Optical control of tissue regeneration through photostimulation of organic semiconducting nanoparticles

Next generation bioengineering strives to identify crucial cues that trigger regeneration of damaged tissues, and to control the cells that execute these programs with biomaterials and devices. Molecular and biophysical mechanisms driving embryogenesis may inspire novel tools to reactivate developmental programs in situ. Here nanoparticles based on conjugated polymers are employed for optical control of regenerating tissues by using an animal with unlimited regenerative potential, the polyp Hydra, as in vivo model, and human keratinocytes as an in vitro model to investigate skin repair. By integrating animal, cellular, molecular, and biochemical approaches, nanoparticles based on poly-3-hexylthiophene (P3HT) are shown able to enhance regeneration kinetics, stem cell proliferation, and biomolecule oxidation levels. Opposite outputs are obtained with PCPDTBT-NPs (Poly[2,6-(4,4-bis-(2-ethylhexyl)-4H-cyclopenta [2,1-b;3,4-b′] dithiophene)-alt-4,7(2,1,3-benzothiadiazole)], causing a beneficial effect on Hydra regeneration but not on the migratory capability of keratinocytes. These results suggest that the artificial modulation of the redox potential in injured tissues may represent a powerful modality to control their regenerative potential. Importantly, the possibility to fine-tuning materials’ photocatalytic efficiency may enable a biphasic modulation over a wide dynamic range, which can be exploited to augment the tissue regenerative capacity or inhibit the unlimited potential of cancerous cells in pathological contexts.C.T. acknowledges Dr. Massimo Rippa (ISASI-CNR, Pozzuoli) for setting up the irradiation system for Hydra and cells. M.M. acknowledges financial support from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 853468), and from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 660228 nd Grant RYC2019-026860-I funded by MCIN/AEI/ 10.13039/501100011033 and “ESF Investing in your future”. M.R.A , F.G. and C.T. as third-party, acknowledge support by the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program “LINCE”, grant agreement n. 803621.Open Access Funding provided by Consiglio Nazionale delle Ricerche within the CRUI-CARE Agreement.Peer reviewe

Phostimulation of semiconducting nanoparticles to control physiological functions in vivo

Resumen del trabajo presentado al 8th Forum on New Materials - CIMTEC, celebrado en Perugia (Italia) del 4 al 14 de junio de 2018.The latest designs of optoelectronic devices powered research on fundamental properties of light and how it interacts with matter. In the context of a living cell, this interaction is hampered by the complexity of the living matter, making technological advancement key aspect of this research. Nanotechnologies may provide unique tools to finely tune biological functions, through development of biocompatible light nanotransducers. Here we show the possibility to modulate cell function by photostimulation of polymer nanoparticles based on poly(3-hexylthiophene) (NP-P3HT), a conjugated polymer widely used in photovoltaic application. By using a freshwater polyp as model organism, presenting photic behaviour despite the absence of proper eyes, we show that NP-P3HT internalized into animal tissue enhances animal photosensitivity. Moreover, the peculiar capabilities of the polyp to regenerate missing parts of amputated body allowed us to investigate the potential of P3HT-NP to enhance tissue regeneration. We observed in treated animals a boost in the regeneration efficiency under light illumination, uncovering a new function of these light nanotransducers in regenerative medicine and opening up new scenario on future therapeutic purposes.Peer Reviewe

Phostimulation of semiconducting nanoparticles to control physiological functions in vivo

Resumen del trabajo presentado al 8th Forum on New Materials - CIMTEC, celebrado en Perugia (Italia) del 4 al 14 de junio de 2018.The latest designs of optoelectronic devices powered research on fundamental properties of light and how it interacts with matter. In the context of a living cell, this interaction is hampered by the complexity of the living matter, making technological advancement key aspect of this research. Nanotechnologies may provide unique tools to finely tune biological functions, through development of biocompatible light nanotransducers. Here we show the possibility to modulate cell function by photostimulation of polymer nanoparticles based on poly(3-hexylthiophene) (NP-P3HT), a conjugated polymer widely used in photovoltaic application. By using a freshwater polyp as model organism, presenting photic behaviour despite the absence of proper eyes, we show that NP-P3HT internalized into animal tissue enhances animal photosensitivity. Moreover, the peculiar capabilities of the polyp to regenerate missing parts of amputated body allowed us to investigate the potential of P3HT-NP to enhance tissue regeneration. We observed in treated animals a boost in the regeneration efficiency under light illumination, uncovering a new function of these light nanotransducers in regenerative medicine and opening up new scenario on future therapeutic purposes.Peer Reviewe

