Enhanced neuroregenerative effects by scaffold for the treatment of a rat spinal cord injury with Wnt3a-secreting fibroblasts.

BACKGROUND: Wnt proteins are bifunctional axon guidance molecules, several of which appear to mediate guidance of corticospinal tract axons along the spinal cord. Here, we studied increasing effect on regeneration by Wnt-containing alginate scaffolds on spinal cord injury (SCI). METHODS: A total of 32 rats were injured at the T7-8 level with an NYU impactor. According to transplantation materials, rats were classified into four groups: a Wnt3a-secreting fibroblast transplantation group (Wnt group, n = 8), a Wnt3a-secreting fibroblast with alginate transplantation group (Wnt + alginate group, n = 8), an alginate transplantation group (alginate group, n = 8), and a contusion-only group (sham group, n = 8). Behavioral tests were performed on the first, second, and third days after injury, and then weekly for 8 weeks. Five of the eight rats from each group were selected for manganese-enhanced magnetic resonance imaging (ME-MRI). Two rats from each group were examined for GAP43 and MAP2 expression using monoclonal and polyclonal primary antibodies, respectively. RESULTS: Seven weeks after transplantation (8 weeks after SCI), Wnt + alginate group rats achieved an average Basso-Beattie-Bresnahan locomotor score of 19.0, which was significantly higher than that of other groups. ME-MRI at 8 weeks after SCI revealed significantly higher relative signal intensities in the Wnt + alginate group. Gap43 and Map2 immunostaining, showed strong positive in the Wnt + alginate group. CONCLUSION: The Wnt + alginate complex exerted significantly enhanced recovery in a rat SCI model compared to alginate or Wnt3a alone. These results suggest that alginate scaffolds facilitate the regeneration of axon working with Wnt3a protein that promotes regeneration of the injured spinal cord.

Analysis of equivalent parameters of two spinal cord injury devices: the New York University impactor versus the Infinite Horizon impactor.

BACKGROUND CONTEXT: The New York University (NYU) impactor and the Infinite Horizon (IH) impactor are used to create spinal cord injury (SCI) models. However, the parameters of these two devices that yield equivalent SCI severity remain unclear. PURPOSE: To identify equivalent parameters, rats with SCIs induced by either device set at various parameters were subjected to behavioral and histologic analyses. STUDY DESIGN: This is an animal laboratory study. METHODS: Groups of eight rats acquired SCIs by dropping a 10 g rod from a height of 25 mm or 50 mm by using the NYU device or by delivering a force of 150 kdyn, 175 kdyn, 200 kdyn, or 250 kdyn by using the IH impactor. All injured rats were tested weekly for 8 weeks by using the Basso, Beattie, and Bresnahan (BBB) test and the ladder rung test. On the 10th week, the lesion volume of each group was measured by using a 9.4 Tesla magnetic resonance imaging (MRI), and the spinal cords were subjected to histologic analysis using anterograde biotinylated dextran amine (BDA) tracing and immunofluorescence staining with an anti-protein kinase C-gamma (PKC-γ) antibody. RESULTS: Basso, Beattie, and Bresnahan test scores between the 25 mm and the 200 kdyn groups as well as between the 50 mm and and 250 kdyn groups were very similar. Although it was not statistically significant, the mean scores of the ladder rung test in the 200 kdyn group were higher than the 25 mm group at all assessment time points. There was a significantly different cavity volume only between the 50 mm and the 200 kdyn groups. Midline sagittal images of the spinal cord on the MRI revealed that the 25 mm group predominantly had dorsal injuries, whereas the 200 kdyn group had deeper injuries. Anterograde tracing with BDA showed that in the 200 kdyn group, the dorsal corticospinal tract of the caudal area of the lesion was labeled. Similar labeling was not observed in the 25 mm group. Immunofluorescence staining of PKC-γ also revealed strong staining of the dorsal corticospinal tract in the 200 kdyn group but not in the 25 mm group. CONCLUSIONS: The 25 mm injuries generated by the NYU impactor are generally equivalent to the 200 kdyn injuries generated by using the IH impactor. However, differences in the ladder rung test scores, MRI images, BDA traces, and PKC-γ staining demonstrate that the two devices exert qualitatively different impacts on the spinal cord.