Evaluation of the osteogenic differentiation of gingiva‑derived stem cells grown on culture plates or in stem cell spheroids: Comparison of two‑ and three‑dimensional cultures

Three‑dimensional cell culture systems provide a convenient in vitro model for the study of complex cell‑cell and cell‑matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva‑derived stem cells cultured in two‑dimensional or three‑dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva‑derived stem cells in monolayer culture to a three‑dimensional culture system with microwells. For three‑dimensional culture, gingiva‑derived stem cells were isolated and seeded into polydimethylsiloxane‑based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two‑dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva‑derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two‑dimensional culture. This highlights the potential of three‑dimensional culture systems using gingiva‑derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.