摘要
目的 利用新型的脱细胞-交联技术制备脱细胞脑组织支架,为脑损伤修复提供一种实用性支架材料.方法 8只健康SD成年大鼠皮层脑组织,完全随机法选取2只用于正常脑基质成分免疫组织化学染色,其余6只经过脱细胞及交联处理,初步制成脱细胞脑组织基质修复支架;光镜及电镜观察支架空间构型,了解支架交联前后失质量率的改变;免疫组织化学对比观察正常脑组织和支架材料中纤维粘连蛋白（FN）、细胞外基质层粘连蛋白（LN）和Ⅲ型胶原（ColⅢ）的含量和分布情况.结果 大鼠皮层脑组织经脱细胞处理后,形态上具有高度仿生的三维立体孔洞结构,孔壁较薄（厚度约2μm）,孔径相近（平均约50～200 μm）,孔隙率约为85%～92%,交联后支架材料失质量率明显变小（15.09比62.50,P=0.001）;免疫组化显示LN、FN和ColⅢ在正常脑组织呈弱阳性表达,在支架材料中LN亦呈弱阳性表达.结论 初步研制的脱细胞脑组织基质修复支架具备了神经修复支架材料的基本物理学条件,具有作为中枢神经系统再生支架材料的良好应用前景.
Objective To prepare an acellular cerebral matrix scaffold using a novel decellularization and crosslinking method, so as to provide a practical biomaterial for repair of brain injuries.Methods The brain tissues of 8 healthy adult rats were randomized into the uncrosslinking group （n=2） for immunohistochemical staining of normal cerebral matrix and the crosslinking group （n=6） for treatment with decellularization and crosslinking procedure to prepare an acellular cerebral matrix scaffold. The scaffold was studied for geometric configurations by light microscope and electron microscopy, analyzed for change in percentage weight loss before and after crosslinking. The contents and distribution of three important extracellular matrix components, fibronectin （FN）, laminin （LN） and collagen （Col） Ⅲ, in uncrosslinking and crosslinking groups were examined by immunohistochemistry. Results After decellularization, the cerebral cortex of rats was shaped into a highly-bionic, three-dimensional porous structure, interspersed by thin-walled pores （2 μm in thickness） of similar size （50 to 200 μm by diameter） with a porosity from 85% to 92%. The percentage weight loss of the scaffold was significantly reduced after cross-linking procedure （15.09 vs 62.50, P=0.001）. Immunohistochemical staining showed weak- positive expressions of three common extracellular matrix elements （LN, FN and Col Ⅲ ） in normal brain tissues and a weak-positive expression of LN in the matrix scaffolds. Conclusion The acellular cerebral matrix scaffold prepared in this preliminary study displays basic physical features as required for nerve- repairing scaffold and may be promising for use in regeneration of central nervous system.
出处
《中华生物医学工程杂志》
CAS
2010年第6期552-556,共5页
Chinese Journal of Biomedical Engineering
基金
国家自然科学基金（30672152）
