fc74e3412ec0b589e2df2f7b0fa16f85 nanomaterials-15-01040.pdf 318450872f9e370b881bcda6881279364232e4f2 nanomaterials-15-01040.pdf 29351a4a623a71b76cbc957752a7aea2f0ae3c412bc877b7833c9fd5a6c958d4 nanomaterials-15-01040.pdf Title: Biological Impact of True-to-Life PET and Titanium-Doped PET Nanoplastics on Human-Derived Monocyte (THP-1) Cells Subject: In the environment, plastic waste degrades into small particles known as microplastics and nanoplastics (MNPLs), depending on their size. Given the potential harmful effects associated with MNPL exposure, it is crucial to develop environmentally representative particles for hazard assessment. These so-called true-to-life MNPLs are generated through in-house degradation of real-world plastic products. In this study, we produced titanium-doped nanoplastics (NPLs) from opaque polyethylene terephthalate (PET) milk bottles, which contain titanium dioxide as a filler. The resulting PET(Ti)-NPLs were thoroughly characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), mass spectrometry (MS), dynamic light scattering (DLS), -potential measurements, transmission electron microscopy (TEM), and Fourier-transform infrared (FTIR) spectroscopy. Human-derived THP-1 monocytes were employed to investigate particle uptake kinetics, dosimetry, and genotoxicity. A combination of flow cytometry and inductively coupled plasma mass spectrometry (ICP-MS) enabled the quantification of internalized particles, while the comet assay assessed DNA damage. The results revealed dose- and time-dependent effects of PET(Ti)-NPLs on THP-1 cells, particularly in terms of internalization. Titanium doping facilitated detection and influenced genotoxic outcomes. This study demonstrates the relevance of using environmentally representative nanoplastic models for evaluating human health risks and underscores the importance of further mechanistic research. Keywords: nanoplastics; polyethylene terephthalate (PET); tru-to life MNPLs Author: Aliro Villacorta, Michelle Morataya-Reyes, Lourdes Vela, Jéssica Arribas Arranz, Joan Martín-Perez, Irene Barguilla, Ricard Marcos and Alba Hernández Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Jul 4 12:54:26 2025 CEST ModDate: Fri Jul 4 12:58:36 2025 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3589993 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JNVRMG+EURM10 Type 1 Builtin yes yes yes 10 0 IAYHVP+VnURWPalladioL Type 1 Custom yes yes yes 15 0 GTBAPX+URWPalladioL-Roma Type 1 Custom yes yes yes 21 0 SATYBQ+URWPalladioL-Bold Type 1 Custom yes yes yes 27 0 KOKMPL+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 XEYJZU+CMSY10 Type 1 Builtin yes yes yes 58 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 81 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 91 0 DFCALH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 101 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 104 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 115 0 DFCFFJ+Arial TrueType WinAnsi yes yes no 125 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 128 0 DFCAMK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 131 0 DFCFFJ+Arial TrueType WinAnsi yes yes no 142 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 145 0 DFCAMK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 148 0 DFCALH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 158 0 DFCALJ+PalatinoLinotype TrueType WinAnsi yes yes no 161 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-09-09 03:10:52 CEST RepresentationInformation: nanomaterials-15-01040.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-09-08 11:57:18 CEST Size: 3589993 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 345 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Material and Methods Destination: section.2 Children: Item: Title: Particle Source and Characteristics Destination: subsection.2.1 Item: Title: Cell Culture Destination: subsection.2.2 Item: Title: Cell Viability Destination: subsection.2.3 Item: Title: Internalization Studies Destination: subsection.2.4 Children: Item: Title: Flow Cytometry Fluorescent Particle Detection Destination: subsubsection.2.4.1 Item: Title: Internalization Kinetics and Cell Complexity Analysis Destination: subsubsection.2.4.2 Item: Title: Confocal Microscopic Imaging Destination: subsubsection.2.4.3 Item: Title: Scanning and Transmission Electron Microscopy Destination: subsubsection.2.4.4 Item: Title: Cell Sorting and Titanium Quantification Destination: subsection.2.5 Item: Title: Effects on ROS Levels Destination: subsection.2.6 Item: Title: Genotoxicity Destination: subsection.2.7 Item: Title: Results and Discussion Destination: section.3 Children: Item: Title: Nanoparticle Characteristics Destination: subsection.3.1 Item: Title: NPL Cytotoxicity Destination: subsection.3.2 Item: Title: Internalization of PET(Ti)-NPLs Destination: subsection.3.3 Item: Title: Cell Sorting and Mass Spectroscopy Destination: subsection.3.4 Item: Title: Reactive Oxygen Species Related to PET(Ti)-NPL and TiO2-NP Exposure Destination: subsection.3.5 Item: Title: DNA Damage Induction Destination: subsection.3.6 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: Biological Impact of True-to-Life PET and Titanium-Doped PET Nanoplastics on Human-Derived Monocyte (THP-1) Cells Author: Aliro Villacorta, Michelle Morataya-Reyes, Lourdes Vela, Jéssica Arribas Arranz, Joan Martín-Perez, Irene Barguilla, Ricard Marcos and Alba Hernández Subject: In the environment, plastic waste degrades into small particles known as microplastics and nanoplastics (MNPLs), depending on their size. Given the potential harmful effects associated with MNPL exposure, it is crucial to develop environmentally representative particles for hazard assessment. These so-called true-to-life MNPLs are generated through in-house degradation of real-world plastic products. In this study, we produced titanium-doped nanoplastics (NPLs) from opaque polyethylene terephthalate (PET) milk bottles, which contain titanium dioxide as a filler. The resulting PET(Ti)-NPLs were thoroughly characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), mass spectrometry (MS), dynamic light scattering (DLS), -potential measurements, transmission electron microscopy (TEM), and Fourier-transform infrared (FTIR) spectroscopy. Human-derived THP-1 monocytes were employed to investigate particle uptake kinetics, dosimetry, and genotoxicity. A combination of flow cytometry and inductively coupled plasma mass spectrometry (ICP-MS) enabled the quantification of internalized particles, while the comet assay assessed DNA damage. The results revealed dose- and time-dependent effects of PET(Ti)-NPLs on THP-1 cells, particularly in terms of internalization. Titanium doping facilitated detection and influenced genotoxic outcomes. This study demonstrates the relevance of using environmentally representative nanoplastic models for evaluating human health risks and underscores the importance of further mechanistic research. 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