Magnetron sputtering formation of Germanium nanoparticles for electrochemical Lithium intercalation.

In the drive towards increased lithium based battery capacity, germanium is an attractive material due to its very high lithium storage capacity, second only to silicon. The persistent down-side is the considerable embrittlement accompanying its remarkable volume expansion of close to 300%. A proven method to accommodate for this lattice expansion is the reduction of the size towards the nanoscale at which the fracturing is prevented by "breathing". In this work we employed a novel magnetron sputtering gas aggregation nanoparticle generator to create unprecedented layers of well-defined germanium nanoparticles with sizes below 20 nm. The electrochemical lithium intercalation was monitored by a suite of techniques under which Raman spectroscopy, which provided clear evidence of the presence of lithium inside the germanium nanoparticles. Moreover, the degree of lattice order was measured and correlated to the initial phases of the lithium-germanium alloy. This was corroborated by electron diffraction and optical absorption spectroscopy, of which the latter provided a strong dielectric change upon lithium intercalation. This study of low lithium concentrations inside layers of well-defined and very small germanium nanoparticles, forms a new avenue towards significantly increasing the lithium battery capacity.

Indoor air pollution impacts cardiovascular autonomic control during sleep and the inflammatory profile.

The present study explores the modifications of cardiovascular autonomic control (CAC) during wake and sleep time and the systemic inflammatory profile associated with exposure to indoor air pollution (IAP) in a cohort of healthy subjects. Twenty healthy volunteers were enrolled. Indoor levels of fine particulate matter (PM2.5), nitrogen dioxide (NO2) and volatile organic compounds (VOCs) were monitored using a portable detector for 7 days. Together, a 7-day monitoring was performed through a wireless patch that continuously recorded electrocardiogram, respiratory activity and actigraphy. Indexes of CAC during wake and sleep time were derived from the biosignals: heart rate and low-frequency to high-frequency ratio (LF/HF), index of sympathovagal balance with higher values corresponding to a predominance of the sympathetic branch. Cyclic variation of heart rate index (CVHRI events/hour) during sleep, a proxy for the evaluation of sleep apnea, was assessed for each night. After the monitoring, blood samples were collected to assess the inflammatory profile. Regression and correlation analyses were performed. A positive association between VOC exposure and the CVHRI (Δ% = +0.2% for 1 µg/m3 VOCs, p = 0.008) was found. The CVHRI was also positively associated with LF/HF during sleep, thus higher CVHRI values corresponded to a shift of the sympathovagal balance towards a sympathetic predominance (r = 0.52; p = 0.018). NO2 exposure was positively associated with both the pro-inflammatory biomarker TREM-1 and the anti-inflammatory biomarker IL-10 (Δ% = +1.2% and Δ% = +2.4%, for 1 µg/m3 NO2; p = 0.005 and p = 0.022, respectively). The study highlights a possible causal relationship between IAP exposure and higher risk of sleep apnea events, associated with impaired CAC during sleep, and a pro-inflammatory state counterbalanced by an increased anti-inflammatory response in healthy subjects. This process may be disrupted in vulnerable populations, leading to a harmful chronic pro-inflammatory profile. Thus, IAP may emerge as a critical and often neglected risk factor for the public health that can be addressed through targeted preventive interventions.

First in-Lab Testing of a Cost-Effective Prototype for PM2.5 Monitoring: The P.ALP Assessment.

The goal of the present research was to assess, under controlled laboratory conditions, the accuracy and precision of a prototype device (named 'P.ALP': Ph.D. Air-quality Low-cost Project) developed for PM2.5 concentration level monitoring. Indeed, this study follows a complementary manuscript (previously published) focusing on the in-field evaluation of the device's performance. Four P.ALP prototypes were co-located with the reference instrument in a calm-air aerosol chamber at the NIOSH laboratories in Pittsburgh, PA (USA), used by the Center for Direct Reading and Sensor Technologies. The devices were tested for 10 monitoring days under several exposure conditions. To evaluate the performance of the prototypes, different approaches were employed. After the data from the devices were stored and prepared for analysis, to assess the accuracy (comparing the reference instrument with the prototypes) and the precision (comparing all the possible pairs of devices) of the P.ALPs, linear regression analysis was performed. Moreover, to find out the applicability field of this device, the US EPA's suggested criteria were adopted, and to assess error trends of the prototype in the process of data acquisition, Bland-Altman plots were built. The findings show that, by introducing ad hoc calibration factors, the P.ALP's performance needs to be further implemented, but the device can monitor the concentration trend variations with satisfying accuracy. Overall, the P.ALP can be involved in and adapted to a wide range of applications because of the inexpensive nature of the components, the small dimensions, and the high data storage capacity.

