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Oxidative stress and trabecular meshwork cells in biomimetic three-dimensional environments: implications for glaucoma
Scherrer, Kathryn A.
Scherrer, Kathryn A.
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2021
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Abstract
Glaucoma is a term for a collective group of degenerative eye diseases which cause damage to the optic nerve, eventually causing a characteristic vision loss. The main risk factor for glaucoma is increased intraocular pressure, often caused by an imbalance of aqueous humor generation or impaired drainage through the trabecular meshwork (TM) outflow tissue. The disfunction or blockage of the TM is frequently linked to cell reactions or apoptosis from oxidative stress. The small size, delicate composition, donor tissue limitations, and high structural complexity of the trabecular meshwork make it difficult to research with good accuracy, as the relationship between cells and their extracellular environment has numerous variables. Thus, a biomaterial-based approach was considered more appropriate for providing a cell culture environment with greater mimicking abilities to the native TM. The 3D models fabricated allowed for studies of oxidation consequences of hydrogen peroxide and transforming growth factor β2, along with drug delivery in vitro to be more representative of the inner eye.In this work, human TM (hTM) cells were cultured on two distinct 3-dimensional matrices to better understand how the cells respond to changes in their surrounding environment. Collagen scaffolds with four varying compositions of glycosaminoglycans were utilized for the known structural similarity to the native TM, while alginate-chitosan hydrogels were fabricated in order to incorporate a drug delivery system into the 3D model. Cellular response was measured by using assays including those of relative cell activity and reactive oxygen species, along with quantitative expression of several fibrotic extracellular proteins. It was found that the hTM cells had significantly different responses to stimuli when grown on the traditional 2D tissue culture plates in comparison to the novel 3D collagen scaffolds, showcasing a greater resistance to cell death and altered levels of fibrotic mRNA and protein expression. Furthermore, the hydrogels were determined to be a viable 3D culture method for hTM cells, with their protein release abilities successfully modeled with myoglobin highlighting potential areas for oxidative agent or antioxidant exposure. Several novel therapeutics were studied for their antioxidant potential to contribute to this aspect, with the greatest promise found with CNPs and peptain-1. This work will help provide insights into the behavior of hTM cells when grown in a biomimetic manner more similar to their native 3D tissue when introducing stimuli to their surrounding microenvironment.
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