硬脑膜体外培养模型的建立及修复机制研究  

A Study about the Construction of an in vitro Model of Dural Cells and its Healing Mechanisms

作　　者：周峰[1] 张建民[1] 陈高[1] 

机构地区：[1]浙江大学医学院附属第二医院神经外科,杭州310009

出　　处：《中国生物医学工程学报》2006年第4期472-475,共4页Chinese Journal of Biomedical Engineering

摘　　要：本研究拟建立一种兔硬脑膜缺损的体外培养模型,观察不同细胞外基质对其愈合过程的影响,研究硬脑膜修复的机制,为以后检测各种硬脑膜移植物提供方便。取1.5cm×1.5cm大小兔硬脑膜在其中央做直径2mm的孔,模仿硬脑膜缺损,放于24孔板中培养。8d^10d后预先铺过胶原的培养板中有细胞从硬脑膜缺损的边缘逸出,增殖,13d^15d硬脑膜缺损愈合;而以多聚赖氨酸及层粘连蛋白铺底的培养板中没有看到细胞的逸出现象。硬脑膜细胞对波形蛋白抗体呈强阳性反应,对第Ⅷ因子抗体则为阴性反应;在电镜下可以看到新生胶原的生成。本研究成功建立一种硬脑膜缺损的体外培养模型,并证实成纤维细胞从硬脑膜创缘的逸出、增殖是硬脑膜缺损愈合的重要机制。To construct an in vitro model of rabbit dural healing and learn the influence of collagen, laminin, polylysine on the migration and proliferation of dural cells and study the healing mechanism of duraplasty. Rabbit dura pieces 1.5cm×1.5cm in size were prepared and were perforated in their central part with a 2mm punch to mimic a dural defect. The dural pieces were cultured in 24-well plates coated with collagen, laminin and polylysine respectively, the effects of different extracellular matrix on the migration and proliferation of dural cells were observed. Cells were subcultured on slides for immunocytochemistry to identify the characteristics of dural cells. The dural healing was observed by scanning electronic microscope. Only the dura pieces cultured in collagen coated wells showed migration of cells into the central defect after a period of 8～10 days and dural defect healing occurred after 13～15 days. Dural cells stained strongly positive with antibodies against vimentin and negative with Ⅷ factor. New collagen fibers were observed in the dural defects. A kind of in vitro model for dural healing was constructed. Experimental results indicated that cellular migration from the dural defect margin was one important mechanism in the process of wound healing after duraplasty.

关 键 词：硬脑膜重建 体外培养 成纤维细胞 细胞外基质 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]