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Noémie Parent, Julie Bérubé, Jean-Michel Bourget, Maxence Mounier, Teodor Veres, Stephanie Proulx; Engineering of a curved corneal stromal substitute. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3911.
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© ARVO (1962-2015); The Authors (2016-present)
The self-assembly approach of tissue engineering allows to reconstruct 3D corneal stromal substitutes (Proulx et al., 2009; Bourget et al., 2016). Presently, these substitutes are flat. The purpose of this study was to engineer a stromal substitute with the curved shape of a native cornea.
Curved plastics were formed using a thermoplastic elastomer (TPE) and sterilized. Their size fitted into a well of 6-well plates. Keratocytes, previoulsy isolated from a human cornea, were seeded on top of the TPE and cultured in the presence of serum and ascorbic acid. Flat 6-well culture plates were used as controls. Cells were cultured for 35 days and formed sheets of extracellular matrix. Two sheets were superposed to form a thicker stromal substitute, and cultured to allow for the sheets to adhere to each other (14 days for curved stromal substitutes; 8 days for flat stromal substitutes). The stromal substitutes were then characterized using macroscopic images. They were then fixed in 3.7% formaldehyde for histology (Masson’s Trichrome staining).
The activated keratocytes secreted extracellular matrix and formed sheets on both the curved TPE and the flat plastic controls. When removed from the curved TPE surface, the stromal substitute maintained a more pronounced curved form. Thickness of the stromal substitutes were calculated using histology cross-sections. The curved stromal subsitutes had a mean thickness of 26.1±0.5µm (n=2 counts) and flat stromal subsitutes 15.4±2.7µm (n=3 counts).
This study demonstrates that the self-assembly approach can be used to reconstruct curved stromal substitute. Therefore, this brings us closer to a reconstructed cornea whose properties are similar to the native tissue.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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