Figure 3

Suppressing inflammation delays hair cell regeneration. (A) Real-time imaging (40×) displays regenerated hair cells in the CuSO₄ and BRS+CuSO₄ group at 24, 48, and 96 hpi. The control group was imaged at the same time points. Scale bar represents 10 μm. (B) The numbers of regenerated hair cells were significantly lower in the BRS+CuSO₄ group compared to the CuSO₄ group at 16 (n ≥ 27, p < 0.01), 24 (n ≥ 21, p < 0.01), and 48 (n ≥ 24, p < 0.001) hpi. At 96 hpi, hair cells in both the CuSO₄ group and the BRS+CuSO₄ group had regenerated to normal levels. Linear analysis in the CuSO₄ group (C) and BRS+CuSO₄ group (D) was conducted on the number of regenerated cells within 48 hpi. The slope in the CuSO₄ group (0.188) was higher than that in the BRS+CuSO₄ group (0.148), and the x-intercept in the CuSO₄ group (4.160) is higher than that in the BRS+CuSO₄ group (8.287). (E) When the time window of inflammatory suppression was delayed, there was no delay in the regeneration of hair cells. BRS-28 was added at the same time as CuSO₄ (CuSO₄+BRS 0 h group), 30 min after the addition of CuSO₄ (CuSO₄+BRS 0.5 h group), or 1 h after the addition of CuSO₄ (CuSO₄+BRS 1 h group) (n ≥ 27 neuromasts at each time point of each group). For (B) and (E), comparisons were performed using two-way ANOVA, with Tukey’ multiple comparisons test. All error bars show the mean ± S.E.M., *** p < 0.001, ** p < 0.01.