Anticoagulants influence the performance of in vitro assays intended for characterization of nanotechnology-based formulations

Edward Cedrone, Barry W. Neun, Jamie Rodriguez, Alison Vermilya, Jeffrey D. Clogston, Scott E. McNeil, Yechezkel Barenholz, J. Szebeni, Marina A. Dobrovolskaia

Research output: Article

3 Citations (Scopus)

Abstract

The preclinical safety assessment of novel nanotechnology-based drug products frequently relies on in vitro assays, especially during the early stages of product development, due to the limited quantities of nanomaterials available for such studies. The majority of immunological tests require donor blood. To enable such tests one has to prevent the blood from coagulating, which is usually achieved by the addition of an anticoagulant into blood collection tubes. Heparin, ethylene diamine tetraacetic acid (EDTA), and citrate are the most commonly used anticoagulants. Novel anticoagulants such as hirudin are also available but are not broadly used. Despite the notion that certain anticoagulants may influence assay performance, a systematic comparison between traditional and novel anticoagulants in the in vitro assays intended for immunological characterization of nanotechnology-based formulations is currently not available. We compared hirudin-anticoagulated blood with its traditional counterparts in the standardized immunological assay cascade, and found that the type of anticoagulant did not influence the performance of the hemolysis assay. However, hirudin was more optimal for the complement activation and leukocyte proliferation assays, while traditional anticoagulants citrate and heparin were more appropriate for the coagulation and cytokine secretion assays. The results also suggest that traditional immunological controls such as lipopolysaccharide (LPS) are not reliable for understanding the role of anticoagulant in the assay performance. We observed differences in the test results between hirudin and traditional anticoagulant-prepared blood for nanomaterials at the time when no such effects were seen with traditional controls. It is, therefore, important to recognize the advantages and limitations of each anticoagulant and consider individual nanoparticles on a case-by-case basis.

Original languageEnglish
Article number12
JournalMolecules
Volume23
Issue number1
DOIs
Publication statusPublished - jan. 1 2018

Fingerprint

anticoagulants
Nanotechnology
nanotechnology
Anticoagulants
Assays
formulations
Hirudins
blood
Blood
heparins
Nanostructures
citrates
Nanostructured materials
Citric Acid
Heparin
In Vitro Techniques
hemolysis
leukocytes
product development
Immunologic Tests

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Cedrone, E., Neun, B. W., Rodriguez, J., Vermilya, A., Clogston, J. D., McNeil, S. E., ... Dobrovolskaia, M. A. (2018). Anticoagulants influence the performance of in vitro assays intended for characterization of nanotechnology-based formulations. Molecules, 23(1), [12]. https://doi.org/10.3390/molecules23010012

Anticoagulants influence the performance of in vitro assays intended for characterization of nanotechnology-based formulations. / Cedrone, Edward; Neun, Barry W.; Rodriguez, Jamie; Vermilya, Alison; Clogston, Jeffrey D.; McNeil, Scott E.; Barenholz, Yechezkel; Szebeni, J.; Dobrovolskaia, Marina A.

In: Molecules, Vol. 23, No. 1, 12, 01.01.2018.

Research output: Article

Cedrone, E, Neun, BW, Rodriguez, J, Vermilya, A, Clogston, JD, McNeil, SE, Barenholz, Y, Szebeni, J & Dobrovolskaia, MA 2018, 'Anticoagulants influence the performance of in vitro assays intended for characterization of nanotechnology-based formulations', Molecules, vol. 23, no. 1, 12. https://doi.org/10.3390/molecules23010012
Cedrone, Edward ; Neun, Barry W. ; Rodriguez, Jamie ; Vermilya, Alison ; Clogston, Jeffrey D. ; McNeil, Scott E. ; Barenholz, Yechezkel ; Szebeni, J. ; Dobrovolskaia, Marina A. / Anticoagulants influence the performance of in vitro assays intended for characterization of nanotechnology-based formulations. In: Molecules. 2018 ; Vol. 23, No. 1.
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