Article Figures & Data
Figures
- Figure 1.
Schematic image of the treatment procedure and the experimental flow. A, Schematic image of the treatment procedure; NS/PCs for transplantation were injected rostrally and caudally from the epicenter, and then a sheet of Sema3Ai or a placebo sheet was placed over the SCI epicenter. B, Schematic image of the experimental flow; SCI models were divided into two groups, ensuring a close match in terms of mean BBB score and body weight up to 46 DPI. A 5 d regimen of conditional training was started on 42 DPI and ended on 46 DPI. The treatment procedure was administered on 49 DPI, followed by rehabilitation training, which began on 53 DPI and continued for 8 weeks.
- Figure 2.
Comparison of fixation and immaturity of transplanted cells. A, Sagittal-slice IHC of the SCI epicenter from a nontransplanted rat at 49 DPI, stained with NG2 and Sema3A. Scale bar, 100 μm. B, Axial slice of the SCI epicenter with H&E staining of TSR group and TR group. Scale bar, 100 μm. Extended Figure 2-1 shows an axial slice of the SCI epicenter of a single rehabilitation group. C, Comparison of the SCI epicenter volume with H&E staining at 110 DPI TSR (n = 4) and TR (n = 4), p = 0.72. D, Axial-slice IHC of the SCI epicenter with HNA and Ki67 at 110 DPI. Scale bar, 100 μm. E, Comparison of the transplanted volume with HNA. TSR (n = 4) and TR (n = 4), p = 0.73. F, Comparison of the transplanted immature volume with Ki67. TSR (n = 4) and TR (n = 4), p = 0.14. All comparisons were tested with a two-sample t test. *p < 0.05, **p < 0.01, ***p < 0.005.
- Figure 3.
Differentiation of transplanted cells and formation of host tissue in the epicenter. A, Axial-slice IHC of the SCI epicenter with HNA and ELAVL3/4. Scale bars: left side, 100 μm; right side, 10 µm. B, Axial-slice IHC of the SCI epicenter with HNA and APC. Scale bars: left side, 100 μm; right side, 10 µm. C, Axial-slice IHC of the SCI epicenter with GFAP and STEM123. Scale bars: left side, 100 μm; right side, 10 µm. D, Comparison of the graft-derived differentiation percentage of neurons with HNA and ELAVL3/4. TSR (n = 4) and TR (n = 4), p = 0.94. E, Comparison of the graft-derived differentiation percentage of oligodendrocytes with HNA and APC. TSR (n = 4) and TR (n = 4), p = 0.98. F, Comparison of the graft-derived astrocyte volume with STEM123. TSR (n = 4) and TR (n = 4), p = 0.29. G, Neuron volume comparison with ELAVL3/4. TSR (n = 4) and TR (n = 4), p = 0.0017, Cohen's d = 3.8. H, Oligodendrocyte volume comparison with APC. TSR (n = 4) and TR (n = 4), p = 0.0038, Cohen's d = 3.2. I, Astrocyte volume comparison with GFAP in the SCI epicenter. TSR (n = 4) and TR (n = 4), p = 0.010, Cohen's d = 2.6. All comparisons were tested with a two-sample t test. *p < 0.05, **p < 0.01, ***p < 0.005.
- Figure 4.
Comparative analysis of axon elongation. A, Axial-slice IHC of the SCI epicenter with pGAP43(S41) and pGAP43(S96). Scale bars: left side, 100 μm; right side, 10 µm. B, Total regenerated axon comparison with pGAP43(S41). TSR (n = 4) and TR (n = 4), p = 0.0018, Cohen's d = 3.8. C, Host-derived regenerated axon comparison with pGAP43(S96). TSR (n = 4) and TR (n = 4), p = 0.034, Cohen's d = 1.9. D, Axial-slice IHC of the SCI epicenter with neurofilament (NF-H) and STEM121. Scale bars: left side, 100 μm; right side, 10 µm. E, Host-derived neurofilament volume comparison with NF-H. TSR (n = 4) and TR (n = 4), p = 0.039, Cohen's d = 1.9. F, NF-H crossing the transplanted field (HNA) in a sagittal section. Scale bars: left side, 100 μm; right side, 5 µm. G, Axon length comparison with NF-H. TSR (n = 4) and TR (n = 4), p = 0.00000081, Cohen's d = 15. H, Axial-slice IHC of the SCI epicenter with 5-HT and STEM121. Scale bars: left side, 100 μm; right side, 10 µm. I, Serotonergic fiber comparison with 5-HT. TSR (n = 4) and TR (n = 4), p = 0.0045, Cohen's d = 3.1. Extended Figure 4-1 shows graft axon expansion with STEM121 at the SCI epicenter. All comparisons were tested with a two-sample t test. *p < 0.05, **p < 0.01, ***p < 0.005.
