Figure 2

Effects of titania nanosurfaces on the expression of polarization markers of macrophages Gene expression of M1 polarization markers (A) inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and (B) M2 polarization markers arginase 1 (Arg1) and cluster of differentiation 206 (CD206) relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) in J774A.1 cells cultured for days 1 and 3 on a polystyrene culture plate or smooth (SM), micro-roughened (MR), or nano-roughened (NR) 1 or 2 surfaces with or without the M1-induction. (C) Representative confocal laser microscopy images of F-actin (red), nuclear (blue), and iNOS (green) staining in the cells. (D) WST-1-based evaluation of the number of adherent cells under different culture conditions on day 1. (E) Concentrations of endotoxins detected on the titanium or titania surfaces without cells. Data represented as mean ± standard deviation (SD; N = 3 in A, B and C) or box plots (N = 6 in E). Different letters indicate statistically significant differences between them (P < 0.05; Bonferroni’s multiple comparison test or Tukey’s honestly significant difference [HSD] test). UT, untreated cells; M1, M1-induced cells; M2, M2-induced cells; n.s., non-significant difference.