PMC

VCAM-1 and ICAM-1 expression by HBOECs and HUVECs seeded on bioartificial tissue and cultured under static or flow conditions for 24 h, in the presence or absence of TNF-α. (A) VCAM-1 expression. (B) ICAM-1 expression. Plots show mean±SE for n=3 tissue samples. *Significant increase over static and flow groups of the same cell type (p<0.05). **Significant increase over all other groups of the same cell type (p<0.05). ICAM-1, intercellular adhesion molecule-1; VCAM-1, vascular cell adhesion molecule-1.

ECM staining of bioartificial tissue. (A–E) Bioartificial tissue stained for ECM components. (F–J) Bioartificial tissue seeded with HBOECs for 7 days before staining. (A, F) Fibrin staining. (B, G) Fibronectin staining. (C, H) Collagen I staining. (D, I) Collagen IV staining. (E, J) Laminin staining. The lumenal surface is on the left side of each image, nuclei counterstained with Hoechst 33342. Scale bar=100 μm. ECM, extracellular matrix. Color images available online at www.liebertonline.com/tea

eNOS expression and NOx production by HBOECs and HUVECs seeded on bioartificial tissue and cultured under static and flow conditions for 24 h. eNOS staining of HBOEC cross sections after (A) static culture or (B) flow exposure. (C) NO production was quantified by measurement of total nitrate and nitrite in the HBOEC and HUVEC culture medium. Values are normalized to the statically cultured HBOEC value. Plot shows mean±SE for n=3 independent experiments. *NO production by HBOECs and HUVECs cultured under flow conditions increased compared to HBOEC static culture (p<0.01). Scale bar=100 μm. eNOs, endothelial nitric oxide synthase. Color images available online at www.liebertonline.com/tea

HBOEC retention under physiological shear stress for short durations. HBOECs were seeded on type I collagen-coated TCP or bioartificial tissue and incrementally exposed to 1, 5, 10, 15, or 25 dyn/cm² for 20 min at which time HBOEC surface coverage was calculated based on the CellTracker Green staining. Plot shows mean±SE for n=3 independent experiments. *Significant difference compared to 1 dyn/cm² samples on the same substrate (p<0.05). ^#Significant difference between HBOEC on TCP and bioartificial at the same shear stress (p<0.05). TCP, tissue culture plastic; SE, standard error of the mean.

Retention, elongation, and alignment of HBOECs and HUVECs seeded on bioartificial tissue and exposed to shear stress. VE-Cadherin staining of (A, B) HBOECs and (C, D) HUVECs. (A, C) Statically cultured cells. (B, D) Cells cultured under 15 dyn/cm² for 24 h. (E) Cell density, (F) elongation, and (G) alignment of HBOECs and HUVECs after exposure to 15 dyn/cm² shear stress or static culture for 24 h. The elongation is given as a shape index where 0 is a line and 1 is a circle. Cell alignment is given on a scale from perfectly aligned (0°) to no alignment (45°). *p<0.05 compared to static conditions. All plots show mean±SE for n=3 independent experiments. Scale bar=50 μm and applies to all images. Color images available online at www.liebertonline.com/tea

HBOECs seeded on bioartificial tissue. (A) Cross-sectional and (B, C) En face images of CellTracker Green-labeled HBOECs seeding on bioartificial tissue show that the HBOECs form a confluent monolayer on the tissue surface. Nuclei of HBOEC and human dermal fibroblast in the tissue are stained blue, and VE-Cadherin staining is pseudo-colored red. (D) HBOECs and (E) HUVECs on the bioartificial tissue express thrombomodulin. (F) Tissue factor expression by HBOEC on unstimulated tissue and (G) after exposure to TNF-α. (B–E) Scale bar=50 μm. (F, G) Scale bar=25 μm. HBOEC, human blood outgrowth endothelial cell; HUVEC, human umbilical vein endothelial cell; TNF, tumor necrosis factor. Color images available online at www.liebertonline.com/tea

Platelet adhesion on HBOEC- and HUVEC-seeded bioartificial tissue preconditioned with 15 dyn/cm² shear stress for 24 h. (A) Normalized platelet coverage. HBOEC-seeded tissue was compared to HBOEC-seeded tissue that was pre-exposed to TNF-α for 4 h or nonseeded tissue. All groups were normalized to the statically cultured HBOEC group. *p<0.001 compared to unstimulated HBOEC. ^#p<0.001 compared to stimulated and nonstimulated HBOEC-seeded tissue. (B) HBOEC- and HUVEC-seeded tissue was statically cultured or exposed to 15 dyn/cm² shear stress for 24 h before whole blood flow exposure. *p<0.05 compared to the statically cultured cells of the same EC type. (C) En face image of platelet localization. Scale bar=25 μm. Cross-sectional views of (D) platelet localization and (E) platelet aggregation on the surface of statically cultured HBOECs. Scale bar=50 μm. Plots show mean±SE for one representative experiment with n=3 tissue samples. Color images available online at www.liebertonline.com/tea