去细胞气管支架复合骨髓间充质干细胞构建组织工程气管补片治疗气管狭窄的实验研究

Severe tracheal stenosis (STS) is an important cause leading to death in infant and particularly in neonate, owing to its significant difficulty in effective management. The tissue engineered trachea (TET) has been an active attempt for treating STS in recent years, but there is no mature model for TET currently. There have been discrepancies in tracheal scaffold, cells seeding and postoperative management of TET transplantation. In our previous rabbit studies in vitro, we had explored the differentiation of tracheal cartilage cells, and established the decellularized tracheal matrix. We will take further researches on their changes in vivo. In this study, we will investigate the efficacy of tracheal reconstruction using decellularized tracheal matrix patch seeded with bone marrow derived mesenchyma stem cells (BMMSCs) in management of STS. We will build the TET patch by seeding the labeled BMMSCs sheet on the inner surface of the decellularized tracheal matrix and a mixture of BMMSCs with differentiated tracheal chondrocytes on its external surface. We will reconstruct the rabbit trachea with the TET patch, and assess the function of the graft before and after implantation from morphology, cytology, molecular biology, ultramicrostructure and engineering perspective respectively. In this study we will establish a new model of TET consisting of decellularized tracheal matrix and BMMSCs sheet. The result of this research will promisingly provide a new idea and theoretical support for the application of TET in the treatment of infant tracheal stenosis.

重度气管狭窄是导致婴幼儿尤其是新生儿死亡的重要原因，目前尚无特效方法。组织工程气管（TET）是近年研究的热点，但缺乏成熟模型，在气管支架选择、种子细胞种植及移植后处理方面均存在差异。我们的前期兔离体研究已探讨了气管软骨细胞的分化,并建立了脱细胞气管支架，现将进一步探讨其在体研究的变化。本研究将探讨去细胞气管支架复合骨髓间充质干细胞（BMMSCs）构建TET补片重建气管治疗气管狭窄的效能。我们将通过把经过示踪标记的BMMSCs膜片种植于去细胞气管支架内面，BMMSCs混合诱导分化的软骨细胞种植于支架外面构建TET补片，在体重建气管，从形态学水平、组织细胞水平、分子生物学水平、超微结构水平及工程学角度分别对体外制备的TET补片及移植后的功能进行评估。本研究将建立复合去细胞支架与种子细胞膜片的新的TET模型，为应用TET治疗婴幼儿气管狭窄提供新思路及理论支持。

重度气管狭窄是导致婴幼儿尤其是新生儿死亡的重要原因，目前尚无特效方法。组织工程气管（TET）是近年研究的热点，但缺乏成熟模型，在气管支架选择、种子细胞种植及移植后处理方面均存在差异。. 本研究将探讨去细胞气管支架复合骨髓间充质干细胞（BMMSCs）构建TET补片重建气管治疗气管狭窄的效能。我们将通过BMMSCs膜片联合去细胞气管支架构建TET补片，在体重建气管，从形态学水平、组织细胞水平、分子生物学水平、超微结构水平及工程学角度分别对体外制备的TET补片及移植后的功能进行评估。. 通过去污剂联合酶消化法，成功构建脱细胞气管支架，在生物力学与细胞相容性中均与新鲜气道无显著差异同时脱细胞气管支架上课共气管软骨细胞生存。通过贴壁法获取骨髓间充质干细胞，并在抗坏血酸诱导下获得干细胞膜片。成功构建了联合脱细胞气管支架与干细胞膜片的组织工程气管补片，在异位与原位动物体内移植手术中，均展示出组织工程气管补片的组织相容性和血管新生性，且在实验终点未见气道狭窄。. 本研究将建立复合去细胞支架与种子细胞膜片的新的TET模型，为应用TET治疗婴幼儿气管狭窄提供新思路及理论支持。