Laryngotracheal Reconstruction with Engineered Cartilage

用工程软骨重建喉气管

• 批准号: 10660455
• 负责人: Riccardo Gottardi
• 金额: $ 58.67万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660455
• 关键词: Acids; Adolescent; Affect; Air Movements; Animals; Autologous; Biochemistry; Biopsy; Cartilage; Cells; Cellular Metabolic Process; Child; Childhood; Chondrocytes; Chondrogenesis; Clinic; Clinical; Cognitive; Collagen; Cultured Cells; DNA; Development; Diagnosis; Diameter; Dimensions; Disease; Ear; Ear Cartilages; Engineering; Ensure; Euthanasia; Exhibits; Face; Family suidae; GAG Gene; Gene Expression; Goals; Harvest; Hormones; Hypertrophy; Hypoxia; Immunohistochemistry; Implant; In Vitro; Incidence; Infection; Intubation; Mechanics; Mesenchymal; Mesenchymal Stem Cells; Metabolic; Microfluidics; Modeling; Modulus; Monitor; Morbidity - disease rate; Mus; Operative Surgical Procedures; Outcome; Oxygen; Patients; Pattern; Phenotype; Physical condensation; Polymers; Porosity; Premature Birth; Procedures; Production; Proliferating; Quality of life; Quantitative Reverse Transcriptase PCR; RNA; Reconstructive Surgical Procedures; Risk; Scientist; Second Look Surgery; Site; Source; Speech; Stenosis; Structure; Surgeon; Technology; Testing; Thick; Thinness; Time; Tissue Engineering; Tissues; Tracheotomy procedure; Translations; Triiodothyronine; United States; Work; adverse outcome; calcification; cartilage implant; cartilage transplantation; cartilaginous; clinical translation; clinically relevant; costal cartilage; efficacy validation; graft failure; implantation; improved; in vivo; innovation; manufacturing scale-up; mechanical properties; meter; minimally invasive; mortality; novel; porcine model; pre-clinical; preclinical safety; premature; reconstruction; response; restenosis; restoration; rib bone structure; safety study; scaffold; stem cells; subcutaneous; success; therapeutically effective; timeline; transcriptome sequencing; vocal cord

PROJECT SUMMARY/ABSTRACT 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 uses a microstructured polymeric scaffold with over 90% porosity and sufficient mechanical properties and with a pore structure uniquely capable of enhancing chondrogenesis of stem cells. 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 the microstructured polymeric scaffold combined with eCPCs will create cartilage implants with suitable mechanical strength, dimensions, and phenotypic stability for personalized, minimally invasive LTR. We propose to identify the microstructure that achieves the maximum chondrogenesis and the specific mechanism of action. The capacity of eCPCs to produce a robust cartilage phenotype, potentially better than that of ear chondrocytes and less prone to calcification, will also be studied. Finally, the engineered cartilage derived from the eCPCs seeded in the microporous scaffold will be test in a porcine LTR model. We expect that our findings will introduce a major innovation in the treatment of subglottic stenosis, laying the basis for long term pre-clinical safety studies and clinical translation in airway reconstructive surgery in children.

项目摘要/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。我们的方法使用具有超过90%孔隙率的微结构聚合物支架， 具有足够的机械性能和独特的能够增强软骨形成的孔结构， 干细胞此外，已经提出软骨祖细胞是一种快速增殖的、高度分化的细胞。 软骨细胞来源。为了利用这些细胞，我们开发了一种微创活检程序， 收获穗Carcinoma祖细胞（eCPC）。我们的总体假设是， 与eCPC组合的聚合物支架将产生具有合适机械强度的软骨植入物， 尺寸和表型稳定性的个性化，微创LTR。我们建议确定 实现最大软骨形成的微观结构和特定的作用机制。的能力 eCPC产生强大的软骨表型，可能优于耳软骨细胞， 容易钙化，也将被研究。最后，将来自接种在 将在猪LTR模型中测试微孔支架。我们希望我们的研究结果将引入一个主要的 声门下狭窄治疗的创新，为长期临床前安全性研究奠定基础， 儿童气道重建手术的临床翻译。