Abstract
Mesothelial cells are specific epithelial cells that are lined in the serosal cavity and internal organs. Nonetheless, few studies have explored the possibility to culture mesothelial cells in a three-dimensional (3D) scaffold for tissue engineering applications. Towards this end, we fabricated macroporous scaffolds from gelatin and gelatin/hyaluronic acid (HA) by cryogelation, and elucidated the influence of HA on cryogel properties and the cellular phenotype of mesothelial cells cultured within the 3D scaffolds. The incorporation of HA was found not to significantly change the pore size, porosity, water uptake kinetics, and swelling ratios of the cryogel scaffolds, but led to a faster scaffold degradation in the collagenase solution. Adding 5% HA in the composite cryogels also decreased the ultimate compressive stress (strain) and toughness of the scaffold, but enhanced the elastic modulus. From the in vitro cell culture, rat mesothelial cells showed quantitative cell viability in gelatin (G) and gelatin/HA (GH) cryogels. Nonetheless, mesothelial cells cultured in GH cryogels showed a change in the cell morphology and cytoskeleton arrangement, reduced cell proliferation rate, and downregulation of the mesothelium specific maker gene expression. The production of key mesothelium proteins E-cadherin and calretinin were also reduced in the GH cryogels. Choosing the best G cryogels for in vivo studies, the cell/cryogel construct was used for the transplantation of allograft mesothelial cells for mesothelium reconstruction in rats. A mesothelium layer similar to the native mesothelium tissue could be obtained 21 days post-implantation, based on hematoxylin and eosin (H&E) and immunohistochemical staining.
Highlights
Mesothelial cells are specific epithelial cells that are lined in the serosal cavity and internal organs
This study demonstrates the feasibly of a cryogel scaffold for a 3D culture of mesothelial cells
There was no change in the pore structure and water absorbing properties, GH showed a faster degradation in the collagenase solution and a higher elastic modulus than G cryogel
Summary
Mesothelial cells are specific epithelial cells that are lined in the serosal cavity (pleura, pericardium, and peritoneum) and internal organs. Their primary function is to provide a non-stick, frictionless protective barrier that facilitates the relative movement of tissues and organs within the serosal cavity [1]. The normal peritoneum is composed of a monolayer of mesothelial cells on the basement membrane as mesothelium, and is supported by a thin layer of connective tissue containing cells, blood vessels, and lymphatic vessels. Cells can be seeded onto a three-dimensional (3D) artificial structure, called a scaffold. The 3D scaffold could act as an artificial extracellular matrix (ECM), and serves as a template to guide cell adhesion, proliferation, and tissue development
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