An advanced method for quantitative measurements of cholesterol crystallization.

Autor: Halkias C; School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria 3083, Australia., Orth A; ARC Centre of Excellence for Nanoscale BioPhotonics (CNBP), School of Science, RMIT University, Melbourne, Victoria 3001, Australia; National Research Council of Canada, Ottawa, Canada., Feltis BN; School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria 3083, Australia., Macrides TA; School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria 3083, Australia., Gibson BC; ARC Centre of Excellence for Nanoscale BioPhotonics (CNBP), School of Science, RMIT University, Melbourne, Victoria 3001, Australia., Wright PFA; School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria 3083, Australia. Electronic address: paul.wright@rmit.edu.au.
Jazyk: angličtina
Zdroj: Biochimica et biophysica acta. Molecular and cell biology of lipids [Biochim Biophys Acta Mol Cell Biol Lipids] 2021 Mar; Vol. 1866 (3), pp. 158872. Date of Electronic Publication: 2020 Dec 23.
DOI: 10.1016/j.bbalip.2020.158872
Abstrakt: Background: Cholesterol crystallization within an atherosclerotic plaque significantly contributes to the acceleration of plaque rupture - a problematic event due to the current lack of specific treatments to prevent such formations. Modelling this pathogenic process is also difficult due to the lack of suitable experimental models that enable quantitative analysis of crystal formation and bioactivity screening of potential therapeutic compounds.
Aim: To develop an in vitro human cell model of cholesterol crystallization combined with an imaging system that incorporates both quantitative analysis and real-time continuous imaging of cholesterol crystal formation.
Methods and Results: An enhanced in vitro model of cholesterol crystallization was developed through the use of acetylated low-density lipoprotein (AcLDL) and 7-ketocholesterol as agents of foam cell induction within a human THP-1 monocytic cell line. Advanced confocal and polarizing microscopies were incorporated into the model so as to allow for quantitation of cholesterol crystallization, with the lipid-loaded group producing significantly greater numbers of cholesterol crystals than the untreated group. The utility of this system was also demonstrated by investigating the effects of the cholesterol-lowering drug lovastatin and therapeutic bile compound ursodeoxycholic acid (UDCA), showing that these drugs influence different aspects of cholesterol crystal formation.
Conclusions: The in vitro human THP-1 monocyte model of cholesterol crystallization provides an effective and efficient means of quantitating cholesterol crystallization in the pre-clinical stage of research. The model also allows for the screening of potentially therapeutic compounds that may be used in attenuating or preventing cholesterol crystallization.
(Copyright © 2020. Published by Elsevier B.V.)
Databáze: MEDLINE
Externí odkaz: