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Differential Aortic and Mitral Valve Interstitial Cell Mineralization and the Induction of Mineralization by Lysophosphatidylcholine In Vitro

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Abstract

Calcific aortic valve disease (CAVD) is a serious condition with vast uncertainty regarding the precise mechanism leading to valve calcification. This study was undertaken to examine the role of the lipid lysophosphatidylcholine (LPC) in a comparison of aortic and mitral valve cellular mineralization. The proportion of LPC in differentially calcified regions of diseased aortic valves was determined using thin layer chromatography (TLC). Next, porcine valvular interstitial cells (pVICs) from the aortic (paVICs) and mitral valve (pmVICs) were cultured with LPC (10−1–105 nM) and analyzed for cellular mineralization, alkaline phosphatase activity (ALPa), proliferation, and apoptosis. TLC showed a higher percentage of LPC in calcified regions of tissue compared to non-calcified regions. In pVIC cultures, with the exception of 105 nM LPC, increasing concentrations of LPC led to an increase in phosphate mineralization. Increased levels of calcium content were exhibited at 104 nm LPC application compared to baseline controls. Compared to pmVIC cultures, paVIC cultures had greater total phosphate mineralization, ALPa, calcium content, and apoptosis, under both a baseline control and LPC-treated conditions. This study showed that LPC has the capacity to promote pVIC calcification. Also, paVICs have a greater propensity for mineralization than pmVICs. LPC may be a key factor in the transition of the aortic valve from a healthy to diseased state. In addition, there are intrinsic differences that exist between VICs from different valves that may play a key role in heart valve pathology.

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Abbreviations

ARS:

Alizarin red S

ALPa:

Alkaline phosphatase activity

AS:

Aortic stenosis

βGP:

Beta-glycerophosphate

CAVD:

Calcific aortic valve disease

LPC:

Lysophosphatidylcholine

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OD:

Optical density

paVIC:

Porcine aortic valve interstitial cell

PKC:

Protein kinase C

pmVIC:

Porcine mitral valve interstitial cell

RFU:

Relative fluorescence units

RLU:

Relative luminescence units

RyR:

Ryanodine receptor

VIC:

Valvular interstitial cell

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Acknowledgments

This research was supported by National Institutes of Health R21 HL104377, T32 HL007812, T32 GM008362, and T32GM007330.

Conflict of interest

Dena Wiltz was supported by NIH training grant T32 GM008362. Richard Han was supported by NIH training grant T32 HL007812. Reid Wilson was supported by NIH training grant T32GM007330. Aditya Kumar declares that he has no conflicts of interest. Joel Morrisett received funding from NIH research grant R21 HL104377 and was the principal investigator of NIH training grant T32 HL007812. Jane Grande-Allen has served as a consultant for Edwards Lifesciences and received funding from NIH research grant R21 HL104377.

Human Subjects Declaration

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000. Informed consent was obtained from all patients for being included in the study.

Animal Studies Declaration

No animal studies were carried out by the authors for this article. Animal tissues were purchased from a commercial abattoir.

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Authors and Affiliations

  1. Department of Bioengineering, Rice University, 6100 Main St., MS 142, Houston, TX, 77005, USA

    Dena C. Wiltz, Richard I. Han, Reid L. Wilson, Aditya Kumar & K. Jane Grande-Allen

  2. Departments of Medicine and Biochemistry, Baylor College of Medicine, Houston, TX, USA

    Richard I. Han & Joel D. Morrisett

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  1. Dena C. Wiltz
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  2. Richard I. Han
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Correspondence to K. Jane Grande-Allen.

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Associate Editor Hanjoong Jo oversaw the review of this article.

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Wiltz, D.C., Han, R.I., Wilson, R.L. et al. Differential Aortic and Mitral Valve Interstitial Cell Mineralization and the Induction of Mineralization by Lysophosphatidylcholine In Vitro . Cardiovasc Eng Tech 5, 371–383 (2014). https://doi.org/10.1007/s13239-014-0197-3

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