The role of mechanotransduction versus hypoxia during simulated orthodontic compressive strain—an in vitro study of human periodontal ligament fibroblasts

During orthodontic tooth movement (OTM) mechanical forces trigger pseudo-inflammatory, osteoclastogenic and remodelling processes in the periodontal ligament (PDL) that are mediated by PDL fibroblasts via the expression of various signalling molecules. Thus far, it is unknown whether these processes are mainly induced by mechanical cellular deformation (mechanotransduction) or by concomitant hypoxic conditions via the compression of periodontal blood vessels. Human primary PDL fibroblasts were randomly seeded in conventional six-well cell culture plates with O-2-impermeable polystyrene membranes and in special plates with gas-permeable membranes (Lumox (R), Sarstedt), enabling the experimental separation of mechanotransducive and hypoxic effects that occur concomitantly during OTM. To simulate physiological orthodontic compressive forces, PDL fibroblasts were stimulated mechanically at 2 g.cm(-2) for 48 h after 24 h of pre-incubation. We quantified the cell viability by MTT assay, gene expression by quantitative real-time polymerase chain reaction (RT-qPCR) and protein expression by western blot/enzyme-linked immunosorbent assays (ELISA). In addition, PDL-fibroblast-mediated osteoclastogenesis (TRAP(+) cells) was measured in a 72-h coculture with RAW264.7 cells. The expression of HIF-1 alpha, COX-2, PGE2, VEGF, COL1A2, collagen and ALPL, and the RANKL/OPG ratios at the mRNA/protein levels during PDL-fibroblast-mediated osteoclastogenesis were significantly elevated by mechanical loading irrespective of the oxygen supply, whereas hypoxic conditions had no significant additional effects. The cellular-molecular mediation of OTM by PDL fibroblasts via the expression of various signalling molecules is expected to be predominantly controlled by the application of force (mechanotransduction), whereas hypoxic effects seem to play only a minor role. In the context of OTM, the hypoxic marker HIF-1 alpha does not appear to be primarily stabilized by a reduced O-2 supply but is rather stabilised mechanically

Streptococcus mutans counts in plaque adjacent to orthodontic brackets bonded with resin-modified glass ionomer cement or resin-based composite

This study investigated the number of Streptococcus mutans CFU (colony forming units) in the saliva and plaque adjacent to orthodontic brackets bonded with a glass ionomer cement - GIC (Fuji Ortho) or a resin-based composite - RC (Concise). Twenty male and female patients, aged 12 to 20 years, participated in the study. Saliva was collected before and after placement of appliances. Plaque was collected from areas adjacent to brackets and saliva was again collected on the 15th, 30th, and 45th day after placement. On the 30th day, 0.4% stannous fluoride gel was applied for 4 minutes. No significant modification in the number of Streptococcus mutans CFU in saliva was observed after placement of the fixed orthodontic appliances. On the 15th day, the percentage of Streptococcus mutans CFU in plaque was statistically lower in sites adjacent to GIC-bonded brackets (mean = 0.365) than in those adjacent to RC-bonded brackets (mean = 0.935). No evidence was found of a contribution of GIC to the reduction of CFU in plaque after the 15th day. Topical application of stannous fluoride gel on the 30th day reduced the number of CFU in saliva, but not in plaque. This study suggests that the antimicrobial activity of GIC occurs only in the initial phase and is not responsible for a long-term anticariogenic property