Biomarkers of exposure and effect in human biomonitoring of metal-based nanomaterials: their use in primary prevention and health surveillance.

Metal-based nanomaterials (MNMs) have gained particular interest in nanotechnology industry. They are used in various industrial processes, in biomedical applications or to improve functional properties of several consumer products. The widescale use of MNMs in the global consumer market has resulted in increases in the likelihood of exposure and risks to human beings. Human exposure to MNMs and assessment of their potential health effects through the concomitant application of biomarkers of exposure and effect of the most commonly used MNMs were reviewed in this paper. In particular, interactions of MNMs with biological systems and the nanobiomonitoring as a prevention tool to detect the early damage caused by MNMs as well as related topics like the influence of some physicochemical features of MNMs and availability of analytical approaches for MNMs testing in human samples were summarized in this review. The studies collected and discussed seek to increase the current knowledge on the internal dose exposure and health effects of MNMs, highlighting the advantages in using biomarkers in primary prevention and health surveillance.

Metals accumulation affects bone and muscle in osteoporotic patients: A pilot study.

Osteoporosis is the most common bone disease, characterized by decreased bone mineral density (BMD) and often associated to decreased muscle mass and function. Metal exposure plays a role in the pathophysiology of osteoporosis and affects also muscle quality. The aim of this study was to assess the association between metal levels in bone and muscle samples and the degeneration of these tissues. A total of 58 subjects (30 male and 28 female) was enrolled and classified in osteoporotic (OP, n = 8), osteopenic (Ope, n = 30) and healthy (CTR, n = 20) subjects, according to BMD measures. Femoral head bone samples and vastus lateralis muscle samples were collected during hip arthroplasty surgeries. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis showed increased levels of Al, Cd and Pb in OP and Ope bone tissue compared to CTR subjects (p = 0.04, p = 0.005 and p = 0.01, respectively). Whereas, increased levels of Co, Cd and Pb were measured in OP and Ope muscle tissues, compared to CTRs (p < 0.001, p = 0.02 and p = 0.01, respectively). In addition, Al, Cd and Pb levels in bone and Cd and Co levels in muscle were negatively correlated with BMD. A negative association among Co, Cd, Cr and Hg levels and muscle fibers diameter was also observed in muscle tissues. This study assessed that metal exposure can affects bone and muscle tissue quality and may contribute to the onset and progression of musculoskeletal diseases such as osteoporosis. Therefore, it is important to implement metal exposure assessment and their impact on disease development, in order to manage and prevent metal accumulation effects on bone and muscle quality.

An Update Overview on Mechanistic Data and Biomarker Levels in Cobalt and Chromium-Induced Neurodegenerative Diseases.

There is increasing evidence that the imbalance of metals as cobalt (Co) and chromium (Cr) may increase the risk of development and progression of neurodegenerative diseases (NDDs). The human exposure to Co and Cr is derived mostly from industry, orthopedic implants, and polluted environments. Neurological effects of Co and Cr include memory deficit, olfactory dysfunction, spatial disorientation, motor neuron disease, and brain cancer. Mechanisms of Co and Cr neurotoxicity included DNA damage and genomic instability, epigenetic changes, mitochondrial disturbance, lipid peroxidation, oxidative stress, inflammation, and apoptosis. This paper seeks to overview the Co and Cr sources, the mechanisms by which these metals induce NDDs, and their levels in fluids of the general population and patients affected by NDDs. To this end, evidence of Co and Cr unbalance in the human body, mechanistic data, and neurological symptoms were collected using in vivo mammalian studies and human samples.

The endocrine disrupting effects of nanoplastic exposure: A systematic review.

