Single particle-inductively coupled plasma mass spectrometry (SP-ICPMS) is a promising technique able to generate the number based-particle size distribution (PSD) of nanoparticles (NPs) in aqueous suspensions. However, SP-ICPMS analysis is not consolidated as routine-technique yet and is not typically applied to real test samples with unknown composition. This work presents a methodology to detect, quantify and characterise the number-based PSD of Ag-NPs in different environmental aqueous samples (drinking and lake waters), aqueous samples derived from migration tests and consumer products using SP-ICPMS. The procedure is built from a pragmatic view and involves the analysis of serial dilutions of the original sample until no variation in the measured size values is observed while keeping particle counts proportional to the dilution applied. After evaluation of the analytical figures of merit, the SP-ICPMS method exhibited excellent linearity (r2>0.999) in the range (1-25) × 104 particlesmL-1 for 30, 50 and 80nm nominal size Ag-NPs standards. The precision in terms of repeatability was studied according to the RSDs of the measured size and particle number concentration values and a t-test (p = 95%) at the two intermediate concentration levels was applied to determine the bias of SP-ICPMS size values compared to reference values. The method showed good repeatability and an overall acceptable bias in the studied concentration range. The experimental minimum detectable size for Ag-NPs ranged between 12 and 15nm. Additionally, results derived from direct SP-ICPMS analysis were compared to the results conducted for fractions collected by asymmetric flow-field flow fractionation and supernatant fractions after centrifugal filtration. The method has been successfully applied to determine the presence of Ag-NPs in: lake water; tap water; tap water filtered by a filter jar; seven different liquid silver-based consumer products; and migration solutions (pure water and sweat simulant) from plasters. Results obtained by SP-ICPMS were supported by transmission electron microscopy and energy dispersive spectroscopy characterisation, suggesting that the proposed methodology can be applied as a positive screening test in the simultaneous quantification and size characterisation of Ag-NPs in samples of environmental interest.

In the era of personalised medicine, novel therapeutic approaches raise increasing hopes to address currently unmet medical needs by developing patient-customised treatments. Three-dimensional (3D) bioprinting is rapidly evolving and has the potential to obtain personalised tissue constructs and overcome some limitations of standard tissue engineering approaches. Bioprinting could support a wide range of biomedical applications, such as drug testing, tissue repair or organ transplantation. There is a growing interest for 3D bioprinting in the orthopaedic field, with remarkable scientific and technical advances. However, the full exploitation of 3D bioprinting in medical applications still requires efforts to anticipate the upcoming challenges in translating bioprinted products from bench to bedside. In this review we summarised current trends, advances and challenges in the application of 3D bioprinting for bone and cartilage tissue engineering. Moreover, we provided a detailed analysis of the applicable regulations through the 3D bioprinting process and an overview of available standards covering bioprinting and additive manufacturing.

Synthetic amorphous silica (SAS) has been used as food additive under the code E551 for decades and the agrifood sector is considered a main exposure vector for humans and environment. However, there is still a lack of detailed methodologies for the determination of SAS' particle size and concentration. This work presents the detection and characterization of NPs in eleven different food-grade SAS samples, following a reasoned and detailed sequential methodology. Dynamic Light Scattering (DLS), Multiangle Light Scattering (MALS), Asymmetric Flow-Field Flow Fractionation (AF4), Inductively Coupled Plasma Mass Spectrometry (ICPMS) and Transmission Electron Microscopy (TEM) were used. The suitability and limitations, information derived from each type of analytical technique and implications related to current EC Regulation 1169/2011 on the provision of food information to consumers are deeply discussed. In general the z-average, AF4 hydrodynamic diameters and root mean square (rms) radii measured were in good agreement. AF4-ICPMS coupling and pre channel calibration with silica NPs standards allowed the reliable detection of NPs below 100nm for ten of eleven samples (AF4 diameters between 20.6 and 39.8nm) and to quantify the mass concentration in seven different samples (at mgL(-1) concentration level). TEM characterisation included the determination of the minimum detectable size and subsequent measurement of the equivalent circle diameter (ECD) of primary particles and small aggregates, which were between 10.3 and 20.3nm. Because of the dynamic size application range is limited by the minimum detectable size, all the techniques in this work can be used only as positive tests.

