Journal: Bioengineering

Article Title: Beyond Decellularization: Remnant Mitochondrial DNA Can Act as Hidden Damage-Associated Molecular Pattern

doi: 10.3390/bioengineering13020193

Figure Lengend Snippet: HpaII treatment of human liver scaffolds reduced macrophage activation and proliferation. Representative images of DAPI-stained cell nuclei of THP-cells on untreated ( A ) and HpaII-treated ( B ) liver ECM, and number of adherent cells per mm 2 . Scalebars: 100 µm. ( C ) Proliferation of M0 macrophages after 4 days of culture with decellularized scaffold, either untreated ( n = 3) ( D ) or treated with HpaII ( n = 3) ( E ). Representative images are shown. Cell nuclei (blue), Ki-67 (red), and F-actin (Phalloidin, green); scalebars: 200 µm. More ki-67 positive cells were detected in cultures with untreated scaffolds compared to HpaII-treated scaffolds. ( F ) Number of Ki-67 + cells per mm 2 plotted as min to max bars with line at mean. ( G – I ) Cytokine production by M0 macrophages after 4 days of culture with decellularized scaffolds. TNF-α, IL-1β, and IL-6 cytokine production was determined using ELISA. Untreated scaffolds ( n = 6) induced production of all cytokines. The HpaII treatment of scaffolds ( n = 6) significantly reduced TNF-α and IL-1β production. * p < 0.05 and ** p < 0.01.

Article Snippet: Adherent THP-1-cells were assessed by staining with DAPI (Vectashield antifade mounting medium with DAPI, Vector Laboratories Inc., Neward, CA, USA) and were visualized with confocal microscopy (Leica SP8 DLS Lightsheet microscope, Wetzlar, Germany) at 20× magnification.

Techniques: Activation Assay, Staining, Enzyme-linked Immunosorbent Assay