Wakimura Yuki, Wang Wei, Itoh Soichiro, Okazaki Mutsumi, Takakuda Kazuo
Tokyo, Japan From Plastic and Reconstructive Surgery and the Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University; and the Department of Orthopedic Surgery, Kawakita General Hospital.
Plast Reconstr Surg. 2015 Sep;136(3):319e-327e. doi: 10.1097/PRS.0000000000001556.
The authors evaluated the efficacy of decellularized nerve as a scaffold for nerve regeneration.
Sciatic nerves harvested from Sprague-Dawley rats were decellularized in combination with sodium dodecyl sulfate and Triton X-100, and examined with scanning electron microscopy and immunofluorescence staining. A graft into the sciatic nerve in Wistar rats was performed with the decellularized Sprague-Dawley rat sciatic nerves [allograft: 10 mm long (n = 3) for short term and 15 mm long (n = 5) for long term]. As a control, a portion of sciatic nerve of Wistar rats was cut, reversed, and resutured in situ [autograft: 10 mm long (n = 3) and 15 mm long (n = 5) for different terms, respectively]. Samples were harvested 4 weeks postoperatively and prepared for immunohistochemistry. Von Frey hair test, static toe spread factor measurement, and electrophysiologic and histomorphologic analyses were carried out to evaluate nerve recovery 24 weeks postoperatively.
Scanning electron microscopic images revealed the honeycomb structure, and immunohistology showed that the three-dimensional structure of the basal lamina column on which cell adhesion molecules are integrated is preserved through the decellularization protocols. Limited ED1-positive macrophage invasion was found through the decellularized sciatic nerves, suggesting that antigenicity remained more or less after this treatment. Nevertheless, NF160-positive axons accompanied by S100-positive Schwann cells penetrated through the decellularized sciatic nerves. Sciatic nerve function had recovered, and there were no significant differences in the electrophysiologic and histomorphologic recovery in the groups.
These results suggest that the decellularized allogeneic nerve is a suitable scaffold to bridge a nerve gap.
作者评估了去细胞神经作为神经再生支架的疗效。
从Sprague-Dawley大鼠获取坐骨神经,用十二烷基硫酸钠和 Triton X-100联合进行去细胞处理,并用扫描电子显微镜和免疫荧光染色进行检测。将去细胞的Sprague-Dawley大鼠坐骨神经移植到Wistar大鼠的坐骨神经中[同种异体移植:短期为10毫米长(n = 3),长期为15毫米长(n = 5)]。作为对照,将Wistar大鼠的一部分坐骨神经切断、翻转并原位重新缝合[自体移植:不同时期分别为10毫米长(n = 3)和15毫米长(n = 5)]。术后4周采集样本并准备进行免疫组织化学检查。术后24周进行von Frey毛发试验、静态足趾展开因子测量以及电生理和组织形态学分析,以评估神经恢复情况。
扫描电子显微镜图像显示出蜂窝状结构,免疫组织学表明,通过去细胞处理方案,整合有细胞粘附分子的基底层柱的三维结构得以保留。在去细胞坐骨神经中发现有限的ED1阳性巨噬细胞浸润,这表明该处理后仍或多或少残留有抗原性。然而,伴有S100阳性雪旺细胞的NF160阳性轴突穿过了去细胞坐骨神经。坐骨神经功能已恢复,各组在电生理和组织形态学恢复方面无显著差异。
这些结果表明,去细胞同种异体神经是桥接神经缺损的合适支架。