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Analysis of cytokines, interleukins and <t>endothelial</t> mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including GM-CSF, MCP-1 and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Analysis of cytokines, interleukins and <t>endothelial</t> mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including GM-CSF, MCP-1 and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Transcriptomic profile of cells and sEVs. (A–C) Transcriptomic profile of undifferentiated MSCs, SH‐MSCs and <t>endothelial</t> cells. (A) endothelial markers; (B) angiogenic growth factors; (C) hypoxia markers. The mRNA expression of 15 genes of interest was quantified using real‐time RT‐qPCR in MSCs (grey box, N = 7, n = 2), SH‐MSCs (blue box, N = 8, n = 2) (Mann–Whitney test, * p < 0.05; ** p < 0.01; *** p < 0.001). For (A) and (B), the level of mRNA expression by HUVECs is shown as a positive control (red box, N = 3, n = 2). (D) Transcriptomic profile of sEVs. mRNA expression of five genes of interest in MSC sEVs (grey box, N = 3, n = 2) and SH‐MSC sEVs (blue box, N = 7, n = 2) (Mann–Whitney test, * p < 0.05).
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PromoCell endothelial cell growth medium supplements
Transcriptomic profile of cells and sEVs. (A–C) Transcriptomic profile of undifferentiated MSCs, SH‐MSCs and <t>endothelial</t> cells. (A) endothelial markers; (B) angiogenic growth factors; (C) hypoxia markers. The mRNA expression of 15 genes of interest was quantified using real‐time RT‐qPCR in MSCs (grey box, N = 7, n = 2), SH‐MSCs (blue box, N = 8, n = 2) (Mann–Whitney test, * p < 0.05; ** p < 0.01; *** p < 0.001). For (A) and (B), the level of mRNA expression by HUVECs is shown as a positive control (red box, N = 3, n = 2). (D) Transcriptomic profile of sEVs. mRNA expression of five genes of interest in MSC sEVs (grey box, N = 3, n = 2) and SH‐MSC sEVs (blue box, N = 7, n = 2) (Mann–Whitney test, * p < 0.05).
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Analysis of cytokines, interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including GM-CSF, MCP-1 and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).

Journal: Redox Biology

Article Title: Fenofibrate attenuates the adverse effects of radiation on endothelial cells through modulation of ROS-NO signalling and inflammation

doi: 10.1016/j.redox.2025.103994

Figure Lengend Snippet: Analysis of cytokines, interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including GM-CSF, MCP-1 and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).

Article Snippet: The HCAECs, primary endothelial cells isolated from the coronary arteries of single donors (Promo Cell, Germany, C-12221) were cultured in ready to use Human Meso Endo Growth Medium (Cell Applications, USA, 212–500) in an incubator with humidified atmosphere at 37 °C with 5 % CO 2 in concentration of 5000–10000 cells/cm 2 .

Techniques: Irradiation, Flow Cytometry, Standard Deviation

Proposed mechanism of action of fenofibrate in irradiated HCAECs in-vitro . Irradiation reduced NO signalling via inactivation of the PI3K–AKT–eNOS pathway, whereas fenofibrate reactivated this pathway and restored NO production (A). Consistent with this, irradiation increased ROS generation, NOX activity, MDA levels, and 3-NT levels, while fenofibrate mitigated this effect (B). Irradiation also triggered an inflammatory response, which was counteracted by fenofibrate (C). The released cytokines contribute to inflammation, changes in cytoskeleton organisation and initiation of EndMT, and fenofibrate effectively attenuated the processes (D). Alterations in the pathways described above play a crucial role in the remodelling of vascular endothelial cells involved in the initiation and progression of atherosclerosis. Fenofibrate acts to reduce or restore the effects of irradiation on these pathways (E–F). Solid lines indicate correlations validated in this study, while dashed lines indicate unvalidated correlations.

Journal: Redox Biology

Article Title: Fenofibrate attenuates the adverse effects of radiation on endothelial cells through modulation of ROS-NO signalling and inflammation

doi: 10.1016/j.redox.2025.103994

Figure Lengend Snippet: Proposed mechanism of action of fenofibrate in irradiated HCAECs in-vitro . Irradiation reduced NO signalling via inactivation of the PI3K–AKT–eNOS pathway, whereas fenofibrate reactivated this pathway and restored NO production (A). Consistent with this, irradiation increased ROS generation, NOX activity, MDA levels, and 3-NT levels, while fenofibrate mitigated this effect (B). Irradiation also triggered an inflammatory response, which was counteracted by fenofibrate (C). The released cytokines contribute to inflammation, changes in cytoskeleton organisation and initiation of EndMT, and fenofibrate effectively attenuated the processes (D). Alterations in the pathways described above play a crucial role in the remodelling of vascular endothelial cells involved in the initiation and progression of atherosclerosis. Fenofibrate acts to reduce or restore the effects of irradiation on these pathways (E–F). Solid lines indicate correlations validated in this study, while dashed lines indicate unvalidated correlations.