- Figure 5.
Image and comparative analysis of MEP. A, Schematic image of MEP. B, Representative MEP comparison. C, MEP comparison with amplitude. TSR (n = 6) and TR (n = 6), p = 0.93. D, MEP comparison with latency. TSR (n = 6) and TR (n = 6), p = 0.62. E, MEP comparison with duration. TSR (n = 6) and TR (n = 6), p = 0.035. All comparisons were tested with a two-sample t test. *p < 0.05, **p < 0.01, ***p < 0.005.
- Figure 6.
Transition and comparative analysis of locomotor abilities. A, BBB score comparison. TSR (n = 15) and TR (n = 15). Repeated-measures ANOVA assuming sphericity. p = 0.018. Tukey’s multiple comparisons of means: 53 DPI, p = 0.013; 60 DPI, p = 0.14; 67 DPI, p = 0.15; 74 DPI, p = 0.20; 81 DPI, p = 0.14; 88 DPI, p = 0.016; 95 DPI, p = 0.0072; 102 DPI, p = 0.07; 109 DPI, p = 0.0031. Extended Figure 6-1 shows a comparison of BBB score with rehabilitation + Sema3Ai and rehabilitation. B, Body weight transition. Body weight comparison TSR (n = 15) and TR (n = 15). Two-sample t test: 109 DPI, p-value = 0.583. C, Schematic image of quantitative walking analysis. D, Quantitative walking analysis with stride length. TSR (n = 15) and TR (n = 13), p = 0.026. E, Quantitative walking analysis with paw angle. TSR (n = 15) and TR (n = 13), p = 0.34. F, Quantitative walking analysis with stride duration time. p = 0.26. G, Gastrocnemius weight/body weight. Two-sample t test p-value = 0.142. Comparisons in D–G were conducted with a two-sample t test. *p < 0.05, **p < 0.01, ***p < 0.005.
Tables
- Table 1.
Correspondence table of BBB for each individual at 46 d when groups were divided and at 109 d when all interventions were completed
ID 46 DPI BBB 109 DPI BBB ID 46 DPI BBB 109 DPI BBB TSR1 5 10 TR1 5.5 7 TSR2 5.5 12.5 TR2 6 7 TSR3 5 9 TR3 5 9 TSR4 4 11 TR4 3.5 5 TSR5 4 6 TR5 3 3 TSR6 5 8 TR6 3.5 6 TSR7 6 10.5 TR7 6 8 TSR8 6 7 TR8 6 8 TSR9 5.5 8.5 TR9 6 5.5 TSR10 6 10 TR10 7 10 TSR11 7 12 TR11 7 7.5 TSR12 6 9 TR12 7 10 TSR13 8 10 TR13 6.5 9 TSR14 7 10 TR14 6.5 8.5 TSR15 9 12.5 TR15 7.5 8.5 - Table 2.
Correspondence table of body weight for each individual at 46 d when groups were divided and at 109 d when all interventions were completed
ID 46 DPI weight 109 DPI weight ID 46 DPI weight 109 DPI weight TSR1 182 202 TR1 166 178 TSR2 174 196 TR2 176 200 TSR3 186 204 TR3 178 192 TSR4 166 178 TR4 172 180 TSR5 180 198 TR5 150 166 TSR6 184 204 TR6 172 178 TSR7 184 200 TR7 192 214 TSR8 194 190 TR8 196 202 TSR9 170 182 TR9 174 198 TSR10 186 210 TR10 180 198 TSR11 166 180 TR11 168 186 TSR12 172 184 TR12 186 198 TSR13 170 184 TR13 188 210 TSR14 166 182 TR14 168 184 TSR15 168 198 TR15 170 190
Figure 2-1
Axial-slice of the SCI epicenter with H&E staining of TR group and single rehabilitation group. Scale bar, 100 μm. Download Figure 2-1, TIF file.
Figure 4-1
Graft axon expansion with STEM121 at the SCI epicenter. A, Graft axon expansion with STEM121 in TSR group at the SCI epicenter.in sagittal direction. Scale bar, 100 μm B, Graft axon expansion comparison with STEM121 volume TSR (n=4) and TR (n=4), p=0.62. The comparison was tested with a two-sample t-test. *p<0.05, **p<0.01, ***p<0.005. Download Figure 4-1, TIF file.
Figure 6-1
Comparison of BBB score with Rehabilitation + Sema3Ai (n=26) and Rehabilitation (n=26). Download Figure 6-1, TIF file.
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