Journal: Mediators of Inflammation
Article Title: Effects of the Highly COX-2-Selective Analgesic NSAID Etoricoxib on Human Periodontal Ligament Fibroblasts during Compressive Orthodontic Mechanical Strain
Figure Lengend Snippet: Apoptosis and necrosis (FACS), cell viability (MTT), and cytotoxicity (LDH) of hPDL fibroblasts incubated with rising etoricoxib concentrations with and without mechanical strain. (a) Representative FACS distributions ( M ± SD, positive cells relative to total cells (%)) of apoptotic (Q4, FITC annexin V), necrotic (Q2, propidium iodide), and viable cells (Q3). N = 3, n = 6. (b) Cell viability as assessed by MTT assay; N = 3, n = 9. (c) Cytotoxicity assessed by LDH assay; N = 3, n = 9. All values given as normalized x-fold induction relative to 0 μ M etoricoxib without mechanical strain. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001. AU: arbitrary units. Bars show mean and standard deviation. PerCP-Cy5-5-H: peridinin-chlorophyll-protein complex channel (propidium iodide, necrosis); FITC-H: fluorescein thiocyanate channel (annexin V, apoptosis); MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid; LDH: lactate dehydrogenase.
Article Snippet: For RT-qPCR analyses as well as LDH/MTT assays, hPDL fibroblasts in six experimental groups with 9 wells (n = 9) on three plates (N = 3) each were, respectively, incubated with either 0 μ M (control), 3.29 μ M, or 5.49 μ M etoricoxib (Etoricoxib VETRANAL™ analytical standard: 32097 FLUKA, Sigma-Aldrich®, Taufkirchen, Germany) for 72 h, corresponding to assumed local concentrations reached in the PDL during normal and subtoxic clinical dosing in man [ , ], with or without (3/3 wells per plate) compressive mechanical strain of 2 g/cm2 for 48 h after a 24 h preincubation phase by means of a glass disc according to a published and well-established protocol for the simulation of compressive orthodontic mechanical strain [ , ] ( ).
Techniques: FACS, MTT Assay, Incubation, Lactate Dehydrogenase Assay, Standard Deviation