Image and flow cytometric analysis of gold nanoparticle uptake by macrophages

Dror Fixler, Rinat Ankri, Ronald Weiss, Anja Grahnert, Susanne Melzer, Attila Tárnok

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

2 Scopus citations

Abstract

Background/Aim: In atherosclerosis stable and vulnerable atherosclerotic plaque types are distinguished that behave differently concerning rupture, thrombosis and clinical events. The stable are rich in M2 macrophages. The unstable are rich in inflammatory M1 macrophages and are highly susceptible to rupture, setting patients at risk for thrombotic events when they undergo invasive diagnosis such as coronary angiography. Therefore, novel approaches for non-invasive detection and classification of vulnerable plaques in vivo are needed. Whereas classical approaches fail to differentiate between both plaque types, a new biophotonic method (combination of the diffusion reflection (DR) method with flow cytometry (FCM) or image cytometry (IC)) to analyze gold nanoparticle (GNP) loading of plaques could overcome this limitation. Methods: Two types of GNP were used three variants of gold nanorods (GNRI with 40x18 nm, II 65x25 nm and III 52x13 nm in size) and gold nanospheres (GNS with an average diameter of 18.5 nm). The GNS had an absorption peak at 520 nm and the GNR at 630 nm. Monocytes were isolated from human buffy blood samples, differentiated into macrophages and their subtypes and labelled with GNR and GNS for 3 and 24 h. GNS and GNR loading were determined by FCM and/or IC. Macrophages within tissue-like phantoms were analyzed by the DR system. Results: After GNR labelling of macrophages the FCM light scatter values increased up to 3.7 fold and the DR slope changed from an average slope of 0.196 (macrophages only) to an average slope of 0.827 (macrophages labelled with GNR). But, GNRIII did not present much higher DR slopes than the control phantoms, indicating that macrophages take up GNRIII in a lower amount than GNRI or II. IC and microscopy showed that all particle variants were taken up by the cells in a heterogeneous fashion. Conclusion and outlook: The combination of FCM and DR measurements provides a potential novel, highly sensitive and non-invasive method for the identification of atherosclerotic vulnerable plaques, aimed to develop a potential tool for in vivo tracking. Further experiments will show, if different macrophage subtypes (M1 or M2) take up the particles differently and may thereby serve to distinguish stable from vulnerable plaques.

Original languageEnglish
Title of host publicationNanoscale Imaging, Sensing, and Actuation for Biomedical Applications XIII
EditorsAlexander N. Cartwright, Dan V. Nicolau
PublisherSPIE
ISBN (Electronic)9781628419559
DOIs
StatePublished - 2016
Externally publishedYes
EventNanoscale Imaging, Sensing, and Actuation for Biomedical Applications XIII - San Francisco, United States
Duration: 15 Feb 201617 Feb 2016

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume9721
ISSN (Print)1605-7422

Conference

ConferenceNanoscale Imaging, Sensing, and Actuation for Biomedical Applications XIII
Country/TerritoryUnited States
CitySan Francisco
Period15/02/1617/02/16

Keywords

  • atherosclerosis
  • cytometry
  • diffusion reflection
  • gold nanoparticle
  • macrophage
  • plaque classification

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