Compressive force regulates cementoblast migration via downregulation of autophagy.

BACKGROUND

Migration of cementoblasts to resorption lacunae is the foundation for repairing root resorption during orthodontic tooth movement. Previous studies reported that autophagy was activated by compression in periodontal ligament cells. The aim of this study was to investigate the migration of cementoblasts and determine whether autophagy is involved in the regulation of cementoblast migration under compressive force.

METHODS

Flow cytometry was employed to examine the apoptosis of murine cementoblasts (OCCM-30) at different compression times (0, 6, 12, and 24 hours) and magnitudes (0, 1.0, 1.5, and 2.0 g/cm2 ). Cell proliferation was examined using the CCK-8 method. Wound healing migration assays and transwell migration assays were performed to compare the migration of cementoblasts. Chloroquine (CQ) and rapamycin were used to inhibit and activate autophagy, respectively. The level of autophagy was determined using western blotting and immunofluorescence staining. The expression of matrix metalloproteinases (MMPs) was assessed using quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blot analysis, and enzyme-linked immunosorbent assay (ELISA).

RESULTS

Cell apoptosis and proliferation did not significantly change in OCCM-30 cells under mechanical compression at magnitude of 1.5 g/cm2 for 12 hours. However, the migration of cementoblasts was significantly inhibited after the application of compressive force. MMP2, MMP9, and MMP13 mRNA expression was decreased, and MMP9 and MMP13 protein expression and secretion level were also decreased. Further, autophagic activity was inhibited in cementoblasts under compressive force. Treatment with chloroquine reduced the cellular migration, and rapamycin partially relieved the inhibition of cementoblast migration induced by the compressive force. MMP9 and MMP13 mRNA expression, protein expression, and secretion levels showed a similar trend.

CONCLUSION

Migration of OCCM-30 cells was inhibited under compressive force partially dependent on the inhibition of MMPs, which was mediated by downregulation of autophagy. The findings provide new insights into the role of autophagy in biological behaviors of cementoblasts under compressive force and a potential therapeutic strategy for reducing external root resorption.