Highlights
- •Free DNA colocalizes with calcifications in human and rat arteries.
- •EDTA calcium chelation allowed to identify free DNA as background of calcification.
- •A calcification model based on intra-aortic free DNA infusion is presented.
- •Free DNA may serve as an anionic platform precipitating calcium phosphate.
Abstract
Background and aims
The arterial wall calcium score and circulating free DNA levels are now used in clinical
practice as biomarkers of cardiovascular risk. Calcium phosphate apatite retention
in the arterial wall necessitates precipitation on an anionic platform. Here, we explore
the role of tissue-free DNA as such a platform.
Methods
The first step consisted of histological observation of samples from human and rat
calcified arteries. Various stains were used to evaluate colocalization of free DNA
with calcified tissue (alizarin red, fluorescent Hoechst, DNA immunostaining and TUNEL
assay). Sections were treated by EDTA to reveal calcification background. Secondly,
a rat model of vascular calcifications induced by intra-aortic infusions of free DNA
and elastase + free DNA was developed. Rat aortas underwent a micro-CT for calcium
score calculation at 3 weeks. Rat and human calcifications were qualitatively characterized
using μFourier Transform Infrared Spectroscopy (μFTIR) and Field Emission-Scanning
Electron Microscopy (FE-SEM).
Results
Our histological study shows colocalization of calcified arterial plaques with free
DNA. In the intra-aortic infusion model, free DNA was able to penetrate into the arterial
wall and induce calcifications whereas no microscopic calcification was seen in control
aortas. The calcification score in the elastase + free DNA group was significantly
higher than in the control groups. Qualitative evaluation with μFTIR and FE-SEM demonstrated
typical calcium phosphate retention in human and rat arterial specimens.
Conclusions
This translational study demonstrates that free DNA could be involved in arterial
calcification formation by precipitating calcium phosphate apatite crystals in the
vessel wall.
Graphical abstract
Graphical Abstract
Keywords
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Article Info
Publication History
Published online: March 04, 2017
Accepted:
March 2,
2017
Received in revised form:
January 29,
2017
Received:
July 19,
2016
Identification
Copyright
© 2017 Elsevier B.V. All rights reserved.
