Analysis of the Capture and Influence of Nanoparticles on the Cytotoxic Effects of 7-Ketocholesterol on Cardiac Cells Investigation by Flow Cytometry and by Dynamic and Spectral Imaging Microscopy Associated with Factor Analysis of Medical Image Sequences

作者：Kahn Edmond^*; Pelloux Sophie; Baarine Mauhamad; Khalfaoui Taoufik; Ragot Kevin; Frouin Frederique; Riedinger Jean Marc; Tourneur Yves; Lizard Gerard

来源：Analytical and Quantitative Cytology and Histology, 2009, 31(6): 380-394.

• confocal microscopy
• FAMIS
• flow cytometry
• nanoparticles
• quantum dots

OBJECTIVE: To evaluate the capture of nanoparticles (quantum dots [QDs], fluorospheres) by nonbeating mouse cardiac cells (HL1-NB) cultured without or with 7-ketocholesterol (7KC) found at an increased level in the plasma of atherosclerotic patients and to simultaneously analyze their cytotoxic, proinflammatory and oxidative properties.
STUDY DESIGN: Flow cytometry (FCM), confocal laser scanning microscopy and subsequent factor analysis image processing were used to characterize the uptake of nanoparticles and to define their cytotoxicity, evaluated by enhanced permeability to SYTOX Green, release of lactate dehydrogenase (LDH) and morphologic nuclear changes determined with Hoechst 33342. Proinflammatory effects were estimated by enzyme linked immunoassay to quantify IL-8 and MCP-1 secretion. The overproduction of reactive oxygen species (ROS) was determined by FCM with hydroethidine.
RESULTS: Whereas the nano particles had no cytotoxic or inflammatory effects, they could stimulate ROS production. QDs were not incorporated. When 7KC was used, LDH release was enhanced and QDs potentialized IL-8 secretion. The incorporation and exit dynamics of nano particles were visualized to differentiate the emission spectra of SYTOX Green and nano particles and to precisely determine the cellular localization of nano particles.
CONCLUSION: The selected nanoparticles, which accumulate at the inner or outer cytoplasmic membrane level, can induce biologic activities and are able to interfere with those of chemically defined molecules such as 7KC. (Anal Quant Cytol Histol 2009;31:380-394)

• 出版日期2009-12