression of the transgene evidenced by the fluorescence signal from the td-Tomato fluorophore fused to the polypeptide. The ability to image noninvasively a neuron-specific CEST-MRI reporter gene could offer valuable insights for further developments of gene therapy for neurological disorders., (© 2024 The Author(s). NMR in Biomedicine published by John Wiley & Sons Ltd.)

Published
2024
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19. Microstructured silk fiber scaffolds with enhanced stretchability.

Author

Viola M, Cedillo-Servin G, van Genderen AM, Imhof I, Vena P, Mihajlovic M, Piluso S, Malda J, Vermonden T, and Castilho M

Subjects
Humans, Tissue Engineering, Bombyx chemistry, Animals, Silk chemistry, Porosity, Cell Line, Biocompatible Materials chemistry, Tissue Scaffolds chemistry, Fibroins chemistry
Abstract

Despite extensive research, current methods for creating three-dimensional (3D) silk fibroin (SF) scaffolds lack control over molecular rearrangement, particularly in the formation of β-sheet nanocrystals that severely embrittle SF, as well as hierarchical fiber organization at both micro- and macroscale. Here, we introduce a fabrication process based on electrowriting of aqueous SF solutions followed by post-processing using an aqueous solution of sodium dihydrogen phosphate (NaH 2 PO 4 ). This approach enables gelation of SF chains via controlled β-sheet formation and partial conservation of compliant random coil structures. Moreover, this process allows for precise architecture control in microfiber scaffolds, enabling the creation of 3D flat and tubular macro-geometries with square-based and crosshatch microarchitectures, featuring inter-fiber distances of 400 μm and ∼97% open porosity. Remarkably, the crosslinked printed structures demonstrated a balanced coexistence of β-sheet and random coil conformations, which is uncommon for organic solvent-based crosslinking methods. This synergy of printing and post-processing yielded stable scaffolds with high compliance (modulus = 0.5-15 MPa) and the ability to support elastic cyclic loading up to 20% deformation. Furthermore, the printed constructs supported in vitro adherence and growth of human renal epithelial and endothelial cells with viability above 95%. These cells formed homogeneous monolayers that aligned with the fiber direction and deposited type-IV collagen as a specific marker of healthy extracellular matrix, indicating that both cell types attach, proliferate, and organize their own microenvironment within the SF scaffolds. These findings represent a significant development in fabricating organized stable SF scaffolds with unique microfiber structures and mechanical and biological properties that make them highly promising for tissue engineering applications.

Published
2024
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20. giRAff: an automated atlas segmentation tool adapted to single histological slices.

Author

Piluso S, Souedet N, Jan C, Hérard AS, Clouchoux C, and Delzescaux T

Abstract

Conventional histology of the brain remains the gold standard in the analysis of animal models. In most biological studies, standard protocols usually involve producing a limited number of histological slices to be analyzed. These slices are often selected into a specific anatomical region of interest or around a specific pathological lesion. Due to the lack of automated solutions to analyze such single slices, neurobiologists perform the segmentation of anatomical regions manually most of the time. Because the task is long, tedious, and operator-dependent, we propose an automated atlas segmentation method called giRAff, which combines rigid and affine registrations and is suitable for conventional histological protocols involving any number of single slices from a given mouse brain. In particular, the method has been tested on several routine experimental protocols involving different anatomical regions of different sizes and for several brains. For a given set of single slices, the method can automatically identify the corresponding slices in the mouse Allen atlas template with good accuracy and segmentations comparable to those of an expert. This versatile and generic method allows the segmentation of any single slice without additional anatomical context in about 1 min. Basically, our proposed giRAff method is an easy-to-use, rapid, and automated atlas segmentation tool compliant with a wide variety of standard histological protocols., Competing Interests: SP and CC were employed by WITSEE. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Piluso, Souedet, Jan, Hérard, Clouchoux and Delzescaux.)

Published
2024
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21. [Caseous calcification of the mitral annulus: case report and review of the literature].

Author

Rodella L, Zanini G, Piluso S, Vaccari A, Triggiani M, Montresor G, Coletti G, and Pasini GF

Subjects
Female, Humans, Aged, 80 and over, Mitral Valve diagnostic imaging, Mitral Valve pathology, Echocardiography, Transesophageal, Lipids, Calcinosis diagnostic imaging, Heart Valve Diseases diagnostic imaging
Abstract

Caseous calcification of the mitral annulus is an uncommon variant of mitral annular calcification. It appears as a round echodense mass containing central areas of echolucencies resembling liquefaction and with no flow in the central zone on color Doppler. In most cases it involves the posterior mitral annulus region, particularly in female subjects. The pathogenesis remains unclear: hypercholesterolemia and the dissolution of lipid-rich macrophages may be implicated in liquefaction necrosis. Transthoracic and transesophageal echocardiography represents the most reliable technique for diagnosis, whereas cardiac magnetic resonance imaging is the choice in doubtful cases. We report the case of an 82-year-old female patient describing different aspects of this particular clinical condition.

Published
2022
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22. Robust gelatin hydrogels for local sustained release of bupivacaine following spinal surgery.

