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1. Poly(3-hexylthiophene) nanoparticles for biophotonics: study of the mutual interaction with living cells

Author

Cosimo D'Andrea, Ilaria Bargigia, Elena Zucchetti, Maria Rosa Antognazza, Mattia Zangoli, Giovanna Barbarella, Francesca Di Maria, Guglielmo Lanzani, Caterina Bossio, Zucchetti, Elena, Zangoli, Mattia, Bargigia, Ilaria, Bossio, Caterina, Di Maria, Francesca, Barbarella, Giovanna, D'Andrea, Cosimo, Lanzani, Guglielmo, and Antognazza, Maria Rosa

Subjects
In situ, inorganic chemicals, Materials science, Photoluminescence, Biocompatibility, education, Biomedical Engineering, Nanoparticle, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Colloid, Calcium imaging, Light Phototrasduction, General Materials Science, Cell Imaging, Poly(3-hexyl)thiophene Nanoparticles, Medicine (all), Chemistry (all), technology, industry, and agriculture, General Chemistry, General Medicine, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Biophotonics, Electrophysiology, Cytosol, Materials Science (all), 0210 nano-technology
Abstract

We report on the mutual interaction between poly(3-hexylthiophene) nanoparticles (P3HT-NPs) and human embryonic kidney (HEK-293) cells. P3HT-NPs, prepared in sterile conditions and efficiently uptaken within the live cells cytosol, show well-ordered morphology, high colloidal stability and excellent biocompatibility. Electrophysiology and calcium imaging experiments demonstrate that physiological functions of live cells are fully preserved in the presence of P3HT-NPs. From a complementary point of view, the photophysical properties of P3HT-NPs are also mainly maintained within the cellular environment, as proven by in situ time-resolved photoluminescence. Interestingly, we detect slight modifications in emission spectra and dynamics, which we ascribe to the contribution from the P3HT-NPs surface, possibly due to conformational changes as the result of the interaction with intracellular proteins or the formation of NPs aggregates. This work demonstrates that P3HT-NPs are excellent candidates for use as light sensitive actuators, due to their remarkable physical properties, optimal biocompatibility and capability of interaction with living cells.

Published
2020
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2. Photocatalytic Activity of Polymer Nanoparticles Modulates Intracellular Calcium Dynamics and Reactive Oxygen Species in HEK-293 Cells

Author

Ilaria Abdel Aziz, Guglielmo Lanzani, Francesca Di Maria, Mattia Zangoli, Gabriele Tullii, Elena Zucchetti, Caterina Bossio, Doriana Debellis, and Maria Rosa Antognazza

Subjects
0301 basic medicine, conjugated polymer, Histology, lcsh:Biotechnology, Cell, Biomedical Engineering, Bioengineering, Context (language use), 02 engineering and technology, Calcium in biology, 03 medical and health sciences, Ca2+ imaging, lcsh:TP248.13-248.65, medicine, Viability assay, bio-organic electronics, Original Research, reactive oxygen species, chemistry.chemical_classification, Reactive oxygen species, organic semiconductor, HEK 293 cells, Bioengineering and Biotechnology, Photocatalytic Activity, Transfection, 021001 nanoscience & nanotechnology, Conjugated Polymer Nanoparticles, Cytosol, 030104 developmental biology, medicine.anatomical_structure, chemistry, Biophysics, Intracellular Calcium Dynamics, photomodulation, light, 0210 nano-technology, cell optical stimulation, Biotechnology
Abstract

Optical modulation of living cells activity by light-absorbing exogenous materials is gaining increasing interest, due to the possibility both to achieve high spatial and temporal resolution with a minimally invasive and reversible technique and to avoid the need of viral transfection with light-sensitive proteins. In this context, conjugated polymers represent ideal candidates for photo-transduction, due to their excellent optoelectronic and biocompatibility properties. In this work, we demonstrate that organic polymer nanoparticles, based on poly(3-hexylthiophene) conjugated polymer, establish a functional interaction with an in vitro cell model (Human Embryonic Kidney cells, HEK-293). They display photocatalytic activity in aqueous environment and, once internalized within the cell cytosol, efficiently generate reactive oxygen species (ROS) upon visible light excitation, without affecting cell viability. Interestingly, light-activated ROS generation deterministically triggers modulation of intracellular calcium ion flux, successfully controlled at the single cell level. In perspective, the capability of polymer NPs to produce ROS and to modulate Ca2+ dynamics by illumination on-demand, at non-toxic levels, may open the path to the study of biological processes with a gene-less approach and unprecedented spatio-temporal resolution, as well as to the development of new biotechnology tools for cell optical modulation.

