Figure 3.

Incubation with the PCN-miR/Col confers robust protection to ROS-exposed cells. (A) Pretreatment of HUVECs with PCN-miR/Col abrogated H₂O₂-induced cell viability loss as indicated by live/dead staining with calcein-AM (green) and propidium iodide (red). Scale bars, 50 μm. (B) Representative CLSM images showing the H₂O₂-induced intracellular ROS accumulation in HUVECs with various pretreatments, using DCF-DA as an ROS indicator. Scale bars, 50 μm. Representative confocal images of (C) JC-1 (a mitochondrial membrane potential-sensitive probe) and (D) γ-H2AX (a marker of DNA double-strand breaks) staining in cells with various pretreatments after exposure to H₂O₂. Scale bars, 20 μm. (E) Lipid peroxidation product MDA and (F) protein carbonylation levels in cells with various pretreatments after exposure to H₂O₂. All results are presented as mean ± SD, *P < 0.05 by two-tailed unpaired Student’s t tests, n = 3. (G) Schematic illustration of the ROS-induced damage responses in cells cultured on collagen-based hydrogels.