Good mechanical properties and low costs have led to a global expansion of plastic production and use. Unfortunately, much of this material can be released into the environment as a waste product and cleaved into micro- and nanoplastics (NPs) whose impact on the environment and human health is still largely unknown. Considering the growing worldwide awareness on exposure to chemicals that can act as endocrine disruptors, a systematic review was performed to assess the impact of NPs on the endocrine function of in vitro and in vivo models. Although a limited number of investigations is currently available, retrieved findings showed that NPs may induce changes in endocrine system functionality, with evident alterations in reproductive and thyroid hormones and gene expression patterns, also with a trans-generational impact. Nanoplastic size, concentration, and the co-exposure to other endocrine disrupting pollutants may have an influencing role on these effects. Overall, although it is still too early to draw conclusions regarding the human health risks derived from NPs, these preliminary results support the need for further studies employing a wider range of plastic polymer types, concentrations, and time points as well as species and life stages to address a great variety of endocrine outcomes and to achieve a broader and shared consensus on the role of NPs as endocrine disruptors.

Occupational Exposure to Metal Engineered Nanoparticles: A Human Biomonitoring Pilot Study Involving Italian Nanomaterial Workers.

Advances in nanotechnology have led to an increased use of engineered nanoparticles (ENPs) and the likelihood for occupational exposures. However, how to assess such exposure remains a challenge. In this study, a methodology for human biomonitoring, based on Single Particle Inductively Coupled Plasma Mass Spectrometry (SP-ICP-MS), was developed as a tool to assess the ENPs exposure of workers involved in nanomaterial activities in two Italian Companies. The method was validated for size and number concentration determination of Ag, Au, In2O3, Ir, Pd, Pt, and TiO2 NPs in urine and blood samples. The results showed the presence of In2O3 NPs in blood of exposed workers (mean, 38 nm and 10,371 particles/mL), but not in blood of controls. Silver, Au, and TiO2 NPs were found in urine (mean, Ag 29 nm and 16,568 particles/mL) or blood (mean, Au 15 nm and 126,635 particles/mL; TiO2 84 nm and 27,705 particles/mL) of workers, though these NPs were found also in controls. The presence of ENPs in both workers and controls suggested that the extra-professional exposure is a source of ENPs that cannot be disregarded. Iridium, Pd, and Pt NPs were not detected neither in blood nor in urine. Overall, the findings provided a rational basis to evaluate the exposure assessment to ENPs in cohorts of workers as part of risk assessment and risk management processes in workplaces.

Human biomonitoring and personal air monitoring. An integrated approach to assess exposure of stainless-steel welders to metal-oxide nanoparticles.

In welding, there is a potential risk due to metal-oxide nanoparticles (MONPs) exposure of workers. To investigate this possibility, the diameter and number particles concentration of MONPs were evaluated in different biological matrices and in personal air samples collected from 18 stainless-steel welders and 15 unexposed administrative employees engaged in two Italian mechanical engineering Companies. Exhaled breath condensate (EBC) and urine were sampled at pre-shift on 1st day and post-shift on 5th day of the workweek, while plasma and inhalable particulate matter (IPM) at post-shift on 5th day and analysed using the Single Particle Mass Spectrometry (SP-ICP-MS) technique to assess possible exposure to Cr2O3, Mn3O4 and NiO nanoparticles (NPs) in welders. The NPs in IPM at both Companies presented a multi-oxide composition consisting of Cr2O3 (median, 871,574 particles/m3; 70 nm), Mn3O4 (median, 713,481 particles/m3; 92 nm) and NiO (median, 369,324 particles/m3; 55 nm). The EBC of welders at both Companies showed Cr2O3 NPs median concentration significantly higher at post-shift (64,645 particles/mL; 55 nm) than at pre-shift (15,836 particles/mL; 58 nm). Significantly lower Cr2O3 NPs median concentration and size (7762 particles/mL; 44 nm) were observed in plasma compared to EBC of welders. At one Company, NiO NPs median concentration in EBC (22,000 particles/mL; 65 nm) and plasma (8248 particles/mL; 37 nm) were detected only at post-shift. No particles of Cr2O3, Mn3O4 and NiO were detected in urine of welders at both Companies. The combined analyses of biological matrices and air samples were a valid approach to investigate both internal and external exposure of welding workers to MONPs. Overall, results may inform suitable risk assessment and management procedures in welding operations.

