Periodic Reporting for period 3 - PLASTICHEAL (Innovative tools to study the impact and mode of action of micro and nanoplastics on human health: towards a knowledge base for risk assessment)
Período documentado: 2024-04-01 hasta 2025-03-31
Humans are exposed to micro and nanoplastics (MNPLs) due to their ubiquitous presence in the environment, but there is only very limited information on the potential impact of this exposure on human health. Therefore, the current regulatory framework cannot ensure that MNPLs present in the air, and in food and beverage products are at safe levels for the population.
In this context, PlasticHeal aims at providing data, methodologies, experimental models and biological endpoints of effect for micro- and nanoplastics (MNPLs) human risk assessment.
The work carried out at CEA within the PlasticHeal Project investigates the effects of MNPLs on the immune system, mostly on B lymphocytes and macrophages, as these two cell types are known to be able to efficiently internalize particulate materials.
Regarding lymphocytes, CEA focused on the impact of PS (50-500 µm), PLA (100-500 µm) and true-to-life PET microparticles on B lymphocytes cells lines. After having demonstrated that these particles did not impact cell viability, DNA integrity, oxidative balance and inflammatory status on RajiB and BCL-1 cell lines work during this period has focused on RPMI1788 cells, which had been demonstrated to secrete significant basal levels of IgM (2-3 µg/mL). Except 50 nm PS-NH2 nanoparticles, none of the tested particles induced any cytotoxicity to the cell line. With respect to PS-NH2, the CI50 at 24 h exposure was close to 25 µg/mL. As for the other cell lines, the tested microparticles did not induce any significant DNA damage, oxidative stress and IL-8 secretion. Exposure also did not further increase IgM secretion.
Regarding macrophages, CEA had initiated proteomics studies on PET particles (produced and provided by the University Autonomous of Barcelona), PLA (poly lactic acid) and PCL (polycaprolactone) particles (from commercial sources), prior to the start of the reporting period. These studies have been finalized and completed by targeted functional studies. This led to a publication in 2024 on the PLA particles (DOI: 10.1039/d4en00335g open access) and an accepted paper on the PET particles (DOI: 10.1039/D4EN01063A open access). The manuscript on PCL particles is currently under revision. The highlights of these papers is the description of cross-toxicities with organic chemicals induced by PLA particles, as well as a decrease in the immune functions of macrophages induced by PET and PCL particles.
In addition, we had launched a series of proteomic studies on macrophages exposed not at a single dose of nanoparticles, as in all of the studies described above, but to repeated doses, which should provide a better appraisal of real-life exposures. The analyses of these proteomic data have been performed and the targeted validation experiments are almost finished. Writing the associated manuscripts will begin soon, with an aim of publication within 12 months after the end of the project.
In this context, PlasticHeal aims at providing data, methodologies, experimental models and biological endpoints of effect for micro- and nanoplastics (MNPLs) human risk assessment.
The work carried out at CEA within the PlasticHeal Project investigates the effects of MNPLs on the immune system, mostly on B lymphocytes and macrophages, as these two cell types are known to be able to efficiently internalize particulate materials.
Regarding lymphocytes, CEA focused on the impact of PS (50-500 µm), PLA (100-500 µm) and true-to-life PET microparticles on B lymphocytes cells lines. After having demonstrated that these particles did not impact cell viability, DNA integrity, oxidative balance and inflammatory status on RajiB and BCL-1 cell lines work during this period has focused on RPMI1788 cells, which had been demonstrated to secrete significant basal levels of IgM (2-3 µg/mL). Except 50 nm PS-NH2 nanoparticles, none of the tested particles induced any cytotoxicity to the cell line. With respect to PS-NH2, the CI50 at 24 h exposure was close to 25 µg/mL. As for the other cell lines, the tested microparticles did not induce any significant DNA damage, oxidative stress and IL-8 secretion. Exposure also did not further increase IgM secretion.
Regarding macrophages, CEA had initiated proteomics studies on PET particles (produced and provided by the University Autonomous of Barcelona), PLA (poly lactic acid) and PCL (polycaprolactone) particles (from commercial sources), prior to the start of the reporting period. These studies have been finalized and completed by targeted functional studies. This led to a publication in 2024 on the PLA particles (DOI: 10.1039/d4en00335g open access) and an accepted paper on the PET particles (DOI: 10.1039/D4EN01063A open access). The manuscript on PCL particles is currently under revision. The highlights of these papers is the description of cross-toxicities with organic chemicals induced by PLA particles, as well as a decrease in the immune functions of macrophages induced by PET and PCL particles.
