Anna Sagnella

A transparent organic transistor structure for bidirectional stimulation and recording of primary neurons

Real-time stimulation and recording of neural cell bioelectrical activity could provide an unprecedented insight in understanding the functions of the nervous system, and it is crucial for developing advanced in vitro drug screening approaches. Among organic materials, suitable candidates for cell interfacing can be found that combine long-term biocompatibility and mechanical flexibility. Here, we report on transparent organic cell stimulating and sensing transistors (O-CSTs), which provide bidirectional stimulation and recording of primary neurons. We demonstrate that the device enables depolarization and hyperpolarization of the primary neuron membrane potential. The transparency of the device also allows the optical imaging of the modulation of the neuron bioelectrical activity. The maximal amplitude-to-noise ratio of the extracellular recording achieved by the O-CST device exceeds that of a microelectrode array system on the same neuronal preparation by a factor of 16. Our organic cell stimulating and sensing device paves the way to a new generation of devices for stimulation, manipulation and recording of cell bioelectrical activity in vitro and in vivo.

Innovative multifunctional silk fibroin and hydrotalcite nanocomposites: a synergic effect of the components.

Novel hybrid functional materials are formed by combining hydrotalcite-like compounds and silk fibroin (SF-HTlc) via an environmental friendly aqueous process. The nanocomposites can be prepared with different weight ratio of the constituting components and preserve the conformational properties of the silk protein and the lamellar structure of hydrotalcites. Optical microscopy, scanning electron microscopy, and atomic force microscopy analyses show a good dispersion degree of the inorganic nanoparticles into the organic silk matrix. A mutual benefit on the stability of both organic and inorganic components was observed in the nanocomposites. SF-HTlc displayed limited dissolution of hydrotalcite in acidic medium, enhanced mechanical properties, and higher protease resistance of silk protein. The transparency, flexibility, and acidic environment resistance of silk fibroin combined to the protective and reinforcing properties of hydrotalcites generate a hybrid material, which is very attractive for applications in recently reported silk based opto-electronic and photonics technologies.

Integration of a silk fibroin based film as a luminescent down-shifting layer in ITO-free organic solar cells

We report here a study on the integration of the silk fibroin (SF) protein in organic solar cells. The intrinsic low toxicity, natural availability, biodegradability, water processing, good film forming properties and capability to be doped with functional molecules of SF biopolymer inspired us to integrate it as a transparent and inert or functional bottom layer in organic solar cells. Water stable, optically transparent, smooth and homogeneous SF thin films (thickness ∼400 nm) were successfully prepared on glass and characterized. Then ITO-free bulk heterojunction (BHJ) solar cells employing P3HT:PC61BM as a standard active layer and a highly conductive PEDOT:PSS formulation as a semi-transparent anode were deposited over the SF films. As a result, the power conversion efficiency (PCE) of all silk-integrated BHJ solar cells was comparable to the references on bare glass. The ability of SF to act as a host matrix for functional moieties was exploited to give to the SF layer the functionality of a Luminescent Down-shifting film (LDS), as confirmed by the spectral response measurements, by using a water soluble stilbene derivative (Stb). The photovoltaic performance of all SF-based devices was significantly stable over time, overcoming the problems of the ITO-based reference cells after 70 days. Finally, flexible SF-integrated ITO-free solar cells were successfully fabricated on PET substrates.

Silk doped with a bio-modified dye as a viable platform for eco-friendly luminescent solar concentrators

The potential of fluorescent silk fibroin (SF) as a fully water-processable platform for an application in luminescent solar concentrators (LSCs) is shown. SF preserves its mechanical properties when doped with a bio-modified dye and the dye shows enhanced fluorescence when embedded in silk. These features, combined with high optical transparency and high refractive index, make SF a viable eco-friendly matrix for LSCs.

A nanostructured conductive bio-composite of silk fibroin–single walled carbon nanotubes

Silk fibroin (SF), a protein core fibre from the silkworm Bombyx mori, has huge potential to become a sustainable, biocompatible, and biodegradable material platform that can pave the way towards the replacement of plastic in the fabrication of bio-derived materials for a variety of technological and biomedical applications. SF has remarkable mechanical flexibility, controllable biodegradability, biocompatibility and is capable of drug/doping inclusion, stabilization and release. However, the dielectric properties of SF limit its potential as a direct bioelectronic interface in biomedical devices intended to control the bioelectrical activity of the cell for regenerative purposes. In this work, a novel wet templating method is proposed to generate nanostructured, conductive Silk Fibroin (SF) composite films. We combine the unusual properties of SF, such as its mechanical properties, its convenience and biocompatibility with the electrical conductivity and stiffness of Single Walled Carbon Nanotubes (SWCNTs). The presented SF-SWCNT composite displays a periodic architecture where SWCNTs are regularly and homogeneously distributed in the SF protein matrix. The morphological and chemo-physical properties of the nanocomposite are analysed and defined by SEM, Raman Spectroscopy, ATR-IR, UFM and contact angle analyses. Notably, the SF-SWCNT composite film is conductive, showing additional functionality compared to the dielectric properties of the bare SF film. Finally, SF-SWCNT is biocompatible and enables the growth of primary rat Dorsal Root Ganglion (DRG) neurons. Collectively our results demonstrate that the nanostructured, conductive, robust and biocompatible SF-SWCNT composite can be fabricated using a wet templating method, paving the way towards the fabrication and development of silk-based electronic devices for use in bioelectronic and biomedical applications.