Published
2018
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3. Conjugated polymers for the optical control of the electrical activity of living cells

Author

Elena Zucchetti, Guglielmo Lanzani, Caterina Bossio, Sebastiano Bellani, Susana Vaquero, Nicola Martino, and Maria Rosa Antognazza

Subjects
chemistry.chemical_classification, Materials science, Absorption spectroscopy, Biocompatibility, Biomedical Engineering, Nanotechnology, 02 engineering and technology, General Chemistry, General Medicine, Polymer, Conjugated system, 010402 general chemistry, 021001 nanoscience & nanotechnology, Electrochemistry, 01 natural sciences, 0104 chemical sciences, Photostimulation, chemistry, General Materials Science, Light emission, 0210 nano-technology, Visible spectrum
Abstract

The possibility to optically excite the electrical activity of living cells by using exogenous absorbers is gaining more and more interest in the neuroscience and biotechnology community. Conjugated polymers, inherently sensitive to visible light, were recently proposed as candidates to this goal. To date, however, only one polymer type, namely regio-regular poly-3-hexylthiophene, has been tested as the active material. In this work four different conjugated polymers, regarded as prototypes of their category, are investigated as photoactive bio-interfaces. The selected materials have different absorption spectra, morphology, light emission efficiency and charge transport properties. We analyze their key-enabling properties, such as electrochemical stability, surface morphology, wettability, sterilization compatibility, interaction with protein adhesion layers and toxicity, throughout all the necessary steps for the realization of an efficient bio-optical interface. We demonstrate that all considered polymers are characterized by good biocompatibility and cell seeding properties, and can optimally sustain thermal sterilization. Conversely, electrochemical stability and cell photostimulation efficacy can vary a lot among different materials, and should be carefully evaluated case by case. Reported results represent the starting point for the implementation of bio-polymer interfaces sensitive to different colors and, in perspective, for the realization of a three-chromatic artificial visual prosthesis.

Published
2016
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4. CONTRIBUTORS