In addition, we had launched a series of proteomic studies on macrophages exposed not at a single dose of nanoparticles, as in all of the studies described above, but to repeated doses, which should provide a better appraisal of real-life exposures. The analyses of these proteomic data have been performed and the targeted validation experiments are almost finished. Writing the associated manuscripts will begin soon, with an aim of publication within 12 months after the end of the project.
The development of the methodologies for MNPLs particle identification and quantification in environmental and biological samples have allowed monitoring of MNPLs in the environment –air, water, and food- and in human exposed populations –occupationally exposed workers-. Concurrently, MNPL representative materials have been prepared to perform a comprehensive hazard assessment of MNPLs using multiple models, test methods, exposure time points, and biological effects. The data generated has been processed by integrative analysis methods to obtain MNPLs' mechanistic insights and to identify key events with the potential to be consolidated as novel biomarkers of MNPLs long-term effects. Finally, all WPs have feed the human risk assessment framework developed in WP7, which has integrated stakeholders' and regulators' input.
Main PlasticHeal achievements
• Obtained a catalogue of real-life MNPLs test materials in the nano range: nPET, nPET-Ti, nPLA. In this last period, the catalogue was complemented with real-life NPLs that were commercially available (PTFE) and available from other Projects (PP-Talc, PVC, PA).
• Developed a PBK model to understand the kinetics (fate, biodistribution) of MNPLs.
• Developed and applied a set of NAMs useful to assess the kinetics and toxicity of MNPLs.
• Identified NPLs hazardous potential: genotoxicity, RT and GIT impairment, gut dysbiosis, inflammasome activation, immune alterations, long-term carcinogenesis.
• Identified NPLs mechanisms of action leading to the observed effects and identified a set of candidate new effect biomarkers. Some of these biomarkers showed useful when applied in a subset of occupationally exposed workers.
• Optimized analytical workflows: FTIR imaging with machine-learning assisted data analysis for MNPL >10 μm, and CRM for MNPL <10 μm (<1 μm). These technologies were successfully applied to the measurement of MNPLs in air and foods (external exposure), and human samples from real-life exposure scenarios (internal exposure).
• Generated exposure and effects human data (textile company, recycling company).
• Developed and applied the risk evaluation framework PlasticRiskCat.
A total of 19 outcomes were defined as Exploitable Results (ERs), with 12 prioritized as Key Exploitable Results (KERs) due to their higher maturity, greater potential for market adoption, and/or significant commercial or societal impact.
In addition, the project has led to the publication of over 50 papers in peer-reviewed journals, demonstrating its strong scientific contribution and visibility within the research community.
Main PlasticHeal achievements
• Obtained a catalogue of real-life MNPLs test materials in the nano range: nPET, nPET-Ti, nPLA. In this last period, the catalogue was complemented with real-life NPLs that were commercially available (PTFE) and available from other Projects (PP-Talc, PVC, PA).
• Developed a PBK model to understand the kinetics (fate, biodistribution) of MNPLs.
• Developed and applied a set of NAMs useful to assess the kinetics and toxicity of MNPLs.
• Identified NPLs hazardous potential: genotoxicity, RT and GIT impairment, gut dysbiosis, inflammasome activation, immune alterations, long-term carcinogenesis.
• Identified NPLs mechanisms of action leading to the observed effects and identified a set of candidate new effect biomarkers. Some of these biomarkers showed useful when applied in a subset of occupationally exposed workers.
• Optimized analytical workflows: FTIR imaging with machine-learning assisted data analysis for MNPL >10 μm, and CRM for MNPL <10 μm (<1 μm). These technologies were successfully applied to the measurement of MNPLs in air and foods (external exposure), and human samples from real-life exposure scenarios (internal exposure).
• Generated exposure and effects human data (textile company, recycling company).
• Developed and applied the risk evaluation framework PlasticRiskCat.
A total of 19 outcomes were defined as Exploitable Results (ERs), with 12 prioritized as Key Exploitable Results (KERs) due to their higher maturity, greater potential for market adoption, and/or significant commercial or societal impact.
In addition, the project has led to the publication of over 50 papers in peer-reviewed journals, demonstrating its strong scientific contribution and visibility within the research community.
PLASTICHEAL has produced data and a risk evaluation framework (PlasticRiskCat) that will impact on EU policies. Key findings from PLASTICHEAL and CUSP have been gathered in Policy brief #3 (https://cusp-research.eu/wp-content/uploads/2025/03/CUSP-Policy-Brief_03.pdf ) which also outlines policy recommendations to mitigate health risks and guide regulatory action.