Micro- and nanotopography with extracellular matrix coating modulate human corneal endothelial cell behavior |
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| Affiliation: | 1. Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore;2. Department of Biomedical Engineering, National University of Singapore, Singapore 117575, Singapore;4. Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore 168751, Singapore;5. Singapore National Eye Centre, Singapore 168751, Singapore;6. Duke-NUS Graduate Medical School, Singapore 169857, Singapore;7. Department of Surgery, National University of Singapore, Singapore 119228, Singapore;1. Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China;2. The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China;1. Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Wake Forest Baptist Medical Center, Medical Center Boulevard, Winston-Salem, NC 27157, USA;2. Department of Ophthalmology, Wake Forest School of Medicine, Wake Forest Baptist Medical Center, Medical Center Boulevard, Winston-Salem, NC 27157, USA;1. Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan;2. Department of Ophthalmology, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan;3. Department of Ophthalmology, Taipei Medical University Hospital, Taipei, Taiwan;4. Department of Ophthalmology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan;1. National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, Suzhou 215021, People''s Republic of China;2. Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA;3. New England Eye Center, Tufts Medical Center, Boston, MA, USA;4. Federal University of São Paulo, School of Medicine, São Paulo, Brazil;5. Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA 15213, USA;1. Department of Mechanical and Mechatronics Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada;2. Department of Systems Design Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada;3. School of Optometry and Vision Science, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada |
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| Abstract: | The human corneal endothelium plays an important role in maintaining corneal transparency. Human corneal endothelial cells have limited regenerative capability in vivo. Consequently, endothelial dysfunction can occur following corneal endothelial trauma or inherited diseases. To restore endothelial function, corneal transplantation is needed. However, there is a worldwide shortage of donor corneas, motivating the development of a tissue-engineered graft alternative using cultivated endothelial cells. To induce in vitro cell proliferation, much effort has been made to improve culture conditions and to mimic the native extracellular microenvironment. We incorporated topographical and biochemical cues in our in vitro culture of human corneal endothelial cell line B4G12 (HCEC-B4G12) and hypothesized that manipulation of the extracellular environment can modulate cell proliferation, morphometry and phenotype. The topographies tested were nanopillars, microwells and micropillars on polydimethylsiloxane, while the biochemical factors were extracellular matrix protein coatings of fibronectin-collagen I (FC), FNC® coating mix (FNC) and laminin-chondroitin sulfate (LC). Cellular morphometry, Na+/K+-ATPase and zona occludens 1 (ZO-1) gene and protein expression were analyzed 3 days after cells had formed a confluent monolayer. The cell circularity on all patterns and coatings was above 0.78. On all coatings, cell area was the lowest on micropillars. The coefficient of variation (CV) of the cell area was the lowest on nanopillars with an LC coating. With an FC coating, micropillars induced a better cellular outcome as the cells had the greatest circularity, smallest cell area and highest Na+/K+-ATPase and ZO-1 gene and protein expression. With the LC coating, HCECs grown on nanopillars resulted in the lowest CV of the cell area and the highest ZO-1 gene expression. Thus, HCEC-B4G12 morphometry and phenotype can be improved using different topographical and biochemical cues. |
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| Keywords: | HCEC-B4G12 Substrate topography Extracellular matrix protein Tissue engineering Cell proliferation |
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