Fig. 1: CSPG digestion enhances immune cell clearance after spinal cord injury.

a, c Experimental design of immune cell recruitment study. b Contusion device impact force and displacement measurements confirm reproducible and consistent injuries between treatment groups. Data were determined as normally distributed by the Shapiro–Wilk test and subsequently analysed using a two-tailed unpaired t test. Data are shown as mean ± SEM. d, e Graphs showing quantification of innate immune cell recruitment, using manual-gated flow cytometry, following spinal cord injury at 1, 3, 7, 14 and 28 dpi with (LV-ChABC) or without (LV-GFP) CSPG digestion. *p < 0.05 versus control (LV-GFP) group. Results were assessed for normality using the Shapiro–Wilk test and analysed using a two-way ANOVA with Bonferroni’s post hoc test (n = 8 per group at 1 dpi; n = 11 at 3 dpi; n = 11 at 7 dpi; n = 6 at 14 dpi; n = 3 at 28 dpi). Data are pooled from at least two independent experiments. Data are shown as mean ± SEM. f t-SNE flow cytometry analysis at 7dpi reveals the presence of 9 major CD45+ cell populations within the injured spinal cord. g Heat map showing the relative expression of extracellular markers in the 9 identified clusters. h–m CSPG digestion reduces the number of microglial cells and monocyte/macrophages at 7 dpi. h tSNE plot highlighting cluster 1 identified as microglia in LV-GFP (left) and LV-ChABC (right) treated rats. i tSNE plot highlighting clusters 2 and 3 identified as monocytes/macrophages in LV-GFP (left) and LV-ChABC (right) treated rats. j Graph showing the number of microglial cells within the injured spinal cord at 7 dpi. Microglial cell number is significantly reduced after CSPG digestion. k Graph showing the number of monocyte/macrophages within the injured spinal cord at 7 dpi. Monocyte/macrophage cell number is significantly reduced after CSPG digestion. l, m LV-ChABC treatment exhibits greater effects in CD43^low monocyte/macrophages. l tSNE plots showing the expression of CD43 in the different clusters. Note that monocytes/macrophages are differentiated in two different subsets by CD43 relative expression. m Graph showing the number of CD43^high and CD43^low monocyte/macrophages within the injured spinal cord at 7 dpi. Both populations are significantly reduced after CSPG digestion. j, k, m *p < 0.05, **p < 0.01 versus control (LV-GFP) group. Results were assessed for normality using the Shapiro–Wilk test and analysed using a two-tailed unpaired t test. Data are shown as mean ± SEM (n = 4 per treatment). Detailed statistics and exact p values are provided in Supplementary Table 8. Source data are provided as a Source Data file.