Journal: bioRxiv

Article Title: Mitochondrial Dysfunction in Endothelial Cells Drives Greater Vascular Impairment in Females with Diabetes-Associated Peripheral Artery Disease

doi: 10.1101/2025.09.22.677648

Figure Lengend Snippet: (A) Schematic illustrating site of tissue collection from below- and above knee amputations. (B) Myography showing increased endothelial dysfunction in female vs male patient arteries. Left , endothelial-dependent relaxation in response to acetylcholine (ACh). Right , No difference in vascular smooth muscle cell (VSMC), sodium nitroprusside (SNP)- mediated relaxation (n=3-5). (C) Left, representative image of microvessels in tibialis anterior (CD31 + SMA + , yellow arrows). Laminin (myocytes, white), CD31 (ECs, red) and SMA (pericytes, green); scale bar 50 μm. Right , quantification (n=5/sex). (D) Myocyte area is unaltered with sex (n=5). (E) NADPH oxidase ( Nox ) mRNA marker expression in muscle measured by qPCR, normalized to β -actin (n=5). (F) 8-isoprostane levels in plasma remain unchanged with sex (n=5). Results are mean±SEM; two-way ANOVA, Student’s t -test or Mann–Whitney U -test; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: Primary human male and female coronary artery ECs were purchased from American Type Culture Collection.

Techniques: Marker, Expressing, Clinical Proteomics, MANN-WHITNEY

Journal: bioRxiv

Article Title: Mitochondrial Dysfunction in Endothelial Cells Drives Greater Vascular Impairment in Females with Diabetes-Associated Peripheral Artery Disease

doi: 10.1101/2025.09.22.677648

Figure Lengend Snippet: (A) Schematic to show vessel collection for myography. (B) Myography showing increased endothelial dysfunction in female vs male arteries. Left , endothelial-dependent relaxation in response to acetylcholine (ACh). Right , sodium nitroprusside (SNP)-mediated relaxation (n=5-7). (C) Myography in non-injured vessels (N=5-7). (D) Plasma nitrite/nitrate levels (n=4-6). Laser Doppler imaging showing reduced blood perfusion over time with diabetes in (E) male and (F) female mice. Left, representative image of blood flow at 14 d. Right, quantification. (n=9-11). (G) Laser doppler perfusion index at 14 days (n=9-11). (H) Microvessel number (CD31+SMA+, <50 µm in diameter) in gastrocnemius tissues normalized to the number of myocytes and control non-ischemic limbs (n=9-11). (I) Tubule formation of male and female murine ECs shows an altered phenotype. (J) Tubulogenesis is reduced in female ECs under diabetic conditions. Left, Wimasis platform for vessel coverage and networks. Right , quantification (n=5/group). Results are mean±SEM; two-way ANOVA, Student’s t -test or Mann–Whitney U -test; * P <0.05, ** P <0.01, *** P <0.001 and **** P <0.0001.

Article Snippet: Primary human male and female coronary artery ECs were purchased from American Type Culture Collection.

Techniques: Clinical Proteomics, Imaging, Control, MANN-WHITNEY

Journal: bioRxiv

Article Title: Co-Therapy with S1P and Heparan Sulfate Derivatives to Restore Endothelial Glycocalyx and Combat Pro-Atherosclerotic Endothelial Dysfunction

doi: 10.1101/2024.11.06.622347

Figure Lengend Snippet: In vitro cellular remodeling as indicated by EC morphology via cell alignment in relation to flow. Representative phase contrast images of HCAECs at 10x are shown for all cohorts, which include HCAECs exposed to: A) static condition with no treatment, B) atheroprone region with no treatment, C) atheroprotective region with no treatment, D) static condition with co-treatment, (E) atheroprone region with co-treatment, and (F) atheroprotective region with co-treatment. All static and dynamic conditions (12 dynes/cm 2 ) were for 12 hours. Cell alignment was analyzed in two ways: (G) Object Orientation Parameter (OOP), which determines cell alignment in relation to flow and (H) Median Pairwise Alignment (MPA), which determines cell alignment in relation to other ECs. Each data point on the graph represents the mean individual OOP or MPA from one flow experiment, in which each cohort as range of n = 16 – 20. Box and whisker plot in (G) for OOP shows the minimum, first quartile, median, third quartile, and maximum values for each condition. MPA in (H) shows the mean and error bars which represents the SEM. Statistical analysis was performed using a two-way ANOVA. Significance is denoted by asterisks: *p<.05, **p<.01, ***p<.001, and ****p<.0001.

Article Snippet: Human coronary arterial ECs (HCAECs) purchased from PromoCell (Heidelberg, Germany) were cultured between passages 4 and 8 in PromoCell Endothelial Cell Growth Medium MV2 supplemented with growth factors, fetal calf serum, and antibiotic penicillin streptomycin.

Techniques: In Vitro, Whisker Assay

Journal: bioRxiv

Article Title: Co-Therapy with S1P and Heparan Sulfate Derivatives to Restore Endothelial Glycocalyx and Combat Pro-Atherosclerotic Endothelial Dysfunction

doi: 10.1101/2024.11.06.622347

Figure Lengend Snippet: Effect of co-treatment on S1PR1 (green) expression in HCAECs after 12 hours of flow at 12 dynes/cm 2 . Shown are representative 63x images of S1PR1 expression in HCAECs amongst different experimental groups. S1PR1, a G-protein coupled receptor found specifically on ECs, was used to determine its involvement in the therapeutic mechanism of action, and its expression increased in the atheroprotective and atheroprone regions when the co-treatment is introduced to the flow system, signifying activation. Blue represents cell nuclei labeled with DAPI. The scale bar is 20 μm. HCAECs in the no treatment group under A) static conditions, B) atheroprone conditions, and C) atheroprotective conditions. HCAECs in the co-treatment group under D) static conditions, E) atheroprone conditions, and F) atheroprotective conditions. G) Graph shows the mean ± SEM of percent area coverage of S1PR1 staining normalized to the control (0 hr static condition; n = 4). Statistical analysis was performed using a two-way ANOVA. Significance is denoted by asterisks: *p<.05, **p<.01, ***p<.001, and ****p<.0001.

Article Snippet: Human coronary arterial ECs (HCAECs) purchased from PromoCell (Heidelberg, Germany) were cultured between passages 4 and 8 in PromoCell Endothelial Cell Growth Medium MV2 supplemented with growth factors, fetal calf serum, and antibiotic penicillin streptomycin.

Techniques: Expressing, Activation Assay, Labeling, Staining, Control