Department of Gastroenterology, People's Hospital of Jiangyin, Wuxi, Jiangsu 214400, China. qianlm0921@ hotmail.com
Chin Med J (Engl). 2009 Sep 5;122(17):2032-40.
Implantation of tissue-engineered scaffolds is one of the most promising therapeutic strategies for inducing nerve regenerations following spinal cord injuries. In this paper, we report a novel bioengineered hybrid scaffold comprised of three major extracellular matrix (ECM) proteins.
ECM-scaffolds (ECM-S) were prepared by gelling fibrinogen, fibronectin and laminin using fresh rat plasma. Olfactory ensheathing cells (OECs) were isolated from fresh rat olfactory mucosa, purified under differential adhesion, and assessed by immunofluorescent staining. OECs were seeded onto ECM-S and cultured. The effects of the scaffolds on the seeded cells were detected using the immunofluorescent staining, Western blotting, scanning electron microscopy and transmission electron microscopy.
Tissue-engineered ECM-S could be easily molded into mat-like or cylindrical shapes and gelled by addition of fresh plasma. Observations by electron microscopy show that the ECM-S forms a stable three-dimensional porous network. Studies on the effects of the ECM-S on the biological behaviors of OECs in vitro indicate that the scaffold can promote OEC adhesion, proliferation and process extensions. Additionally, OECs seeded on the scaffold maintained the expression of nerve growth factor, matrix metalloproteinase-3 and matrix metalloproteinase-9.
We developed a biosynthetic hybrid gel which could be used as a scaffold for OEC transplantation; this gel can promote nerve regeneration following spinal cord injuries.
组织工程支架的植入是脊髓损伤后诱导神经再生的最有前途的治疗策略之一。在本文中,我们报告了一种由三种主要细胞外基质(ECM)蛋白组成的新型生物工程混合支架。
使用新鲜大鼠血浆将纤维蛋白原、纤维连接蛋白和层粘连蛋白凝胶化,制备 ECM 支架(ECM-S)。从新鲜大鼠嗅粘膜中分离出嗅鞘细胞(OECs),通过差速贴壁法进行纯化,并通过免疫荧光染色进行评估。将 OEC 接种到 ECM-S 上进行培养。通过免疫荧光染色、Western blot、扫描电子显微镜和透射电子显微镜检测支架对接种细胞的影响。
组织工程 ECM-S 可以很容易地模制成类似垫子或圆柱形,并通过添加新鲜血浆凝胶化。电子显微镜观察表明,ECM-S 形成了稳定的三维多孔网络。体外研究 ECM-S 对 OEC 生物学行为的影响表明,支架可以促进 OEC 的黏附、增殖和突起延伸。此外,接种在支架上的 OEC 保持了神经生长因子、基质金属蛋白酶-3 和基质金属蛋白酶-9 的表达。
我们开发了一种可作为 OEC 移植支架的生物合成混合凝胶,该凝胶可促进脊髓损伤后的神经再生。