Natural history and predictors for progression in pediatric keratoconus.

We studied the demographic and clinical predictors associated with keratoconus progression in a pediatric population. Retrospective cohort study. We evaluated 305 eyes without previous surgeries from 168 patients, 9 to < 18 years old, and with a minimum 36-month follow-up in a hospital corneal ambulatory. We used Kaplan-Meyer survival curves; the dependent variable (main outcome measure) was the interval time (months) until the event, defined as an increase of 1.5 D in the maximum keratometry (Kmax), obtained with Pentacam. We evaluated the predictors: age (< or ≥ 14 years), sex, keratoconus familial history, allergy medical history, and the baseline tomographic parameters: mean keratometry (Km), Kmax (< or ≥ 55 D); and thinnest pachymetry (TP). We used log-rank tests and compared median survival times for right (RE)/left eyes (LE) and better (BE)/worse eyes (WE). A p value < 0.05 was considered significant. The patients' mean ± SD age was 15.1 ± 2.3 years old; 67% were boys, 30% were < 14 years, 15% had keratoconus familial history, and 70% were allergic. The general Kaplan-Meyer curves showed no differences between RE/LE or BE/WE. RE with allergy and LE with Kmax ≥ 55 D had smaller survival times ((95%CI 9.67-32.1), p 0.031 and (95%CI 10.1-44.1), p 0.042, respectively). For BE and WE, Kmax ≥ 55 D had smaller survival times ((95% CI 6.42- ), p 0.031 and (95%CI 8.75-31.8), p 0.043, respectively). Keratoconus progression was similar between RE/LE and BE/WE. Steepest corneas are predictors of faster progression. Allergy is also a predictor of keratoconus progression in RE.

Clinical, Demographic, and Tomographic Aspects Related to Iris Mammillations Among Patients With Keratoconus: A Cross-Sectional Study.

PURPOSE: Iris mammillations (IM) were previously described in patients with keratoconus, but the clinical implications of this finding have never been studied. Our aim was to investigate demographic, tomographic, and clinical characteristics potentially associated with the presence of IM among patients with keratoconus. METHODS: This was a cross-sectional study performed among patients with keratoconus in a public-affiliated university hospital. All patients under follow-up were considered eligible to participate in the study. Participants were evaluated by 2 trained ophthalmologists and submitted to corneal tomography (Pentacam). Selected demographic, clinical, and tomographic characteristics were assessed and compared among participants with IM (IM group) and without IM (No-IM group) using the Wilcoxon test or 2-tailed Fisher exact test, as appropriate. RESULTS: The study population consisted of 106 subjects and 19 (17.9%) presented with IM. The median age and interquartile range were 18 years old (14-24) in the IM group and 20 years old (17-24) in the No-IM group (P = 0.135). The female proportion was 47.3% in the IM group and 52.8% in the No-IM group (P = 0.801). Median (interquartile ranges) pachymetric values of the right eyes were 498 (466-525) for the IM group and 459 (421-482) for the No-IM group (P = 0.005). For the left eyes, the values were 490 (456-523) in the IM group and 450 (418-485) in the No-IM group (P = 0.024). CONCLUSIONS: Subjects with keratoconus presenting with IM have thicker corneas than those without IM. Follow-up studies should be performed to evaluate the clinical implications of this finding.

Autoradiographic study on the regenerative capability of the epithelium lining the center of the cornea after multiple debridements of its peripheral region.

BACKGROUND: The epithelium lining the center of the cornea is assumed to lack stem cells.The purpose is to investigate by autoradiography the regenerative capability of the epithelium lining the central region of the rabbit cornea following seven scrapings of its peripheral lining, during several months. METHODS: After marking the center of the cornea with a 6 mm-diameter trephine, the epithelium outside this area was scraped until reaching the corneoscleral zone. This procedure was repeated seven times on the same eye at intervals of 20 days. One day after the last scraping, (3)H-thymidine was injected intravitreally and the corneas processed for autoradiography. RESULTS: At 2 days after injection, the corneal surface was entirely lined by an epithelium made up by two layers of squamous cells, most of them being labeled with the DNA precursor. A multilayered epithelium was visualized at the center with most of its basal cells also labeled. The limbal epithelium had at least two of its layers labeled with the precursor. At 9 days, the multilayered central unscraped epithelium exhibited labeled cells not only in the basal but also in its suprabasal layers. The labeling index (labeled nuclei/100 cells) for its basal stratum was very close to 100%. A similar feature was observed at 16 days, except that the mutilayered central epithelium was seen lining a larger area when compared to the precedent interval and that it exhibited evidences for vertical renewal. CONCLUSIONS: The epithelium lining the central region of the cornea--where it was assumed that stem cells do not exist--exhibited capability for regeneration and self-renewal in spite of seven consecutive debridements of its periphery. No evidence was found for transposition of limbal epithelial cells to the center of the cornea during the early merger of the epithelial sliding fronts.

