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 严重的亚平瓣狭窄，即声音褶皱下方的气道的狭窄，是为了回应 在美国，插管近20,000个早产的近10％。严重的案件需要 喉气管重建（LTR），其中外科医生将cricoid分开，并添加一块自体患者 - 派生的软骨以扩大气道并恢复适当的气流。但是，在儿童中，成功率是 低至50％的重新狭窄事件需要修订手术。移植失败直接与缺乏有关 儿童中足够尺寸的自体软骨，并提出了组织工程来发展 小儿LTR的替代嫁接选择。一些方法，包括我们以前的一些工作， 有效地生产功能软骨，但是结构匹配所需的整体时间框架 天然软骨的机械性能（> 24周）与临床翻译不兼容（<8周）。 此外，当前的细胞源，例如扩展的自体软骨细胞和间质干细胞 经常导致肥厚和计算的组织。我们的目标是设计一种新型的软骨 由患者细胞填充的植入物，机械可行，适用于临床内的LTR 相关时间范围。我们的方法使用微观结构的聚合物支架，孔隙率超过90％和 足够的机械性能和孔结构具有独特的能力 干细胞。此外，已经提出软骨祖细胞作为一种快速增殖，高度 软骨细胞来源。为了利用这些细胞，我们开发了一种微创的活检程序 收获耳软骨祖细胞（ECPC）。我们的总体假设是微结构 聚合物支架与ECPC结合会产生软骨，并具有合适的机械强度， 个性化，微创LTR的尺寸和表型稳定性。我们建议确定 实现最大软骨发生和特定作用机理的微观结构。容量 ECPC产生健壮的软骨表型，可能比早期软骨细胞更好，更少 容易钙化，也将进行研究。最后，源自播种的ECPC的工程软骨 微孔支架将在猪LTR模型中进行测试。我们希望我们的发现将引入专业 在治疗亚平狭窄的治疗方面的创新，为长期临床前安全研究的基础奠定了基础 儿童气道重建手术中的临床翻译。