High glucose impairs human periodontal ligament cells migration through lowered microRNAs 221 and 222

Abstract Objective To investigate the effects of miR‐221 and miR‐222 and high glucose on human periodontal ligament (PL) cells morphology, cytoskeleton, adhesion, and migration. Background Chronic hyperglycemia is common in uncontrolled diabetes mellitus (DM) and plays a central role in long‐term DM complications, such as impaired periodontal healing. We have previously shown that high glucose increases apoptosis of human PL cells by inhibiting miR‐221 and miR‐222 and consequently augmenting their target caspase‐3. However, other effects of miR‐221/222 downregulation on PL cells are still unknown. Methods Cells from young humans' premolar teeth were cultured for 7 days under 5 or 30 mM glucose. Directional and spontaneous migration on fibronectin were studied using transwell and time‐lapse assays, respectively. F‐actin staining was employed to study cell morphology and the actin cytoskeleton. MiR‐221 and miR‐222 were inhibited using antagomiRs, and their expressions were evaluated by real‐time RT‐PCR. Results High glucose inhibited PL cells early adhesion, spreading, and migration on fibronectin. Cells exposed to high glucose showed reduced polarization, velocity, and directionality. They formed several simultaneous unstable and short‐lived protrusions, suggesting impairment of adhesion maturation. MiR‐221 and miR‐222 inhibition also reduced migration, decreasing cell directionality but not significantly cell velocity. After miR‐221 and miR‐222 downregulation cells showed morphological resemblance with cells exposed to high glucose. Conclusion High glucose impairs human PL cells migration potentially through a mechanism involving reduction of microRNA‐221 and microRNA‐222 expression. These effects may contribute to the impairment of periodontal healing, especially after surgery and during guided regeneration therapies.