Exposure to airborne particulate matter in working from office and working from home employees.

The main aim of this study is to quantitatively evaluate the differences, in terms of exposure to PM (particulate matter), between WFO (working-from-office) and WFH (working-from-home) conditions. Two measurement surveys were performed: a long-term and a short-term campaign, focused on the monitoring of personal exposure to size-fractionated PM in these different working conditions. Results of the long-term campaign show that the WFH subject is exposed to higher (up to 4 times) PM concentration, compared to the WFO subject. Specific activities performed by the subjects impacted their exposure concentrations, even if the most relevant contribution to total exposure was made by desk work. Results of the short-term campaign indicate that the subjects can be divided into two groups: subjects most exposed during the WFH mode (HE_H - Higher_Exposure_Home) and subjects most exposed during the WFO mode (HE_O - Higher_Exposure_Office). HE_H group is exposed to levels of pollutants up to 4 times higher in the domestic than in the office environment, during the moment of desk work. The HE_O group is exposed to higher (double) concentration levels during desk work during the WFO day. Considering the possible growing trend towards remote work it is important to evaluate these "new domestic offices" comprehensively.

Exposure Assessment and Monitoring of Antiblastic Drugs Preparation in Health Care Settings: A Systematic Review.

Several antiblastic drugs (ADs) are classified as carcinogenic, mutagenic, and/or toxic for reproduction. Despite established guidelines and safe handling technologies, ADs contamination of the work environments could occur in healthcare settings, leading to potential exposure of healthcare staff. This systematic review aims to investigate the main techniques and practices for assessing ADs occupational exposure in healthcare settings. The reviewed studies unveil that workplace contamination by ADs appears to be a still-topical problem in healthcare settings. These issues are linked to difficulties in guaranteeing: (i) the adherence to standardized protocols when dealing with ADs, (ii) the effective use of personal protective equipment by operators involved in the administration or management of ADs, (iii) a comprehensive training of the healthcare personnel, and (iv) a thorough health surveillance of exposed workers. A "multi-parametric" approach emerges as a desirable strategy for exposure assessment. In parallel, exposure assessment should coincide with the introduction of novel technologies aimed at minimizing exposure (i.e., risk management). Assessment must consider various departments and health operators susceptible to ADs contamination, with a focus extended beyond worst-case scenarios, also considering activities like surface cleaning and logistical tasks related to ADs management. A comprehensive approach in ADs risk assessment enables the evaluation of distinct substance behaviors and subsequent exposure routes, affording a more holistic understanding of potential risks.

The glycocalyx affects the mechanotransductive perception of the topographical microenvironment.

The cell/microenvironment interface is the starting point of integrin-mediated mechanotransduction, but many details of mechanotransductive signal integration remain elusive due to the complexity of the involved (extra)cellular structures, such as the glycocalyx. We used nano-bio-interfaces reproducing the complex nanotopographical features of the extracellular matrix to analyse the glycocalyx impact on PC12 cell mechanosensing at the nanoscale (e.g., by force spectroscopy with functionalised probes). Our data demonstrates that the glycocalyx configuration affects spatio-temporal nanotopography-sensitive mechanotransductive events at the cell/microenvironment interface. Opposing effects of major glycocalyx removal were observed, when comparing flat and specific nanotopographical conditions. The excessive retrograde actin flow speed and force loading are strongly reduced on certain nanotopographies upon strong reduction of the native glycocalyx, while on the flat substrate we observe the opposite trend. Our results highlight the importance of the glycocalyx configuration in a molecular clutch force loading-dependent cellular mechanism for mechanosensing of microenvironmental nanotopographical features.

Evolution and Applications of Recent Sensing Technology for Occupational Risk Assessment: A Rapid Review of the Literature.

Over the last decade, technological advancements have been made available and applied in a wide range of applications in several work fields, ranging from personal to industrial enforcements. One of the emerging issues concerns occupational safety and health in the Fourth Industrial Revolution and, in more detail, it deals with how industrial hygienists could improve the risk-assessment process. A possible way to achieve these aims is the adoption of new exposure-monitoring tools. In this study, a systematic review of the up-to-date scientific literature has been performed to identify and discuss the most-used sensors that could be useful for occupational risk assessment, with the intent of highlighting their pros and cons. A total of 40 papers have been included in this manuscript. The results show that sensors able to investigate airborne pollutants (i.e., gaseous pollutants and particulate matter), environmental conditions, physical agents, and workers' postures could be usefully adopted in the risk-assessment process, since they could report significant data without significantly interfering with the job activities of the investigated subjects. To date, there are only few "next-generation" monitors and sensors (NGMSs) that could be effectively used on the workplace to preserve human health. Due to this fact, the development and the validation of new NGMSs will be crucial in the upcoming years, to adopt these technologies in occupational-risk assessment.

