Department of Surgery, Division of Otolaryngology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, U.S.A.
Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, Pennsylvania, U.S.A.
Laryngoscope. 2024 Feb;134(2):807-814. doi: 10.1002/lary.31017. Epub 2023 Sep 2.
Severe subglottic stenosis develops as a response to intubation in 1% of the >200,000 neonatal intensive care unit infants per year and may require laryngotracheal reconstruction (LTR) with autologous hyaline cartilage. Although effective, LTR is limited by comorbidities, severity of stenosis, and graft integration. In children, there is a significant incidence of restenosis requiring revision surgery. Tissue engineering has been proposed to develop alterative grafting options to improve outcomes and eliminate donor-site morbidity. Our objective is to engineer a decellularized, channel-laden xenogeneic cartilage graft, that we deployed in a proof-of-concept, neonatal porcine LTR model.
Meniscal porcine cartilage was freeze-thawed and washed with pepsin/elastase to decellularize and create microchannels. A 6 × 10-mm decellularized cartilage graft was then implanted in 4 infant pigs in an anterior cricoid split. Airway patency and host response were monitored endoscopically until sacrifice at 12 weeks, when the construct phenotype, cricoid expansion, mechanics, and histomorphometry were evaluated.
The selective digestion of meniscal components yielded decellularized cartilage with cell-size channels. After LTR with decellularized meniscus, neonatal pigs were monitored via periodic endoscopy observing re-epithelization, integration, and neocartilage formation. At 12 weeks, the graft appeared integrated and exhibited airway expansion of 4 mm in micro-CT and endoscopy. Micro-CT revealed a larger lumen compared with age-matched controls. Finally, histology showed significant neocartilage formation.
Our neonatal porcine LTR model with a decellularized cartilage graft is a novel approach to tissue engineered pediatric LTR. This pilot study sets the stage for "off-the-shelf" graft procurement and future optimization of MEND for LTR.
NA Laryngoscope, 134:807-814, 2024.
每年有超过 20 万名新生儿重症监护病房的婴儿中，有 1%会因插管而出现严重的声门下狭窄，可能需要进行自体透明软骨的喉气管重建（LTR）。尽管 LTR 有效，但它受到合并症、狭窄严重程度和移植物整合的限制。在儿童中，存在显著的再狭窄发生率，需要进行修正手术。组织工程学已经被提出用于开发替代移植选择，以改善结果并消除供体部位的发病率。我们的目的是设计一种脱细胞、有通道的异种软骨移植物，并将其应用于新生猪 LTR 模型的概念验证研究中。
冷冻-解冻半月板猪软骨，并用胃蛋白酶/弹性蛋白酶进行清洗以脱细胞并创建微通道。然后，将 6×10mm 的脱细胞软骨移植物植入 4 只新生猪的前环状软骨切开处。通过内窥镜监测气道通畅性和宿主反应，直到 12 周处死，此时评估构建体表型、环状软骨扩张、力学和组织形态计量学。
半月板成分的选择性消化产生了具有细胞大小通道的脱细胞软骨。在 LTR 中使用脱细胞半月板后，通过定期内窥镜检查监测新生猪的再上皮化、整合和新软骨形成。在 12 周时，移植物显示出整合，并在 micro-CT 和内窥镜下显示气道扩张 4mm。micro-CT 显示与年龄匹配的对照组相比，管腔更大。最后，组织学显示出显著的新软骨形成。
我们的新生猪 LTR 模型与脱细胞软骨移植物是组织工程学小儿 LTR 的一种新方法。这项初步研究为“现成”移植物采购和未来优化用于 LTR 的 MEND 奠定了基础。
无喉镜，134：807-814，2024。