Article Snippet: The HCAECs, primary endothelial cells isolated from the coronary arteries of single donors (Promo Cell, Germany, C-12221) were cultured in ready to use Human Meso Endo Growth Medium (Cell Applications, USA, 212–500) in an incubator with humidified atmosphere at 37 °C with 5 % CO 2 in concentration of 5000–10000 cells/cm 2 .

Techniques: Irradiation, In Vitro, Activity Assay

Transcriptomic profile of cells and sEVs. (A–C) Transcriptomic profile of undifferentiated MSCs, SH‐MSCs and endothelial cells. (A) endothelial markers; (B) angiogenic growth factors; (C) hypoxia markers. The mRNA expression of 15 genes of interest was quantified using real‐time RT‐qPCR in MSCs (grey box, N = 7, n = 2), SH‐MSCs (blue box, N = 8, n = 2) (Mann–Whitney test, * p < 0.05; ** p < 0.01; *** p < 0.001). For (A) and (B), the level of mRNA expression by HUVECs is shown as a positive control (red box, N = 3, n = 2). (D) Transcriptomic profile of sEVs. mRNA expression of five genes of interest in MSC sEVs (grey box, N = 3, n = 2) and SH‐MSC sEVs (blue box, N = 7, n = 2) (Mann–Whitney test, * p < 0.05).

Journal: Journal of Extracellular Biology

Article Title: Angiogenic Potential of Small Extracellular Vesicles Produced by Stimulated Mesenchymal Stromal Cells Under Hypoxic Conditions

doi: 10.1002/jex2.70105

Figure Lengend Snippet: Transcriptomic profile of cells and sEVs. (A–C) Transcriptomic profile of undifferentiated MSCs, SH‐MSCs and endothelial cells. (A) endothelial markers; (B) angiogenic growth factors; (C) hypoxia markers. The mRNA expression of 15 genes of interest was quantified using real‐time RT‐qPCR in MSCs (grey box, N = 7, n = 2), SH‐MSCs (blue box, N = 8, n = 2) (Mann–Whitney test, * p < 0.05; ** p < 0.01; *** p < 0.001). For (A) and (B), the level of mRNA expression by HUVECs is shown as a positive control (red box, N = 3, n = 2). (D) Transcriptomic profile of sEVs. mRNA expression of five genes of interest in MSC sEVs (grey box, N = 3, n = 2) and SH‐MSC sEVs (blue box, N = 7, n = 2) (Mann–Whitney test, * p < 0.05).

Article Snippet: Cells were cultured in Endothelial Cell Growth Medium (Promocell) supplemented with 5% sEV‐depleted FBS and 1% antibiotic‐antimycotic for 24 h. sEV‐depleted FBS was obtained after centrifugation at 100,000 × g for 1 h to exclude sEVs.

Techniques: Expressing, Quantitative RT-PCR, MANN-WHITNEY, Positive Control

Phenotypic characterisation of MSCs and SH‐MSCs by FC. (A) MSC (positive) markers: CD105, CD90, CD73 and CD44; negative markers: CD34, CD45, CD11b, CD19 and HLA‐DR. (B) Endothelial cell markers: CD31 (PECAM1), CD144 (VE‐Cadherin), CD309 (VEGF‐R2), intracellular NOS3, VEGF‐R1, CD146 (MCAM), CD106 (VCAM1) and intracellular vWF. (C) Cell surface receptors of interest: CD49b (α2‐subunit integrin), CD49e (α5‐subunit integrin), CD29 (β1‐subunit integrin), CD140a (PDGF‐Ra) and CD140b (PDGF‐Rb). Negative controls (light grey). MFI ratios are indicated.

Journal: Journal of Extracellular Biology

Article Title: Angiogenic Potential of Small Extracellular Vesicles Produced by Stimulated Mesenchymal Stromal Cells Under Hypoxic Conditions

doi: 10.1002/jex2.70105

Figure Lengend Snippet: Phenotypic characterisation of MSCs and SH‐MSCs by FC. (A) MSC (positive) markers: CD105, CD90, CD73 and CD44; negative markers: CD34, CD45, CD11b, CD19 and HLA‐DR. (B) Endothelial cell markers: CD31 (PECAM1), CD144 (VE‐Cadherin), CD309 (VEGF‐R2), intracellular NOS3, VEGF‐R1, CD146 (MCAM), CD106 (VCAM1) and intracellular vWF. (C) Cell surface receptors of interest: CD49b (α2‐subunit integrin), CD49e (α5‐subunit integrin), CD29 (β1‐subunit integrin), CD140a (PDGF‐Ra) and CD140b (PDGF‐Rb). Negative controls (light grey). MFI ratios are indicated.

Article Snippet: Cells were cultured in Endothelial Cell Growth Medium (Promocell) supplemented with 5% sEV‐depleted FBS and 1% antibiotic‐antimycotic for 24 h. sEV‐depleted FBS was obtained after centrifugation at 100,000 × g for 1 h to exclude sEVs.

Techniques: