Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Three-Layered Silk Fibroin Tubular Scaffold for the Repair and Regeneration of Small Caliber Blood Vessels: From Design to in vivo Pilot Tests

doi: 10.3389/fbioe.2019.00356

Figure Lengend Snippet: Time dependence of the absolute cell numbers of HAAF (A) , HASMC (B) , and HCAEC (C) cells cultured on SilkGraft and on polystyrene. Absolute cell numbers were lower on SilkGraft than on polystyrene because the available surface area was reduced due to the use of the steel ring which kept the silk substrate under water. Total cell growth differences between cells cultured on SilkGraft or on polystyrene are expressed by the areas under the corresponding curves, the statistical levels of significance of which are: for HAAFs, P < 0.001; for HASMCs, P < 0.01; and for HCAECs, P < 0.001.

Article Snippet: Adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) were provided by ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Three-Layered Silk Fibroin Tubular Scaffold for the Repair and Regeneration of Small Caliber Blood Vessels: From Design to in vivo Pilot Tests

doi: 10.3389/fbioe.2019.00356

Figure Lengend Snippet: Living cells density/mm 2 of apparent surface area after 21 days of in vitro culture.

Article Snippet: Adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) were provided by ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: In Vitro

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Three-Layered Silk Fibroin Tubular Scaffold for the Repair and Regeneration of Small Caliber Blood Vessels: From Design to in vivo Pilot Tests

doi: 10.3389/fbioe.2019.00356

Figure Lengend Snippet: Cumulative consumption of glucose and glutamine and release of lactate. Results were normalized per 10 3 cells. (A–C) The cumulative glucose consumption was higher for HAAFs ( P < 0.05) and HCAECs ( P < 0.001) seeded on SilkGraft, whereas it showed only marginal differences between the two substrates for HASMCs ( P > 0.05). (D–F) Glutamine consumption was lower for HAAFs ( P < 0.05) seeded on SilkGraft, similar for HASMCs ( P > 0.05) cultured on the two substrates, and significantly larger for HCAECs ( P < 0.001) grown on the silk substrate. (G,H) The cumulative amount of lactate released by HAAFs and HASMCs was the same whichever the substrate ( P > 0.05). Lactate release could not be assessed for HCAECs because the released lactate was re-uptaken and used for metabolic purposes. The statistical analysis of these data is shown in .

Article Snippet: Adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) were provided by ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Three-Layered Silk Fibroin Tubular Scaffold for the Repair and Regeneration of Small Caliber Blood Vessels: From Design to in vivo Pilot Tests

doi: 10.3389/fbioe.2019.00356

Figure Lengend Snippet: Comparison of metabolic parameters ^* of the different cell types cultured on SilkGraft and polystyrene.

Article Snippet: Adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) were provided by ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Comparison, Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Three-Layered Silk Fibroin Tubular Scaffold for the Repair and Regeneration of Small Caliber Blood Vessels: From Design to in vivo Pilot Tests

doi: 10.3389/fbioe.2019.00356

Figure Lengend Snippet: Relevant cytokines and chemokines secreted by each cell type cultured between 18 and 20 days on SilkGraft and polystyrene: HAFFs (A) , HCAECs (B) , and HASMCs (C) . Results of immunofluorescence intensities were normalized to 10 3 cells. IL-6: Interleukin-6; MCP-1: Monocyte chemoattractant protein-1; TIMP-2: Tissue inhibitor of metal proteinases-2; IP-10: Interferon gamma-induced protein-10; MCP-2: Monocyte chemoattractant protein-2; Eotaxin-1; RANTES: Regulated on activation normal T cell expressed and secreted; MIP-1β: Macrophage inflammatory protein-1β; TNF-β: Tumor necrosis factor-β; GM-CSF: Granulocyte-macrophage colony stimulating factor; IL-1α: Interleukin-1α; IL-1β: Interleukin-1β; ICAM-1: Intercellular adhesion molecule-1. The bars are the mean values of three independent experiments corrected for cell numbers. * P < 0.01; ** P < 0.001. SEMs, not shown, ranged between 5 and 10% of corresponding mean values.

Article Snippet: Adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) were provided by ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Cell Culture, Immunofluorescence, Activation Assay

Journal: Cellular signalling

Article Title: PAI-1 contributes to homocysteine-induced cellular senescence

doi: 10.1016/j.cellsig.2019.109394

Figure Lengend Snippet: Cultures of EA.hy926 endothelial cells (A) and HCAEC (B, C) were pretreated with TM5441 (10 μM) (A, B) or TM5A15 (10 μM) (C) in triplicate followed by Homocysteine (Hcy) treatment for 4–5 days. Whole cell extracts were prepared and equal amount of pooled proteins from three wells were subjected to Western blot analysis for senescence markers and regulators using specific antibodies as indicated (A–C). Bar represents mean ± sem. Quantitative data are shown on the right (A’-C’). The levels of at least 2–3 senescence markers were determined in repeat experiments. D, E. Whole cell extracts (HCAEC) were prepared from two separate experiments and equal amount of pooled proteins from three wells were subjected to Western blot analysis for senescence markers and regulators p53 and pERK1/2 (D), integrin β3 and PAI-1 (E) using specific antibodies. Quantitative data in the lower panel showing the levels of each regulator relative to loading control Actin (D’, E’).