Author

Steverink JG, van Tol FR, Oosterman BJ, Vermonden T, Verlaan JJ, Malda J, and Piluso S

Subjects
Analgesics, Opioid adverse effects, Animals, Cadaver, Delayed-Action Preparations chemistry, Humans, Hydrogels chemistry, Pain, Postoperative drug therapy, Riboflavin, Sheep, Tyramine, Bupivacaine pharmacology, Gelatin chemistry
Abstract

Adequate treatment of pain arising from spinal surgery is a major clinical challenge. Opioids are the mainstay of current treatment methods, but the frequency and severity of their side effects display a clear need for opioid-free analgesia. Local anesthetics have been encapsulated into sustained-release drug delivery systems to provide postoperative pain relief. However, these formulations are limited by rapid diffusion out of the surgical site. To overcome this limitation, we synthesized ring-shaped hydrogels incorporating bupivacaine, designed to be co-implanted with pedicle screws during spinal surgery. Hydrogels were prepared by riboflavin-mediated crosslinking of gelatin functionalized with tyramine moieties. Additionally, oxidized β-cyclodextrin was introduced into the hydrogel formulation to form dynamic bonds with tyramine functionalities, which enables self-healing behavior and resistance to shear. Feasibility of hydrogel implantation combined with pedicle screws was qualitatively assessed in cadaveric sheep as a model for instrumented spinal surgery. The in-situ crystallization of bupivacaine within the hydrogel matrix provided a moderate burst decrease and sustained release that exceeded 72 hours in vitro. The use of bupivacaine crystals decreased drug-induced cytotoxicity in vitro compared to bupivacaine HCl. Thus, the presented robust hydrogel formulation provides promising properties to enable the stationary release of non-opioid analgesics following spinal surgery. STATEMENT OF SIGNIFICANCE: Currently, postoperative pain following spinal surgery is mainly treated with opioids. However, the use of opioids is associated with several side effects including addiction. Here we developed robust and cytocompatible gelatin hydrogels, prepared via riboflavin-mediated photocrosslinking, that can withstand orthopedic implantation. The implantability was confirmed in cadaveric instrumented spinal surgery. Further, hydrogels were loaded with bupivacaine crystals to provide sustained release beyond 72 hours in vitro. The use of crystallized bupivacaine decreased cytotoxicity compared to bupivacaine HCl. The present formulation can aid in enabling opioid-free analgesia following instrumented spinal surgery., Competing Interests: Declaration of Competing Interest SP, JS, BO, and JJV are inventors of (U.S.) patent WO2020249695A1 covering hydrogel for in vivo release of medication. JS, FT, SP, BO and JJV are employed by and own stock in SentryX B.V. TV and JM report no conflicts of interest., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Published
2022
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23. The Importance of Interfaces in Multi-Material Biofabricated Tissue Structures.

Author

Viola M, Piluso S, Groll J, Vermonden T, Malda J, and Castilho M

Subjects
Printing, Three-Dimensional, Tissue Engineering, Tissue Scaffolds, Bioprinting
Abstract

Biofabrication exploits additive manufacturing techniques for creating 3D structures with a precise geometry that aim to mimic a physiological cellular environment and to develop the growth of native tissues. The most recent approaches of 3D biofabrication integrate multiple technologies into a single biofabrication platform combining different materials within different length scales to achieve improved construct functionality. However, the importance of interfaces between the different material phases, has not been adequately explored. This is known to determine material's interaction and ultimately mechanical and biological performance of biofabricated parts. In this review, this gap is bridged by critically examining the interface between different material phases in (bio)fabricated structures, with a particular focus on how interfacial interactions can compromise or define the mechanical (and biological) properties of the engineered structures. It is believed that the importance of interfacial properties between the different constituents of a composite material, deserves particular attention in its role in modulating the final characteristics of 3D tissue-like structures., (© 2021 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

Published
2021
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24. Comparison of in vitro and in vivo Toxicity of Bupivacaine in Musculoskeletal Applications.

Author

Steverink JG, Piluso S, Malda J, and Verlaan JJ

Abstract

The recent societal debate on opioid use in treating postoperative pain has sparked the development of long-acting, opioid-free analgesic alternatives, often using the amino-amide local anesthetic bupivacaine as active pharmaceutical ingredient. A potential application is musculoskeletal surgeries, as these interventions rank amongst the most painful overall. Current literature showed that bupivacaine induced dose-dependent myo-, chondro-, and neurotoxicity, as well as delayed osteogenesis and disturbed wound healing in vitro . These observations did not translate to animal and clinical research, where toxic phenomena were seldom reported. An exception was bupivacaine-induced chondrotoxicity, which can mainly occur during continuous joint infusion. To decrease opioid consumption and provide sustained pain relief following musculoskeletal surgery, new strategies incorporating high concentrations of bupivacaine in drug delivery carriers are currently being developed. Local toxicity of these high concentrations is an area of further research. This review appraises relevant in vitro , animal and clinical studies on musculoskeletal local toxicity of bupivacaine., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Steverink, Piluso, Malda and Verlaan.)

Published
2021
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25. 3D bioprinting of molecularly engineered PEG-based hydrogels utilizing gelatin fragments.

