Tissue Engineering the Oesophagus

Abstract

Current treatments for oesophageal cancer and oesophageal atresia, that involves the repair of the entire thickness of the oesophagus, presents various complications and challenges due to the lack of functional oesophageal replacement tissue. Tissue engineering, through the combination of cells, scaffolds, and biologically active molecules, presents an innovative approach to develop constructs that can mimic the multi-layered architecture of the oesophagus. This study utilises the combination of a novel biomaterial in the form of self-assembling peptide hydrogel, PeptiGels, and a traditional biomaterial, polycaprolactone (PCL) scaffolds to fabricate a proof-of-concept composite substrate that mimics the multi-layered architecture of the oesophageal wall. The findings of this study showed that plasma-treated and PeptiGel-coated PCL scaffolds were capable of supporting the survival and proliferation of primary human oesophageal smooth muscle cells (HuOesSMC) up to day 21. Furthermore, the HuOesSMCs were aligned in the direction of the fibres and had retained their smooth muscle cell markers up to day 21. This indicated the suitability of aligned PCL fibres to mimic the aligned architecture of the muscle layer of the oesophagus. Similarly, primary human oesophageal fibroblasts cultured in 3D within PeptiGels were capable of proliferation, ECM protein synthesis and keratinocyte growth factor (KGF) release up to day 22. Cell proliferation and KGF secretion at day 22 was enhanced in PeptiGel Alpha4Plus compared to other PeptiGels. Likewise, 2D culture of primary human oesophageal epithelial cells (HuOesEpiC) on PeptiGels, showed that PeptiGels Alpha4, Alpha4RGD and Alpha4Plus were capable of facilitating the survival, proliferation, and monolayer formation of HuOesEpiCs by day 20. Introduction of air-liquid interface to the 2D PeptiGel-HuOesEpiC culture system facilitated the formation of multiple layers of epithelial cells on PeptiGels Alpha4, Alpha4RGD and Alpha4Plus. This study was able to demonstrate that each biomaterial, selected in this study, has the capability to support the proliferation of oesophageal cells and has also shown the potential to provide an environment that allows the cells to function and behave as desired. Therefore, a combination of PeptiGel and PCL scaffolds could potentially provide a viable approach to tissue engineer a multi-layered substrate for full thickness repair of the oesophagus.

Date of Award	6 Jan 2025
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Alberto Saiani (Supervisor), Julie Gough (Supervisor) & Sarah Herrick (Supervisor)