Carotenoids co-localize with hydroxyapatite, cholesterol, and other lipids in calcified stenotic aortic valves. Ex vivo Raman maps compared to histological patterns

  • A. Bonetti University of Udine, Italy.
  • A. Bonifacio University of Trieste, Italy.
  • A. Della Mora University of Udine, Italy.
  • U. Livi University of Udine, Italy.
  • M. Marchini University of Udine, Italy.
  • F. Ortolani | fulvia.ortolani@uniud.it University of Udine, Italy.

Abstract

Unlike its application for atherosclerotic plaque analysis, Raman microspectroscopy was sporadically used to check the sole nature of bioapatite deposits in stenotic aortic valves, neglecting the involvement of accumulated lipids/lipoproteins in the calcific process. Here, Raman microspectroscopy was employed for examination of stenotic aortic valve leaflets to add information on nature and distribution of accumulated lipids and their correlation with mineralization in the light of its potential precocious diagnostic use. Cryosections from surgically explanted stenotic aortic valves (n=4) were studied matching Raman maps against specific histological patterns. Raman maps revealed the presence of phospholipids/triglycerides and cholesterol, which showed spatial overlapping with one another and Raman-identified hydroxyapatite. Moreover, the Raman patterns correlated with those displayed by both von-Kossa-calcium- and Nile-blue-stained serial cryosections. Raman analysis also provided the first identification of carotenoids, which co-localized with the identified lipid moieties. Additional fit concerned the distribution of collagen and elastin. The good correlation of Raman maps with high-affinity staining patterns proved that Raman microspectroscopy is a reliable tool in evaluating calcification degree, alteration/displacement of extracellular matrix components, and accumulation rate of different lipid forms in calcified heart valves. In addition, the novel identification of carotenoids supports the concept that valve stenosis is an atherosclerosis-like valve lesion, consistently with their previous Raman microspectroscopical identification inside atherosclerotic plaques.

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Author Biographies

A. Bonetti, University of Udine
Department of Experimental and Clinical MedicineAlois
A. Bonifacio, University of Trieste
Department of Engineering and Architecture
A. Della Mora, University of Udine
Department of Experimental and Clinical Medicine
U. Livi, University of Udine
Department of Experimental and Clinical Medicine
M. Marchini, University of Udine
Department of Experimental and Clinical Medicine
F. Ortolani, University of Udine
Department of Experimental and Clinical Medicine
Published
2015-04-20
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Original Papers
Keywords:
Valve calcification, stenosis, carotenoids, lipids, Raman microspectroscopy.
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How to Cite
Bonetti, A., Bonifacio, A., Della Mora, A., Livi, U., Marchini, M., & Ortolani, F. (2015). Carotenoids co-localize with hydroxyapatite, cholesterol, and other lipids in calcified stenotic aortic valves. Ex vivo Raman maps compared to histological patterns. European Journal of Histochemistry, 59(2). https://doi.org/10.4081/ejh.2015.2505