Effects of TAV and BAV WSS on tissue structure and cell viability.

<p>H&E stain (A), mean tissue thickness measurements (B), TUNEL assay (C) and quantitative TUNEL results (D) in porcine aortic valve leaflets subjected to TAV and BAV WSS (F: fibrosa; V: ventricularis, green: apoptotic cells, blue: cell nuclei).</p

Effects of TAV and BAV WSS on valvular remodeling.

<p>MMP-2, MMP-9, TIMP-2, cathepsin L and cathepsin S immunostaining (A), semi-quantitative results (B) and TIMP-2/MMP-2 ratio (C) in porcine aortic valve leaflets subjected to TAV and BAV WSS (F: fibrosa; green: positively stained cells; blue: cell nuclei; *: p<0.05 vs. fresh control; #: p<0.05 vs. TAV WSS).</p

Double-sided shear stress bioreactor setup.

<p>Setup schematic (A) and tissue isolation and mounting (B) (R: radial tissue direction; C: circumferential tissue direction).</p

Effects of TAV and BAV WSS on endothelial activation.

<p>ICAM-1 and VCAM-1 immunostaining in porcine aortic valve leaflets subjected to TAV and BAV WSS (F: fibrosa; V: ventricularis; green: positively stained cells; blue: cell nuclei).</p

Effects of TAV and BAV WSS on α-SMA and Runx2 expressions.

<p>α-SMA and Runx2 immunoblotting (A) and densitometric results (B) in porcine aortic valve leaflets subjected to TAV and BAV WSS.</p

Effects of TAV and BAV WSS on valvular osteogenesis.

<p>Osteocalcin immunostaining (A), immunoblotting (B) and densitometric results (C) in porcine aortic valve leaflets subjected to TAV and BAV WSS (F: fibrosa; green: positively stained cells; blue: cell nuclei; *: p<0.05 vs. fresh control; #: p<0.05 vs. TAV WSS; +: p<0.05 vs. NC-BAV WSS).</p

Effects of TAV and BAV WSS on paracrine signaling.

<p>BMP-4 and TGF-β1 immunostaining (A), immunoblotting (B) and densitometric results (C) in porcine aortic valve leaflets subjected to TAV and BAV WSS (F: fibrosa; green: positively stained cells; blue: cell nuclei; *: p<0.05 vs. fresh control; #: p<0.05 vs. TAV WSS; +: p<0.05 vs. NC-BAV WSS).</p

Effects of FSS abnormalities on paracrine signaling.

<p>Immunostaining for BMP-4 (A) and TGF-β1 (B) after FSS conditioning for 48 hours, and Western blot results with densitometric assessment (C) (F: fibrosa, blue: cell nuclei, green: positively stained cells, * p<0.05 vs. fresh control, # p<0.05 vs. physiologic FSS).</p

Time-dependence of pro-inflammatory and ECM remodeling biomarker expression following tissue exposure to supra-physiologic FSS magnitude under normal frequency for 24, 48 and 72 hours.

<p>Immunostaining (A) and semi-quantitative results (B) (F: fibrosa, V: ventricularis, blue: cell nuclei; green: positively stained cells, * p<0.05 vs. fresh control, # p<0.05 vs. 24 h).</p

Effects of FSS abnormalities on elastolytic protease expression.

<p>Immunostaining for cathepsin-L (A) and cathepsin-S (B) after FSS conditioning for 48 hours and semi-quantitative results (C) (F: fibrosa, blue: cell nuclei; green: positively stained cells, * p<0.05 vs. fresh control, # p<0.05 vs. physiologic FSS).</p