Fig. 1: Morphological and bioactivity assessment of developed sheep uterus scaffolds.

Hematoxylin and eosin-stained sections from normal and decellularized uterus showed that the extracellular matrix was well-preserved and that all hematoxylin positive nuclei had been removed by the different decellularization protocols (A–D; scale bar = 100 μm). Scanning electron microscopy pictures obtained at ×3000 magnification (E–H; scale bar = 8 μm) showed that protocol (P) 1 generated scaffolds resulted in an organized porous structure (F) while P2 and P3 derived scaffolds resulted in a more compact arrangement with less organized fiber structure. Higher magnification (×12,000; I–L; scale bar = 2 μm) showed that collagen bundles remained intact in all three scaffold types. The dorsal root ganglion (DRG) assay and the chorioallantioc membrane (CAM) assay showed that all scaffolds were functionally bioactive (M–V). The DRGs regenerated axons in wells coated with collagen that was given nerve growth factor supplement in the culture medium (positive control; M), but not when this growth factor was omitted (negative control, N). Scaffolds derived from the three different decellularization protocols all stimulated axonal regeneration (O–Q; scale bar = 500 µm).