Article Snippet: Endothelial cell culture: treatment with Hcy and small molecule inhibitors of PAI-1 Primary cultures of Human Coronary Artery Endothelial Cells (HCAEC) (Cell Applications; Cat # 300–05a) and EA.hy926 (ATCC cat #CRL-2922) were grown in MesoEndo Cell Growth Media and Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum and 1% penicillin and streptomycin respectively and maintained at 37 °C in a 5% CO 2 incubator.

Techniques: Western Blot, Control

Journal: Journal of Translational Medicine

Article Title: Quinic acid regulated TMA/TMAO-related lipid metabolism and vascular endothelial function through gut microbiota to inhibit atherosclerotic

doi: 10.1186/s12967-024-05120-y

Figure Lengend Snippet: QA improved TMAO-induced inflammatory lesions and endothelial dysfunction in HCAECs. (A) CCK-8 was applied to detect the toxicity of QA on HCAECs. (B) CCK-8 was used to detect HCAECs proliferation. (C) The expression of COX-2, IL-6, E-selectin, ICAM-1, HMGB1 was detected by RT-qPCR. (D) The expression of p-P65, p-MAPK14 protein was detected by western blot. (E) HMGB1 levels were detected by ELISA. (F) The expression of ZO-2, VE-Cadherin and Occludin were detected by western blot. * P < 0.05 vs. Control, # P < 0.05 vs. TMAO

Article Snippet: To investigate the cytotoxicity of QA, human coronary artery endothelial cells (HCAECs, HUM-iCell-c006, iCell) were treated with 1, 2.5, 5, 10 and 20 μM QA.

Techniques: CCK-8 Assay, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: Post-ischemic inactivation of HIF prolyl hydroxylases in endothelium promotes maladaptive kidney repair by inducing glycolysis

doi: 10.1101/2023.10.03.560700

Figure Lengend Snippet: ( A ) Experimental scheme for HPAECs subjected to 0.5% O 2 for 18 hours in the presence of syrosingopine (5 µM) or MCT4 siRNA followed by reoxygenation for 8 hours in the presence of IL-1β (1 ng/ml). ( B ) mRNA levels of VCAM1 and ICAM1 in syrosingopine- vs vehicle-treated HPAECs, that were activated by Hypoxia/Reoxygenation and IL-1β. ( C ) THP1 monocyte adhesion to inflamed ECs. THP1 monocyte cells, labeled with Green CMFDA Dye, were introduced on a monolayer of HPAECs that had been subjected to the indicated experimental conditions. Following a 90-minute incubation period, floating cells were washed away and adhered THP1 cells were visualized using a fluorescent microscope and subsequently quantified. Representative images of fluorescent THP1 cells attached to ECs in different experimental groups are presented. Scale bar, 200 μm. ( D ) mRNA expression of VCAM1 and ICAM1 in HPAECs transfected with control or MCT4 siRNA and subjected to Hypoxia/Reoxygenation and IL-1β ( E ) THP1 monocyte adhesion to inflamed ECs. Labeled THP1 cells were introduced on a monolayer of HPAECs that had been subjected to the indicated experimental conditions. Following incubation, adhered THP1 cells were visualized quantified as in C . Representative images of fluorescent THP1 cells attached to ECs in different experimental conditions are presented. Scale bar, 200 μm. Data are represented as mean ± SEM. Statistics were determined by one-way ANOVA with Sidak correction for multiple comparisons. n=3-4; *, P < 0.05; **, P <0.01; ***, P <0.001; ****, P < 0.0001; ns, not statistically significant. Nx, Normoxia; Hx, Hypoxia/Reoxygenation; Veh, vehicle; Syro, syrosingopine; C, negative control siRNA; MCT4si, MCT4siRNA.

Article Snippet: Human primary pulmonary artery ECs (HPAECs) were obtained from ATCC and grown on gelatin-coated dishes in Endothelial Cell Basal Medium-2 (Lonza, Catalog # CC-3156) supplemented with EGM-2 SingleQuots Supplements (Lonza, Catalog # CC-4176).

Techniques: Labeling, Incubation, Microscopy, Expressing, Transfection, Control, Negative Control