In single particle inductively coupled plasma mass spectrometry (spICP-MS), the transport efficiency is fundamental for the correct determination of both particle number concentration and size. In the present study, transport efficiency was systematically determined on three different days with six carefully characterised gold nanoparticle (AuNP) suspensions and in seven European and US expert laboratories using different ICP-MS instruments and spICP-MS software. Both particle size—(TES)—and particle frequency—(TEF)—methods were applied. The resulting transport efficiencies did not deviate much under ideal conditions. The TEF method however systematically resulted in lower transport efficiencies. The extent of this difference (0–300% rel. difference) depended largely on the choice and storage conditions of the nanoparticle suspensions used for the determination. The TES method is recommended when the principal measurement objective is particle size. If the main aim of the measurement is the determination of the particle number concentration, the TEF approach could be preferred as it might better account for particle losses in the sample introduction system.

Defining the best combination of cells and biomaterials is a key challenge for the development of tendon tissue engineering (TE) strategies. Adipose-derived stem cells (ASCs) are ideal candidates for this purpose. In addition, controlled cell-based products adherent to good manufacturing practice (GMP) are required for their clinical scale-up. With this aim, in this study, ASC 3D bioprinting and GMP-compliant tenogenic differentiation were investigated. In detail, primary human ASCs were embedded within a nanofibrillar-cellulose/alginate bioink and 3D-bioprinted into multi-layered square-grid matrices. Bioink viscoelastic properties and scaffold ultrastructural morphology were analyzed by rheology and scanning electron microscopy (SEM). The optimal cell concentration for printing among 3, 6 and 9 × 10⁶ ASC/mL was evaluated in terms of cell viability. ASC morphology was characterized by SEM and F-actin immunostaining. Tenogenic differentiation ability was then evaluated in terms of cell viability, morphology and expression of scleraxis and collagen type III by biochemical induction using BMP-12, TGF-β3, CTGF and ascorbic acid supplementation (TENO). Pro-inflammatory cytokine release was also assessed. Bioprinted ASCs showed high viability and survival and exhibited a tenocyte-like phenotype after biochemical induction, with no inflammatory response to the bioink. In conclusion, we report a first proof of concept for the clinical scale-up of ASC 3D bioprinting for tendon TE.

Limonene oxidation products (LOPs) have gained interest on their harmful health effects over time. Recently, studies have shown that the selected LOPs: 4-oxopentanal (4-OPA), 3-isopropenyl-6-oxo-heptanal (IPOH) and 4-acetyl-1-methylcyclohexene (4-AMCH) have sensory irritation effects in mice and inflammatory effects in human lung cells. This study was therefore undertaken to investigate the potential capacity of 4-OPA, IPOH and 4-AMCH to cause cell membrane damage, oxidative stress and inflammation in human bronchial (16HBE14o-) and alveolar (A549) epithelial cell lines. Overall results suggest that 4-OPA, IPOH have cytotoxic effects on human lung cells that might be mediated by ROS: the highest concentration applied of IPOH [500 μM] enhanced ROS generation by 100-fold ± 7.7 (A549) and 230-fold ± 19.9 (16HBE14o-) compared to the baseline. 4-OPA [500 μM] increased ROS levels by 1.4-fold ± 0.3 (A549) and by 127-fold ± 10.5 (16HBE14o-), while treatment with 4-AMCH [500 μM] led to 0.9-fold ± 0.2 (A549) and 49-fold ± 12.8 (16HBE14o-) increase. IPOH [500 μM] caused a decrease in the thiol-state balance (e.g. after 2 h, GSH:GSSG was reduced by 37% compared to the untreated 16HBE14o-cells). 4-OPA [500 μM] decreased the GSH:GSSG by 1.3-fold change in A549 cells and 1.4-fold change in 16HBE14o-cells. No statistically significant decrease in the GSH:GSSG in A549 and 16HBE14o-cell lines was observed for 4-AMCH [500 μM]. In addition, IPOH and 4-OPA [31.2 μM] increased the amount of the inflammatory markers: RANTES, VEGF and EGF. On the other hand, 4-AMCH [31.2 μM] did not show inflammatory effects in A549 or 16HBE14o-cells. The 4-OPA, IPOH and 4-AMCH treatment concentration and time-dependently induce oxidative stress and/or alteration of inflammatory markers on human bronchial and alveolar cell lines.