Figure 6.
The MMP inhibitor doxycycline enhances collagen I and elastin accumulation in both of the engineered SMCs tissues. Biochemical assay of total a) collagen and b) elastin revealed higher assembly of collagen within the backside tissues and higher elastin content within the frontside tissues, but the effect of low‐velocity flow reduced in the presence of doxycycline (10 µg mL−1, yellow‐shaded bars) after 3 weeks of culture. Comparative immunostaining of c) collagen I (magenta) and d) elastin (red) in the presence and absence of doxycycline. Fibronectin was visualized not by immunostaining, but by supplementing labeled Alexa Flour 488 human plasma fibronectin (green) to the cell culture medium.[ 38 ] Relative signal intensity analysis of e) collagen I and f) elastin showed the same trend as found in biochemical assays. Number of biological replicates was 3; ≥5 images were analyzed. All tissues have been treated according to the timeline given with the Figure 4e. Scale bars 50 µm. See also Figures S10–S12, Supporting Information, for single‐channel confocal images of collagen I, III, and elastin, respectively.