Author

Piluso S, Skvortsov GA, Altunbek M, Afghah F, Khani N, Koç B, and Patterson J

Subjects
Biocompatible Materials, Gelatin, Hydrogels, Printing, Three-Dimensional, Tissue Engineering, Tissue Scaffolds, Bioprinting
Abstract

Three-dimensional (3D) bioprinting is an additive manufacturing process in which the combination of biomaterials and living cells, referred to as a bioink, is deposited layer-by-layer to form biologically active 3D tissue constructs. Recent advancements in the field show that the success of this technology requires the development of novel biomaterials or the improvement of existing bioinks. Polyethylene glycol (PEG) is one of the well-known synthetic biomaterials and has been commonly used as a photocrosslinkable bioink for bioprinting; however, other types of cell-friendly crosslinking mechanisms to form PEG hydrogels need to be explored for bioprinting and tissue engineering. In this work, we proposed micro-capillary based bioprinting of a novel molecularly engineered PEG-based bioink that transiently incorporates low molecular weight gelatin (LMWG) fragments. The rheological properties and release profile of the LMWG fragments were characterized, and their presence during hydrogel formation had no effect on the swelling ratio or sol fraction when compared to PEG hydrogels formed without the LMWG fragments. For bioprinting, PEG was first functionalized with cell-adhesive RGD ligands and was then crosslinked using protease-sensitive peptides via a Michael-type addition reaction inside the micro-capillary. The printability was assessed by the analysis of extrudability, shape fidelity, and printing accuracy of the hydrogel filaments after the optimization of the gelation conditions of the PEG-based bioink. The LMWG fragments supplemented into the bioink allowed the extrusion of smooth and uniform cylindrical strands of the hydrogel and improved shape fidelity and printing accuracy. Encapsulated cells in both bioprinted and non-bioprinted PEG-based hydrogels showed high viability and continued to proliferate over time in culture with a well-defined cell morphology depending on the presence of the cell adhesive peptide RGD. The presented micro-capillary based bioprinting process for a novel PEG-based bioink can be promising to construct complex 3D structures with micro-scale range and spatiotemporal variations without using any cytotoxic photoinitiator, UV light, or polymer support., (© 2021 IOP Publishing Ltd.)

Published
2021
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26. Hydrogel-Based Bioinks for Cell Electrowriting of Well-Organized Living Structures with Micrometer-Scale Resolution.

Author

Castilho M, Levato R, Bernal PN, de Ruijter M, Sheng CY, van Duijn J, Piluso S, Ito K, and Malda J

Subjects
Printing, Three-Dimensional, Tissue Engineering, Tissue Scaffolds, Bioprinting, Hydrogels
Abstract

Bioprinting has become an important tool for fabricating regenerative implants and in vitro cell culture platforms. However, until today, extrusion-based bioprinting processes are limited to resolutions of hundreds of micrometers, which hamper the reproduction of intrinsic functions and morphologies of living tissues. This study describes novel hydrogel-based bioinks for cell electrowriting (CEW) of well-organized cell-laden fiber structures with diameters ranging from 5 to 40 μm. Two novel photoresponsive hydrogel bioinks, that is, based on gelatin and silk fibroin, which display distinctly different gelation chemistries, are introduced. The rapid photomediated cross-linking mechanisms, electrical conductivity, and viscosity of these two engineered bioinks allow the fabrication of 3D ordered fiber constructs with small pores (down to 100 μm) with different geometries ( e.g. , squares, hexagons, and curved patterns) of relevant thicknesses (up to 200 μm). Importantly, the biocompatibility of the gelatin- and silk fibroin-based bioinks enables the fabrication of cell-laden constructs, while maintaining high cell viability post printing. Taken together, CEW and the two hydrogel bioinks open up fascinating opportunities to manufacture microstructured constructs for applications in regenerative medicine and in vitro models that can better resemble cellular microenvironments.

Published
2021
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27. Rapid and cytocompatible cell-laden silk hydrogel formation via riboflavin-mediated crosslinking.

Author

Piluso S, Flores Gomez D, Dokter I, Moreira Texeira L, Li Y, Leijten J, van Weeren R, Vermonden T, Karperien M, and Malda J

Subjects
Animals, Bombyx chemistry, Cell Line, Cell Survival, Cross-Linking Reagents chemistry, Goats, Humans, Biocompatible Materials chemistry, Fibroins chemistry, Hydrogels chemistry, Riboflavin chemistry
Abstract

Bioactive hydrogels based on naturally-derived polymers are of great interest for regenerative medicine applications. Among naturally-derived polymers, silk fibroin has been extensively explored as a biomaterial for tissue engineering due to its unique mechanical properties. Here, we demonstrate the rapid gelation of cell-laden silk fibroin hydrogels by visible light-induced crosslinking using riboflavin as a photo-initiator, in presence of an electron acceptor. The gelation kinetics were monitored by in situ photo-rheometry. Gelation was achieved in minutes and could be tuned owing to its direct proportionality to the electron acceptor concentration. The concentration of the electron acceptor did not affect the elastic modulus of the hydrogels, which could be altered by varying the polymer content. Further, the biocompatible riboflavin photo-initiator combined with sodium persulfate allowed for the encapsulation of cells within silk fibroin hydrogels. To confirm the cytocompatibility of the silk fibroin formulations, three cell types (articular cartilage-derived progenitor cells, mesenchymal stem cells and dental-pulp-derived stem cells) were encapsulated within the hydrogels, which associated with a viability >80% for all cell types. These results demonstrated that fast gelation of silk fibroin can be achieved by combining it with riboflavin and electron acceptors, which results in a hydrogel that can be used in tissue engineering and cell delivery applications.

