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Karl E. Kador, Ramon Montero, Benjamin Yungher, Jonathan Hertz, Erin B. Lavik, Fotios M. Andreopoulos, Jeffrey L. Goldberg; Radial Scaffold Design For Proper Ganglion Cell Orientation. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6656.
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The inability of the adult mammalian retina to replace or regenerate retinal ganglion cells (RGCs) following injury motivates the study of cell and tissue engineering approaches. However, it is not sufficient to merely create a scaffold supporting cell growth, it is also necessary to orient axon growth. Here we describe an electrospinning (ES) method and a novel collector to create radially aligned ES scaffolds that direct axon growth towards a central point, mimicking retinal axon growth towards the optic nerve head.
Radial aligned scaffolds were spun from a polylactic acid (PLA) solution using a grounding collector with a center pole surrounded by a conducting rim. Scaffold fiber diameter and alignment were determined by scanning electron microscopy. RGCs dissected from postnatal Sprague-Dawley rats were purified to 99.9% homogeneity by immunopanning. Scaffolds were sterilized and coated with laminin, seeded, and cultured in a defined serum-free media for 2-3 days. Following culture, cells were fixed and neurites stained with a monoclonal β3 tubulin antibody to quantify radial alignment.
ES scaffolds under various flow rates and electric potentials produced PLA fibers ranging from 191 to 423 nm in diameter. Alignment Analysis showed directional coherency ranging from 50-78% based upon spinning conditions; control (random) alignment had a coherency of 10-14%. RGC axon growth showed a significantly higher orientation of axon growth along the radial orientation of the scaffold fibers compared to that in control cultures.
Seeding RGCs onto a radially aligned scaffold directs axon growth towards a central point, potentially creating both a model to study optic nerve formation and also a translatable tissue implant for the injured inner retina.
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