Bone Marrow Mesenchymal Stem Cells in a Three-Dimensional Gelatin Sponge Scaffold Attenuate Inflammation, Promote Angiogenesis, and Reduce Cavity Formation in Experimental Spinal Cord Injury

Abstract

Three-dimensional (3D) gelatin sponge (GS) scaffolds were constructed by ensheathing GS with a thin film of poly-(lactide-co-glycolide) (PLGA). Rat bone marrow-derived mesenchymal stem cells (MSCs) were isolated, cultured, and then seeded to the scaffolds. Distribution of cells and cell growth, survival, and proliferation within the scaffolds were then determined. Immunofluorescence and Western blot analysis were employed to detect the deposition of fibronectin to the scaffolds on day 3 and day 7 of culture. Scaffolds with or without MSCs were then transplanted into the transected rat spinal cord. One or 8 weeks following transplantation, cavity areas, activated macrophages/microglia, expression of TNF-α and IL-1β, and neovascularization within the grafts were examined and quantified. Deposition of fibronectin (FN) and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) as potential inducing factors for angiogenesis were also examined. Results showed that 3D GS scaffolds allowed MSCs to adhere, survive, and proliferate and also FN to deposit. In vivo transplantation experiments demonstrated that these scaffolds were biocompatible, and MSCs seeded to the scaffolds played an important role in attenuating inflammation, promoting angiogenesis, and reducing cavity formation. Therefore, the GS scaffolds with MSCs may serve as promising supporting transplants for repairing spinal cord injury.

Autocrine fibronectin from differentiating mesenchymal stem cells induces the neurite elongationin vitroand promotes nerve fiber regeneration in transected spinal cord injury

“…Although RT‐PCR showed an obvious reduction in FN expression, the amount of FN protein within the scaffold increased with time. This observation was consistent with our previous finding that gelatin could accumulate FN21 due to their binding via the GBD domain 22. Since this binding is relatively strong as it even resists normal detergents,23 almost the entire surface of gelatin sheet was covered by FN fibrils.…”

“…4(F)]. We reported previously that deposited FN was detected on GS scaffold at least one week after implantation in the spinal cord 21. In this study over a protracted time period in the spinal cord, autocrine FN was detected at 4 weeks after implantation.…”

Mesenchymal Stem Cell Therapy for Apoptosis After Spinal Cord Injury

“…Even though the present data are promising, further research is needed to establish whether bone marrow cell treatments can serve as a safe and efficacious autologous source for the treatment of the injured SCI (Wright et al, 2011). Downregulation of TNF-α expression in macrophages/microglia was observed at an early stage after SCI in rats transplanted with a gelatin sponge scaffold impregnated with rat bone marrow-derived mesenchymal stem cells at the site of injury (Zeng et al, 2011).…”

Graft of a Tissue-Engineered Neural Scaffold Serves as a Promising Strategy to Restore Myelination after Rat Spinal Cord Transection

“…The transfection efficiency with these viral vectors has been well documented previously by us [13,21]. Three-dimensional GS scaffolds were prepared as previously described [22]. A total of 5 · 10 5 cells (NSCs and SCs were mixed in 1:1 ratio) in 10 mL culture medium were seeded to each scaffold.…”