Comparison of chondrogenic differentiation of mesenchymal stromal cells from human amniotic fluid and human adipose-derived tissue in chitosan-xanthan gum scaffolds

For AF-hSCs, cells positive for CD117 (ckit) were selected, and SCs from both sources followed the same cell expansion steps, with characterization based on adherence, specific markers in flow cytometry, and differentiation into three mesenchymal lineages: adipogenesis, chondrogenesis, and osteogenesis. SCs were placed in CX scaffolds and cultured under TGF-β3 stimulation for 21 days. To confirm and assess chondrogenesis, scanning electron microscopy (SEM), histology, immunohistochemistry, and immunofluorescence were performed.

SCs from both sources demonstrated rapid and high expansion capacity, adherent culture with fibroblast-like morphology, positivity for mesenchymal lineage markers, pluripotency, chondrogenic potential, and negativity for hematopoietic markers. Differentiation into adipogenic, chondrogenic, and osteogenic mesenchymal lineages was also observed. Chondrogenesis was confirmed by histological staining with Hematoxylin and Eosin, Alcian Blue, Picrosirius Red, and Masson's Trichrome, indicating matrix production, and presence of collagen and proteoglycans.

Immunohistochemical analysis showed positive labeling for collagen II and aggrecan, as well as in immunofluorescence. SEM analysis revealed an intense collagen fiber network in the scaffold. Cells from both sources proved effective in inducing chondrogenesis.