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。 在儿童体内制作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 我们的方法使用孔隙率超过 90% 的微结构聚合物支架。 足够的机械性能和独特的孔结构能够增强软骨形成 此外，软骨祖细胞被认为是一种快速增殖、高度增殖的细胞。 为了利用这些细胞，我们开发了一种微创活检程序来 收获耳朵软骨祖细胞（eCPC）我们的首要假设是微结构。 聚合物支架与 eCPC 相结合将产生具有适当机械强度的软骨植入物， 我们建议确定个性化微创 LTR 的尺寸和表型稳定性。 实现最大软骨形成的微观结构和具体作用机制。 eCPC 产生强大的软骨表型，可能比耳软骨细胞更好，并且更少 最后，还将研究源自 eCPC 的工程软骨。 微孔支架将在猪 LTR 模型中进行测试，我们预计我们的研究结果将带来重大影响。 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转化。