Figure 5.

Differential mRNA levels and comparative immunostainings of the fibrin revealed that pulsatile high‐velocity flow on frontside decreased the overall MMP activity in engineered SMCs tissues by inhibiting their activity via TIMPs after 21 days in culture and doxycycline inhibit the fibrin remodeling by inhibition of MMPs. a) Relative mRNA expression of metalloproteinase inhibitors (TIMP1, 2, and 3) and MMP (MMP 2, 9, 12, 14, and 15) at the frontside versus backside. b) Comparative confocal microscopy and c) pixel‐by‐pixel intensity analysis of images showed delayed degradation of fibrin in the tissue's front versus backside in the absence or presence of doxycycline (10 µg mL⁻¹). Fibronectin was visualized not by immunostaining, but by supplementing labeled Alexa Flour 488 human plasma fibronectin (green) to the cell culture medium.^{[ 38 ]} Number of biological replicates was 3, ≥5 images were analyzed per replicate. Flow diagram shows how the samples have been treated during the culturing period. Scale bars 50 µm. See also Figures S8 and S9, Supporting Information, for respective single channel and cross‐sectional confocal images.