Reproducibility of non-invasive assessment of skin endothelial function using laser Doppler flowmetry and laser speckle contrast imaging

Cyril Puissant, Pierre Abraham, Sylvain Durand, Anne Humeau-Heurtier, Sébastien Faure, Georges Lefthériotis, Pascal Rousseau, Guillaume Mahé, Cyril Puissant, Pierre Abraham, Sylvain Durand, Anne Humeau-Heurtier, Sébastien Faure, Georges Lefthériotis, Pascal Rousseau, Guillaume Mahé

Abstract

Background: Endothelial dysfunction precedes atherosclerosis. Vasodilation induced by acetylcholine (ACh) is a specific test of endothelial function. Reproducibility of laser techniques such as laser-Doppler-flowmetry (LDF) and Laser-speckle-contrast-imaging (LSCI) to detect ACh vasodilation is debated and results expressions lack standardization. We aimed to study at a 7-day interval (i) the inter-subject reproducibility, (ii) the intra-subjects reproducibility, and (iii) the effect of the results expressions over variability.

Methods and results: Using LDF and LSCI simultaneously, we performed two different ACh-iontophoresis protocols. The maximal ACh vasodilation (peak-ACh) was expressed as absolute or normalized flow or conductance values. Inter-subject reproducibility was expressed as coefficient of variation (inter-CV,%). Intra-subject reproducibility was expressed as within subject coefficients of variation (intra-CV,%), and intra-class correlation coefficients (ICC). Fifteen healthy subjects were included. The inter-subject reproducibility of peak-ACh depended upon the expression of the results and ranged from 55% to 162% for LDF and from 17% to 83% for LSCI. The intra-subject reproducibility (intra-CV/ICC) of peak-ACh was reduced when assessed with LSCI compared to LDF no matter how the results were expressed and whatever the protocol used. The highest intra-subject reproducibility was found using LSCI. It was 18.7%/0.87 for a single current stimulation (expressed as cutaneous vascular conductance) and 11.4%/0.61 for multiple current stimulations (expressed as absolute value).

Conclusion: ACh-iontophoresis coupled with LSCI is a promising test to assess endothelial function because it is reproducible, safe, and non-invasive. N°: NCT01664572.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Typical recording using laser Doppler…
Figure 1. Typical recording using laser Doppler flowmetry (LDF) and laser speckle contrast imaging (LSCI) during protocol 1 and protocol 2.
Figure 2. Mean vasodilation patterns obtained with…
Figure 2. Mean vasodilation patterns obtained with ACh iontophoresis between D1 and D7 for each technique (LDF: laser Doppler flowmetry and LSCI: laser speckle contrast imaging) using a single current stimulation (0.1 mA and 30 s).
Figure 3. Mean vasodilation patterns obtained with…
Figure 3. Mean vasodilation patterns obtained with ACh iontophoresis between D1 and D7 for each technique (LDF: laser Doppler flowmetry and LSCI: laser speckle contrast imaging) using multiple current stimulations (0.1 mA and 10 s separated with a 2-minute interval).

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