ae0f0d00afd5124390d83f711caf56aa pmc_33467414.pdf 6e0583b45c079ac5dcf22f656f2b428efc3b6396 pmc_33467414.pdf 7d60305e10a2ac02f89c8d09246c8631f2f814d22447a35763a74b37ab201ca5 pmc_33467414.pdf Title: Interaction between Macrophages and Nanoparticles: In Vitro 3D Cultures for the Realistic Assessment of Inflammatory Activation and Modulation of Innate Memory Subject: Understanding the modes of interaction between human monocytes/macrophages and engineered nanoparticles is the basis for assessing particle safety, in terms of activation of innate/inflammatory reactions, and their possible exploitation for medical applications. In vitro assessment of nanoparticle-macrophage interaction allows for examining the response of primary human cells, but the conventional 2D cultures do not reproduce the three-dimensional spacing of a tissue and the interaction of macrophages with the extracellular tissue matrix, conditions that shape macrophage recognition capacity and reactivity. Here, we have compared traditional 2D cultures with cultures on a 3D collagen matrix for evaluating the capacity gold nanoparticles to induce monocyte activation and subsequent innate memory in human blood monocytes in comparison to bacterial LPS. Results show that monocytes react to stimuli almost in the same way in 2D and 3D cultures in terms of production of TNF and IL-6, but that notable differences are found when IL-8 and IL-1Ra are examined, in particular in the recall/memory response of primed cells to a second stimulation, with the 3D cultures showing cell activation and memory effects of nanoparticles better. In addition, the response variations in monocytes/macrophages from different donors point towards a personalized assessment of the nanoparticle effects on macrophage activation. Keywords: monocytes; macrophages; gold nanoparticles; in vitro models; innate immunity; inflammation; innate memory; 2D cultures; 3D cultures Author: Benjamin J. Swartzwelter, Alessandro Verde, Laura Rehak, Mariusz Madej, Victor. F. Puntes, Anna Chiara De Luca, Diana Boraschi and Paola Italiani Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Jan 15 14:09:38 2021 CET ModDate: Fri Jan 15 14:16:16 2021 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1928601 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HJZZXQ+VnURWPalladioL Type 1 Custom yes yes no 10 0 VVSYYB+EURM10 Type 1 Builtin yes yes no 15 0 HHULRL+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 CMMYZZ+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 SVWVIY+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 NPLVPL+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 36 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 58 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 62 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 66 0 OADALT+CMSY10 Type 1 Builtin yes yes no 81 0 BAFHAJ+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 98 0 BAFHBJ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 104 0 BAFHCK+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 110 0 BAFHCL+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 116 0 BAFHCM+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 119 0 BAFHCN+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 122 0 BOKHCC+PalatinoLinotype TrueType WinAnsi yes yes no 132 0 ZMACFQ+MSAM10 Type 1 Builtin yes yes no 139 0 MXURFW+wasy9 Type 1 Builtin yes yes no 143 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 147 0 BAFHAJ+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 158 0 BAFKML+Wingdings-Regular CID TrueType Identity-H yes yes yes 164 0 BAFHBJ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 170 0 BAFHCK+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 176 0 BAFHCL+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 182 0 BAFHCM+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 185 0 BAFHCN+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 188 0 BAFKMI+Wingdings2 CID TrueType Identity-H yes yes yes 191 0 BAFKMJ+Webdings CID TrueType Identity-H yes yes yes 197 0 BAFKMK+Wingdings3 CID TrueType Identity-H yes yes yes 203 0 BAFHAJ+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 221 0 BAFHBJ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 227 0 BAFHCK+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 233 0 BAFHCL+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 239 0 BAFHCM+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 242 0 BAFHCN+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 245 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 03:34:44 CEST RepresentationInformation: pmc_33467414.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:08:36 CEST Size: 1928601 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 437 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Synthesis and Characterization of AuNP Destination: subsection.2.1 Children: Item: Title: AuNP Synthesis and Purification Destination: subsubsection.2.1.1 Item: Title: Nanoparticle Characterization Destination: subsubsection.2.1.2 Item: Title: LAL Assay Destination: subsection.2.2 Item: Title: Human Monocyte Isolation and Culture Destination: subsection.2.3 Item: Title: Cell Activation and Induction of Innate Memory Destination: subsection.2.4 Item: Title: Cytokine Analysis Destination: subsection.2.5 Item: Title: Statistical Analysis Destination: subsection.2.6 Item: Title: Results Destination: section.3 Children: Item: Title: Gold Nanoparticle Characterisation Destination: subsection.3.1 Item: Title: In Vitro Development of Macrophage Innate Memory in 2D vs. 3D Cultures Destination: subsection.3.2 Item: Title: Differences in the Primary Monocyte Reactivity in 2D vs. 3D Cultures Destination: subsection.3.3 Item: Title: Extinction of Inflammatory Activation in Macrophages Six Days after Priming Destination: subsection.3.4 Item: Title: Secondary Response of Unprimed Macrophages Destination: subsection.3.5 Item: Title: Memory Response of LPS-Primed Macrophages Destination: subsection.3.6 Item: Title: Memory Response of AuNP-Primed Macrophages Destination: subsection.3.7 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Interaction between Macrophages and Nanoparticles: In Vitro 3D Cultures for the Realistic Assessment of Inflammatory Activation and Modulation of Innate Memory Author: Benjamin J. Swartzwelter, Alessandro Verde, Laura Rehak, Mariusz Madej, Victor. F. Puntes, Anna Chiara De Luca, Diana Boraschi and Paola Italiani Subject: Understanding the modes of interaction between human monocytes/macrophages and engineered nanoparticles is the basis for assessing particle safety, in terms of activation of innate/inflammatory reactions, and their possible exploitation for medical applications. In vitro assessment of nanoparticle-macrophage interaction allows for examining the response of primary human cells, but the conventional 2D cultures do not reproduce the three-dimensional spacing of a tissue and the interaction of macrophages with the extracellular tissue matrix, conditions that shape macrophage recognition capacity and reactivity. Here, we have compared traditional 2D cultures with cultures on a 3D collagen matrix for evaluating the capacity gold nanoparticles to induce monocyte activation and subsequent innate memory in human blood monocytes in comparison to bacterial LPS. Results show that monocytes react to stimuli almost in the same way in 2D and 3D cultures in terms of production of TNF and IL-6, but that notable differences are found when IL-8 and IL-1Ra are examined, in particular in the recall/memory response of primed cells to a second stimulation, with the 3D cultures showing cell activation and memory effects of nanoparticles better. In addition, the response variations in monocytes/macrophages from different donors point towards a personalized assessment of the nanoparticle effects on macrophage activation. Keywords: monocytes; macrophages; gold nanoparticles; in vitro models; innate immunity; inflammation; innate memory; 2D cultures; 3D cultures Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Jan 15 14:09:38 CET 2021 ModDate: Fri Jan 15 14:16:16 CET 2021 ID: 0x801d49a1eac3d36de39f90c8576d96a2, 0x801d49a1eac3d36de39f90c8576d96a2 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: OADALT+CMSY10 FontSubset: true FirstChar: 0 LastChar: 21 FontDescriptor: FontName: OADALT+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: HHULRL+URWPalladioL-Ital FontSubset: true FirstChar: 3 LastChar: 122 FontDescriptor: FontName: HHULRL+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: MPUDFL+PazoMath FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: MPUDFL+PazoMath Flags: Symbolic FontBBox: -40, -283, 878, 946 FontFile: true Font: 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