An improved in vitro model simulating the feto-maternal interface to study developmental effects of potentially toxic compounds: The example of titanium dioxide nanoparticles.

The study of developmental effect of xenobiotics in humans is limited and often relies on epidemiological data. Whether and to which extent potentially toxic compounds may cross the placental barrier, and whether adverse effects on embryo development are the consequence of direct or indirect placental-mediated action is debated. The availability of in vitro models simulating the feto-maternal interface could contribute to elucidate this issue. Here, we report the development of a novel in vitro model using murine blastocyst derived trophoblast stem cells (TSC) to mimic the placental barrier and mouse embryoid bodies (EBs) to represent the embryonic tissues. We demonstrate that this model can be used for translocation studies, as well as embryotoxicity assessment of titanium dioxide nanoparticles (TiO2NPs). By evaluating trans-epithelial electrical resistance, translocation of fluorescein isothiocyanate-dextran beads and expression of junctional complex proteins, we show that TSCs cultured on transwell inserts under differentiating condition form syncytia. We also show that TiO2NPs administered in the upper transwell compartment are able to reach the lower compartment and interfere with EB differentiation when no TSC are cultured on the insert. On the contrary, when TSC are present, NPs translocate to a lesser extent and do not affect EB development. These results indicate that the proposed in vitro model is suitable to study the correlation between translocation and toxicity of TiO2NPs and suggest a direct effect of the particles on EB development. We propose that this model could be exploited to study developmental effect of other xenobiotics.

E-WASTE threatens health: The scientific solution adopts the one health strategy.

The aggressively extractive advanced technology industry thrives on intensive use of non-renewable resources and hyper-consumeristic culture. The environmental impact of its exponential growth means extreme mining, hazardous labour practices including child labour, and exposure burden to inorganic and organic hazardous chemicals for the environment and current and future human generations. Globally, processes such as in-country reduce, reuse and recycle have so far received less attention than outer-circle strategies like the uncontrolled dumping of e-waste in countries that are unprotected by regulatory frameworks. Here, in the absence of infrastructures for sound hazardous e-waste management, the crude recycling, open burning and dumping into landfills of e-waste severely expose people, animal and the environment. Along with economic, political, social, and cultural solutions to the e-waste global problem, the scientific approach based on risk analysis encompassing risk assessment, risk management and risk communication can foster a technical support to resist transgenerational e-waste exposure and health inequalities. This paper presents the latest public health strategies based on the use of integrated human and animal biomonitoring and appropriate biomarkers to assess and manage the risk of e-waste embracing the One Health approach. Advantages and challenges of integrated biomonitoring are described, along with ad-hoc biomarkers of exposure, effect and susceptibility with special focus on metals and metalloids. Indeed, the safe and sustainable management of novel technologies will benefit of the integration and coordination of human and animal biomonitoring.

In-house validation of AF4-MALS-UV for polystyrene nanoplastic analysis.

The suitability of asymmetric flow field-flow fractionation (AF4) coupled on-line to multi-angle light scattering (MALS) and UV diode array (UV-DAD) detectors was tested to simultaneously detect polystyrene nanoplastics (PS-NPs) and collect information about their size. A mixture of four sizes of PS-NPs at 20 nm, 60 nm, 100 nm and 200 nm was prepared by dilution with ultrapure deionized water and gentle mixing and was used as test sample for a polydisperse nanoplastic system. The AF4 method separated each single size of PS-NP mixture in a total time of 48 min by using 0.2% SDS as carrier solution. Then, the PS-NPs were sized and detected by following their MALS (90° scattering angle) and UV (215 nm) signals. Quality control (QC) performances as linearity, between-day repeatability, resolution factor, trueness/recovery, limit of detection (LoD) and selectivity were calculated, according to the ISO/TS 21362:2018. Method uncertainty was calculated following the ISO/TS 21748:2002 by summing between-day repeatability and trueness or recovery uncertainties. In-house validation results demonstrated good peak resolution and selectivity, R2 linearity of 0.998-0.999 in the range 50-1000 µg/mL, between-day repeatability of ca. 10%, trueness/recovery above 90% and LoD between 15 µg/mL (20 nm) and 33 µg/mL (200 nm). Expanded uncertainty was 16.1-17.9% on PS-NP size between 60 and 200 nm and 10.4-14.7% on PS-NP concentration between 100 and 1000 µg/mL. Compared to traditional single-technique analysis, this hyphenated method offers great promise for separating and analysing diverse populations of PS-NPs present in real matrices, which is critical for health and risk assessment studies and any regulatory action.