Published
2020
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28. Printability and Shape Fidelity of Bioinks in 3D Bioprinting.

Author

Schwab A, Levato R, D'Este M, Piluso S, Eglin D, and Malda J

Subjects
Humans, Bioprinting, Ink, Printing, Three-Dimensional, Tissue Engineering
Abstract

Three-dimensional bioprinting uses additive manufacturing techniques for the automated fabrication of hierarchically organized living constructs. The building blocks are often hydrogel-based bioinks, which need to be printed into structures with high shape fidelity to the intended computer-aided design. For optimal cell performance, relatively soft and printable inks are preferred, although these undergo significant deformation during the printing process, which may impair shape fidelity. While the concept of good or poor printability seems rather intuitive, its quantitative definition lacks consensus and depends on multiple rheological and chemical parameters of the ink. This review discusses qualitative and quantitative methodologies to evaluate printability of bioinks for extrusion- and lithography-based bioprinting. The physicochemical parameters influencing shape fidelity are discussed, together with their importance in establishing new models, predictive tools and printing methods that are deemed instrumental for the design of next-generation bioinks, and for reproducible comparison of their structural performance.

Published
2020
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29. Engineered Three-Dimensional Microenvironments with Starch Nanocrystals as Cell-Instructive Materials.

Author

Piluso S, Labet M, Zhou C, Seo JW, Thielemans W, and Patterson J

Subjects
Animals, Cell Line, Cell Line, Tumor, Cell Proliferation drug effects, Cell Shape drug effects, Compressive Strength, Hydrogels pharmacology, Mice, Wettability, Hydrogels chemistry, Nanocomposites chemistry, Starch analogs & derivatives, Tissue Scaffolds chemistry
Abstract

Naturally, cells reside in three-dimensional (3D) microenvironments composed of biopolymers that guide cellular behavior via topographical features as well as through mechanical and biochemical cues. However, most studies describing the influence of topography on cells' behavior are performed on rigid and synthetic two-dimensional substrates. To design systems that more closely resemble native microenvironments, herein we develop 3D nanocomposite hydrogels consisting of starch nanocrystals (SNCs) embedded in a gelatin matrix. The incorporation of different concentrations of SNCs (0.05, 0.2, and 0.5 wt %) results in an increase of compressive modulus when compared to hydrogels without SNCs, without affecting the swelling ratio, thus providing a tunable system. Confirming the cytocompatibility of the novel composites, the viability of encapsulated L929 fibroblasts is >90% in all hydrogels. The cellular metabolic activity and DNA content are similar for all formulations and increase over time, indicating that the fibroblasts proliferate within the hydrogels. After 4 d of culture, Live/Dead staining and F-actin/nuclei staining show that the encapsulated fibroblasts develop an elongated morphology in the hydrogels. On the other hand, encapsulated chondrogenic progenitor ATDC5 cells also maintain a viability around 90% but display a round morphology, especially in the hydrogels with SNCs, indicating a potential application of the materials for cartilage tissue engineering. We believe that topographical and mechanical cues within 3D microenvironments can be a powerful tool to instruct cells' behavior and that the developed gelatin/SNC nanocomposite warrants further study.

Published
2019
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30. Mimicking the Articular Joint with In Vitro Models.

Author

Piluso S, Li Y, Abinzano F, Levato R, Moreira Teixeira L, Karperien M, Leijten J, van Weeren R, and Malda J

Subjects
Animals, Biomimetics, Bioprinting, Bioreactors, Cartilage, Articular physiopathology, Cell Culture Techniques methods, Cells, Cultured, Coculture Techniques, Humans, Joint Capsule physiology, Lab-On-A-Chip Devices, Organ Culture Techniques, Osteoarthritis physiopathology, Tissue Culture Techniques, Cartilage, Articular physiology, In Vitro Techniques methods
Abstract

Treating joint diseases remains a significant clinical challenge. Conventional in vitro cultures and animal models have been helpful, but suffer from limited predictive power for the human response. Advanced models are therefore required to mimic the complex biological interactions within the human joint. However, the intricate structure of the joint microenvironment and the complex nature of joint diseases have challenged the development of in vitro models that can faithfully mimic the in vivo physiological and pathological environments. In this review, we discuss the current in vitro models of the joint and the progress achieved in the development of novel and potentially more predictive models, and highlight the application of new technologies to accurately emulate the articular joint., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Published
2019
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31. Cytocompatible carbon nanotube reinforced polyethylene glycol composite hydrogels for tissue engineering.

