FIGURE 1.

In vitro validation of anti‐DKK1 treatment and viability of cultured human ASCs after seeding on a composite osteoconductive scaffold. A‐D, Effects of anti‐DKK1 on human adipose derived stem/stromal cell (ASC) osteogenic differentiation. A, Alizarin red staining after 7 days of osteogenic differentiation (2 μg/mL anti‐DKK1 or IgG isotype control). B‐D, Gene expression after 3 days of anti‐DKK1 treatment during osteogenic induction, including (B) RUNX2 (Runt related transcription factor 2), (C) ALP (alkaline phosphatase), and (D) COL1A1 (Collagen Type I Alpha 1). E‐H, Wnt signaling gene expression with anti‐DKK1 treatment for 3 days, including (E) AXIN2 (Axis Inhibition Protein 2), (F) CCND1 (Cyclin D1), (G) WNT5A (Wnt Family Member 5A), and (H) DKK1 (Dickkopf‐1). I‐K, In vitro validation of cell seeding on HA‐PLGA [hydroxyapatite coated poly(lactic‐co‐glycolic acid)] composite scaffolds. I, Distribution of DAPI labeled ASCs (appearing blue) on seeded scaffolds after 1 hour. Differential interference contrast used. J, Live‐dead staining of ASC seeded scaffolds after 1 hour. Live cells appear green while dead cells are red. K, Adhesion of seeded ASCs to the scaffold confirmed with scanning electron microscopy. K′, High power of cell membrane filopodia interacting with the scaffold surface. All experiments were performed with an appropriate isotype IgG control and in at least experimental and biological triplicate. Error bars represent 1 SD. White scale bars = 20 μm; yellow scale bars = 2 μm. *P < .01. ASCs, adipose‐derived stem cells