Silver and gold nanoparticles characterization by SP-ICP-MS and AF4-FFF-MALS-UV-ICP-MS in human samples used for biomonitoring.

The rapid increase in the use of silver (Ag) and gold (Au) nanoparticles (NPs) can be a potential risk to humans. Ag and Au NPs may enter the blood, accumulate in organs and be cleared from the body. It is therefore necessary to develop detection and quantification methods for Ag and Au NPs in human matrices. To this end, the inductively coupled plasma mass spectrometry was used as single particle detector (SP-ICP-MS) and coupled on-line with asymmetric flow field flow fractionation (AF4-FFF-ICP-MS), multi-angle scattering (MALS) and UV. Both methods enabled the qualitative and quantitative measurement of mixtures of Ag NPs (20, 60 and 100 nm) and Au NPs (5, 20, 40 and 60 nm) in human urine, blood and serum. Methods were validated by estimating linearity, limit of detection, resolution, repeatability, recovery and stability of Ag and Au NPs measurements in fluids. The SP-ICP-MS showed concentration limits for Ag and Au NPs lower than AF4-FFF-ICP-MS (pg/mL vs. ng/mL, respectively), while AF4-FFF-ICP-MS could detect smaller sized NPs (2-5 nm vs. 7-14 nm for SP-ICP-MS) with good resolution between monodispersed NPs fractions. In addition, MALS detector was more promising respect to higher sizes of Ag and Au NPs (>40 nm), while UV for lower sized particle (<20 nm). The observed performances will allow to use ICP-MS-based methods, also coupled to other detectors, to carry out human biomonitoring campaigns dedicated to the analysis of metallic NPs in the general population and in exposed subjects.

Quantitative analysis of metals and metal-based nano- and submicron-particles in tattoo inks.

Exposure to metals and metal-based nano- (NPs, 1-100 nm) and submicron-particles (SPs, 0.1-1 µm) contained in tattoo inks and related health safety is currently receiving a great deal of interest. Twenty inks of different brands and colours were sampled in Italy in 2019. The SemiQuant Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis allowed quantifying the concentration of 18 metals (Al, As, Ba, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Sn, Ti, Zn) in inks. The Single Particle ICP-MS was used to detect the diameters and concentration of NPs and SPs of 9 metals (Al, Co, Cr, Cu, Hg, Ni, Pb, Ti and Zn). Concentration of metals in tattoo inks were below the recommended concentrations reported in the Resolution ResAP (2008)1 indicating ink production have shifted to purer materials and best manufacturing practices. Regarding particles, Al was found at nano- (62-80 nm) and submicron-sizes (105-140 nm). Sizes of Cr, Cu, Pb and Zn were in the intervals 42-62 nm, 44-96 nm, 26-28 nm and 26-59 nm, respectively. Titanium was at submicron-diameters (166-383 nm). In addition, Cr and Ti particles accounted for the 47% and 80% of their total concentration, respectively. Tattooing practice exposed humans to metal-based NPs and SPs and the presence of a combination of particles of different metals and/or their dynamics (e.g., dissolution) may change their bioavailability and toxicity.