Author

Van den Broeck L, Piluso S, Soultan AH, De Volder M, and Patterson J

Subjects
Animals, Cell Proliferation drug effects, Cell Survival drug effects, Cells, Cultured, Fibroblasts drug effects, Hydrogels pharmacology, Mice, Polymers chemistry, Tissue Engineering methods, Biocompatible Materials chemistry, Hydrogels chemistry, Nanocomposites chemistry, Nanotubes, Carbon chemistry, Polyethylene Glycols chemistry, Tissue Scaffolds chemistry
Abstract

Hydrogels are attractive materials for stimulating 3D cell growth and tissue regeneration, and they provide mechanical support and physical cues to guide cell behavior. Herein, we developed a robust methodology to increase the stiffness of polyethylene glycol (PEG) hydrogels by successfully incorporating carbon nanotubes (CNTs) within the polymer matrix. Interestingly, hydrogels containing pristine CNTs showed a higher stiffness (1915 ± 102 Pa) than both hydrogels without CNTs (1197 ± 125 Pa) and hydrogels incorporating PEG-grafted CNTs (867 ± 103 Pa) (p < 0.005). The swelling ratio was lower for hydrogels with pristine CNTs (45.4 ± 3.5) and hydrogels without CNTs (46.7 ± 5.1) compared to the hydrogels with PEG-grafted CNTs (62.8 ± 2.6). To confirm that the CNT-reinforced hydrogels were cytocompatible, the viability, proliferation, and morphology of encapsulated L929 fibroblasts was investigated. All hydrogel formulations supported cell proliferation, and the addition of pristine CNTs increased initial cell viability (83.3 ± 10.7%) compared to both pure PEG hydrogels (51.9 ± 8.3%) and hydrogels with PEG-CNTs (63.1 ± 10.9%) (p < 0.005). Altogether, these results demonstrate that incorporation of CNTs could effectively reinforce PEG hydrogels and that the resulting cytocompatible nanocomposites are promising scaffolds for tissue engineering., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Published
2019
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32. Molecularly Engineered Polymer-Based Systems in Drug Delivery and Regenerative Medicine.

Author

Piluso S, Soultan AH, and Patterson J

Subjects
Drug Delivery Systems, Polymers chemistry, Regenerative Medicine
Abstract

Background: Polymer-based systems are attractive in drug delivery and regenerative medicine due to the possibility of tailoring their properties and functions to a specific application., Methods: The present review provides several examples of molecularly engineered polymer systems, including stimuli responsive polymers and supramolecular polymers., Results: The advent of controlled polymerization techniques has enabled the preparation of polymers with controlled molecular weight and well-defined architecture. By using these techniques coupled to orthogonal chemical modification reactions, polymers can be molecularly engineered to incorporate functional groups able to respond to small changes in the local environment or to a specific biological signal. This review highlights the properties and applications of stimuli-responsive systems and polymer therapeutics, such as polymer-drug conjugates, polymer-protein conjugates, polymersomes, and hyperbranched systems. The applications of polymeric membranes in regenerative medicine are also discussed., Conclusion: The examples presented in this review suggest that the combination of membranes with polymers that are molecularly engineered to respond to specific biological functions could be relevant in the field of regenerative medicine., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

Published
2017
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33. Design of Decorin-Based Peptides That Bind to Collagen I and their Potential as Adhesion Moieties in Biomaterials.

Author

Federico S, Pierce BF, Piluso S, Wischke C, Lendlein A, and Neffe AT

Subjects
Biocompatible Materials, Cell Adhesion Molecules chemistry, Collagen Type I chemistry, Decorin chemistry, Peptides chemistry
Abstract

Mimicking the binding epitopes of protein-protein interactions by using small peptides is important for generating modular biomimetic systems. A strategy is described for the design of such bioactive peptides without accessible structural data for the targeted interaction, and the effect of incorporating such adhesion peptides in complex biomaterial systems is demonstrated. The highly repetitive structure of decorin was analyzed to identify peptides that are representative of the inner and outer surface, and it was shown that only peptides based on the inner surface of decorin bind to collagen. The peptide with the highest binding affinity for collagen I, LHERHLNNN, served to slow down the diffusion of a conjugated dye in a collagen gel, while its dimer could physically crosslink collagen, thereby enhancing the elastic modulus of the gel by one order of magnitude. These results show the potential of the identified peptides for the design of biomaterials for applications in regenerative medicine., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Published
2015
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34. Site-specific, covalent incorporation of Tus, a DNA-binding protein, on ionic-complementary self-assembling peptide hydrogels using transpeptidase Sortase A as a conjugation tool†Dedicated to the memory of Joachim H. G. Steinke.‡Electronic supplementary information (ESI) available: Further experimental data. See DOI: 10.1039/c3sm00131hClick here for additional data file.

Author

Piluso S, Cassell HC, Gibbons JL, Waller TE, Plant NJ, Miller AF, and Cavalli G

Abstract

The site-specific conjugation of DNA-binding protein (Tus) to self-assembling peptide FEFEFKFKK was demonstrated. Rheology studies and TEM of the corresponding hydrogels (including PNIPAAm-containing systems) showed no significant variation in properties and hydrogel morphology compared to FEFEFKFKK. Critically, we demonstrate that Tus is accessible within the gel network displaying DNA-binding properties.