A lysinated thiophene-based semiconductor as a multifunctional neural bioorganic interface.

Lysinated molecular organic semiconductors are introduced as valuable multifunctional platforms for neural cells growth and interfacing. Cast films of quaterthiophene (T4) semiconductor covalently modified with lysine-end moieties (T4Lys) are fabricated and their stability, morphology, optical/electrical, and biocompatibility properties are characterized. T4Lys films exhibit fluorescence and electronic transport as generally observed for unsubstituted oligothiophenes combined to humidity-activated ionic conduction promoted by the charged lysine-end moieties. The Lys insertion in T4 enables adhesion of primary culture of rat dorsal root ganglion (DRG), which is not achievable by plating cells on T4. Notably, on T4Lys, the number on adhering neurons/area is higher and displays a twofold longer neurite length than neurons plated on glass coated with poly-l-lysine. Finally, by whole-cell patch-clamp, it is shown that the biofunctionality of neurons cultured on T4Lys is preserved. The present study introduces an innovative concept for organic material neural interface that combines optical and iono-electronic functionalities with improved biocompatibility and neuron affinity promoted by Lys linkage and the softness of organic semiconductors. Lysinated organic semiconductors could set the scene for the fabrication of simplified bioorganic devices geometry for cells bidirectional communication or optoelectronic control of neural cells biofunctionality.

Bio-doping of regenerated silk fibroin solution and films: a green route for biomanufacturing

Silk fibroin (SF) is a natural biocompatible material that can be integrated in a variety of photonic systems and optoelectronics: i.e. organic lasing from dye-doped nano-structured silk film. In this context, biological incorporation of doping molecules into SF by means of feeding silk worms with dyes to their diet could be an innovative and eco-sustainable approach to obtain doped SF substrates, thus avoiding additional chemical processes and post-treatments of the protein solution. In the present work, we demonstrated that SF regenerated solutions and films containing rhodamine B (RhB) could be successfully obtained from the cocoons of Bombyx mori fed with a RhB-added diet (RhB-md-SF). Comparative analyses of optical and vibration characteristics of the RhB-md-SF solution and films with those of white SF blended with RhB (RhB-d-SF) revealed significant differences, suggesting that the silkworms metabolism could be involved in the binding mechanism of SF with RhB. In conclusion, we observed that the doping diet is a promising method for the green fabrication of SF-based optically active materials, and it opens novel routes for silk-based biophotonics.

Effect of different fabrication methods on the chemo-physical properties of silk fibroin films and on their interaction with neural cells

In this study, we investigated the influence of processing methods on the chemo-physical properties of silk fibroin (SF) film and on their interaction with neural cells. Structural, thermal and morphological analysis revealed a strong correlation between the conformation, stability and texture of silk films and the fabrication conditions. An increase in temperature, methanol treatment and the use of a microfluidic approach led to an improvement in SF film stability in terms of β-sheet content, mechanical resistance, dissolution and enzymatic degradation. An effect on the interaction of SF films with neural cells, through a modulation of the surface properties, was also observed. In particular, hydrophobic surfaces induce proliferation of astrocytes and neuron adhesion whereas hydrophilic surfaces promote a remarkable neurite outgrowth. A detailed knowledge of the effect of manufacturing parameters on SF film properties can facilitate and extend the applications of silk-based biomaterials in tissue engineering and drug release systems.

Selective MW-assisted surface chemical tailoring of hydrotalcites for fluorescent and biocompatible nanocomposites

ZnAl based hydrotalcite nanoparticles (ZnAl-HTlc NPs) were covalently modified by an organic oligothiophene fluorescent compound (T4Si) by using direct microwave (MW)-assisted silylation. Morphological and optical characterization proved that the MW-assisted method enables efficient grafting of the target fluorescent dye on the nanoparticles (NPs) surface in a few minutes with a predefined loading ratio only depending by the MW irradiation time. Moreover, the presented approach preserved the HTlc interlayer region, allowing further functionalization. Filmability, fluorescent properties, and biocompatibility of the silylated compound was also demonstrated highlighting the potential of the so-obtained lamellar NPs in applications broadening from diagnostic biomedical tools to photonics and sensing.

A Nanoscale Interface Promoting Molecular and Functional Differentiation of Neural Cells.

Potassium channels and aquaporins expressed by astrocytes are key players in the maintenance of cerebral homeostasis and in brain pathophysiologies. One major challenge in the study of astrocyte membrane channels in vitro, is that their expression pattern does not resemble the one observed in vivo. Nanostructured interfaces represent a significant resource to control the cellular behaviour and functionalities at micro and nanoscale as well as to generate novel and more reliable models to study astrocytes in vitro. However, the potential of nanotechnologies in the manipulation of astrocytes ion channels and aquaporins has never been previously reported. Hydrotalcite-like compounds (HTlc) are layered materials with increasing potential as biocompatible nanoscale interface. Here, we evaluate the effect of the interaction of HTlc nanoparticles films with primary rat neocortical astrocytes. We show that HTlc films are biocompatible and do not promote gliotic reaction, while favouring astrocytes differentiation by induction of F-actin fibre alignment and vinculin polarization. Western Blot, Immunofluorescence and patch-clamp revealed that differentiation was accompanied by molecular and functional up-regulation of both inward rectifying potassium channel Kir 4.1 and aquaporin 4, AQP4. The reported results pave the way to engineering novel in vitro models to study astrocytes in a in vivo like condition.