Detection of clustered circulating tumour cells in early breast cancer.

Circulating tumour cell (CTC) clusters have been proposed to be major players in the metastatic spread of breast cancer, particularly during advanced disease stages. Yet, it is unclear whether or not they manifest in early breast cancer, as their occurrence in patients with metastasis-free primary disease has not been thoroughly evaluated. In this study, exploiting nanostructured titanium oxide-coated slides for shear-free CTC identification, we detect clustered CTCs in the curative setting of multiple patients with early breast cancer prior to surgical treatment, highlighting their presence already at early disease stages. These results spotlight an important aspect of metastasis biology and the possibility to intervene with anti-cluster therapeutics already during the early manifestation of breast cancer.

Features and Practicability of the Next-Generation Sensors and Monitors for Exposure Assessment to Airborne Pollutants: A Systematic Review.

In the last years, the issue of exposure assessment of airborne pollutants has been on the rise, both in the environmental and occupational fields. Increasingly severe national and international air quality standards, indoor air guidance values, and exposure limit values have been developed to protect the health of the general population and workers; this issue required a significant and continuous improvement in monitoring technologies to allow the execution of proper exposure assessment studies. One of the most interesting aspects in this field is the development of the "next-generation" of airborne pollutants monitors and sensors (NGMS). The principal aim of this review is to analyze and characterize the state of the art and of NGMS and their practical applications in exposure assessment studies. A systematic review of the literature was performed analyzing outcomes from three different databases (Scopus, PubMed, Isi Web of Knowledge); a total of 67 scientific papers were analyzed. The reviewing process was conducting systematically with the aim to extrapolate information about the specifications, technologies, and applicability of NGMSs in both environmental and occupational exposure assessment. The principal results of this review show that the use of NGMSs is becoming increasingly common in the scientific community for both environmental and occupational exposure assessment. The available studies outlined that NGMSs cannot be used as reference instrumentation in air monitoring for regulatory purposes, but at the same time, they can be easily adapted to more specific applications, improving exposure assessment studies in terms of spatiotemporal resolution, wearability, and adaptability to different types of projects and applications. Nevertheless, improvements needed to further enhance NGMSs performances and allow their wider use in the field of exposure assessment are also discussed.

Solid-Like Ordering of Imidazolium-Based Ionic Liquids at Rough Nanostructured Oxidized Silicon Surfaces.

The investigation of ionic liquids (ILs) confined in a solid porous matrix is of particular interest considering that these substances are increasingly used as an electrolyte in devices employing nanostructured nanoporous materials for the electrodes. Furthermore, the confinement of the ILs into a porous matrix would allow overcoming the difficulties of their packaging, leakage, and portability. In order to support the applications, a deeper understanding of the interaction of ILs with the nanoporous solid material and its increased interface is required. In this work, we report on the modification of morphological and mechanical properties of the imidazolium-based [Bmim][NTf2] IL upon surface spatial confinement on a cluster-assembled, nanostructured, rough, oxidized silicon (ns-SiOx) surface. An atomic force microscopy investigation revealed that upon the interaction with the ns-SiOx film, [Bmim][NTf2] locally rearranges into ordered, layered, stiff, and poorly conducting solid-like domains, coexisting with, and embedded into, the liquid IL film. The observed interfacial layering of [Bmim][NTf2] deposited on ns-SiOx suggests that the behavior of the IL-electrode interface in photoelectrochemical devices employing nanostructured nanoporous materials can be far more complex than expected under the hypothesis of an IL-based electrolyte in the stable liquid phase. The observed effects reported in this work could in principle also occur inside the bulk nanoporous matrix, where they could be further amplified by the extreme spatial confinement.

Electrostatic Double-Layer Interaction at the Surface of Rough Cluster-Assembled Films: The Case of Nanostructured Zirconia.