Published
2013
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35. Hyaluronic acid-based hydrogels crosslinked by copper-catalyzed azide-alkyne cycloaddition with tailorable mechanical properties.

Author

Piluso S, Hiebl B, Gorb SN, Kovalev A, Lendlein A, and Neffe AT

Subjects
Adhesiveness, Animals, Cell Line, Cell Survival, Diazonium Compounds chemistry, Elastic Modulus, Fibroblasts drug effects, Hyaluronic Acid toxicity, Mice, Molecular Structure, Rheology, Stilbenes chemistry, Sulfonic Acids chemistry, Alkynes chemistry, Azides chemistry, Biocompatible Materials, Copper Sulfate chemistry, Cross-Linking Reagents chemistry, Hyaluronic Acid chemistry, Hydrogels
Abstract

Biopolymers of the extracellular matrix are attractive starting materials for providing degradable and biocompatible biomaterials. In this study, hyaluronic acid-based hydrogels with tunable mechanical properties were prepared by the use of copper- catalyzed azide-alkyne cycloaddition (known as "click chemistry"). Alkyne-functionalized hyaluronic acid was crosslinked with linkers having two terminal azide functionalities, varying crosslinker density as well as the lengths and rigidity of the linker molecules. By variation of the crosslinker density and crosslinker type, hydrogels with elastic moduli in the range of 0.5-4 kPa were prepared. The washed materials contained a maximum of 6.8 mg copper per kg dry weight and the eluate of the gel crosslinked with diazidostilbene did not show toxic effects on L929 cells. The hyaluronic acid-based hydrogels have potential as biomaterials for cell culture or soft tissue regeneration applications.

Published
2011
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36. Oocyte maturation in Xenopus laevis is blocked by the hormonal herbicide, 2,4-dichlorophenoxy acetic acid.

Author

Stebbins-Boaz B, Fortner K, Frazier J, Piluso S, Pullen S, Rasar M, Reid W, Sinclair K, and Winger E

Subjects
Animals, Cytoplasm drug effects, Cytoplasm metabolism, Cytoskeleton drug effects, Female, In Vitro Techniques, MAP Kinase Signaling System drug effects, Oocytes growth & development, Oocytes metabolism, Progesterone pharmacology, RNA, Messenger metabolism, Xenopus laevis, 2,4-Dichlorophenoxyacetic Acid toxicity, Herbicides toxicity, Oocytes drug effects
Abstract

Oocyte maturation is dependent on a complex program of morphological, ultrastructural, and biochemical signaling events, and if disrupted could lead to decreased fertility and population decline. The in vitro sensitivity of amphibian oocytes and oocyte maturation to plant growth factor and widely used hormonal herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), was examined in this study to determine its potential impact on early development and possible contribution to the global amphibian decline. Progesterone, which acts through a membrane receptor, triggers meiotic maturation in full grown (stage VI) Xenopus oocytes, characterized by cytoskeletal reorganization, nuclear dissolution, chromosome condensation, and spindle formation. Biochemically, the Mos/MAPK/MPF signaling pathway is activated, in part dependent on translational activation of specific maternal mRNAs such as c-Mos. Light microscopy revealed unusual asymmetric morphotypes in oocytes exposed to 2,4-D alone characterized by a white spot and bulge, termed coning, in the animal pole where the germinal vesicle (nucleus) persisted intact. Treatment of oocytes with cytochalasin B, a microfilament inhibitor, blocked these morphotypes but nocodazole, a microtubule depolymerizing agent, did not. Confocal microscopy showed that 2,4-D, itself, caused substantial depolymerization of perinuclear microtubules. Importantly, 2,4-D blocked progesterone-induced maturation as measured by the lack of nuclear breakdown, confirmed by the lack of Mos expression, MPF activation, and cytoplasmic polyadenylation of cyclin B1 mRNA. However, Western blot analysis and U0126 inhibitor studies showed that 2,4-D, either alone or in the presence of progesterone, induced MAPK phosphorylation through MAPKK. These results show that 2,4-D disrupts oocyte cytoskeletal organization and blocks maturation while stimulating an independent MAPK signaling pathway., (Copyright 2004 Wiley-Liss, Inc.)

Published
2004
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37. Oral lesions among HIV-infected hemophiliacs. A study of 54 patients.

Author

Ficarra G, Chiodo M, Morfini M, Longo G, Orsi A, Piluso S, and Rafanelli D

Subjects
Humans, Male, Mouth Diseases epidemiology, Prevalence, Retrospective Studies, HIV Seropositivity complications, Hemophilia A complications, Mouth Diseases complications, Transfusion Reaction
Abstract

