【摘要】 目的探索壳聚糖复合支架经透明质酸(HA)及多聚赖氨酸(PLL)修饰后,神经元在体外复合支架黏附情况,以及将不同复合支架植入鼠脑,观察支架对大鼠脑组织炎症反应、胶质纤维表达的差异。方法孔隙为(70.32±15.33)μm壳聚糖复合支架分别用质量浓度0.05mg/mLHA和质量浓度2mg/mLPLL进行表面涂镀法修饰,继而在上述两组及未修饰复合支架上进行小鼠神经细胞培养,1d后对支架细胞固定行扫描电子显微镜形态学观察,记录两组各30例支架高倍视野下细胞黏附数量;另将复合支架植入大鼠脑皮层损伤区,术后不同时间点分别取脑组织支架行苏木精-伊红染色、免疫组织化学观察炎性细胞及胶质纤维的表达情况。数据用SAS9.1.3统计软件处理,P<0.05为差异有统计学意义。结果 HA修饰的壳聚糖复合支架上细胞黏附较多[扫描电子显微镜,×1700,(24.9±4.5)/高倍镜],而经PLL修饰的壳聚糖复合支架上细胞黏附相对较少[扫描电子显微镜,×1700,(19.2±3.2)/高倍镜],未修饰支架上细胞碎片较多,并且细胞聚集,细胞总数少;在体内用HA修饰的壳聚糖复合支架组与其他组相比,支架周围炎症反应轻、支架周围胶质纤维酸性蛋白(GFAP)阳性细胞数少。所获得数据分析后差异均有显著统计学意义(P<0.01)。结论 HA修饰的壳聚糖复合支架能够提高体外神经元黏附,并且在体内观察炎性细胞,胶质纤维表达均优越于PLL修饰的壳聚糖复合支架、单纯壳聚糖复合支架。HA修饰的壳聚糖复合支架在神经组织工程应用中更具有研发前景。更多还原

【Abstract】 Objective To investigate the neuronal adherence in vitro between chitosan scaffold and the scaffold with modified hyaluronic acid(HA) and poly-L-lysine(PLL),and observe the differences in the expression of glial fibrillary after stent implantation in rat brain in vivo.Methods The surface-coated method was used to modify chitosan scaffolds(pore size 70.32 μm±15.33 μm) with 0.05 % W/V HA and 2 mg/mL PLL.Then rat neurons were cultured on these scaffolds and unmodified scaffolds,and investigated morphologically by scanning electron microscopy(SEM) after fixed 1 day.The number of cell adhesion at high-power microscope of 30 cases in each group was recorded.The brain tissues in the damage zone of rat cerebral cortex were stained by hematoxylin-eosin(HE) and immunohistochemistry,then observed inflammation and glial fibers expression.The result treated with SAS 9.1.3 statistical software,and P<0.05 was statistically significance.Results There were more cells adhered on HA scaffold[SEM,×1700,(24.9±4.5)/high-power] than that on PLL scaffold[SEM,×1700,(19.2±3.2)/high-power],more cellular debris,less cell numbers and aggregation on unmodified scaffolds.In vivo compared with other groups the HA modified chitosan scaffold group showed slight inflammation and less GFAP-positive cells around the scaffolds.The differences were statistically significant(P<0.01).Conclusion It is demonstrated that scaffolds of chitosan modified with HA could elevate the cell adhesion in vitro and showed superiority in aspect of inflammatory cells and glial fibres expression compared with unmodified scaffold and chitosan scaffold modified by PLL.The HA modified chitosan scaffolds has the value of research and development prospect in nervous tissue engineering.更多还原