Outline of experiment procedure. MLO‐Y4 cells were seeded to collagen coated glass slides and cultured for 48 hours (A), before being transferred to parallel plate flow chambers for dynamic (OFF, 1 Pa, 1 Hz, 2 hours) or static culture. The slides were then transferred to culture dishes and 2.5 mL of serum free medium was applied, with a control group being present with collagen coated glass slides without cells. The serum‐free medium was collected and centrifuged to remove debris. One milliliter of each sample was collected, and proteins were precipitated and digested in solution before being purified via C18 stage tips (B). Samples were analyzed via liquid chromatography‐mass spectrometry and tandem mass spectrometry (LC‐MS/MS), and label‐free quantification was carried out in MaxQuant before a bioinformatic analysis was completed in Perseus (&&&
P < .001 vs Medium using one‐way analysis of variance (ANOVA) and Bonferroni's multiple comparison post‐test) (C). Pearson correlations between technical replicates, biological replicates, and sample groups were determined, with correlations between biological replicates with combined technical replicates shown (D)