Background: HIV-infected individuals develop a large variety of oral manifestations. This study was designed to assess the prevalence and types of oral lesions among HIV-positive hemophiliacs., Materials and Methods: A study population of 54 hemophiliacs was evaluated from February, 1987 to March, 1992 in order to analyze types, prevalence and relationships to clinical stages of HIV-related oral lesions. Thirty-six (67%) of the group of patients were HIV seropositive. The remaining 18 tested negative to HIV during the observation period., Results: The majority of patients suffered from hemophilia A. One patient was also bisexual and two were also intravenous drug abusers. Analysis of patient stage revealed that half had a CD4+ T-lymphocyte count over 0.5 x 10(9)/L cells, 10 between 0.2 and 0.499 x 10(9)/L and 8 showed a count lower than 200 x 10(9)/L. Oral lesions were recorded in 18 (50%) HIV-seropositive hemophiliacs. No oral lesions were observed among the HIV-seronegative hemophiliacs. Advanced stage of immunosuppression and presence of oral lesions were significantly associated (p = 0.040). Candidiasis was the most common disturbance, followed by hairy leukoplakia. Oral herpes simplex infection, necrotizing gingivitis and facial herpes zoster were found in a small number of patients. Those with oral lesions showed a lower median CD4+ T lymphocyte count (0.209 x 10(9)/L cells; range 0.008 to 0.615) when compared to the ones without oral lesions (median CD4+ count was 0.539 x 10(9)/L cells; range 0.042 to 1.180; p = 0.002)., Conclusions: HIV-seropositive hemophiliacs may develop oral lesions during the course of their disease. Candidiasis and hairy leukoplakia are among the most common manifestations. A careful oral examination should be included in the clinical evaluation of all HIV-infected hemophiliacs.

Published
1994

38. Unusual clinical aspects of oral non-Hodgkin lymphomas in patients with HIV infection.

Author

Piluso S, Di Lollo S, Baroni G, Leoncini F, Gaglioti D, Saccardi A, and Ficarra G

Subjects
Adult, DNA, Viral analysis, Herpesvirus 4, Human isolation & purification, Homosexuality, Male, Humans, Lymphoma, AIDS-Related virology, Male, Necrosis, Oral Ulcer, Palatal Neoplasms pathology, Palatal Neoplasms virology, Precursor Cell Lymphoblastic Leukemia-Lymphoma virology, Tongue Neoplasms pathology, Tongue Neoplasms virology, HIV Seropositivity, Lymphoma, AIDS-Related pathology, Mouth Neoplasms pathology, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology
Abstract

Patients with HIV infection are at greater risk of developing malignancies. We report two HIV-seropositive patients with primary oral manifestation of a B-cell non-Hodgkin lymphoma (NHL). Localisations of tumours were the palate, with an unusual bifocal origin, and the tongue which is rarely the primary site of NHL. Ulcerations and extensive tissue necrosis were observed. Histologically both cases were high grade malignant lymphomas, immunoblastic. Epstein-Barr virus DNA was detected with in situ hybridisation in 1 patient.

Published
1994
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39. White lichenoid lesions of the buccal mucosa in patients with HIV infection.

Author

Ficarra G, Flaitz CM, Gaglioti D, Piluso S, Milo D, Adler-Storthz K, and Eversole LR

Subjects
Adult, Female, HIV Infections drug therapy, Humans, Lichen Planus, Oral complications, Lichen Planus, Oral pathology, Male, HIV Infections complications, Ketoconazole adverse effects, Lichen Planus, Oral chemically induced, Mouth Mucosa drug effects, Zidovudine adverse effects
Abstract

We report on eight patients who developed white lichenoid lesions of the buccal mucosa during the course of human immunodeficiency virus infection. In five patients the lesions appeared after the administration of zidovudine, in two after the intake of both zidovudine and ketoconazole, and in one after ketoconazole. In the majority of cases, lesions presented as bilateral reticular keratosis or atrophic changes of the buccal mucosa. Three patients manifested lichenoid atrophic changes of the dorsum of the tongue. The histopathologic features were hyperkeratosis, epithelial atrophy, basal cell liquefaction, and the presence in the lamina propria of either a patchy or diffuse lymphocytic infiltrate. All specimens tested negative for Epstein-Barr virus and human papillomavirus. Our study suggests that lichenoid lesions of the buccal mucosa, similar to what has been described as lichenoid drug reactions or idiopathic lichen planus, can be observed during human immunodeficiency virus infection and that administration of zidovudine and ketoconazole should be considered as a possible cause.

Published
1993
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40. Clinical aspects and microbiology of HIV-associated periodontal lesions.

Author

Piluso S, Ficarra G, Orsi A, Gaglioti D, Pierotti P, and Orlando S

Subjects
AIDS-Related Opportunistic Infections microbiology, Adult, Bacteria isolation & purification, Candida albicans isolation & purification, Female, HIV Infections microbiology, HIV Seronegativity, HIV Seropositivity diagnosis, HIV Seropositivity microbiology, Humans, Male, Middle Aged, Periodontal Diseases microbiology, Substance Abuse, Intravenous complications, AIDS-Related Opportunistic Infections diagnosis, HIV Infections diagnosis, HIV-1 immunology, Periodontal Diseases diagnosis
Abstract

This study examines the clinical aspects of HIV-associated periodontal lesions and the prevalence of periodontal pathogens. Subgingival plaque samples were taken from 55 subjects in six study groups: 1) HIV-seropositive patients with gingivitis, 2) necrotizing gingivitis, 3) periodontitis or 4) with health periodontium, 5) patients with rapidly progressive periodontitis or 6) periodontally healthy in whom there was no evidence of HIV infection. Among HIV-positive patients there was a majority (66%) of intravenous drug users. We detected more Bacteroides intermedius, B. buccae and B. oralis in HIV-infected patients with periodontal lesions and in HIV-negative subjects with rapidly progressive periodontitis than in the other groups. High levels of Spirochetes were recovered in both HIV associated necrotizing gingivitis and periodontitis. The results indicate that there is a similarity in the microbiological profile of HIV-associated gingivitis, necrotizing gingivitis, periodontitis and rapidly progressive periodontitis of HIV-negative subjects although significant differences in the clinical aspects of the lesions and in the immune status of the host have been observed.

