Periodontal ligament (PDL) cells are regarded as the cell type with the highest potential for periodontal regeneration. Biophysical cues of the culture substrate are increasingly identified as vital parameters to affect cell behavior. Compared to traditional tissue culture polystyrene (TCPS), polydimethylsiloxane (PDMS) substrates corroborate more closely the elastic modulus values of the physiological environment. Consequently, the aim of this study was to evaluate the effect of PDMS-based substrates with different stiffness on cellular responses of human PDL cells. PDMS substrates with different stiffness were fabricated by varying the ratio of base to curing component. The influence of PDMS substrates on PDL cell spreading and cytoskeletal morphologies, motility, proliferation, stemness gene expression, and osteogenic differentiation was evaluated and compared to that on conventional TCPS. PDL cells cultured on PDMS substrates exhibited a smaller cell size and more elongated morphology, with less spreading area, fewer focal adhesions, and faster migration than cells on TCPS. Compared to TCPS, PDMS substrates promoted the rapid in vitro expansion of PDL cells without interfering with their self-renewal ability. In contrast, the osteogenic differentiation ability of PDL cells cultured on PDMS was lower in comparison to cells on TCPS. PDL cells on PDMS exhibited similar cell morphology, motility, proliferation, and self-renewal gene expression. The stiffer PDMS substrate increased the osteogenic gene expression of PDL cells compared to the soft PDMS group in one donor. These data indicate that PDMS-based substrates have the potential for the efficient PDL cell expansion.
Keywords: cell culture; periodontal ligament cells; polydimethylsiloxane; proliferation; stiffness.
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