Research activities

Surface-acoustic-wave biosensors and microfluidics

Surface acoustic waves (SAWs) are acoustic waves that travel along the surface of an elastic material, with an amplitude that typically decays exponentially with depth into the substrate. Given their very superficial nature, SAWs are highly sensitive to surface perturbations of the substrate along which they propagate. For example, they can interact with liquid droplets or streams inducing macroscopic fluid manipulations or, in a different configuration, be exploited for sensing applications. The interest of the NeuroSens group in this field is to explore and study novel SAW-driven microfluidic phenomena, and apply this new knowledge to the fields of biosensing and cell biology.

A Rayleigh surface acoustic wave (R-SAW) resonator biosensor based on positive and negative reflectors with sub-nanomolar limit of detection

M. Agostini, G. Greco, M. Cecchini. Sensors and Actuators B: Chemical 254, 1–7 (2018).  
A label-free sub-nanomolar Rayleigh surface acoustic wave (R-SAW)-based resonator biosensor is demonstrated for biomolecular detection in liquid after drying. The biosensor comprises two interdigital transducers for R-SAW generation and two positive and negative reflectors to confine the acoustic energy in the sensitive area. We benchmark this biosensor against biotin-streptavidin binding, which is a stan- dard, well-known model for a variety of biosensing processes. The experiments demonstrate a limit of detection of 104 pM and a normalized sensitivity of −296 m^2 kg^−1. As a comparison with similar acoustic- wave based systems, both sensitivity and limit of detection are better than that of standard commercial gravimetric sensors (i.e., quartz-crystal-microbalances) and generally better than that of more common Love-SAW biosensors. Our biosensor has a dynamic range potentially compatible with several health- and safety-related assays, among all cancer biomarker detection.

Full-SAW Microfluidics-Based Lab-on-a-Chip for Biosensing

M. Agostini, G. Greco and M. Cecchini, IEEE Access, DOI: 10.1109/ACCESS.2019.2919000 (2019). 

Many approaches to diagnostic testing remain decades old. Well-established biosensing technologies (e.g., enzyme linked immunosorbent assays, radio-immunoassays) typically cannot fulfill the requirements of portability and ease of use necessary for point-of-care purposes. Several alternatives have been proposed (e.g., quartz-crystal-microbalances, electrochemical sensors, cantilevers, surface-plasmon-resonance sensors) but often lack high performance or still necessitate bulk ancillary instruments to operate. Here we present a highly sensitive, versatile and easily integrable microfluidic lab-on-a-chip (LoC) for biosensing, fully based on surface acoustic waves (SAWs). By using ultra-high-frequency resonator-biosensors, we show that it is possible to perform highly sensitive assays in complex media. This all-electrical readout platform is benchmarked with the biotin-streptavidin binding in presence of non-specific binding proteins (serum albumin) at physiological concentration. The benchmark experiments were performed with the idea of mimicking a biological fluid, in which other molecular species at high concentration are present together with the analytes. We demonstrate that this LoC can detect sub-nanomolar concentrations of analytes in complex media. As a comparison with similar acoustic-wave based systems, this full-SAW platform outperforms the standard commercial gravimetric sensors (i.e., quartz-crystal-microbalances) and the more common Love-SAW biosensors. This full-SAW LoC could be further developed for the detection of biomarkers in biological fluids.































































Nanotechnologies for the nervous system

The nervous system (NS) is in some ways the most complex and fascinating organ of the human body. Unfortunately, NS pathologies that lead to tissue loss are dramatically difficult to treat because of the negligible regenerative potential of the central nervous system (CNS) from one side, and of the very slow and ineffective repair mechanisms of peripheral nervous system (PNS) from the other side. The interest of the NeuroSens group in this field is to develop biocompatible nanostructured materials to help the heal of the PNS and cure and study CNS diseases. In particular, at the moment our attention is focused on textured surfaces for helping nerve regeneration and on nanotechnological methods for treating Globoid Cell Leukodystrophy (or Krabbe disease; OMIM #245200).

Dysregulated autophagy as a new aspect of the molecular pathogenesis of Krabbe disease

A. Del Grosso, L. Angella, I. Tonazzini, A. Moscardini, N. Giordano, M. Caleo, S. Rocchiccioli, M. Cecchini. Neurobiology of Disease 129, 195-207 (2019).  
Krabbe disease (KD) is a childhood leukodystrophy with no cure currently available. KD is due to a deficiency of a lysosomal enzyme called galactosyl-ceramidase (GALC) and is characterized by the accumulation in the nervous system of the sphingolipid psychosine (PSY), whose cytotoxic molecular mechanism is not fully known yet. Here, we study the expression of some fundamental autophagy markers (LC3, p62, and Beclin-1) in a KD murine model [the twitcher (TWI) mouse] by immunohistochemistry and Western blot. Moreover, the autophagy molecular process is also shown in primary fibroblasts from TWI and WT mice, with and without PSY treatment. Data demonstrate that large p62 cytoplasmic aggregates are present in the brain of both early and late symptomatic TWI mice. p62 expression is also upregulated in TWI sciatic nerves compared to that measured for WT nerves. In vitro data suggest that this effect might not be fully PSY-driven. Finally, we investigate in vitro the capability of autophagy inducers (Rapamycin, RAP and Resveratrol, RESV) to reinstate the WT phenotype in TWI cells. We show that RAP administration can partially restore the autophagy markers levels, while RESV cannot, indicating a line along which new therapeutic approaches can be developed.

Hierarchical thermoplastic rippled nanostructures regulate Schwann cell adhesion, morphology and spatial organization

C. Masciullo*, R. Dell’Anna*, I. Tonazzini, R. Böttger, G. Pepponi, M. Cecchini. Nanoscale, 9 (39), 14861-14874 (2017).  

Periodic ripples are a variety of anisotropic nanostructures that can be realized by ion beam irradiation on a wide range of solid surfaces. Only a few authors have investigated these surfaces for tuning the response of biological systems, probably because it is challenging to directly produce them in materials that well sustain long-term cellular cultures. Here, hierarchical rippled nanotopographies with a lateral periodicity of ∼300 nm are produced from a gold-irradiated germanium mold in polyethylene terephthalate (PET), a biocompatible polymer approved by the US Food and Drug Administration for clinical applications, by a novel three-step embossing process. The effects of nano-ripples on Schwann Cells (SCs) are studied in view of their possible use for nerve-repair applications. The data demonstrate that nano-ripples can enhance short-term SC adhesion and proliferation (3–24 h after seeding), drive their actin cytoskeleton spatial organization and sustain long-term cell growth. Notably, SCs are oriented perpendicularly with respect to the nanopattern lines. These results provide information about the possible use of hierarchical nano-rippled elements for nerve-regeneration protocols.