Published
1993

41. Oral Crohn's disease and pyostomatitis vegetans. An unusual association.

Author

Ficarra G, Cicchi P, Amorosi A, and Piluso S

Subjects
Abscess pathology, Crohn Disease pathology, Female, Granuloma pathology, Humans, Intestinal Absorption, Middle Aged, Mouth Mucosa pathology, Stomatitis drug therapy, Stomatitis pathology, Ulcer pathology, Zinc metabolism, Zinc therapeutic use, Crohn Disease complications, Deficiency Diseases complications, Stomatitis etiology, Zinc deficiency
Abstract

Oral features of Crohn's disease include ulcers, lip fissuring, cobblestone plaques, angular cheilitis, polypoid lesions, and perioral erythema. Pyostomatitis vegetans is a rare eruption of the oral mucosa characterized by tiny yellow pustules. It is considered a marker for inflammatory bowel disease. We describe a 45-year-old woman with a 6-month history of painful sores in her mouth, diarrhea, weight loss, and cutaneous lesions. Oral examination revealed cobblestone plaques and indentation on the tongue and friable vegetating pustules on the labial commissures. Staphylococcus simulans was isolated from the pustules. Laboratory studies revealed leucocytosis, eosinophilia, and low hemoglobin and zinc levels. Histologic study of the labial lesions revealed hyperplastic epithelium with intraepithelial clefts that contain eosinophils and neutrophils. Tongue lesions showed chronic inflammation with noncaseating granulomas. Later, colonoscopy and biopsy demonstrated Crohn's disease of the anorectal region. Pyostomatitis vegetans lesions regressed after oral zinc supplementation. Prednisone treatment resulted in healing of the tongue lesions. In our patient, pyostomatitis vegetans appeared to be related to zinc deficiency that may have been caused by malabsorption. The pathogenetic interrelationship between pyostomatitis vegetans and Crohn's disease is discussed.

Published
1993
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42. [Benign lymphoepithelial cysts of the parotid in a patient with HIV infection].

Author

Ficarra G, Panzoni E, Piluso S, Pierleoni F, and Adler-Storthz K

Subjects
Adult, Biopsy, Cytomegalovirus genetics, DNA, Viral genetics, HIV Infections complications, HIV Infections pathology, Herpesvirus 4, Human genetics, Humans, Lymphocele complications, Lymphocele pathology, Male, Nucleic Acid Hybridization, Parotid Diseases complications, Parotid Diseases pathology, Parotid Gland pathology, HIV Infections diagnosis, HIV-1, Lymphocele diagnosis, Parotid Diseases diagnosis
Abstract

We report on a HIV-positive patient presenting bilateral enlargement of the parotid gland. The ultrasound examination demonstrated multiple lesions with cystic aspect. A partial surgical excision of the parotid gland was performed, and histologic examination confirmed the diagnosis of a cystic benign lymphoepithelial lesion. The specimens which were tested for Epstein-Barr virus and cytomegalovirus by in situ hybridization proved negative. The clinical and pathologic features of this unusual salivary gland lesion which affects HIV-positive patients are discussed.

Published
1992

43. [Peripheral facial paralysis and HIV infection].

Author

Piluso S, Ficarra G, Gaglioti D, and Pierleoni F

Subjects
AIDS Dementia Complex complications, AIDS Dementia Complex diagnosis, AIDS-Related Complex diagnosis, Acquired Immunodeficiency Syndrome diagnosis, Adult, Brain diagnostic imaging, Facial Paralysis diagnosis, Female, Humans, Male, Neurologic Examination, Substance-Related Disorders complications, Tomography, X-Ray Computed, AIDS-Related Complex complications, Acquired Immunodeficiency Syndrome complications, Facial Paralysis etiology, HIV-1
Abstract

Two cases of peripheral facial paralysis associated with infection by the human immunodeficiency virus in young intravenous drug abuser patients are reported. One patient manifested AIDS-related complex (ARC) and the other fulfilled the Center for Disease Control (CDC) criteria for AIDS. Clinical symptoms and signs occurred as facial paralysis and headache. One patient presented progressive motor slowing and the computerized tomography (CT) of the brain showed a wide hypodense lesion in the left temporal-anterior region and the involvement of the left basal ganglia. In this patient the impairment of the facial nerve was associated with a HIV-related encephalitis. In the second patient the CT did not show cerebral lesions and the etiology of the paralysis was unknown.

Published
1991

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