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Lauren Cornell, Jennifer McDaniel, Brian Lund, David O Zamora; Primary human corneal endothelial cell transplantation in an ex vivo model. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1377.
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© ARVO (1962-2015); The Authors (2016-present)
The corneal endothelium is responsible for maintaining corneal clarity. However, this cell layer poses great challenges for clinicians due to its lack of regenerative potential and reducing cell population with age. This study investigates the potential of human corneal endothelial cells (HCEC), loaded with iron-based nanoparticles, to be magnetically-directed to injured regions of the cornea.
Primary cultures of human HCEC from CellProgen were maintained in human endothelial serum free media containing 10 ng/ml FGF-2 and plated at 75,000 cells in a 48 well plate for 24 hours. Cells were then exposed to 50nm dextran-coated biotin conjugated super paramagnetic iron oxide nanoparticles (SPIONP) at 37°C for up to 48 hours in serum free media. Western blot, PCR, and Prussian Blue staining were utilized to evaluate uptake of nanoparticles and cellular response by SPIONP dose. Mathematic modeling based upon stokes law, gravity, and magnetic field strength as well as fluid flow dynamics and outflow of the aqueous chamber were used to determine optimum SPIONP cell loading in relation to magnetic field strength for induced cellular movement within the aqueous chamber. Mathematical modeling efficacy was evaluated by injecting SPIONP loaded HCECs onto a denuded human corneal endothelium in the presence of an applied magnetic field.
HCEC were successfully cultured and maintained their in-vivo marker expression of Glypican 4. PCR revealed there was dose dependent impact on cell pump expression with SPIONP loading and an optimized loading dose was determined. When SPIONP loaded-HCEC were placed in solution with the denuded cornea, up to 1 million cells/mL, the cells showed targeted movement through the solution towards the externally applied magnetic field of 1.23 Tesla.
These studies show that HCEC maintained their lineage after readily incorporating SPIONPs. Proof of concept studies performed here indicate that cells with internally-loaded SPIONP can be directed and manipulated through an aqueous solution to a predetermined area when a magnetic field is applied. Mathematical modeling of the cell loading capacity and magnetic strength needed for this movement to occur can be an effective tool for tailoring specific ocular therapeutic needs for patients. Results of this study may lead to the development of a non-surgical technique to replenish this vital cell layer.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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