Title

去细胞同种异体移植面神经材料的制备

Translated Titles

Acellular Scaffold for Tissue Engineered Facial Nerve Allograft

Authors

刘洪飞(Hong-Fei Liu);胡敏(Min Hu);刘洪臣(Hong-Chen Liu);孙明学;王新

Key Words

面神经 ; 组织工程 ; 化学萃取 ; Facial nerves ; Tissue engineering ; Chemical extraction

PublicationName

口腔醫學研究

Volume or Term/Year and Month of Publication

21卷1期(2005 / 02 / 28)

Page #

8 - 11

Content Language

簡體中文

Chinese Abstract

目的:探讨如何构建异体组织工程化面神经,提供具有仿生结钩的支架。方法:取8段长5cm、直径2mm兔的面神经,其中6段用3%三硝基甲苯和4%脱氧胆酸钠分别萃取1、2和3次,在萃取神经和末萃取神经的中段取材,行HE染色、Masson染色和S-100免疫组化染色,在光镜和电镜下观察神经萃取前后形态学的变化结果:用只硝基甲苯和脱氧胆酸钠萃取后,面神经内的细胞消失,而纤维队支架结构与未经萃取的神经相仿,电镜下可见萃取后的神经由空的神经基底膜管和管之间的胶原纤维构成,随着萃取次数的增加,神经内残留的S-100蛋白显著减少,但反复萃取后神经的支架结构受破坏。结论:用二硝基甲苯和脱氧胆酸钠萃取2次可去除兔面神经的细胞而保留完整的神经基底膜管和纤维支架结构,是制备具有仿生结构的组织工程化面神经支架较为理想的方法。

English Abstract

Objective: To net clap a procedure in xx bob Schwann cells and myelin in the facial nerve could be removed while the basal lamina tubes remained intact, anti to obtain an acellular scaffold for fabricating tissue-engineered facial allograft in rabbits. Methods: eight facial nerves with 5.0 cm in length 2.0 mm in diameter were excised from rabbits anti cleaned from external debris. The nerves were treated with a solution of Triton X-100 and a solution of sodium deoxycholate at room temperature. After a finial wash in water, the nerves were stored in phosphate-buffered saline (PBS, pH 7.2) at 4 degrees. HE, luxol fast blue and fibrin staining were performed to visualize cells, myelin and basal membranes respondent, in and immunohistocbemical staining was performed to visualize the presence of laminin, a Schwann cell lamina component, in both fresh and acellular wae segments. To revcal overall structure better, methylene blue-fuchsin staining was performed in semithin section. The ultrastructure of acellular and fresh nerves were observed and photographed in a transmission electron microscope. Results: The acellular facial nerve was white long cylinder with well elasticity and ductility. HE, myelin and fibrin staining revealed that cells, axons and myelin sheath were removed and basal membrane was preserved after extraction procedure. Staining for the presence of laminin showed that the Schwann cell basal lamina component were present in the nerves after obemical treatment. Methylene blue-fuchsin staining and transmission electron microscopy showed that the myelin sheaths were absent in the extracted nerve segments anti empty basal lamina tubes remained in the endoneurium. Conclusions: We developed an extracted procedure with the detergents of Triton X-100 and deoxycholate, by which cells, axons and myelin sheaths could be removed from the rabbit facial nerve while the basal lamina tubes remained intact anti an aeellular nerve allograft was obtained. The laminin, a Schwann cell basal lamina component, can be preserved in the acellular nerve. Chemical extraction is an ideal method to prepare nerve scaffold for fabricating tissue-engineered nerve in human.

Topic Category 醫藥衛生 > 牙科與口腔科