Bioengineering fluorescent conductive microfibrils in vivo

Resumen del trabajo presentado al 8th Forum on New Materials - CIMTEC, celebrado en Perugia (Italia) del 4 al 14 de junio de 2018.Conductive polymers are very attractive for biomedical applications. Their responsiveness to electrical stimulation found application in nerve regeneration strategies, enhanced neuronal growth or tissue regeneration. Often their broad use is hampered by low biocompatibility,lack of recognition elements for endogenous components, thus alternative strategies may rely on improvement of fibrillar proteins properties in situ. In this direction, the fluorophore dithienothiophene-S,S-dioxide (DTTO) is able to spontaneously enter human fibroblasts and become incorporated into collagen quaternary structures giving rise to fluorescent and conducting fibrils. In order to translate to an in vivo system, we employed the freshwater Hydra vulgaris, which structural simplicity resembles a living tissue, without organs. By challenging polyps with DTTO we demonstrated the incorporation of this thiophene into supramolecular structures giving rise to microfibrils. Electronic force microscopy showed electrical conductivity and circular dichroism confirmed the presence of proteins within the fibrils. Besides the possibility of engineering endogenous components with synthetic moieties, this approach opens the path to a sort of ‘live animal factory’ for the production of innovative biomaterials.Peer Reviewe

Real time dynamics of β-catenin expression during hydra development, regeneration and wnt signalling activation

Understanding the dynamic cellular behaviours driving morphogenesis and regeneration is a long-standing challenge in biology. Live imaging, together with genetically encoded reporters, may provide the necessary tool to address this issue, permitting the in vivo monitoring of the spatial and temporal expression dynamics of a gene of interest during a variety of developmental processes. Canonical Wnt/β-catenin signalling controls a plethora of cellular activities during development, regeneration and adulthood throughout the animal kingdom. Several reporters have been produced in animal models to reveal sites of active Wnt signalling. In order to monitor in vivo Wnt/β-catenin signalling activity in the freshwater polyp Hydra vulgaris, we generated a β-cat-eGFP transgenic Hydra, in which eGFP is driven by the Hydra β-catenin promoter.We characterized the expression dynamics during budding, regeneration and chemical activation of the Wnt/β-cat signalling pathway using light sheet fluorescence microscopy. Live imaging of the β-cat-eGFP lines recapitulated the previously reported endogenous expression pattern of β-catenin and revealed the dynamic appearance of novel sites of Wnt/β-catenin signalling, that earlier evaded detection by mean of in situ hybridization. By combining the Wnt activity read-out efficiency of the β-catenin promoter with advanced imaging, we have created a novel model system to monitor in real time the activity of Hydra β-cat regulatory sequences in vivo, and open the path to reveal β-catenin modulation in many other physiological contexts

2019 Dataset of Participatory Budgeting in Italy

The dataset provides data on Italian Municipalities’ policies pertaining to participatory budgeting (PB). The dataset comprises information on municipalities with a population of over 2,000 inhabitants, and is composed as follows: Municipality ISTAT Code (ID), Municipality Name (MN) Province, Region, Population (2011 Census), Researcher (IR), Mapping randomized process (MR), Mapping Method (MM), twenty dummy variables identifying Municipalities running PB in a given year (y1994, y2002 to y2020), Number of years with PB (TOT). Overall, the dataset comprises 4517 units (with 467 missing), for a total of 4050 mapped municipalities

Risk of Guillain-Barr\ue9 syndrome after 2010-2011 influenza vaccination

Influenza vaccination has been implicated in Guillain Barr\ue9 Syndrome (GBS) although the evidence for this link is controversial. A case-control study was conducted between October 2010 and May 2011 in seven Italian Regions to explore the relation between influenza vaccination and GBS. The study included 176 GBS incident cases aged 6518 years from 86 neurological centers. Controls were selected among patients admitted for acute conditions to the Emergency Department of the same hospital as cases. Each control was matched to a case by sex, age, Region and admission date. Two different analyses were conducted: a matched case-control analysis and a self-controlled case series analysis (SCCS). Case-control analysis included 140 cases matched to 308 controls. The adjusted matched odds ratio (OR) for GBS occurrence within 6 weeks after influenza vaccination was 3.8 (95 % CI: 1.3, 10.5). A much stronger association with gastrointestinal infections (OR = 23.8; 95 % CI 7.3, 77.6) and influenza-like illness or upper respiratory tract infections (OR = 11.5; 95 % CI 5.6, 23.5) was highlighted. The SCCS analysis included all 176 GBS cases. Influenza vaccination was associated with GBS, with a relative risk of 2.1 (95 % CI 1.1, 3.9). According to these results the attributable risk in adults ranges from two to five GBS cases per 1,000,000 vaccinations