Article
Impact of amino-PVA coated nanoparticles on viability and cytokine secretion of human immune cells obtained from healthy donors and patients with rheumatoid arthritis
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Authors
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Cindy Strehl
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
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Timo Gaber
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
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Manuela Jakstadt
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
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Martin Hahne
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
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Paula Hoff
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
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Heinrich Hofmann
- École polytechnique fédérale de Lausanne EPFL, Institute of Materials Powder Technology Laboratory, Lausanne, Switzerland
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Gerd-Rüdiger Burmester
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
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Frank Buttgereit
- Charité - Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Berlin
| Published: | September 12, 2014 |
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Outline
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Background: Nanotechnology has developed into a key technology of the 21st century. Over the recent years, the number of nanotechnical products has received an enormous boost. Great efforts are currently being done to use this technology in rheumatology for diagnostic and therapeutic purposes. Therefore, crucial questions concern the safety aspects. We analysed the effects of PVA-coated-SPION on cell viability and cell activation with regard to cytokine secretion in a whole-blood-assay.
Methods: Venous blood, obtained from 18 healthy donors (HD) or 19 patients suffering from RA was collected. Whole blood was diluted 1:1 with RPMI 1640 culture medium in deep-well-plates, and cells were stimulated with LPS (1µg/ml), PHA (5µg/ml), PVA-SPION (1µg/ml, 10µg/ml, 100µg/ml, 1000µg/ml) or left untreated for 20h at 37°C. Cells were analysed for viability by flow cytometry, and supernatants were collected for quantification of cytokines via Bio-Plex®. Cell viability was analysed by a combined staining for cell specific surface markers (CD3; CD14; CD15; CD19) and apoptotic and dead cells using Annexin V and 7AAD. As a second approach, intracellular IL1β was analysed to identify the main producers of IL1β.
Results: For both HD and RA, we did not find any significant influence on cell viability at PVA-SPION concentrations <1000µg/ml. However, when analysing the impact of PVA-SPION on cytokine secretion, we demonstrated a significant dose dependent increase of several secreted cytokines in the cell culture supernatant (such as IL1β, IL4, IL6, IL8 or MIP1b for both HD and RA samples). When analysing IL1β secretion in more detail, we found a PVA-SPION concentration dependent increase in the percentage of IL1β positive cells. Furthermore, we observed that the major cell populations producing IL1β after incubation with PVA-SPION are granulocytes as well as monocytes.
Conclusion: PVA-SPION at concentrations up to 1000µg/ml do not increase the frequencies of apoptotic or dead human immune cells but do induce a PVA-SPION dose dependent cytokine secretion. We conclude that PVA-SPION represent a promising diagnostic and possibly also a therapeutic tool in rheumatic diseases, but prior to clinical use the impact of PVA-SPION on other human immune cells and the induction of cytokine secretion has to be further evaluated.
