Corneal femtosecond laser keratotomy results in isolated stromal injury and favorable wound-healing response.
OBJECTIVE
To examine the corneal repair response after intrastromal femtosecond (fs) laser keratotomy.


METHODS
Twelve rabbits underwent monocular intrastromal keratotomy performed with an fs laser at a preoperatively determined corneal depth of 160 to 200 microm. The fs laser-induced corneal repair response was compared with that of nonoperated control eyes and eyes treated with photorefractive keratectomy (PRK). Follow-up examinations were performed 1, 3, 7, and 28 days after surgery. Corneas were evaluated using slit lamp, in vivo confocal microscopy, and light microscopy. The extracellular matrix components fibronectin and tenascin were located using immunofluorescence staining. Anti-Thy-1 and anti-alpha-SMA antibodies and phalloidin were used to identify repair fibroblasts. Cell proliferation and nuclear DNA fragmentation were detected using an anti-Ki-67 antibody and the TUNEL assay, respectively.


RESULTS
Intrastromal fs keratotomy resulted in a hypocellular stromal scar discernible as a narrow band of increased reflectivity on slit lamp examination. Deposition of fibronectin and tenascin as well as death and subsequent proliferation of keratocytes were observed. No differentiation of keratocytes into Thy-1- or alpha-SMA-positive fibroblasts could be detected. In contrast, after PRK, which causes epithelial and stromal wounding, all markers for repair fibroblasts were found in subepithelial stromal layers. On slit lamp examination, a fibrotic scar and a corneal haze were revealed.


CONCLUSIONS
Isolated stromal injury using an fs laser avoids epithelial injury and is associated with a favorable wound-healing response preserving corneal transparency. Thus, fs laser keratotomy is a highly selective laser treatment that can be useful for the treatment of refractive errors.
Meltendorf
Christian
C
Department of Ophthalmology, Johann Wolfgang Goethe-University, Frankfurt am Main, Germany. meltendorf@em.uni-frankfurt.de
Burbach
Guido J
GJ
Bühren
Jens
J
Bug
Reinhold
R
Ohrloff
Christian
C
Deller
Thomas
T
D000199:Actins / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation; D003319:Corneal Stroma / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D053938:DNA Fragmentation; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005500:Follow-Up Studies; D020287:In Situ Nick-End Labeling; D007518:Isoantibodies / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D053685:Laser Therapy / Q000379:methods*; D054018:Lasers, Excimer; D018613:Microscopy, Confocal; D018903:Photorefractive Keratectomy; D011817:Rabbits; D019063:Tenascin / Q000378:metabolism; D014945:Wound Healing / Q000502:physiology*
D000199:Actins; D005353:Fibronectins; D007518:Isoantibodies; D019394:Ki-67 Antigen; D019063:Tenascin; C035678:anti-Thy antibody