Accelerated construction of an in vitro model of human periodontal ligament tissue: vacuum plasma combined with fibronectin coating and a polydimethylsiloxane matrix

Tying shape memory wires to crowded teeth causes the wires to deform according to the dental arch. This deformation results in a resilient force that is delivered to the tooth. The appropriate amount of force can activate the osteogenetic and osteoclastic ability of the periodontal ligament (PDL) and the tooth can be moved. This is the biological basis of orthodontic treatment. To achieve further insight into the mechanisms underlying orthodontic treatment, we examined whether accelerated construction of an in vitro human PDL fibroblast (HPdLF) stretching model can be achieved by combining fibronectin coating and vacuum plasma treatment with polydimethylsiloxane (PDMS) cell-culture chambers. Each chamber was randomly assigned to a no-surface modification (NN), fibronectin coating (FN), vacuum plasma treatment (PN), or vacuum plasma treatment followed by a fibronectin coating (PF) treatment protocol. The physical and chemical features and ability to promote cellular proliferation of the PDMS chamber surfaces were evaluated. Cellular adhesion of four materials were evaluated and two best-proliferated groups were considered as better model-constructing surfaces and used in subsequent experiments and used in subsequent experiments. HPdLFs were cultured on these two kinds of chambers without stretching for 3 days, then with stretching for 7 days. Time-course gene expression cellular morphology were evaluated. Chambers in the PN group had high wettability and surface component changes. The FN and PF chambers had high cellular proliferation ability. They were selected into subsequent experiments. After 3 days of culturing HPdLFs on the PF and PN chambers, the cells in the PF chambers had significantly higher levels of runt-related transcription factor 2 (Runx-2) and osteocalcin (OCN) gene expression compared with the cells in the PN chambers. After cyclic stretch application to the cells in the PN and PF chambers, expression of the type-3 collagen (COL-3) gene in PF group continued to increase for 7 days and was significantly higher than that in the PN group from day 5 onwards. The HPdLFs in the PF group showed parallel alignment from days 3 to 7 after imposition of cyclic stretch, while those in the PN group aligned in parallel from day 5 on. Our results suggested that applying a fibronectin coating to a PDMS chamber after plasma treatment can accelerate establishment of an in vitro PDL stretching model.