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K Y Chan, M Järveläinen, J H Chang, M J Edenfield; A cryodamage model for studying corneal nerve regeneration.. Invest. Ophthalmol. Vis. Sci. 1990;31(10):2008-2021.
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This study used a transcorneal freezing technique to produce a 2-mm circular, central wound in the rabbit cornea for investigating corneal nerve regeneration. All the corneal cells, nerves, and associated Schwann cells were dead inside the wound, but the extracellular matrix components remained intact. The destroyed epithelium and endothelium were replaced in 1 and 5-7 days, respectively. The necrotic keratocytes and stromal and subepithelial nerves were removed completely in 1-3 days by invading macrophage-like cells. The wounded stroma was repopulated centripetally by migrating keratocytes between days 1-5. Two types of nerve growth were identified in the stroma. The first type was novel sprouting of straight, long neurites between days 2-21, initially from the undamaged, periwound nerves and later from regenerated stromal nerves inside the wound. These small-caliber neurites proliferated in a random and disorderly pattern both inside and outside the wound and sometimes terminated on stationary, stellate keratocytes. The second type was genuine regrowth of stromal and subepithelial nerves in a centripetal direction between days 3-7. Schwann cells appeared on the newly formed nerves starting on day 4 or 5. A near-normal pattern and size of the nerves were established in the wound as early as day 10. In the epithelium, transient, wound-oriented neurites (days 1-3), single nerves, and semileashes (days 4-10) appeared. A near-normal leash pattern was restored between days 10-21 only at the wound periphery. Thus, in this model, the major groundwork of nerve regeneration occurred between days 3-10, simultaneously, at all three levels of nerve organization. These data suggest that nerve-Schwann cell interaction contributes to the restoration of stromal and subepithelial nerves, whereas a reparative epithelium deficient in trophic activity may account for the incomplete regrowth of epithelial nerves. The cryodamage model offers an efficient and multifaceted system for the experimental study of corneal nerve regeneration.
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