ae5089aa9f8899ed719adbedf8299358 pmc_33673220.pdf ea8ed9dc9ef7f467334ffd89580c23a4fdf20163 pmc_33673220.pdf 5fe79fe1368955ffb350fcbb2698f1849b992ae13f7dbb764a70bc56f8d45395 pmc_33673220.pdf Title: Functional and Morphological Changes Induced in Mytilus Hemocytes by Selected Nanoparticles Subject: Nanoparticles (NPs) show various properties depending on their composition, size, and surface coating, which shape their interactions with biological systems. In particular, NPs have been shown to interact with immune cells, that represent a sensitive surveillance system of external and internal stimuli. In this light, in vitro models represent useful tools for investigating nano-bio-interactions in immune cells of different organisms, including invertebrates. In this work, the effects of selected types of NPs with different core composition, size and functionalization (custom-made PVP-AuNP and commercial nanopolystyrenes PS-NH2 and PS-COOH) were investigated in the hemocytes of the marine bivalve Mytilus galloprovincialis. The role of exposure medium was evaluated using either artificial seawater (ASW) or hemolymph serum (HS). Hemocyte morphology was investigated by scanning electron microscopy (SEM) and different functional parameters (lysosomal membrane stability, phagocytosis, and lysozyme release) were evaluated. The results show distinct morphological and functional changes induced in mussel hemocytes depending on the NP type and exposure medium. Mussel hemocytes may represent a powerful alternative in vitro model for a rapid pre-screening strategy for NPs, whose utilization will contribute to the understanding of the possible impact of environmental exposure to NPs in marine invertebrates. Keywords: hemocytes; Mytilus; in vitro; scanning electron microscopy; immune response Author: Manon Auguste, Craig Mayall, Francesco Barbero, Matej Hočevar, Stefano Alberti, Giacomo Grassi, Victor F. Puntes, Damjana Drobne and Laura Canesi Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Feb 12 12:34:49 2021 CET ModDate: Fri Feb 12 13:41:32 2021 CET 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: 8362496 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZFDPUM+VnURWPalladioL Type 1 Custom yes yes no 10 0 JKFAQZ+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 TFQVKY+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 GNZBJW+URWPalladioL-BoldItal Type 1 Custom yes yes no 24 0 VVGXIB+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 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no 374 0 PCOAOA+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 390 0 PCOBMM+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 396 0 PCOBNM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 402 0 PCOBON+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 408 0 PCOBOO+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 411 0 PCOBOP+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 414 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 03:28:15 CEST RepresentationInformation: pmc_33673220.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:09:06 CEST Size: 8362496 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 650 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: Nanoparticle Synthesis and Characterization Destination: subsection.2.1 Item: Title: Mussels, Hemolymph Collection, Preparation of Hemolymph Serum and Hemocyte Monolayers and Exposure Conditions Destination: subsection.2.2 Item: Title: Hemocyte Morphology Using Scanning Electron Microscopy (SEM) Destination: subsection.2.3 Item: Title: Hemocyte Functional Assays Destination: subsection.2.4 Item: Title: Statistics Destination: subsection.2.5 Item: Title: Results Destination: section.3 Children: Item: Title: NP Characterization Destination: subsection.3.1 Item: Title: Effects of NPs on Hemocyte Morphology: SEM Destination: subsection.3.2 Item: Title: Hemocyte Functional Parameters Destination: subsection.3.3 Children: Item: Title: PVP-AuNP Destination: subsubsection.3.3.1 Item: Title: Amino-Modified Nanopolystyrene PS-NH2 Destination: subsubsection.3.3.2 Item: Title: Carboxy-Modified Nanopolystyrene PS-COOH Destination: subsubsection.3.3.3 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Functional and Morphological Changes Induced in Mytilus Hemocytes by Selected Nanoparticles Author: きちのはの〠ぁふでふびぴづ〬〠ぃひちどで〠きちべちぬぬ〬〠うひちのっづびっは〠あちひぢづひは〬〠きちぴづな〠えはㄍづぶちひ〬〠こぴづてちのは〠ぁぬぢづひぴど〬〠ぇどちっはねは〠ぇひちびびど〬〠ざどっぴはひ〠う〮〠ぐふのぴづび〬〠いちねなちのち〠いひはぢのづ〠ちのつ〠がちふひち〠ぃちのづびど Subject: Nanoparticles (NPs) show various properties depending on their composition, size, and surface coating, which shape their interactions with biological systems. In particular, NPs have been shown to interact with immune cells, that represent a sensitive surveillance system of external and internal stimuli. In this light, in vitro models represent useful tools for investigating nano-bio-interactions in immune cells of different organisms, including invertebrates. In this work, the effects of selected types of NPs with different core composition, size and functionalization (custom-made PVP-AuNP and commercial nanopolystyrenes PS-NH2 and PS-COOH) were investigated in the hemocytes of the marine bivalve Mytilus galloprovincialis. The role of exposure medium was evaluated using either artificial seawater (ASW) or hemolymph serum (HS). Hemocyte morphology was investigated by scanning electron microscopy (SEM) and different functional parameters (lysosomal membrane stability, phagocytosis, and lysozyme release) were evaluated. The results show distinct morphological and functional changes induced in mussel hemocytes depending on the NP type and exposure medium. Mussel hemocytes may represent a powerful alternative in vitro model for a rapid pre-screening strategy for NPs, whose utilization will contribute to the understanding of the possible impact of environmental exposure to NPs in marine invertebrates. 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