Background Spinal cord injury (SCI) causes a rapid loss of motor neurons, leading to weakness and paralysis. resulted in an enhanced neuronal repair and motor behavioral outcome compared to the normal NSCs transplanted group. VEGF-NSCs increased the release of neurotrophic factors and reduced the expression of TRPV1. Conclusions Recombinant VEGF-NSCs transplantation following SCI is more efficacious compared to normal NSC transplantation. This might also be related to a reduced pain in the process of recovery due to reduced TRPV1 expression. Western blot analyses, with a greater VEGF protein level in VEGF-NSC transgene-induced rats (Physique 1B, 1C). NSC-transplanted rats produced a significant increase in VEGF mRNA Selumetinib distributor compared to PBS-treated and sham-operated rats (control), while the increase in VEGF protein level was not significant. VEGF is usually endogenously upregulated following SCI and the expression was evidently upregulated in the VNSC transplanted experimental group. Immunohistochemical analyses also supported the results showing the rats transplanted with VNSCs produced more VEGF compared to NSC-transplanted rats (Physique 1D). VEGF-positive staining was quantified in all groups (Physique 1E). Open in a separate window Physique 1 VEGF expression following spinal cord injury in rats. SC C sham-operated controls, PBS C PBS transplantation, NSCs C normal NSC transplantation, VNSC C recombinant VEGF-NSCs transplantation. (A) VEGF mRNA expression post-transplantation, where VNSC exhibit maximum VEGF expression. (B) Western blot analyses of VEGF expression at protein level. (C) Quantification of VEGF protein expression. (D) Immunohistochemical staining of VEGF and CD34 to determine the post-transplantation effect of VEGF on spinal cord sections (400). (E) Quantification of VEGF-positive cells using Image-Proplus v 6.0 software. The quantification was done by taking 10 photo-micrographs, and the number of positive VEGF stained cells was measured. The total number of the positively stained cells was expressed as percentages relative to the total number of cells in the analyzed section. Values are expressed as mean SEM, * p 0.05 compared to sham-operated controls; # p 0.05 compared to NSC. Selumetinib distributor Increase in the motor Selumetinib distributor neurons following VNSC transgene transplantation To determine the effect of VEGF-NSC transgene transplantation on the loss of motor neurons, the number of motor neurons was counted in the spinal cord sections. Rats transplanted with VNSC had 45% greater density of motor neurons compared to NSC-transplanted rats (Physique 2). These results strongly support that VEGF-induced NSCs restore the loss of neurons following SCI. Open in a separate window Physique 2 Motor neuron regeneration post-transplantation. (A) Positive Nissl staining exhibited the number of motor neurons. There was a significant increase in the number of neurons following VNSC transplantation compared to NSC-transplanted groups. PBS transplanted groups had the greatest neuronal loss. (B) Quantification of number of neurons. The quantification was performed by counting the number of positively stained neurons (with intact cell body) in each group and is expressed as percentages relative to the total number of cells in the examined tissue section. Values are represented as mean SEM, * p 0.05 compared to PBS transplanted group; # p 0.05 compared to normal NSC-transplanted group. VNSC transplantation is also directly related to the increased release of BDNF, GDNF, and NT-3 ELISA evaluated the release of growth factors following VEGF-NSC transgene transplantation and compared it with the normal NSC transplantation. The release of these factors was significantly increased in NSC-VEGF transplanted rats compared to normal NSC transplantation following SCI (Physique 3AC3C). The stimulated release of these neurotrophic factors and neurite outgrowth-promoting factor positively enhanced the neuronal repair. This adds to the evidence that VEGF recombinant NSC transplantation is usually a better approach than the transplantation of normal Rabbit Polyclonal to ABCC3 NSCs alone. Open in a separate window Physique 3 Quantification of release of GDNF, BDNF, and NT-3 post-transplantation. (A).