Author

Judith A. Aberg, Amy P. Abernethy, Janet L. Abrahm, Michael Adolph, Michael Aherne, K. Allsopp, Rogelio Altisent, Carmen Fernandez Alvarez, Pablo Amigo, Wendy G. Anderson, Sik Kim Ang, Tiziana Antonelli, John Armstrong, Wendy S. Armstrong, Robert M. Arnold, Pilar Arranz, Koen Augustyns, Isabel Barreiro-Meiro Sáenz-Diez, Pilar Barreto, Debra Barton, Ursula Bates, Maria B. Fernandez-Creuchet Santos, Jacinto Bátiz, Costantino Benedetti, Nabila Bennani-Baiti, Michael I. Bennett, Kevin Berger, Mamta Bhatnagar, Lesley Bicanovsky, Lynda Blue, Barton Bobb, Jean-Jacques Body, Gian Domenico Borasio, Claudia Borreani, Federico Bozzetti, Valentina Bozzetti, Jason Braybrooke, William Breitbart, Barry Bresnihan, Bert Broeckaert, Eduardo Bruera, Kay Brune, Bradley Buckhout, Phyllis N. Butow, Ira Byock, Anthony Byrne, Clare Byrne, Beryl E. Cable-Williams, Sarah E. Callin, David Casarett, David Casper, Eric J. Cassell, Barrie Cassileth, Emanuele Castagno, Carlos Centeno, Walter Ceranski, Lucas Ceulemans, Meghna Chadha, Bruce H. Chamberlain, Eric L. Chang, Victor T. Chang, Harvey Max Chochinov, Edward Chow, Grace Christ, Katherine Clark, Stephen Clarke, Josephine M. Clayton, James F. Cleary, Lawrence J. Clein, Katri Elina Clemens, Libby Clemens, Robert Colebunders, Steven R. Connor, Viviane Conraads, Colm Cooney, Massimo Costantini, Azucena Couceiro, Holly Covington, John D. Cowan, Patrick Coyne, Garnet Crawford, Brian Creedon, Hilary Cronin, Garret Cullen, Jennifer E. Cummings, David C. Currow, Paul J. Daeninck, Pamela Dalinis, Prajnan Das, Mellar P. Davis, Sara N. Davison, Catherine Deamant, Liliana de Lima, Conor P. Delany, Peter Demeulenaere, Lena Dergham, Noël Derycke, Rajeev Dhupar, Mario Dicato, Edwin D. Dickerson, Andrew Dickman, Maria Dietrich, Pamela Dixon, Philip C. Dodd, James T. D'Olimpio, Per Dombernowsky, Michael Dooley, Deborah Dudgeon, Geoffrey P. Dunn, David Dunwoodie, Jane Eades, Badi El Osta, Katja Elbert-Avila, John Ellershaw, Bassam Estfan, Louise Exton, Alysa Fairchild, Matthew Farrelly, Konrad Fassbender, Jason Faulhaber, Kenneth C.H. Fearon, Lynda E. Fenelon, Peter F. Ferson, Petra Feyer, Marilene Filbet, Pam Firth, Susan F. FitzGerald, Hugh D. Flood, Francesca Crippa Floriani, Paul J. Ford, Barry Fortner, Darlene Foth, Bridget Fowler, Karen Frame, Thomas G. Fraser, Fred Frost, Michael J. Fulham, Pierre R. Gagnon, Lisa M. Gallagher, Maureen Gambles, Subhasis K. Giri, Paul Glare, Cynthia R. Goh, Xavier Gómez-Batiste, Leah Gramlich, Luigi Grassi, Phyllis A. Grauer, Claire Green, Gareth Griffiths, Yvona Griffo, Hunter Groninger, David A. Gruenewald, Jyothirmai Gubili, Terence L. Gutgsell, Elizabeth Gwyther, Paul S. Haber, Achiel Haemers, Mindi C. Haley, Mazen A. Hanna, Janet R. Hardy, Jodie Haselkorn, Katherine Hauser, Cathy Heaven, Michael Herman, Jørn Herrstedt, Stephen Higgins, Irene J. Higginson, Joanne M. Hilden, Kathryn L. Hillenbrand, Burkhard Hinz, Jade Homsi, Kerry Hood, Juliet Y. Hou, Guy Hubens, Peter Hudson, John G. Hughes, John Hunt, Craig A. Hurwitz, James Ibinson, Nora Janjan, Birgit Jaspers, Thomas Jehser, A. Mark Joffe, Laurence John, Jennie Johnstone, J. Stephen Jones, Javier R. Kane, Matthew T. Karafa, Andrew P. Keaveny, Dorothy M.K. Keefe, Catherine McVearry Kelso, Rose Anne Kenny, Martina Kern, Dilara Seyidova Khoshknabi, Jordanka Kirkova, Kenneth L. Kirsh, David W. Kissane, Eberhard Klaschik, Seref Komurcu, Kandice Kottke-Marchant, Kathryn M. Kozell, Sunil Krishnan, Deborah Kuban, Damian A. Laber, Ruth L. Lagman, Rajesh V. Lalla, Deforia Lane, Philip J. Larkin, Wael Lasheen, Peter Lawlor, Susan B. LeGrand, Vincent Lens, Dona Leskuski, Pamela Levack, Marcia Levetown, Jeanne G. Lewandowski, William R. Lewis, S. Lawrence Librach, Wendy G. Lichtenthal, J. Norelle Lickiss, Stefano Lijoi, Edward Lin, Arthur G. Lipman, Jean-Michel Livrozet, Mari Lloyd-Williams, Richard M. Logan, Francisco López-Lara Martín, Charles L. Loprinzi, John Loughnane, Michael Lucey, Laurie Lyckholm, Carol Macmillan, Frances Mair, Stephen N. Makoni, Bushra Malik, Kevin Malone, Marco Maltoni, Aruna Mani, Lucille R. Marchand, Darren P. Mareiniss, Anna L. Marsland, Joan Marston, Julia Romero Martinez, Isabel Martínez de Ubago, Lina M. Martins, Timothy S. Maughan, Catriona Mayland, Susan E. McClement, Ian McCutcheon, Michael F. McGee, Neil McGill, Stephen McNamara, Mary Lynn McPherson, Henry McQuay, Regina McQuillan, Robert E. McQuown, Michelle Meiring, Sebastiano Mercadante, Elaine C. Meyer, Randy D. Miller, Yvonne Millerick, Roberto Miniero, Armin Mohamed, Busi Mooka, Helen M. Morrison, J. Cameron Muir, Fiona Mulcahy, Hugh E. Mulcahy, Monica Muller, H. Christof Müller-Busch, Scott A. Murray, Friedemann Nauck, Katherine Neasham, Busisiwe Nkosi, Simon Noble, Antonio Noguera, Anna K. Nowak, Juan Nuñez-Olarte, Eugenie A.M.T. Obbens, Tony O'Brien, Megan Olden, Norma O'Leary, David Oliver, David Oliviere, Aurelius G. Omlin, Kaci Osenga, Diarmuid O'Shea, Christophe Ostgathe, Faith D. Ottery, Michel Ouellette, Edgar Turner Overton, Moné Palacios, Robert Palmer, Teresa Palmer, Carmen Paradis, Armida G. Parala, Antonio Pascual-López, Steven D. Passik, Timothy M. Pawlik, Malcolm Payne, Sheila Payne, Silvia Paz, José Pereira, George Perkins, Karin Peschardt, Hayley Pessin, Douglas E. Peterson, Vinod K. Podichetty, Robin Pollens, Eliza Pontifex, Susan Poole, Josep Porta-Sales, Graeme Poston, Ruth D. Powazki, William Powderly, Leopoldo Pozuelo, Eric Prommer, Christina M. Puchalski, Lukas Radbruch, David F.J. Raes, Jane Read, Anantha Reddy, Steven I. Reger, Susan J. Rehm, Stephen G. Reich, Javier Rocafort, Adam Rosenblatt, Cynda Hylton Rushton, K. Mitchell Russell, Karen Ryan, Lisa A. Rybicki, Paola Sacerdote, Vinod Sahgal, Mary Ann Sammon, Dirk Sandrock, Mark Sands, Denise L. Schilling, Valerie Nocent Schulz, Lisa N. Schum, Peter Selwyn, Joshua Shadd, Charles L. Shapiro, Aktham Sharif, Helen M. Sharp, Kirk V. Shepard, J. Timothy Sherwood, Nabin K. Shrestha, Richard J.E. Skipworth, Howard S. Smith, Mildred Z. Solomon, Diego Soto de Prado Otero, Denise Wells Spencer, Ron Spice, David Spiegel, Manish Srivastava, John N. Staffurth, Randall Starling, Grant D. Stewart, Jan Stjernswärd, Florian Strasser, Edna Strauss, Imke Strohscheer, Brett Taylor Summey, Graham Sutton, Nigel P. Sykes, Alan J. Taege, Marcello Tamburini, Yoko Tarumi, Davide Tassinari, Martin H.N. Tattersall, Karl S. Theil, Keri Thomas, Adrian Tookman, María P. Torrubia, Anna Towers, Daphne Tsoi, Rodney O. Tucker, James A. Tulsky, Rachel A. Tunick, Claire Turner, Martha L. Twaddle, Marie Twomey, Christina Ullrich, Catherine E. Urch, Mary L.S. Vachon, Bart Van den Eynden, Antonio Vigano, Erika Vlieghe, Angelo E. Volandes, Raymond Voltz, Paul W. Walker, Sharon Watanabe, Michael A. Weber, Elizabeth Weinstein, Sharon M. Weinstein, Kathryn L. Weise, Sherri Weisenfluh, John Welsh, Clare White, Donna M. Wilson, Joanne Wolfe, Tugba Yavuzsen, Albert J.M. Yee, Lisa M. Yerian, and Elena Zucchetti

