胶原海绵支架促进Moyamoya颅骨钻孔术后侧支新生及其机制

Multiple burr hole is an option in the treatment of Moyamoya. The curative effect can be impaired due to the airspace between the dura and cortex and the relatively deficiency of EPCs in both quality and quality in patient with Moyamoya. As a result, the amount of collateral angiogenesis is decreased and the speed of collateral formation is slowed down. In clinical practice we found implantation of collagen disc under the dura incision could enhance the collateral angiogenesis both in quality and in quatity after multiple burr hole. We speculate the mechanism may be related to disc bridging the dura and the cortex and the disc recruiting EPCs and EPCs paracrining angiogenesis related factores. In this research, we plan to implant collagen disc under dura incision in Moyamoya animal model and detect focal EPCs and angiogenesis related factors. The aim is to explore the mechanism of collagen disc implantation enhancing collateral angiogenesis. Then we further implant the collagen disc which has been cultivated with EPCS in vitro until the capillary-like vessel has appeared, or dropwise EPCS after simple collagen disc implantation. These may further enhance collateral angiogenesis in both quality and quatity. Related research has not been reported and the results may provide clue to improve the curative effect of multiple burr hole in the treatment of Moyamoya.

颅骨钻孔血运重建是Moyamoya治疗常用术式，但由于硬膜与脑皮层间的空隙，以及Moyamoya患者内皮祖细胞（EPC）质和量相对不足，影响了侧支形成的速度和数量。课题组发现，术中于硬膜缘植入胶原海绵支架能促进侧支形成。我们推测其机制可能与支架连接硬膜与脑皮层的桥梁作用和胶原海绵促进局部EPC富集以及富集的EPC释放血管新生因子有关。课题拟通过动物模型观察支架植入对EPC的招募作用及局部血管新生相关因子的变化，探讨胶原海绵支架植入促进Moyamoya侧支形成机制，并进一步提出支架加EPC悬液接种以及EPC-支架体外共培养，待支架内出现丰富毛细血管样结构后植入两种方法，以直接在手术部位添加EPC，省去体内EPC在局部富集、增殖的过程，以进一步提高侧支形成的速度和数量。本研究在Moyamoya治疗中无类似研究，将为Moyamoya治疗提供全新的思路。

Moyamoya是神经外科常见脑血管疾病，颅骨钻孔血运重建是Moyamoya治疗常用术式，但由于硬膜与脑皮层间的空隙，以及Moyamoya患者内皮祖细胞（EPCs）质和量相对不足，影响了术后侧支形成的速度和数量。课题组在既往临床工作中发现，术中于硬膜缘植入胶原海绵支架能促进侧支形成的速度和数量。我们推测其机制可能与支架连接硬膜与脑皮层的桥梁作用和胶原海绵促进局部EPC富集有关。以此为基础，本课题拟通过在Moyamoya动物模型中模拟颅骨钻孔血运重建手术，在硬膜切开部位分别植入胶原海绵支架以及EPCs复合胶原海绵支架，通过对术后局部侧支血管、EPCs以及血管新生情况的动态检测，观察EPCs复合胶原海绵支架植入这一治疗Moyamoya的新术式是否能改善手术效果，并探讨可能机制。为此，课题按计划进行了：1、体外培养扩增同源性F344大鼠骨髓来源EPCs，并与胶原海绵共培养形成EPCs-胶原海绵复合体。2、采用双侧颈内动脉同期结扎法建立慢性脑缺血同源性F344大鼠动物模型。模型建立后第3天，行颅骨钻孔间接血运重建并于骨孔处植入EPCs-胶原海绵复合体。3、激光多普勒血流仪检测局部脑血流变化，荧光示踪体外扩增EPCs，免疫组化分析钻孔处新生血管重建情况。结果发现：颅骨钻孔血运重建术前，EPCs-胶原海绵复合体、胶原海绵及对照组三组间脑血流变化无统计学差异(p>0.05)。血运重建术后第21天，EPCs-胶原海绵组和胶原海绵组的局部脑血流量较对照组升高，且EPCs-胶原海绵复合体组骨孔处新生血管数量明显高于其余两组。标记的EPC融合入钻孔处的血管内皮。本研究初步证明钻孔部EPCs复合胶原海绵支架植入能促进血管新生的数量和速度。其科学意义在于这为Moyamoya间接血运重建提供了新的手术思路和理论依据。