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1.
Acta Biomater ; 10(12): 4939-4946, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25194930

ABSTRACT

Cell transplantation therapies to treat diseases related to dysfunction of retinal ganglion cells (RGCs) are limited in part by an inability to navigate to the optic nerve head within the retina. During development, RGCs are guided by a series of neurotrophic factors and guidance cues; however, these factors and their receptors on the RGCs are developmentally regulated and often not expressed during adulthood. Netrin-1 is a guidance factor capable of guiding RGCs in culture and relevant to guiding RGC axons toward the optic nerve head in vivo. Here we immobilized Netrin-1 using UV-initiated crosslinking to form a gradient capable of guiding the axonal growth of RGCs on a radial electrospun scaffold. Netrin-gradient scaffolds promoted both the percentage of RGCs polarized with a single axon, and also the percentage of cells polarized toward the scaffold center, from 31% to 52%. Thus, an immobilized protein gradient on a radial electrospun scaffold increases RGC axon growth in a direction consistent with developmental optic nerve head guidance, and may prove beneficial for use in cell transplant therapies for the treatment of glaucoma and other optic neuropathies.


Subject(s)
Guided Tissue Regeneration/instrumentation , Nerve Growth Factors/chemistry , Nerve Growth Factors/pharmacokinetics , Retinal Ganglion Cells/cytology , Retinal Ganglion Cells/physiology , Tissue Engineering/instrumentation , Tissue Scaffolds , Tumor Suppressor Proteins/chemistry , Tumor Suppressor Proteins/pharmacokinetics , Adsorption , Animals , Animals, Newborn , Axons/physiology , Axons/ultrastructure , Cell Polarity/physiology , Cell Proliferation/physiology , Cells, Cultured , Coated Materials, Biocompatible/chemical synthesis , Coated Materials, Biocompatible/pharmacokinetics , Equipment Design , Equipment Failure Analysis , Mice , Netrin-1
2.
Biomaterials ; 34(17): 4242-50, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23489919

ABSTRACT

Retinal degenerative diseases, such as glaucoma and macular degeneration, affect millions of people worldwide and ultimately lead to retinal cell death and blindness. Cell transplantation therapies for photoreceptors demonstrate integration and restoration of function, but transplantation into the ganglion cell layer is more complex, requiring guidance of axons from transplanted cells to the optic nerve head in order to reach targets in the brain. Here we create a biodegradable electrospun (ES) scaffold designed to direct the growth of retinal ganglion cell (RGC) axons radially, mimicking axon orientation in the retina. Using this scaffold we observed an increase in RGC survival and no significant change in their electrophysiological properties. When analyzed for alignment, 81% of RGCs were observed to project axons radially along the scaffold fibers, with no difference in alignment compared to the nerve fiber layer of retinal explants. When transplanted onto retinal explants, RGCs on ES scaffolds followed the radial pattern of the host retinal nerve fibers, whereas RGCs transplanted directly grew axons in a random pattern. Thus, the use of this scaffold as a cell delivery device represents a significant step towards the use of cell transplant therapies for the treatment of glaucoma and other retinal degenerative diseases.


Subject(s)
Nerve Fibers/physiology , Retinal Ganglion Cells/cytology , Tissue Engineering/methods , Animals , Axons/physiology , Cell Survival , Electrophysiological Phenomena , Mice , Rats , Rats, Sprague-Dawley , Tissue Scaffolds/chemistry
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