Histological Study of Human Mesenchymal Stem Cells Behavior Cultured on Decellularized Human Gingival Matrix in the Presence of Hyaluronic Acid

Abstract

Collagen bioscaffolds show promising results in oral mucosa engineering due to the high biocompatibility and biodegradability of collagen. Because of the high collagen content of gingiva, it can be used to prepare a natural collagen scaffold. Hyaluronic acid (HA) is one of the major components of extracellular matrix that regulates cellular behavior. In the present study, the effects of HA on the behavior of adipose-derived mesenchymal stem cells (Ad-MSCs) cultured on a decellularized gingival matrix were investigated by histological methods. Human gingival tissue (5×5×5 mm) was decellularized using physical methods as well as treatment with sodium dodecyl sulfate and Triton X-100, followed by washing and sterilization procedures. Scaffolds were divided into two groups including soaked scaffolds in HA (0.3% solution) and scaffolds without HA, which served as controls. In the next step, 3×105 Ad-MSCs were seeded on each scaffold in both groups, and histological studies were performed after 1, 2, 3 and 4 weeks of culture. Histological studies revealed effective cell removal and also preservation of collagen fibers in the decellularized gingival connective tissue. In vitro analysis of cellular behaviors showed adhesion and also migration of human Ad-MSCs towards connective tissue matrix, as well as formation of epithelial-like structures. Overall, statistical analyses indicated significant differences in cell density (P<0.01), migration (P<0.001) and nuclear size (P<0.01) at week one in HA-treated scaffolds versus the control group. The results of the present study demonstrated that the extracellular matrix of decellularized human gingival stroma can induce cellular behaviors such as adhesion, proliferation and migration of human Ad-MSCs.