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Jose A. Gomes, Gustavo T. Grottone, Renata R. Loureiro, Priscila Cristovam, Gisela T. Oh; Ex-vivo Cornea Model For Magnetic Attraction Assay On Ocular Cultured Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):427.
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© ARVO (1962-2015); The Authors (2016-present)
Validate a cheap and useful cornea/aqueous humor experimental model, testing the attraction of ocular cells impregnated with magnetic nanoparticles.
Two different ocular derived cell lines were tested in our experiment(ARPE-19 and human corneal endothelial cells). These cells were impregnated with 0.21mg Fe/ml Ferumoxsil nanoparticles for 24 hours. Cells were dissociated and became aggregates with a 2mg/ml collagenase A solution after 12 hours incubation. Corneas tested were exposed to nitrogen gas for 7 days in a sealed container for acellularization. Then they were dissected to cover the external vertical wall of a Labtek Permanox 8 well chamber slide and left to dry and attach to the plastic for 4 hours at the laminar flow cabinet. A 3500 gauss neodymium magnet was fixed at the external side of the same well containing the cornea previously dissected. Digested aggregates were poured slowly at the well and left for 24 hours before any other manipulation. After 48 hours the slides were checked for the presence of cell at the surface of the chamber slide or the cornea attached to the vertical wall of the tested well.
The accelularization process with nitrogen provided corneas with a nice transparecy and few cells on stroma. After pouring the cells at the wells, cells impregnated with magnetic nanoparticles promptly adhered to the cornea against gravity forces. In the other hand,cell aggregates not containing magnetic nanoparticles were not able to adhere the corneas displaced vertically, and kept adhered at the bottom of the cornea ex-vivo model or the chamber slide surface.
The experimental model used at this project was able to simulate the interaction of magnetized cells, magnetic field, and gravity forces. The concentration used to both cell types was enough to provide attraction of cell aggregates towards the magnetic field. Aggregates with no magnetic nanoparticles laid at the surface of the slide without any interaction with the magnetic field.
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