Olson Heather E, Rooney Gemma E, Gross LouAnn, Nesbitt Jarred J, Galvin Katherine E, Knight Andrew, Chen BingKun, Yaszemski Michael J, Windebank Anthony J
Department of Neurology, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA.
Tissue Eng Part A. 2009 Jul;15(7):1797-805. doi: 10.1089/ten.tea.2008.0364.
Biodegradable polymer scaffolds provide an excellent approach to quantifying critical factors necessary for restoration of function after a transection spinal cord injury. Neural stem cells (NSCs) and Schwann cells (SCs) support axonal regeneration. This study examines the compatibility of NSCs and SCs with the poly-lactic-co-glycolic acid polymer scaffold and quantitatively assesses their potential to promote regeneration after a spinal cord transection injury in rats. NSCs were cultured as neurospheres and characterized by immunostaining for nestin (NSCs), glial fibrillary acidic protein (GFAP) (astrocytes), betaIII-tubulin (immature neurons), oligodendrocyte-4 (immature oligodendrocytes), and myelin oligodendrocyte (mature oligodendrocytes), while SCs were characterized by immunostaining for S-100. Rats with transection injuries received scaffold implants containing NSCs (n=17), SCs (n=17), and no cells (control) (n=8). The degree of axonal regeneration was determined by counting neurofilament-stained axons through the scaffold channels 1 month after transplantation. Serial sectioning through the scaffold channels in NSC- and SC-treated groups revealed the presence of nestin, neurofilament, S-100, and betaIII tubulin-positive cells. GFAP-positive cells were only seen at the spinal cord-scaffold border. There were significantly more axons in the NSC- and SC- treated groups compared to the control group. In conclusion, biodegradable scaffolds with aligned columns seeded with NSCs or SCs facilitate regeneration across the transected spinal cord. Further, these multichannel biodegradable polymer scaffolds effectively serve as platforms for quantitative analysis of axonal regeneration.
可生物降解的聚合物支架为量化脊髓横断损伤后功能恢复所需的关键因素提供了一种极佳的方法。神经干细胞(NSCs)和雪旺细胞(SCs)支持轴突再生。本研究检测了NSCs和SCs与聚乳酸-乙醇酸聚合物支架的相容性,并定量评估了它们在大鼠脊髓横断损伤后促进再生的潜力。将NSCs培养为神经球,并通过对巢蛋白(NSCs)、胶质纤维酸性蛋白(GFAP)(星形胶质细胞)、βIII微管蛋白(未成熟神经元)、少突胶质细胞-4(未成熟少突胶质细胞)和髓鞘少突胶质细胞(成熟少突胶质细胞)进行免疫染色来鉴定,而SCs则通过对S-100进行免疫染色来鉴定。患有横断损伤的大鼠接受含有NSCs(n = 17)、SCs(n = 17)和无细胞(对照)(n = 8)的支架植入物。移植1个月后,通过计数穿过支架通道的神经丝染色轴突来确定轴突再生的程度。对NSC和SC治疗组的支架通道进行连续切片,发现存在巢蛋白、神经丝、S-100和βIII微管蛋白阳性细胞。仅在脊髓-支架边界处可见GFAP阳性细胞。与对照组相比,NSC和SC治疗组的轴突明显更多。总之,接种了NSCs或SCs的具有排列柱的可生物降解支架有助于脊髓横断处的再生。此外,这些多通道可生物降解聚合物支架有效地作为轴突再生定量分析的平台。