Okuda Akinori, Horii-Hayashi Noriko, Sasagawa Takayo, Shimizu Takamasa, Shigematsu Hideki, Iwata Eiichiro, Morimoto Yasuhiko, Masuda Keisuke, Koizumi Munehisa, Akahane Manabu, Nishi Mayumi, Tanaka Yasuhito
Department of Orthopaedic Surgery, Nara Medical University, Kashihara.
Department of Anatomy and Cell Biology, Faculty of Medicine, Nara Medical University, Kashihara.
J Neurosurg Spine. 2017 Mar;26(3):388-395. doi: 10.3171/2016.8.SPINE16250. Epub 2016 Nov 25.
OBJECTIVE Transplantation of bone marrow stromal cells (BMSCs) is a theoretical potential as a therapeutic strategy in the treatment of spinal cord injury (SCI). Although a scaffold is sometimes used for retaining transplanted cells in damaged tissue, it is also known to induce redundant immunoreactions during the degradation processes. In this study, the authors prepared cell sheets made of BMSCs, which are transplantable without a scaffold, and investigated their effects on axonal regeneration, glial scar formation, and functional recovery in a completely transected SCI model in rats. METHODS BMSC sheets were prepared from the bone marrow of female Fischer 344 rats using ascorbic acid and were cryopreserved until the day of transplantation. A gelatin sponge (GS), as a control, or BMSC sheet was transplanted into a 2-mm-sized defect of the spinal cord at the T-8 level. Axonal regeneration and glial scar formation were assessed 2 and 8 weeks after transplantation by immunohistochemical analyses using anti-Tuj1 and glial fibrillary acidic protein (GFAP) antibodies, respectively. Locomotor function was evaluated using the Basso, Beattie, and Bresnahan scale. RESULTS The BMSC sheets promoted axonal regeneration at 2 weeks after transplantation, but there was no significant difference in the number of Tuj1-positive axons between the sheet- and GS-transplanted groups. At 8 weeks after transplantation, Tuj1-positive axons elongated across the sheet, and their numbers were significantly greater in the sheet group than in the GS group. The areas of GFAP-positive glial scars in the sheet group were significantly reduced compared with those of the GS group at both time points. Finally, hindlimb locomotor function was ameliorated in the sheet group at 4 and 8 weeks after transplantation. CONCLUSIONS The results of the present study indicate that an ascorbic acid-induced BMSC sheet is effective in the treatment of SCI and enables autologous transplantation without requiring a scaffold.
目的 骨髓基质细胞（BMSCs）移植作为脊髓损伤（SCI）治疗策略具有理论潜力。尽管有时使用支架来将移植细胞保留在受损组织中，但也已知其在降解过程中会引发多余的免疫反应。在本研究中，作者制备了由BMSCs制成的细胞片，其可在无支架的情况下移植，并在大鼠完全横断性SCI模型中研究了它们对轴突再生、胶质瘢痕形成和功能恢复的影响。方法 使用抗坏血酸从雌性Fischer 344大鼠的骨髓中制备BMSC片，并冷冻保存至移植当天。将明胶海绵（GS）作为对照或BMSC片移植到T-8水平脊髓的2毫米大小缺损处。分别在移植后2周和8周通过使用抗Tuj1和胶质纤维酸性蛋白（GFAP）抗体的免疫组织化学分析评估轴突再生和胶质瘢痕形成。使用Basso、Beattie和Bresnahan评分评估运动功能。结果 BMSC片在移植后2周促进了轴突再生，但在细胞片移植组和GS移植组之间Tuj1阳性轴突数量没有显著差异。移植后8周，Tuj1阳性轴突穿过细胞片伸长，细胞片组中的轴突数量明显多于GS组。在两个时间点，细胞片组中GFAP阳性胶质瘢痕的面积均明显小于GS组。最后，移植后4周和8周时细胞片组的后肢运动功能得到改善。结论 本研究结果表明，抗坏血酸诱导的BMSC片在SCI治疗中有效，并且无需支架即可进行自体移植。
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