Here, we investigated the influence of the nanoscale surface morphology on the electrostatic double layer at corrugated surfaces in aqueous electrolytes. For this purpose, we have produced cluster-assembled nanostructured zirconium dioxide (ns-ZrO x, x ≈ 2) films with controlled morphological properties by supersonic cluster beam deposition (SCBD) and measured the double-layer interaction by atomic force microscopy with colloidal probes. SCBD allowed tuning the characteristic widths of the corrugated interface (root-mean-square roughness, correlation length) across a wide range of values, matching the width of the electrostatic double layer (Debye length) and the typical size of nanocolloids (proteins, enzymes, and catalytic nanoparticles). To accurately characterize the surface charge density in the high-roughness regime, we have developed a two-exponential model of the electrostatic force that explicitly includes roughness and better accounts for the roughness-induced amplification of the interaction. We were then able to observe a marked reduction of the isoelectric point of ns-ZrO x surfaces of increasing roughness. This result is in good agreement with our previous observations on cluster-assembled nanostructured titania films and demonstrates that the phenomenon is not limited to a specific material, but more generally depends on peculiar nanoscale morphological effects, related to the competition of the characteristic lengths of the system.

An Amperometric Sensor for Thiocholine Based on Cluster-Assembled Zirconia Modified Electrodes.

Here we report on the fabrication and characterization of cluster-assembled nanostructured zirconia electrodes for the electrochemical detection of enzymatically produced thiocholine. Zirconia nanostructures are produced by Supersonic Cluster Beam Deposition on thin gold films. This technique enables nanoscale control of the deposited film surface morphology, providing high active surface area for electrochemical detection of the analyte, along with high double-layer capacitance and suitable charge transfer resistance of the system. The electrochemical behavior of the electrodes has been characterized in the presence of the Potassium ferricyanide/Potassium ferrocyanide redox couple and the system performance showed to be enhanced starting from a thickness of the deposited layer of 60 nm. The electrochemical response for the oxidation of an enzymatic product was assessed by means of cyclic voltammetry, electrochemical impedance spectroscopy and chronoamperometry. The nanostructured zirconia film yields a good electrochemical detection of thiocholine. The limit of detection for thiocholine under working potential of 0.810-0.820 V versus reference was found to be comprised between 0.25 µM and 1.3 µM. Nanostructured electrodes, combining gold and zirconia nanoparticles can be implemented as functional transducers in biosensing devices, for example based on Acetylcholinesterase for electrochemical detection of polluting agents.

Precision and Accuracy of a Direct-Reading Miniaturized Monitor in PM2.5 Exposure Assessment.

The aim of this study was to evaluate the precision, accuracy, practicality, and potential uses of a PM2.5 miniaturized monitor (MM) in exposure assessment. These monitors (AirBeam, HabitatMap) were compared with the widely used direct-reading particulate matter monitors and a gravimetric reference method for PM2.5. Instruments were tested during 20 monitoring sessions that were subdivided in two different seasons to evaluate the performance of sensors across various environmental and meteorological conditions. Measurements were performed at an urban background site in Como, Italy. To evaluate the performance of the instruments, different analyses were conducted on 8-h averaged PM2.5 concentrations for comparison between direct-reading monitors and the gravimetric method, and minute-averaged data for comparison between the direct-reading instruments. A linear regression analysis was performed to evaluate whether the two measurement methods, when compared, could be considered comparable and/or mutually predictive. Further, Bland-Altman plots were used to determine whether the methods were characterized by specific biases. Finally, the correlations between the error associated with the direct-reading instruments and the meteorological parameters acquired at the sampling point were investigated. Principal results show a moderate degree of agreement between MMs and the reference method and a bias that increased with an increase in PM2.5 concentrations.

Exposure to airborne particles associated with the handling of graphene nanoplatelets.

BACKGROUND: It is recognized that engineering control measures are needed to reduce occupational exposure to engineered nanomaterials (NMs): of these, fume hoods are among the most widespread collective protection equipment used while handling NMs in occupational settings.  It is known that in some circumstances, handling NMs in fume hoods can result in a significant release of NMs. OBJECTIVE: To assess the effectiveness of fume hoods in reducing exposure while handling graphene nanoplatelets and to define the conditions that result in a lower dispersion of particles and thus less operator exposure. METHODS: An experimental protocol was established to monitor the variations of airborne particle concentrations while handling graphene in fume hoods (transferring and pouring). The measurement locations were at the laboratory, inside the hood and at operator's breathing zone. Handling tasks were performed under different operating conditions: the variable factors included hood face velocity and sash height. RESULTS: Results of this study indicate that the handling of graphene nanoplatelets may pose a potential risk of contamination of the work environment and hence exposure of the involved operators, if adequate control measures are not taken. In fact, when inadequate or not sufficiently cautionary operational conditions were utilized, non-negligible increases in airborne graphene particle concentrations during the nanomaterial manipulation phases were observed. CONCLUSIONS: Some operating conditions (e.g., face velocity, sash height) can be adjusted to avoid relevant personal exposure conditions and contamination of the work environment by NMs, thus ensuring safer conditions.

