SiO 2 nanoparticles (NPs), in addition to their widespread utilization in consumer goods, are also being engineered for clinical use. They are considered to exert low toxicity both in vivo and in vitro, but the mechanisms involved in the cellular responses activated by these nanoobjects, even at non toxic doses, have not been characterized in detail.This is of particular relevance for their interaction with the nervous system: silica NPs are good candidates for nanoneuromedicine applications. Here, by using the GT1-7 neuronal cell line, derived from gonadotropin hormone releasing hormone (GnRH) neurons, we describe the mechanisms involved in the perturbation of calcium signaling, a key controller of neuronal function. At the non toxic dose of 20 µg mL -1 , 50nm SiO 2 NPs induce long lasting but reversible calcium signals, that in most cases show a complex oscillatory behavior. Using fluorescent NPs, we show that these signals do not depend on NPs internalization, are totally ascribable to calcium influx and are dependent in a complex way from size and surface charge. We provide evidence of the involvement of voltage-dependent and transient receptor potential-vanilloid 4 (TRPV4) TRPV4 channels.

The adsorption of bovine serum albumin (BSA) on two types of silica nanoparticles (NPs), one pyrolytic (P−SiO 2 ; namely AOX50 by Evonik) and the other colloidal (lab-made by using inverse micelles microemulsion, M−SiO 2 ), is studied. Both materials are characterized in terms of size of primary particles (by transmission electron microscopy), amounts (by thermogravimetry) and distribution of silanols (IR spectroscopy in controlled atmosphere, augmented by H/D isotopic exchange and reaction with VOCl 3 , to distinguish silanols actually located at the surface of nanoparticles), water contact angle, ζ−potential and dispersion state in water, PBS buffer and BSA solutions in PBS (by dynamic light scattering, DLS). Proteins are found to act as dispersing agent toward the large aggregates formed by both types of NPs in PBS buffer, although monodispersion was not attained in the conditions investigated. The problem of the determination of the silica surface actually available in NPs agglomerates for protein adsorption is addressed, and a model based on the external area of the agglomerates determined by DLS is proposed, supported by the trend of ζ−potential in dependence on the amount of adsorbed BSA and by the UV circular dichroism spectra of adsorbed proteins. The spectra reveal the occurrence of protein-protein interactions for BSA on P−SiO 2 , where multilayers of irreversibly adsorbed BSA molecules (i.e. a so called protein hard corona) are proposed to be formed. Conversely, the model indicates the formation of a sub-monolayer protein hard corona on M−SiO 2 . The difference in protein coverage appears to be related to differences in the distribution of surface silanols, more than to differences in ζ−potential.

Highly bright and photostable cyanine dye-doped silica nanoparticles, IRIS Dots, are developed, which can efficiently label human mesenchymal stem cells (hMSCs). The application procedure used to label hMSCs is fast (2 h), the concentration of IRIS Dots for efficient labeling is low (20 μg mL(-1) ), and the labeled cells can be visualized by flow cytometry, confocal microscopy, and transmission electron microscopy. Labeled hMSCs are unaffected in their viability and proliferation, as well as stemness surface marker expression and differentiation capability into osteocytes. Moreover, this is the first report that shows nonfunctionalized IRIS Dots can discriminate between live and early-stage apoptotic stem cells (both mesenchymal and embryonic) through a distinct external cell surface distribution. On the basis of biocompatibility, efficient labeling, and apoptotic discrimination potential, it is suggested that IRIS Dots can serve as a promising stem cell tracking agent.

Tumor resistance to chemotherapy represents an important challenge in modern oncology. Although platinum (Pt)-based drugs have demonstrated excellent therapeutic potential, their effectiveness in a wide range of tumors is limited by the development of resistance mechanisms. One of these mechanisms includes increased cisplatin sequestration/efflux by the copper-transporting ATPase, ATP7B. However, targeting ATP7B to reduce Pt tolerance in tumors could represent a serious risk because suppression of ATP7B might compromise copper homeostasis, as happens in Wilson disease. To circumvent ATP7B-mediated Pt tolerance we employed a high-throughput screen (HTS) of an FDA/EMA-approved drug library to detect safe therapeutic molecules that promote cisplatin toxicity in the IGROV-CP20 ovarian carcinoma cells, whose resistance significantly relies on ATP7B. Using a synthetic lethality approach, we identified and validated three hits (Tranilast, Telmisartan, and Amphotericin B) that reduced cisplatin resistance. All three drugs induced Pt-mediated DNA damage and inhibited either expression or trafficking of ATP7B in a tumor-specific manner. Global transcriptome analyses showed that Tranilast and Amphotericin B affect expression of genes operating in several pathways that confer tolerance to cisplatin. In the case of Tranilast, these comprised key Pt-transporting proteins, including ATOX1, whose suppression affected ability of ATP7B to traffic in response to cisplatin. In summary, our findings reveal Tranilast, Telmisartan, and Amphotericin B as effective drugs that selectively promote cisplatin toxicity in Pt-resistant ovarian cancer cells and underscore the efficiency of HTS strategy for identification of biosafe compounds, which might be rapidly repurposed to overcome resistance of tumors to Pt-based chemotherapy.