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 严重的声门下狭窄，即声带下方的气道狭窄，是对以下症状的反应： 在美国，每年超过20，000例早产儿中有近10%使用插管。严重病例需要 喉气管重建（LTR），其中外科医生分裂环状软骨，并添加一块自体患者- 衍生的软骨，以扩大气道和恢复适当的气流。然而，在儿童中，成功率为 低至50%，需要翻修手术的再狭窄发生率高。移植失败与缺乏 足够大的自体软骨在孩子身上，组织工程已经被提出来发展 儿科LTR的替代移植选择。一些方法，包括我们以前的一些工作， 在生产功能性软骨方面是有效的，但是构建匹配所需的总体时间表 天然软骨（>24周）的机械性能与临床平移（<8周）不相容。 此外，目前的细胞来源，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。我们的目标是制造一种新型软骨 植入物填充有患者细胞，具有机械活性并适合在临床上 相关时间表。我们的方法是利用血管和弹性蛋白纤维，这是唯一存在于 半月板的纤维弹性软骨形成微通道，用于酶促后的有效再细胞化 去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的机械性能。 此外，软骨祖细胞被认为是一种快速增殖、高度成软骨的细胞 源头为了利用这些细胞，我们开发了一种微创活检程序， Carcinoma祖细胞（eCPC）。我们的总体假设是，MEND和eCPC可以结合起来， 制造具有合适的机械强度、尺寸和表型稳定性的软骨植入物， 个性化微创LTR我们建议使用用eCPC再细胞化的MEND来工程化 具有与天然软骨相匹配的组织特性的软骨。然后，我们将验证MEND工程 在小型猪LTR模型中的软骨。我们希望我们的研究结果将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， 我们会继续进行长期的研究，以最终应用这项技术来恢复儿童的气道。