Conversion of nanoscale topographical information of cluster-assembled zirconia surfaces into mechanotransductive events promotes neuronal differentiation.

BACKGROUND: Thanks to mechanotransductive components cells are competent to perceive nanoscale topographical features of their environment and to convert the immanent information into corresponding physiological responses. Due to its complex configuration, unraveling the role of the extracellular matrix is particularly challenging. Cell substrates with simplified topographical cues, fabricated by top-down micro- and nanofabrication approaches, have been useful in order to identify basic principles. However, the underlying molecular mechanisms of this conversion remain only partially understood. RESULTS: Here we present the results of a broad, systematic and quantitative approach aimed at understanding how the surface nanoscale information is converted into cell response providing a profound causal link between mechanotransductive events, proceeding from the cell/nanostructure interface to the nucleus. We produced nanostructured ZrO2 substrates with disordered yet controlled topographic features by the bottom-up technique supersonic cluster beam deposition, i.e. the assembling of zirconia nanoparticles from the gas phase on a flat substrate through a supersonic expansion. We used PC12 cells, a well-established model in the context of neuronal differentiation. We found that the cell/nanotopography interaction enforces a nanoscopic architecture of the adhesion regions that affects the focal adhesion dynamics and the cytoskeletal organization, which thereby modulates the general biomechanical properties by decreasing the rigidity of the cell. The mechanotransduction impacts furthermore on transcription factors relevant for neuronal differentiation (e.g. CREB), and eventually the protein expression profile. Detailed proteomic data validated the observed differentiation. In particular, the abundance of proteins that are involved in adhesome and/or cytoskeletal organization is striking, and their up- or downregulation is in line with their demonstrated functions in neuronal differentiation processes. CONCLUSION: Our work provides a deep insight into the molecular mechanotransductive mechanisms that realize the conversion of the nanoscale topographical information of SCBD-fabricated surfaces into cellular responses, in this case neuronal differentiation. The results lay a profound cell biological foundation indicating the strong potential of these surfaces in promoting neuronal differentiation events which could be exploited for the development of prospective research and/or biomedical applications. These applications could be e.g. tools to study mechanotransductive processes, improved neural interfaces and circuits, or cell culture devices supporting neurogenic processes.

Biomechanics of Macrophages on Disordered Surface Nanotopography.

Macrophages are involved in every stage of the innate/inflammatory immune responses in the body tissues, including the resolution of the reaction, and they do so in close collaboration with the extracellular matrix (ECM). Simplified substrates with nanotopographical features attempt to mimic the structural properties of the ECM to clarify the functional features of the interaction of the ECM with macrophages. We still have a limited understanding of the macrophage behavior upon interaction with disordered nanotopography, especially with features smaller than 10 nm. Here, we combine atomic force microscopy (AFM), finite element modeling (FEM), and quantitative biochemical approaches in order to understand the mechanotransduction from the nanostructured surface into cellular responses. AFM experiments show a decrease of macrophage stiffness, measured with the Young's modulus, as a biomechanical response to a nanostructured (ns-) ZrOx surface. FEM experiments suggest that ZrOx surfaces with increasing roughness represent weaker mechanical boundary conditions. The mechanical cues from the substrate are transduced into the cell through the formation of integrin-regulated focal adhesions and cytoskeletal reorganization, which, in turn, modulate cell biomechanics by downregulating cell stiffness. Surface nanotopography and consequent biomechanical response impact the overall behavior of macrophages by increasing movement and phagocytic ability without significantly influencing their inflammatory behavior. Our study suggests a strong potential of surface nanotopography for the regulation of macrophage functions, which implies a prospective application relative to coating technology for biomedical devices.

An In-Field Assessment of the P.ALP Device in Four Different Real Working Conditions: A Performance Evaluation in Particulate Matter Monitoring.

