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Transfer studies of polystyrene nanoparticles in the ex vivo human placenta perfusion model: key sources of artifacts

Grafmueller, Stefanie, Manser, Pius, Diener, Liliane, Maurizi, Lionel, Diener, Pierre-André, Hofmann, Heinrich, Jochum, Wolfram, Krug, Harald F., Buerki-Thurnherr, Tina, von Mandach, Ursula, Wick, Peter·2015·Science and Technology of Advanced Materials
DOI 10.1088/1468-6996/16/4/044602PMC PMC5090179
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Figure 1. (a) TEM micrographs of PS bead suspensions in DD water with low and high magnification (upper left corner). (b) Size distribution of PS beads in DD water and PM was determined by nanoparticle-tracking analysis.
Figure 1. (a) TEM micrographs of PS bead suspensions in DD water with low and high magnification (upper left corner). (b) Size distribution of PS beads in DD water and PM was determined by nanoparticle-tracking analysis.CC BY · PubMed Central
Figure 2. (a), (b) Perfusion profiles after ex vivo human placental perfusion of amine-modified PS beads N1 (n = 1), N3 (n = 4), N4 (n = 3), N5, N6, N7 (all n = 3), N8 (n = 1). (c) Perfusion profiles of plain and carboxylate-modified PS beads P1, P2, C1 and C2 of at least 3 independent experiments. Data are expressed as mean particle concentration (μg mL–1) in the fetal (F) and maternal (M) circulation determined by fluorescence measurement at the indicated time points ± SD. The red line indicates the initially added concentration of PS beads (25 μg mL−1). (d) Percentage of plain and carboxylate-modified PS beads (P1, P2, C1, C2) in the fetal circulation after 6 h of perfusion compared to the initially added particle amount (mean ± SD of at least 3 independent experiments). (∗) p < 0.05, (∗∗) p < 0.01.
Figure 2. (a), (b) Perfusion profiles after ex vivo human placental perfusion of amine-modified PS beads N1 (n = 1), N3 (n = 4), N4 (n = 3), N5, N6, N7 (all n = 3), N8 (n = 1). (c) Perfusion profiles of plain and carboxylate-modified PS beads P1, P2, C1 and C2 of at least 3 independent experiments. Data are expressed as mean particle concentration (μg mL–1) in the fetal (F) and maternal (M) circulation determined by fluorescence measurement at the indicated time points ± SD. The red line indicates the initially added concentration of PS beads (25 μg mL−1). (d) Percentage of plain and carboxylate-modified PS beads (P1, P2, C1, C2) in the fetal circulation after 6 h of perfusion compared to the initially added particle amount (mean ± SD of at least 3 independent experiments). (∗) p < 0.05, (∗∗) p < 0.01.CC BY · PubMed Central
Figure 3. To assess the presence of amino groups on the surface of PS beads N5, N6 and N7, a ninhydrin assay was performed. The amino acid glycine served as positive control. Absorption was determined at 550 nm and mean ± SD of 2 technical replicates is displayed.
Figure 3. To assess the presence of amino groups on the surface of PS beads N5, N6 and N7, a ninhydrin assay was performed. The amino acid glycine served as positive control. Absorption was determined at 550 nm and mean ± SD of 2 technical replicates is displayed.CC BY · PubMed Central
Figure 4. (a) Stability of the fluorescent signal of PS beads in PM at 37 °C over 72 h. The figure displays the percentage of initial fluorescence intensity at several time points compared to the initial fluorescence intensity (mean ± SD; n = 3). b) Leakage of the fluorescent dye from the large PS beads (>150 nm) during 72 h at 37 °C in PM. Data are expressed as percentage of free fluorescence compared to the total fluorescence intensity (mean ± SD; n = 3). (c) Elution of fluorescent dye of the large PS beads after 3 and 6 h of ex vivo placental perfusion. Data are expressed as percentage of free fluorescence compared to the total fluorescence intensity (mean ± SD of at least 2 independent experiments).
Figure 4. (a) Stability of the fluorescent signal of PS beads in PM at 37 °C over 72 h. The figure displays the percentage of initial fluorescence intensity at several time points compared to the initial fluorescence intensity (mean ± SD; n = 3). b) Leakage of the fluorescent dye from the large PS beads (>150 nm) during 72 h at 37 °C in PM. Data are expressed as percentage of free fluorescence compared to the total fluorescence intensity (mean ± SD; n = 3). (c) Elution of fluorescent dye of the large PS beads after 3 and 6 h of ex vivo placental perfusion. Data are expressed as percentage of free fluorescence compared to the total fluorescence intensity (mean ± SD of at least 2 independent experiments).CC BY · PubMed Central
Figure 5. Fluorescence microscopy of placental tissue after 6 h of ex vivo perfusion. Displayed are images from representative placenta perfusion experiments with two amine-modified PS beads (N3 and N4), which showed fluorescent dye leakage, and one amine-modified PS particle representative for an NP without fluorescence leakage (N6). As control placental perfusion without addition of particles was performed. Micrographs show PS beads (green) and DAPI-stained nuclei (blue). Due to autofluorescence, erythrocytes appear also in green. Red arrows indicate accumulated PS beads.
Figure 5. Fluorescence microscopy of placental tissue after 6 h of ex vivo perfusion. Displayed are images from representative placenta perfusion experiments with two amine-modified PS beads (N3 and N4), which showed fluorescent dye leakage, and one amine-modified PS particle representative for an NP without fluorescence leakage (N6). As control placental perfusion without addition of particles was performed. Micrographs show PS beads (green) and DAPI-stained nuclei (blue). Due to autofluorescence, erythrocytes appear also in green. Red arrows indicate accumulated PS beads.CC BY · PubMed Central

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Metadata

Authors
Grafmueller, S., Manser, P., Diener, L., Maurizi, L., Diener, P.-A., Hofmann, H., Jochum, W., Krug, H. F., Buerki-Thurnherr, T., von Mandach, U., & Wick, P.
Venue
Science and Technology of Advanced Materials
Publisher
Informa UK Limited
Published
2015-07-16
Volume
16
Issue
4
Pages
044602
ISSN
  • 1468-6996
  • 1878-5514
DOI
10.1088/1468-6996/16/4/044602
PMC
PMC5090179
Cited by
48
References
0
Language
en
Funders
European Union’s Seventh Framework Programme for research (263215, 309329)

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Grafmueller, S., Manser, P., Diener, L., Maurizi, L., Diener, P.-A., Hofmann, H., Jochum, W., Krug, H. F., Buerki-Thurnherr, T., von Mandach, U., & Wick, P. (2015). Transfer studies of polystyrene nanoparticles in the ex vivo human placenta perfusion model: key sources of artifacts. Science and Technology of Advanced Materials. https://doi.org/10.1088/1468-6996/16/4/044602