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01.12.2018 | Research | Ausgabe 1/2018 Open Access

Journal of Inflammation 1/2018

Anti-inflammatory cellular targets on neutrophils elucidated using a novel cell migration model and confocal microscopy: a clinical supplementation study

Zeitschrift:
Journal of Inflammation > Ausgabe 1/2018
Autoren:
T. Smith, L. Engelbrecht, C. Smith
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1186/​s12950-017-0177-0) contains supplementary material, which is available to authorized users.

Abstract

Background

In vivo studies have shown grape seed-derived polyphenols (GSP) to benefit in recovery from muscle injury by modulation of neutrophil infiltration into damaged tissue, thereby reducing secondary damage, as well as by facilitating an early anti-inflammatory macrophage phenotype shift. The current study aimed to provide data in this context from human models and to elucidate specific molecular targets of GSP.
Using a placebo-controlled, double-blind study design, eighteen normally healthy volunteers between the ages of 18–35 years old (13 female and 5 male) were orally supplemented with 140 mg/day of GSP for 2 weeks.
Blood samples (days 0 and 14) were comprehensively analysed for in vitro neutrophil chemokinetic capacity towards a chemotaxin (fMLP) using a novel neutrophil migration assay, in combination with live cell tracking, as well as immunostaining for neutrophil polarisation factors (ROCK, PI3K) at migration endpoint. Macrophage phenotype marker expression was assessed using flow cytometry.

Results

fMLP induced significant chemokinesis (P < 0.01), validating our model. GSP did not exert a significant effect on neutrophil chemokinesis in this non-compromised population, but tended to decrease overall ROCK expression in fMLP-stimulated neutrophils (P = 0.06). Macrophage phenotype markers CD274 and MPO – indicators of a pro-inflammatory M1 phenotype – seemed to be normalised relative to baseline expression levels after GSP treatment.

Conclusions

Current data suggest that GSP may have a modulatory effect on the ROCK-PI3K-PTEN system, but results in this normal population is not conclusive and should be confirmed in a larger, more inflamed population. Potential modulation of macrophage phenotype by GSP should be investigated further.
Zusatzmaterial
Additional file 1: Clinical trial consort diagram and basic comparative subject demographics. (DOCX 34 kb)
12950_2017_177_MOESM1_ESM.docx
Additional file 2: Full blood counts and clotting profiles performed on day 7 of supplementation. (DOCX 14 kb)
12950_2017_177_MOESM2_ESM.docx
Additional file 3: Representative flow cytometry scatter plots and fluorescence graphs for the analysis of CD66b, ICAM-1 and VCAM-1. (DOCX 62 kb)
12950_2017_177_MOESM3_ESM.docx
Additional file 4: Representative flow cytometry scatter plots for the analysis of macrophage phenotype marker expression. (DOCX 203 kb)
12950_2017_177_MOESM4_ESM.docx
Literatur
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