Tissue Engineering of a Neotrachea

新气管的组织工程

• 批准号: 7787630
• 负责人: JAMES E DENNIS
• 金额: $ 20.14万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2010-09-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7787630
• 关键词: Abdomen; Attention; Autologous; Biochemical; Biocompatible; Biological Assay; Biomechanics; Bioreactors; Bone Tissue; Breathing; Cartilage; Cells; Cheek structure; Chondrocytes; Chondrogenesis; Clinical; Collagen Type I; Custom; Defect; Development; Diffusion; Dimensions; Ear; Elastin; Endoscopes; Engineering; Epithelial Cells; Fascia; Fibrosis; Future; Glycosaminoglycans; Goals; Growth Factor; Harvest; Histologic; Histology; Human; Immunohistochemistry; Implant; In Vitro; Laboratories; Longitudinal Studies; Mechanics; Methodology; Methods; Modeling; Mucous Membrane; Muscle; Nose; Nude Rats; Operative Surgical Procedures; Oryctolagus cuniculus; Palpation; Phase; Postoperative Period; Production; Property; Shoulder; Silicones; Site; Source; Staging; Staining method; Stains; Structure; Structure of respiratory epithelium; Surface; Surgical Flaps; System; Technology; Testing; Thick; Tissue Engineering; Tissues; Trachea; Translating; Translations; Transplantation; Tube; articular cartilage; base; bone; clinical application; clinically relevant; cytokine; design; in vivo; in vivo Model; pre-clinical; public health relevance; reconstruction; repaired; research study; scaffold; sound; tool

DESCRIPTION (provided by applicant): Currently, there is no adequate treatment available for the replacement of tracheal segments larger than 6 cm. The goal of these studies is to develop and fabricate a functional trachea replacement in a rabbit model using tissue engineering principles, and to also fabricate a human-sized neotrachea using human cells in an athymic rat model. This laboratory has developed the methodology to produce large scaffold-free cartilage sheets, which when implanted in vivo and surrounded by fascia or muscle, produces a vascularized neotracheal construct. Rabbit ear and shoulder chondrocytes will be tested and optimized for cartilage sheet production and will be tested for their long-term stability and function in a trachea segmental defect model. The sheets are first fabricated in a custom double diffusion bioreactor and then implanted proximal to the future segmental repair site in the trachea where it is allowed to mature for approximately 12 weeks prior to segmental tracheal reconstruction. The neotrachea is transplanted along its vasculature into segmental tracheal defect and will be harvested at 4, 8 and 12 weeks postimplantation. Three distinct surgical formats will be tested including the direct transplant alone, transplant with a T-tube (to assist breathing and promote repair), and transplant with T-tube along with cheek mucosal free transplant. The harvested neotracheas are assessed by histologic and immunohistochemical methods including staining for glycosaminoglycans, and immunohistochemistry for collagens type I, II, and X, and elastin, examined, histologically, for structure, tissue integration, epithelialization, and are evaluated for biomechanical strength. Particular attention will be paid to the fibrosis, the formation of a mucous membrane, and also whether bone forms within the neotrachea. Alternative approaches to apply epithelial cells to the engineered tissue may be employed. The long-term goals are to develop the methodology to produce a functional trachea to repair segmental defects in rabbits, and to translate this technology to human derived cells. To this end, in parallel with the optimization study of rabbit chondrocytes, human chondrocytes will be obtained from the trachea, ear, nose and from articular surfaces and will be optimized for cartilage sheet formation by first testing a bank of growth factors and cytokines using a chondrogenesis aggregate culture assay system. Human chondrocyte-derived sheets will then be tested in an athymic rat model for their ability to form a human-sized neotrachea in vivo, and will be assessed by the histologic, biochemical and mechanical assays. PUBLIC HEALTH RELEVANCE: Currently, there is no adequate treatment for large (6 cm or greater) trachea defects. The focus of this project is to use tissue engineering principles to fabricate a neo-trachea that can repair segmental defects in rabbits and to then apply these principles to fabricate a neo-trachea using human chondrocytes. The long-term goal this project seeks to develop a tissue engineering method to repair damaged tracheas in people.

描述（由申请人提供）：目前，没有足够的治疗方法可用于更换大于6厘米的气管段。这些研究的目的是使用组织工程原理在兔模型中开发和制造功能性气管替代，并在无胸腺大鼠模型中使用人类细胞制造人类大小的新刺激性。该实验室已经开发了生产大型无脚手架软骨片的方法，当体内植入并被筋膜或肌肉包围时，它会产生一种血管化的新型气管结构。兔耳朵和肩部软骨细胞将经过测试和优化用于软骨片的产生，并将在气管分段缺陷模型中测试其长期稳定性和功能。这些板首先是在定制的双扩散生物反应器中制造的，然后植入气管中未来的分段维修位点的近端，允许在节段气管重建前大约12周成熟。沿其脉管系统移植到节段性气管缺陷中，并将在植入后4、8和12周收获。将测试三种不同的手术格式，包括单独进行直接移植，用T管移植（有助于呼吸和促进修复），并使用T型管移植以及无脸颊粘膜无粘膜移植。通过组织学和免疫组织化学方法评估收获的新刹车，包括糖胺聚糖染色，以及I型胶原蛋白，II，II和X型胶原蛋白的免疫组织化学，以及在组织学上检查的，组织学，结构，组织整合，上皮力和生物机械强度的结构。特别关注纤维化，粘膜的形成，以及在新牙本质中是否形成骨骼。可以采用将上皮细胞应用于工程组织的替代方法。 长期目标是开发方法，以产生功能性气管来修复兔子中的分段缺陷，并将这项技术转化为人类衍生细胞。为此，与兔软骨细胞的优化研究并行，将从气管，耳朵，鼻子和关节表面获得人类软骨细胞，并通过首先使用软骨基因培养物培养物分析系统来测试生长因子和细胞因子。然后，人类软骨细胞衍生的片将在无性大鼠模型中进行测试，以便其在体内形成人尺寸的新型新刺激性，并将通过组织学，生化和机械测定法进行评估。 公共卫生相关性：目前，对于大（6厘米或更高）的气管缺陷尚无适当的治疗。该项目的重点是使用组织工程原理来制造可修复兔子中分段缺陷的新通道，然后将这些原理应用以使用人软骨细胞制造新疗法。该项目旨在开发一种组织工程方法来修复人员中受损的气管的长期目标。