Tissue Engineering of Neotrachea

新气管组织工程

• 批准号: 8956927
• 负责人: JAMES E DENNIS
• 金额: $ 39.57万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8956927
• 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; Health; 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; 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.

描述（由申请方提供）：目前，没有足够的治疗方法可用于替换大于6 cm的气管段。这些研究的目的是开发和制造一个功能性的气管替代兔模型使用组织工程原理，并在无胸腺大鼠模型中使用人类细胞制造一个人类大小的新气管。该实验室已经开发出生产大型无支架软骨片的方法，当植入体内并被筋膜或肌肉包围时，产生血管化的新气管结构。将对兔耳和肩软骨细胞进行测试和优化，用于软骨片生产，并将测试其在气管节段性缺损模型中的长期稳定性和功能。首先在定制的双扩散生物反应器中制造片材，然后将其植入气管中未来节段性修复部位的近端，在节段性气管重建之前使其成熟约12周。将新生气管沿着其脉管系统移植到节段性气管缺损中，并在植入后4、8和12周收获。将测试三种不同的手术形式，包括单独的直接移植、T型管移植（以辅助呼吸和促进修复）和T型管移植沿着面颊粘膜游离移植。通过组织学和免疫组织化学方法评估收获的新气管，包括糖胺聚糖染色和I型、II型和X型胶原和弹性蛋白的免疫组织化学，在组织学上检查结构、组织整合、上皮形成，并评价生物力学强度。将特别注意纤维化，粘膜的形成，以及新气管内是否形成骨。可以采用将上皮细胞应用于工程化组织的替代方法。 长期的目标是开发出一种方法来生产功能性气管，以修复兔的节段性缺损，并将这种技术转化为人源性细胞。为此，与兔软骨细胞的优化研究平行，将从气管、耳、鼻和关节表面获得人软骨细胞，并通过首先使用软骨形成聚集体培养测定系统测试生长因子和细胞因子库来优化软骨片形成。然后将在无胸腺大鼠模型中测试人软骨细胞衍生的片层在体内形成人大小的新气管的能力，并将通过组织学、生物化学和机械测定进行评估。

项目成果

Tissue engineering of a composite trachea construct using autologous rabbit chondrocytes.

使用自体软骨细胞的复合气管构建体的组织工程。

• DOI: 10.1002/term.2523
• 发表时间: 2018-03
• 期刊: Journal of tissue engineering and regenerative medicine
• 影响因子: 3.3
• 作者: Dennis JE;Bernardi KG;Kean TJ;Liou NE;Meyer TK
• 通讯作者: Meyer TK

Hyaluronan-based scaffolds to tissue-engineer cartilage implants for laryngotracheal reconstruction.

基于透明质酸的支架用于组织工程软骨植入物以进行喉气管重建。

• DOI: 10.1097/mlg.0b013e31811434ae
• 发表时间: 2007
• 期刊: The Laryngoscope
• 作者: Weidenbecher,Mark;Henderson,JamesH;Tucker,HarveyM;Baskin,JonathanZ;Awadallah,Amad;Dennis,JamesE
• 通讯作者: Dennis,JamesE

Cartilage repair: past and future--lessons for regenerative medicine.

• DOI: 10.1111/j.1582-4934.2009.00789.x
• 发表时间: 2009-05
• 期刊: Journal of cellular and molecular medicine
• 影响因子: 5.3
• 作者: van Osch GJ;Brittberg M;Dennis JE;Bastiaansen-Jenniskens YM;Erben RG;Konttinen YT;Luyten FP
• 通讯作者: Luyten FP

Tissue-engineered trachea for airway reconstruction.

• DOI: 10.1002/lary.20700
• 发表时间: 2009-11
• 期刊: The Laryngoscope
• 作者: Weidenbecher M;Tucker HM;Gilpin DA;Dennis JE
• 通讯作者: Dennis JE