壳聚糖支架与大鼠施万细胞体外组织相容性研究壳聚糖支架与大鼠许旺细胞的体外组织相容性☆

背景：与聚乳酸、胶原等材料相比，壳聚糖作为一种新的生物材料研究时间较短，在神经修复方面的工作开展的较少。 目的：介绍一种简单的壳聚糖支架制备方法并分析其与许旺细胞的生物组织相容性。 设计、时间及地点：体外对比观察实验，于2008-11/2009-07在包头医学院生物医学中心实验室完成。 材料：Wistar 乳鼠10只用于分离培养许旺细胞。医用级壳聚糖粉(脱乙酰度为99%)由浙江澳兴生物科技有限公司提供。 方法：以6%壳聚糖乙酸水溶胶制备壳聚糖支架，取乳鼠坐骨神经采用酶消化后组织块贴壁培养的方法培养许旺细胞。实验分为正常对照组和实验组，实验组将壳聚糖支架平铺于孔底，超净工作台内干燥，然后将许旺细胞接种到培养板内培养，24 h后培养液中加入4,6-二氨基-2-苯基吲哚孵育过夜，去除未与细胞结合的4,6-二氨基-2-苯基吲哚继续培养1周。正常对照组不加任何干预，同样在37 ℃和体积分数为5%CO2的条件下培养1周。 主要观察指标：培养第1，3，5，7天分别选择细胞总数不低于100和200个的视野进行照像，应用图像分析软件(Image pro plus 6.0)分析，计算凋亡细胞的百分率。 结果：壳聚糖支架表面光滑，透明状，质地均匀，厚度0.15 mm。正常对照组和实验组在第1，3，5，7天凋亡细胞的百分率比较，差异无显著性意义(P > 0.05)。 结论：该壳聚糖支架所需材料少，制作方法简单，与许旺细胞具有良好的组织相容性。 关键词：壳聚糖；许旺细胞；组织相容性

In vitro histocompatibility of chitosan scaffolds and Schwann cells in rats

BACKGROUND: Compared to polylactic acid and collagen, chitosan is a newly material in biomaterial field with little understanding in nerve reconstruction. OBJECTIVE: To introduce a simple preparation method of chitosan scaffolds and to analyze its biological tissue compatibility with Schwann cells. DESIGN, TIME AND SETTING: In vitro comparative study experiment. The experiment was conducted in the Laboratory of Medical Center, Baotou Medical College, between November 2008 and July 2009. MATERIALS: A total of 10 wistar neonatal rats were used to prepare Schwann cells. Medical grade chitosan bulk powder with 99% deacetylation was provided by Zhejiang Aoxing Biotechnology Co., ltd. METHODS: Chitosan scaffolds were prepared by 6% chitosan based hydrolgels produced from acetic acid and chitosan. Schwan cell of neonatal rat sciatic nerves was cultured using enzymatic digestion adherent method. All cells were assigned into experimental and control groups. In the experimental group, chitosan membrane was placed at the hole, dried with clean benches. Then Schwann cells was cultured in the culture plate, 24 hours later, DAPI was added, stayed overnight, cultured with renewed DAPI for 1 week. Cells in the control group were cultured under the same conditions for 1 week without any interference. MAIN OUTCOME MEASURES: Photographs of cells (more than 100 or 200 in a visual field) were obtained and analyzed with Image pro plus 6.0 to calculated percentage of apoptotic cells at days 1, 3, 5 and 7 after culture. RESULTS: The surface of chitosan scaffold was smooth and glossy with 0.15 mm thickness. The difference of apoptotic cells percentage between the control group and experimental group had no significance at days 1, 3, 5 and 7 after culture (P > 0.05). CONCLUSION: Chitosan scaffold is simple to prepare, which has good histocompatibility of Schwann cells.