Published
2009
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6. Photothermal cellular stimulation in functional bio-polymer interfaces

Author

Nicola Martino, Elena Zucchetti, Maria Rosa Antognazza, Matteo Porro, Fabio Benfenati, Guglielmo Lanzani, Caterina Bossio, Diego Ghezzi, and Paul Feyen

Subjects
chemistry.chemical_classification, Membrane potential, Materials science, Multidisciplinary, Light, Cell Polarity, Depolarization, Nanotechnology, Polymer, Thiophenes, Photothermal therapy, Conjugated system, Article, 3. Good health, Membrane Potentials, Organic semiconductor, Membrane, Biopolymers, HEK293 Cells, chemistry, Humans, Thin film
Abstract

Hybrid interfaces between organic semiconductors and living tissues represent a new tool for in-vitro and in-vivo applications, bearing a huge potential, from basic researches to clinical applications. In particular, light sensitive conjugated polymers can be exploited as a new approach for optical modulation of cellular activity. In this work we focus on light-induced changes in the membrane potential of Human Embryonic Kidney (HEK-293) cells grown on top of a poly(3-hexylthiophene) (P3HT) thin film. On top of a capacitive charging of the polymer interface, we identify and fully characterize two concomitant mechanisms, leading to membrane depolarization and hyperpolarisation, both mediated by a thermal effect. Our results can be usefully exploited in the creation of a new platform for light-controlled cell manipulation, with possible applications in neuroscience and medicine.

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