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CHAPTER 4 4.4.4 Red light reduces astrocyte reactivity after 1-dpi in the ipsilateral spinal cord Both astrocytes and microglia/macrophages have been associated with the development and the maintenance of NP (Chiang et al., 2012), and it has been previously demonstrated reductions in activated microglia/macrophage following 670 nm treatment in spinal cord injured rats following 7 days of spinal cord injury recovery (Hu et al., 2016). Therefore it was worthwhile to examine astrocytic involvement in the spinal cord response to injury and red light treatment. Astrocyte activation was quantified as the percentage area of GFAP positive immunofluorescence (Figure 4.6) across 6 regions of the spinal cord (Figure 4.6a). Examples of GFAP+ staining for SCI and SCI+670 groups are shown in Figure 4.6b. Ipsilateral regions were significantly increased (p = 6.3e-14, LMER) compared to the contralateral side at all levels, particularly at 1-dpi (p < 2e-16) and 7-dpi (p = 0.027, LMER). GFAP+ staining was similar across the three funiculi. There was an overall significant reduction of GFAP+ area in the 670 nm light-treated animals throughout the spinal cord (p = 0.009, LMER). In the dorsal regions (Figure 4.6c-d), the contralateral side contained significantly less GFAP staining compared to the ipsilateral side (p = 2.6e-07, LMER), and red light treatment significantly reduced GFAP expression throughout (p = 0.003, LMER). In the SCI group, approximately 10% of the contralateral dorsal region of the spinal cord was GFAP+, which was maintained throughout the recovery period, while SCI+670 group showed significantly reduced GFAP+ staining (Figure 4.6c; p = 0.020; LMER). In the SCI group in the ipsilateral dorsal region (Figure 4.6d), GFAP+ staining was approximately 20-30% of the area, while this elevation was significantly reduced in the SCI+670 group (p = 0.0013), which was notable starting from 3-dpi. In the lateral regions (Figure 4.6e-f), the contralateral side was again significantly reduced compared to the ipsilateral side (p = 6.879e-05, LMER) but there was no overall red light treatment effect across both sides of the cord. In the lateral region on the contralateral side of the spinal cord (Figure 4.6e), both groups showed similar levels of GFAP+ staining throughout the recovery period. However, on the ipsilateral side (Figure 4.6f), GFAP+ staining was only similar at 1-dpi, as the SCI+670 group displayed significantly less GFAP from 3 to 7-dpi (p = 0.043, LMER). In the ventral region of the spinal cord (Figure 4.6g-h), the contralateral side was significantly reduced compared to the ipsilateral side (p = 7.694e-06, LMER) and there was an overall red light treatment effect across both sides of the cord (p = 0.046). While the two groups showed similar GFAP+ staining again on the contralateral side (Figure 4.6g), red light treatment significantly reduced GFAP+ staining on the ipsilateral side from 3 to 7-dpi (Figure 4.6h, p = 0.017). 94PDF Image | Effects of Red Light Treatment on Spinal Cord Injury
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