Cyclic Stretching Triggers Cell Orientation and Extracellular Matrix Remodeling in a Periodontal Ligament 3D In Vitro Model

During orthodontic tooth movement (OTM), the periodontal ligament (PDL) plays a crucial role in regulating the tissue remodeling process. To decipher the cellular and molecular mechanisms underlying this process in vitro, suitable 3D models are needed that more closely approximate the situation in vivo. Here, a customized bioreactor is developed that allows dynamic loading of PDL-derived fibroblasts (PDLF). A collagen-based hydrogel mixture is optimized to maintain structural integrity and constant cell growth during stretching. Numerical simulations show a uniform stress distribution in the hydrogel construct under stretching. Compared to static conditions, controlled cyclic stretching results in directional alignment of collagen fibers and enhances proliferation and spreading ability of the embedded PDLF cells. Effective force transmission to the embedded cells is demonstrated by a more than threefold increase in Periostin protein expression. The cyclic stretch conditions also promote extensive remodeling of the extracellular matrix, as confirmed by increased glycosaminoglycan production. These results highlight the importance of dynamic loading over an extended period of time to determine the behavior of PDLF and to identify in vitro mechanobiological cues triggered during OTM-like stimulus. The introduced dynamic bioreactor is therefore a useful in vitro tool to study these mechanisms. This study establishes an in vitro model using a custom-made bioreactor to investigate the behavior of PDLFs. PDLFs are encapsulated into 3D hydrogel blends and cultivated dynamically to study the effect of mechanical stretching. This approach helps to bridge the gap between in vitro and in vivo research, in particular to improve the understanding of periodontal tissue dynamics.image