Transplantation of a 3D-printed tracheal graft combined with iPS cell-derived MSCs and chondrocytes

In Gul Kim,Su A Park,Seong Keun Kwon,Sang Jin Lee,Shin-Hyae Lee,Hana Cho,Ji Suk Choi,Yoo-Wook Kwon

doi:10.1038/s41598-020-61405-4

Abstract

For successful tracheal reconstruction, tissue-engineered artificial trachea should meet several requirements, such as biocompatible constructs comparable to natural trachea, coverage with ciliated respiratory mucosa, and adequate cartilage remodeling to support a cylindrical structure. Here, we designed an artificial trachea with mechanical properties similar to the native trachea that can enhance the regeneration of tracheal mucosa and cartilage through the optimal combination of a two-layered tubular scaffold and human induced pluripotent stem cell (iPSC)-derived cells. The framework of the artificial trachea was fabricated with electrospun polycaprolactone (PCL) nanofibers (inner) and 3D-printed PCL microfibers (outer). Also, human bronchial epithelial cells (hBECs), iPSC-derived mesenchymal stem cells (iPSC-MSCs), and iPSC-derived chondrocytes (iPSC-Chds) were used to maximize the regeneration of tracheal mucosa and cartilage in vivo. After 2 days of cultivation using a bioreactor system, tissue-engineered artificial tracheas were transplanted into a segmental trachea defect (1.5-cm length) rabbit model. Endoscopy did not reveal granulation ingrowth into tracheal lumen. Alcian blue staining clearly showed the formation of ciliated columnar epithelium in iPSC-MSC groups. In addition, micro-CT analysis showed that iPSC-Chd groups were effective in forming neocartilage at defect sites. Therefore, this study describes a promising approach for long-term functional reconstruction of a segmental tracheal defect.

Highlights

The trachea is a rigid fibroelastic organ composed of C-shaped hyaline cartilage, ciliated columnar epithelium, and connective tissue[1]
Further bioactivity for re-epithelialization is conferred to this scaffold through the seeding of human bronchial epithelia cells onto the inner layers of the scaffold. induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells and iPSC-MSC-derived chondrocytes were seeded onto the outer region of the scaffold using Matrigel to promote the regeneration of cartilage tissue (Fig. 1B)
We investigated whether co-transplantation of iPSC-MSCs and chondrocytes within the artificial trachea led to the enhanced regeneration of tracheal cartilage and tracheal mucosa in vivo

Summary

Introduction

The trachea is a rigid fibroelastic organ composed of C-shaped hyaline cartilage, ciliated columnar epithelium, and connective tissue[1]. Rapid remodeling of the mucosa layer on the inner surface of artificial trachea may be a critical strategy[11] To this end, we have previously reported enhanced mucosal regeneration using a laminin-coated asymmetrical polymer membrane[12]. Cartilage tissue commonly has a low capacity for spontaneous healing owing to its avascularity and low cellularity[15,16] From this perspective, cell-based therapy has been recognized as a potentially effective approach to healing tracheal cartilage. We evaluated the effect of a new artificial trachea which can enhance the regeneration of mucosal layer and cartilage through analyzing the most optimal combination of iPSC-derived precursor cells, epithelial cells and 3D-printed tubular scaffold in long circumferential tracheal defect (1.5 cm). We investigated whether co-transplantation of iPSC-MSCs and chondrocytes within the artificial trachea led to the enhanced regeneration of tracheal cartilage and tracheal mucosa in vivo

Methods

Results

Discussion

Conclusion