Material Characterisation and Stratification of Conjunctival Epithelial Cells on Electrospun Poly(ε-Caprolactone) Fibres Loaded with Decellularised Tissue Matrices.

Lucy A Bosworth,Catalina Pineda Molina,Kyle G Doherty,James D Hsuan,Rachel L Williams,Stephen F Badylak,Samuel P Cray,Raechelle A D’Sa

doi:10.3390/pharmaceutics13030318

Abstract

The conjunctiva, an under-researched yet incredibly important tissue, plays key roles in providing protection to the eye and maintaining homeostasis of its ocular surface. Multiple diseases can impair conjunctival function leading to severe consequences that require surgical intervention. Small conjunctival defects can be repaired relatively easily, but larger defects rely on tissue grafts which generally do not provide adequate healing. A tissue engineering approach involving a biomaterial substrate capable of supporting a stratified epithelium with embedded, mucin-secreting goblet cells offers a potential solution. As a first step, this study aimed to induce stratification of human conjunctival epithelial cells cultured on electrospun scaffolds composed from poly(ε-caprolactone) (PCL) and decellularised tissue matrix (small intestinal submucosa (SIS) or urinary bladder matrix (UBM)) and held at the air/liquid interface. Stratification, up to 5 cell layers, occurred more frequently on scaffolds containing PCL + UBM. Incorporation of these decellularised tissue matrices also impacted material properties, with significant changes occurring to their fibre diameter, tensile properties, and chemical composition throughout the scaffold structure compared to PCL alone. These matrix containing scaffolds warrant further long-term investigation as a potential advanced therapy medicinal product for conjunctiva repair and regeneration.

Highlights

The conjunctiva is a mucous membrane that has an important role in maintaining a normal ocular surface and motility of the eye and eyelids
The inclusion of decellularised tissue powders to the PCL, demonstrated numerous droplets throughout the collected area, which increased in number with greater dECM content
Increasing the quantity of dECM to 10% demonstrated a further decrease in fibre diameter for small intestinal submucosa (SIS) (median 0.22 μm (IQR 0.15–0.38 μm)), but a similar distribution of fibre diameters for urinary bladder matrix (UBM) (median 0.22 μm (IQR 0.16–0.31 μm))

Summary

Introduction

The conjunctiva is a mucous membrane that has an important role in maintaining a normal ocular surface and motility of the eye and eyelids. The palpebral conjunctiva lines the posterior surfaces of the eyelids and is reflected in the fornices to become the bulbar conjunctiva overlying the anterior sclera until it merges with the cornea. It has a stratified epithelium up to six layers thick with numerous goblet cells, and an underlying basement membrane. The conjunctiva serves to protect the eye and contributes to a healthy ocular surface and tear film through mucin production by the goblet cells. These stabilise the Pharmaceutics 2021, 13, 318.

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