Cultivation of human corneal limbal stem cells in Mebiol gel--A thermo-reversible gelation polymer.

BACKGROUND & OBJECTIVES: Cultivated limbal stem cell transplantation is being used as a current treatment modality for limbal stem cell deficiency. However, use of allogenic biological material as substrate is associated with risks of transmission of certain diseases and allograft rejection. Therefore development of non-toxic biodegradable synthetic polymers is important. We undertook this study to evaluate the use of a synthetic polymer Mebiol gel as a substrate for the growth of limbal phenotype cells and cornea phenotype cells from limbal explants. METHODS: Human cadaveric limbal explants cells were cultivated on Mebiol gel. The proliferative capacity of cultivated cells was analyzed with thymidine incorporation studies. Immunostaining for presumed limbal stem cell association markers and cornea differentiation markers was performed and confirmed with reverse transcription (RT-PCR). RESULTS: The limbal explants underwent proliferation in vitro. The cultivated cells expressed the presumed limbal stem cell association markers (ABCG2 and p63), the transient amplifying cell markers (connexin 43, integrin alpha9) and the cornea differentiation marker (K3). RT PCR confirmed the immunohistochemical data. INTERPRETATION & CONCLUSION: Our findings showed that the synthetic polymer Mebiol gel was able to support limbal explant proliferation. The cultured cells expressed presumed limbal stem cell association markers, transient amplifying cells and cornea phenotype markers. Mebiol Gel can be used as a scaffold for growing limbal explants.

Nocardia Asteroides keratitis: report of seven patients and literature review.

PURPOSE: To describe clinical features and treatment outcomes in patients with advanced Nocardia asteroides keratitis. METHODS: Retrospective review of case records of 7 patients with culture-proven Nocardia keratitis. RESULTS: Corneal infection occurred after corneal trauma in two patients, cataract surgery in three patients, penetrating keratoplasty in one patient and was associated with a silicone buckle element infection in one patient. Mean duration of infection at presentation was 33.4 days (7-75 days), and five patients had received prior treatment with corticosteroids. Six of seven patients had deep corneal suppuration at the time of presentation, clinically suggestive of mycotic keratitis. In two patients who had received prolonged corticosteroid therapy (> or = 45 days), the eyes could not be salvaged. Complete resolution of infection was achieved in all 4 eyes treated with topical fortified cefazolin eye drops (50 mg/ml).

Partial flap during laser in-situ keratomileusis: pathogenesis and timing of retreatment.

PURPOSE: To report the timing of retreatment and clinical outcomes in patients with a partial corneal flap during laser in-situ keratomileusis (LASIK), and to describe the causes of this complication. METHODS: Retrospective review of case records of four patients (4 eyes) who had a partial corneal flap during LASIK. RESULTS: The mean age of the four patients was 23 +/- 4.7 years, and mean preoperative spherical equivalent (SE) refraction was -9.1 +/- 3.1 D (range, -5.5 to -13 D). A 160 microns corneal flap was attempted during the initial treatment. Retreatment with a 180 microns corneal flap was performed at a mean of 5.1 +/- 1.6 weeks (range, 4 to 7.5 weeks) after the initial procedure. There were no intraoperative complications during retreatment. Post-LASIK mean SE refraction was -1.0 +/- 1.1 D (range, +0.38 to -2.0 D), after a mean follow up of 19 +/- 15.7 weeks (range, 7 to 42 weeks). Best spectacle-corrected visual acuity decreased in one eye from 6/5 to 6/6 and was maintained in the others. CONCLUSION: LASIK retreatment can be performed as early as one month after a partial flap, if the refraction is stable and a thicker corneal flap is created.

Fellow eye treatment in excimer photo refractive keratectomy.

PURPOSE: To describe symmetry of response in fellow eyes of patients undergoing photorefractive keratectomy (PRK) for myopia, analyse the risk factors leading to asymmetry in response and to determine if delayed treatment of the second eye increases safety and predictability of PRK. METHODS: Retrospective review of case records of 133 patients who underwent bilateral myopic PRK and had a minimum follow up of 6 months in both eyes. RESULTS: Postoperative uncorrected visual acuity, spherical equivalent (SE) refraction within +/- 1 D of emmetropia, best-corrected visual acuity (BCVA) and corneal haze were not significantly different in fellow eyes of patients undergoing PRK for myopia. Of 87 eyes in group 1 (myopia < 6 D), 96.6% had uncorrected visual acuity > or = 6/12, 89.7% were within +/- 1 D of emmetropia, none lost > or = 1 line BCVA, and none had haze > or = grade 3. Similar results for 98 eyes in group II (myopia 6 to 9.9 D) were 75.6%, 55.1%, 2.0% and 2.0% respectively. For 81 eyes in group III (myopia > or = 10 D) the results were 42.7%, 33.3%, 8.6%, and 4.9% respectively. Among 84 patients with similar preoperative myopia in both eyes, 54 (64.3%) patients had a postoperative SE difference < or = 1 D in fellow eyes. Risk factors for asymmetric response among fellow eyes included increasing preoperative myopia (p < 0.001) and dissimilar treatment technique in the two eyes (p = 0.03). Corneal haze did not increase significantly after the third postoperative month. CONCLUSION: This study demonstrates that considerable symmetry of response exists in fellow eyes of patients undergoing myopic PRK. Early PRK in the fellow eye of patients with < 6 D myopia is safe and allows quick visual rehabilitation of the patient. In patients with myopia > or = 6 D, a 3-month interval before treating the second eye may improve the safety of the procedure.

Conjunctival-limbal autografts for primary and recurrent pterygia: technique and results.

Our technique of pterygium excision with conjunctival-limbal autografting is described and the safety and efficacy of the procedure in India is analysed. Case records of 51 consecutive patients (53 eyes) who underwent surgery at our institute between November 1992 and September 1994 were retrospectively analysed. Recurrence was defined as fibrovascular tissue crossing the corneoscleral limbus onto clear cornea in the area of previous pterygium excision. 2 (3.8%) of the 53 pterygia (primary 36; recurrent 17) recurred, after a mean follow up of 18.9 +/- 12.1 months (range: 1.5-43 months). Both recurrences occurred within a year of follow up, in patients who were < or = 40 years of age. No major operative or postoperative complications were encountered. The inclusion of limbal tissue in conjunctival autografts following pterygium excision appears to be essential to ensure low recurrence rates. The technique is safe, simple and inexpensive and is recommended for the management of both primary and recurrent pterygia in Indian eyes.

Evaluation of laboratory tests for diagnosis of chlamydial infections in conjunctival specimens.

We evaluated the chlamydial direct diagnostic tests and McCoy cell culture methods for detection of Chlamydia trachomatis in 234 conjunctival scrapings from 127 patients. Direct (rapid) diagnostic methods of Giemsa stain cytology, fluorescent antibody test (FAT) and immunoperoxidase (IPO) stain and McCoy cell culture isolation method using IPO staining and FAT for detection of C. trachomatis antigen were carried out. McCoy cell culture isolation using FAT antigen detection method was considered as the gold standard. The specificity of all the direct diagnostic tests were above 90 per cent but sensitivity of Giemsa cytology, FAT and IPO stain were 36, 54 and 53 per cent respectively showing that as direct diagnostic tests Giemsa stain cytology was inefficient and FAT and IPO were useful. Specificity and sensitivity of McCoy cell culture using IPO stain for detection C. trachomatis antigen were above 90 per cent and it is suggested that this test can be used as a routine diagnostic method.