Decellularized cartilage and progenitor cells for laryngotracheal reconstruction

用于喉气管重建的脱细胞软骨和祖细胞

• 批准号: 10704303
• 负责人: Riccardo Gottardi
• 金额: $ 61.67万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10704303
• 关键词: Address; Adolescent; Affect; Air Movements; Allogenic; Animals; Autologous; Back; Benchmarking; Biopsy; Blood Vessels; Cadaver; Cartilage; Cell Differentiation process; Cells; Child; Childhood; Chondrocytes; Clinic; Clinical; Cognitive; Cytoplasmic Granules; Deposition; Development; Diagnosis; Digestion; Dimensions; Disease; Ear; Ear Cartilages; Elastic Cartilage; Elastin Fiber; Endoscopy; Engineering; Ensure; Event; Exhibits; Face; Family suidae; Gene Expression; Goals; Hand; Harvest; Heart Valves; Histology; Hormones; Human; Immunohistochemistry; Implant; In Vitro; Incidence; Infection; Intubation; Joints; Larynx; Legal patent; Longitudinal Studies; Longterm Follow-up; Mechanics; Meniscus structure of joint; Mesenchymal Stem Cells; Modeling; Monitor; Morbidity - disease rate; Mus; Operative Surgical Procedures; Oryctolagus cuniculus; Outcome; Patients; Phenotype; Pilot Projects; Premature Birth; Procedures; Production; Proliferating; Property; Quality of life; Risk; Shapes; Site; Source; Speech; Stenosis; Surgeon; Technology; Testing; Time; TimeLine; Tissue Engineering; Tissues; Trachea; Tracheotomy procedure; Translations; Triiodothyronine; United States; Work; adverse outcome; base; calcification; cartilage cell; cartilage implant; cartilage repair; cartilage transplantation; cartilaginous; clinical translation; clinically relevant; costal cartilage; efficacy validation; extracellular; graft failure; implantation; improved; in vivo; innovation; mechanical properties; minimally invasive; pediatric patients; porcine model; pre-clinical; preclinical safety; premature; reconstruction; response; restenosis; restoration; rib bone structure; safety study; scaffold; stem cells; subcutaneous; success; therapeutically effective; tool; vocal cord

PROJECT SUMMARY/ABSTRACTS Severe subglottic stenosis, the narrowing of the airway just below the vocal folds, develops as a response to intubation in close to 10% of the > 20,000 premature births per year in the United States. Severe cases require laryngotracheal reconstruction (LTR), in which surgeons split the cricoid and add a piece of autologous patient- derived cartilage to expand the airway and restore proper airflow. However, in children, the success rate is as low as 50% with a high incidence of restenosis requiring revision surgery. Graft failure is tied directly to the lack of sufficiently sized autologous cartilage in the child, and tissue engineering has been proposed to develop alterative grafting options for pediatric LTR. Some approaches, including some of our previous work, have been effective in producing functional cartilage, but the overall timeframe required for the construct to match the mechanical properties of native cartilage (>24 weeks) is not compatible with clinical translation (<8 weeks). Furthermore, current cell sources such as expanded autologous chondrocytes and mesenchymal stem cells frequently result in hypertrophic and calcified tissue. Our objective is to engineer a new type of cartilage implant that is populated with patients’ cells, mechanically viable and suitable for LTR within a clinically relevant timeframe. Our approach is to exploit the blood vessels and elastin fibers that are uniquely present in the fibro-elastic cartilage of the meniscus to form microchannels for effective recellularization after enzymatic decellularization. Our patent-pending Meniscal Decellularized scaffold (MEND) technology can indeed be easily recellularized and has mechanical properties of the same order as native tracheal cartilage. Furthermore, cartilage progenitor cells have been proposed as a rapidly proliferating, highly chondrogenic cell source. To harness these cells, we have developed a minimally invasive biopsy procedure to harvest ear Cartilage Progenitor Cells (eCPCs). Our overarching hypothesis is that MEND and eCPCs can be combined to create cartilage implants with suitable mechanical strength, dimensions, and phenotypic stability for personalized, minimally invasive LTR. We propose to use MEND recellularized with eCPCs to engineer cartilage with tissue properties matching those of native cartilage. We will then validate the MEND-engineered cartilage in a miniature pig LTR model. We expect that our findings will provide strong pre-clinical evidence of functional laryngotracheal cartilage repair by our innovative eCPC-MEND technology and will thereby prompt follow up long term studies to eventually apply this technology to restore children’s airway.

项目概要/摘要 严重声门下狭窄，即声带下方气道变窄，是对以下疾病的反应： 在美国，每年超过 20,000 例早产儿中，有近 10% 的重症病例需要插管。 喉气管重建（LTR），其中外科医生将环状软骨分开并添加一块自体患者 - 然而，在儿童中，成功率却很低。 低至 50%，且需要修复手术的再狭窄发生率很高，这与移植失败直接相关。 为儿童提供足够大小的自体软骨，并建议开发组织工程 儿科 LTR 的替代移植选择 一些方法，包括我们之前的一些工作，已经取得了进展。 能够有效地生产功能性软骨，但构建体所需的总体时间框架相匹配 天然软骨的机械特性（>24周）与临床转化（<8周）不相容。 此外，目前的细胞来源，例如扩增的自体软骨细胞和间充质干细胞 经常导致组织肥大和钙化，我们的目标是设计一种新型软骨。 充满患者细胞的植入物，机械上可行并且适合临床上的 LTR 我们的方法是利用独特存在的血管和弹性蛋白纤维。 半月板的纤维弹性软骨形成微通道，在酶处理后进行有效的再细胞化 我们正在申请专利的半月板脱细胞支架 (MEND) 技术确实可以 易于再细胞化，并具有与天然气管软骨相同数量级的机械性能。 此外，软骨祖细胞被认为是一种快速增殖、高度软骨形成的细胞 为了利用这些细胞，我们开发了一种微创活检程序来采集耳朵。 我们的首要假设是 MEND 和 eCPC 可以结合起来。 创建具有适当机械强度、尺寸和表型稳定性的软骨植入物 我们建议使用带有 eCPC 的 MEND 进行再细胞化来设计。 然后我们将验证 MEND 工程设计的软骨的组织特性。 我们预计我们的研究结果将为小型猪 LTR 模型中的软骨提供强有力的临床前证据。 通过我们创新的 eCPC-MEND 技术进行功能性喉气管软骨修复，从而促进 后续长期研究最终应用这项技术来恢复儿童的呼吸道。