目的:探讨纤维蛋白支架对人嗅鞘细胞髓鞘相关蛋白表达的影响。方法:将体外培养的成人鼻粘膜嗅鞘细胞种植于纤维蛋白支架,用多聚赖氨酸修饰的玻片作为对照底物,培养14天后用扫描电镜观察细胞的形态学变化并对细胞内髓鞘形成相关蛋白2,3-环核苷3-磷酸二酯酶(2′,3′cyclic nucleotide 3′phosphodiesterase,CNPase)和髓鞘碱性蛋白(myelin basic protein,MBP)进行免疫荧光染色定位和免疫印迹半定量测定,分析纤维蛋白支架对人嗅鞘细胞CNPase和MBP表达的影响。结果:人嗅鞘细胞种植于纤维蛋白支架后,其形态为长条带状,细胞成束排列,方向一致。细胞内CNPase和MBP表达水平明显高于对照组。结论:纤维蛋白支架与人嗅鞘细胞具有很好的生物相容性并可促进细胞表达髓鞘相关蛋白CNPase和MBP;将该组织工程支架用于移植治疗脊髓损伤,有利于再生神经纤维的髓鞘形成。
Objective: To investigate the effects of fibrin scaffold on the myelin associated proteins expression of the human olfactory ensheathing cells (hOECs) derived from olfactory mucosa. Methods: The hOECs were cultured and seeded separately onto the fibrin scaffolds as experimental group and the glass slides modified with poly-L-lysine(PLL) as control group. After cultured 14 days, the morph of the hOECs was observed by scanning electron microscopy. To investigate the effects of fibrin scaffold on the myelin associated proteins expression, the hOECs were immunofluorescence stained with antibodies against 2′,3′ cyclic nucleotide 3′ phosphodiesterase (CNPase) and myelin basic protein (MBP). The relative contents of CNPase and MBP were detected by Western blotting. Results: The hOECs growthed on the fibrin scaffold showed ribbon-like and arranged in alignment. Some hOECs had migrated into the scaffolds. The results of immunofluorescence staining and Western blotting indicated that the expressions of CNPase and MBP of hOECs grew on the fibrin scaffold were higher than that in the control group. Conclusion: The fibrin scaffold possesses a good biocompatibility with hOECs and could promote expressions of CNPase and MBP in the hOECs seeded in the scaffold. The tissue engineering composed of hOECs and fibrin could be used for implantation to repair the injured spinal cord.