组织工程环杓后肌的构建与神经再支配研究

One of the key problems for laryngeal function restore after total laryngectomy is to re-innervate the posterior cricoarytenoid muscle.Tissue engineering is an effective way to solve the problem but remains muscle cell quantity not sufficient and not mature.The most improtant reason is that the tissue engineered muscle constuct lacks of re-innervation.Documents and our previous work have proved that muscle satellite is the main cell source for muscle regeneration and the bone marrow mesenchymal cells can paracrine multiple growth factors to regulate the surrounding cell growth.In this study, we try to co-culture the MSCs and muscle satellite cells on the decellulalized matrix to form a muscle contruct and then transplant the nerve-muscle pedicle in it to stimulate the construct mature and finally fabricate a tissue engineed cricoarytenoid muscle that has a nerve innervation.RT-PCR, Western blosts and immunofluorenscence are used to observe the muturity of the muscle construct. Muscle electrophysiology and histomorphology are used to evaluate the nerve-muscle function. This study will elucidate the principles of combined implantation of MSC and nerve-muscle pedicle to the function restore of the allogenic posterior cricoarytenoid muscle.

全喉切除后重建喉功能的关键之一是建立有神经支配的环杓后肌。组织工程构建肌肉是解决这一难题的有效途径，但存在肌细胞数量少和不成熟问题。重要原因是现有组织工程方法构建的肌肉移植体内后缺乏有效生长因子的长期刺激，特别是缺乏在体神经支配所产生的持续营养和刺激作用。文献和申请者前期工作已证实，肌卫星细胞是肌再生的主要细胞来源，骨髓间质干细胞移植修复肌损伤的机制是可持续旁分泌多种生长因子调节周围细胞生长。本课题拟将骨髓间质干细胞和肌卫星细胞共培养于异体喉环杓后肌去细胞基质上并将其移植体内，同时移植自体神经肌蒂，以促使肌卫星细胞增殖和成熟分化，进而构建出有神经支配的组织工程环杓后肌。从分子、细胞、肌电生理和组织形态学多水平评价肌细胞成熟及神经肌肉间的功能联系。阐明骨髓间质干细胞、肌卫星细胞与神经肌蒂共移植对环杓后肌再细胞化与功能整合作用的机制，为组织工程技术再造全喉提供理论和实验依据。