Figure 4

C5aR or LPS-stimulated DPSCs express more BDNF than BM-MSCs. (A–L) Magnified pictures from the respective boxes of Figs. 2 and 3 in subsequent order. Groups: Resting (A,G), C5aR agonist (B,H), C5aR antagonist (C,I), LPS (D,J), LPS + C5aR agonist (E,K), LPS + C5aR antagonist (F,L). Double fluorescence immunohistochemistry with anti-BDNF (green) and anti-NGF (red) deposit’s reaction product in punctae of both stem cells. BDNF and NGF immune intensities are significantly higher in the C5aR agonist or LPS + C5aR-agonist treatment group (B,E & H,K) compared to resting (A,G) and C5aR antagonist treatment (C,I) groups. (M,N) Bar graphs showing the fluorescence intensity of BDNF and NGF in various treatment groups normalized to untreated control of resting cells. C5aR stimulation by C5aR-agonist or co-stimulation with LPS specifically increases BDNF and NGF (E) expression in DPSCs undergoing differentiation. (D1) DPSCs negative control. (M1) BM-MSCs negative control. Values for resting control were set at 100 as a reference. *p < 0.05, **p < 0.01 and ***p < 0.001 vs untreated control of resting cells (n = 3). Scale bar: 50 μm.