Regeneration of the corneal epithelium after debridement of its central region: an autoradiographic study on rabbits.

PURPOSE: To investigate the proliferative behavior of the corneal and limbal epithelia after debridement on the central region of the rabbit cornea. METHODS: After scraping a circular epithelial area, 5 mm in diameter, in the center of the cornea, ([3]) H-thymidine ( ([3]) H-TdR) was injected intravitreally, and the rabbits killed from 1 to 49 days afterward. The cornea, together with the adjacent conjunctiva, was processed for autoradiography. RESULTS: The regenerating epithelium at the center of the cornea exhibited high frequencies of labeled nuclei when compared to controls. The mitotic indexes for the limbus were comparable in experimental and control eyes. The unique basal stratum of the limbal epithelium exhibited quick proliferation and vertical migration in all eyes. Cells that remained labeled for four weeks or more were observed throughout the corneal epithelium, including its basal stratum, and this did not depend on epithelial damage. CONCLUSION: Corneal epithelium wounds are healed by sliding and proliferation of cells surrounding the epithelial gap without any evidence for the participation of the limbal epithelium. Daughter cells labeled with ([3]) H-TdR were visualized in all layers of the corneal epithelium up to 7 weeks after the DNA precursor injection. However, at this long interval, the only labeled cells in the limbus were in the suprabasal layers.

Proliferation of the vascular endothelium of the iris following total debridement of the corneal epithelium and limbal excision of rabbits.

BACKGROUND: Damage to the corneal epithelium causes not only a reaction for its repair but also affects other parts of the cornea as well as different components of the anterior segment of the eye. The purpose of this investigation was to analyze the consequences, following epithelial and limbal damage, to the iris of rabbits (Oryctolagus cuniculus). METHODS: The corneal epithelium was thoroughly scraped followed by surgical excision of the limbus. Next, (3)H-thymidine ((3)H-TdR) was injected intravitreally both into the right (experimental) and left (control) eyes which had their anterior segments processed for autoradiography at intervals of 2, 7 and 21 days after surgery (three rabbits per interval). The irises were also examined with scanning-electron and confocal microscopy after Evans blue injection. RESULTS: There was a high frequency of labeling in the cells of the iris blood vessels in the experimental eye, particularly the endothelial ones. The ratio of labeled cells between experimental and control irises was 40:1, with a population of nuclei increasing by 25% and remaining labeled up to 21 days. There was also an increase in the volume of the iris vasculature as shown by confocal microscopy. The high labeling frequencies of the vascular cells were observed throughout the iris from the ciliary to the pupillary regions. CONCLUSIONS: The lesions on the corneal epithelium elicit proliferation of the iris vascular cells, mainly its endothelium, as well as an early breakdown of the blood-aqueous barrier. The daughter cells resulting from the damage to the eye surface were detected up to 21 days after a single injection of (3)H-TdR, most likely due to their slow turnover. As a consequence of this proliferation, the vasculature of the iris increased in volume.

Morphological and autoradiographic studies on the corneal and limbal epithelium of rabbits.

The investigation was centered on the morphological features of the conjunctiva-cornea transition (limbus) of the rabbit eye and the proliferative behavior of its epithelium. The eyes were processed for examination with light and electron microscopy, as well as for autoradiography after intravitreal injection of [(3)H]thymidine ([(3)H]TdR). At the sites of extraocular muscle insertion, the vascularization of the stroma extended to the peripheral cornea, and the limbal epithelium was thin with its basal stratum made up by clear cuboidal cells. In between the muscle insertions, the cuboidal clear cells, as well as the stroma blood vessels, were scarce. At the light microscope level, the basement membrane was distinct in the cornea but not in the limbus or the conjunctiva. Autoradiographs demonstrated that, at the limbus, the basal cells migrated very quickly to the suprabasal region and remained there up to the 28-day interval. Labeled cells were identified in all epithelial layers of the cornea, including the basal one, at 21 and 28 days but not in the limbal basal clear cells. The rate of renewal of conjunctival epithelium was similar to that observed for the transition with scarce clear cells. The high-resolution autoradiographs demonstrated that the basal cuboidal clear limbal cells exhibit a quick renewal and that they are not label-retaining cells. These latter ones were detected all over the corneal epithelium and in the suprabasal layers of the limbus up to 28 days, in physiological conditions, without the need of stimulation by damage to the corneal epithelium.