Purchase this article with an account.
David O Zamora, Mauris N DeSilva, Lauren E Cornell, Randolph D Glickman, Heuy-Ching Hetty Wang, Anthony James Johnson; Characterization of Magnetic Nanoparticle Loaded Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1442.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Current methods to replace diseased or traumatized endothelium involve a full thickness keratoplasty or a Descemet’s Stripping Automated Endothelial Keratoplasty procedure to replace this targeted region of the cornea. We hypothesize magnetic nanoparticles (MNPs) can be pre-loaded into endothelial cells and used to deliver them to the posterior of the cornea after injection into the anterior chamber, while in the presence of a magnetic field.
Bovine corneal endothelial cells (Astarte Biologics) were cultured in DMEM medium, containing 10% FBS. Twenty-thousand cells per well were seeded into 48 well plates (Passage 6-8) and maintained in culture for 48 hrs. Nanomag -D-Spio (biotin coated; Micromod, Rostock, Germany) were added to the cells in the following bead to cell ratios: 0:1, 100:1, 1,000:1, and 10,000:1. Phase contrast and fluorescence microscopy were used to evaluate the cell morphology of the treated cells. MTT viability assays were used to evaluate the effects of the MNPs on endothelial cells after being exposed to them for up to a 48 hr period. Assays were each performed in triplicate.
Exposure of the MNPs to the endothelium, at any of the concentrations analyzed, did not appear to affect the confluency or morphology of the monolayer of cells, as determined by phase contrast microscopy. Furthermore, the F-actin cytoskeleton of the cells did not appear to re-arrange or disassemble after these treatments. Interestingly, endothelial cell MTT metabolism OD values slightly decreased (0.95, 0.92, 0.89, 0.89) as MNP bead:cell ratios increased (0:1, 100:1, 1000:1, 10000:1), but was not statistically significant.
MNPs are an FDA-approved technology for use in medical procedures in humans. Here we have demonstrated that bovine endothelial cells can be cultured in the presence of MNPs and incorporated into these cultures apparently with minimal toxicity. The initial proof of concept data described here will be used to further develop this technology using human corneal endothelial cells, in order to target these cells to the back of the cornea for repairing injuries to the endothelium.
This PDF is available to Subscribers Only