This study aimed to assess the performance, in terms of precision and accuracy, of a prototype (called "P.ALP"-Ph.D. Air Quality Low-cost Project) developed for monitoring PM2.5 concentration levels. Four prototypes were co-located with reference instrumentation in four different microenvironments simulating real-world and working conditions, namely (i) office, (ii) home, (iii) outdoor, and (iv) occupational environments. The devices were evaluated for a total of 20 monitoring days (approximately 168 h) under a wide range of PM2.5 concentrations. The performances of the prototypes (based on the light-scattering working principle) were tested through different statistical methods. After the data acquisition and data cleaning processes, a linear regression analysis was performed to assess the precision (by comparing all possible pairs of devices) and the accuracy (by comparing the prototypes against the reference instrumentation) of the P.ALP. Moreover, the United States Environmental Protection Agency (US EPA) criteria were applied to assess the possible usage of this instrumentation, and to evaluate the eventual error trends of the P.ALP in the data storage process, Bland-Altman plots were also adopted. The outcomes of this study underlined that the P.ALP performed differently depending on the microenvironment in which it was tested and, consequently, on the PM2.5 concentrations. The device can monitor PM2.5 variations with acceptable results, but the performance cannot be considered satisfactory at extremely low and remarkably high PM2.5 concentrations. Thanks to modular components and open-source software, the tested device has the potential to be customized and adapted to better fit specific study design needs, but it must be implemented with ad hoc calibration factors depending on the application before being used in field.

Interaction of bacterial cells with cluster-assembled nanostructured titania surfaces: an atomic force microscopy study.

The nanoscale interaction of bacterial cells with solid surfaces is a key issue in biomedicine because it constitutes the first pathogenic event in the complex series of biofilm development on prosthetic devices. We report on an Atomic Force Microscopy study of the interaction of Escherichia coli and Pseudomonas aeruginosa bacterial cells with nanostructured titania thin films with controlled and reproducible nanometer-scale morphology, produced by assembling Ti clusters from the gas phase in a Supersonic Cluster Beam Deposition apparatus. The results demonstrate that bacterial adhesion and biofilm formation are significantly influenced by a pure physical stimulus, that is, the nanoscale variation of surface topography. The increase of nanoscale film roughness promotes bacterial adhesion with respect to flat substrates; remarkably, Pseudomonas aeruginosa cells lose their flagella on nanostructured TiO2 thin films upon adhesion, as opposed to same bacteria onto reference smooth glass substrates. Further, we have observed increased cell biovolume and other biofilm properties on nanostructured substrates in comparison with smooth glasses. These findings suggest that the design of innovative biomaterials with a suitable patterning of biomaterials surfaces can be an effective approach to control the adhesion of microorganisms to in vivo implant surfaces with active biological functionalities.

Engineering the structural and electrical interplay of nanostructured Au resistive switching networks by controlling the forming process.

Networks of random-assembled gold clusters produced in the gas phase show resistive switching (RS) activity at room temperature and they are suitable for the fabrication of devices for neuromorphic data processing and classification. Fully connected cluster-assembled nanostructured Au films are characterized by a granular structure rich of interfaces, grain boundaries and crystalline defects. Here we report a systematic characterization of the electroforming process of the cluster-assembled films demonstrating how this process affects the interplay between the nano- and mesoscale film structure and the neuromorphic characteristics of the resistive switching activity. The understanding and the control of the influence of the resistive switching forming process on the organization of specific structures at different scales of the cluster-assembled films, provide the possibility to engineer random-assembled neuromorphic architectures for data processing task.

Nanotopography and Microconfinement Impact on Primary Hippocampal Astrocyte Morphology, Cytoskeleton and Spontaneous Calcium Wave Signalling.

Astrocytes' organisation affects the functioning and the fine morphology of the brain, both in physiological and pathological contexts. Although many aspects of their role have been characterised, their complex functions remain, to a certain extent, unclear with respect to their contribution to brain cell communication. Here, we studied the effects of nanotopography and microconfinement on primary hippocampal rat astrocytes. For this purpose, we fabricated nanostructured zirconia surfaces as homogenous substrates and as micrometric patterns, the latter produced by a combination of an additive nanofabrication and micropatterning technique. These engineered substrates reproduce both nanotopographical features and microscale geometries that astrocytes encounter in their natural environment, such as basement membrane topography, as well as blood vessels and axonal fibre topology. The impact of restrictive adhesion manifests in the modulation of several cellular properties of single cells (morphological and actin cytoskeletal changes) and the network organisation and functioning. Calcium wave signalling was observed only in astrocytes grown in confined geometries, with an activity enhancement in cells forming elongated agglomerates with dimensions typical of blood vessels or axon fibres. Our results suggest that calcium oscillation and wave propagation are closely related to astrocytic morphology and actin cytoskeleton organisation.