Speaker
Description
Introduction: Fibroblast populated collagen matrices (FPCMs) are well established as a model for assessing the behaviour of fibroblasts. When used on a fixed substrate, FPCMs have also proven to be a reliable model for the study of tissue healing. Until now, such techniques have only been used with dermal fibroblasts to examine skin healing. Our work will develop a fixed (tethered) FPCM seeded with rectus sheath fibroblasts that will act as an in vitro model of rectus sheath healing. We will use this model to test the effects of hernia mesh upon fascial healing in vitro. We hope this will mark the earliest iteration of an evolving model of rectus sheath healing that may standardize the process of pre-clinical biomaterial testing.
Methods: FPCMs will be formed using rat-tail collagen seeded with fibroblasts taken from human rectus sheath. FPCMs will be formed in custom 3D-printed rectangular moulds. Such moulds will fix the collagen gel at each far end - establishing a uniaxial tension across the gel. This tension will mimic that which lies perpendicular to a fascial wound closure –representing an in vitro model of early wound healing/ contraction and fascial granulation. During culture, 5 different polypropylene hernia meshes will be suspended mid-way through the collagen gel so that they may be incorporated into the tissue model. After 3-5 days, samples will be plastic compressed – leaving a dense collagen structure analogous to human rectus sheath. Each mesh will be tested at two perpendicular orientations for different culture lengths. Once formed, collagen/ mesh constructs will be assessed for their collagen alignment and their tensile strength.
Results: Thus far, our custom moulds have established a reliable ability to form contracting FPCMS which stimulate fibroblast and collagen alignment when formed at 1.5x106 cells/ml. We are able to successfully incorporate different hernia meshes into our model and test them through fluorescence microscopy and uniaxial tensile strength testing.
Conclusion: This new methodology shows promising early results. In the long term such tethered FPCMs may help develop a new generation of in vitro models for the testing of medical biomaterials
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