tretinoin and Corneal-Diseases

The purpose of the present study is to investigate the expression of aldehyde dehydrogenases (ALDHs) in rabbit corneas with limbal stem cell deficiency (LSCD) and corneas treated with cultured autologous oral mucosa epithelial cell sheet CAOMECS designed to reconstruct the ocular surface with LSCD.. New Zealand white rabbit autologous oral mucosal epithelial cells were isolated from a buccal biopsy and cultured to be grafted back onto corneas of rabbit model of LSCD. Immunofluorescent staining and Western blot analysis were used to compare the expression of ALDH1A1 and ALDH1A3 in healthy, LSCD-diseased, CAOMECS treated corneas. Human oral mucosal and corneal epithelial cells (OMECS and CECs) were cultured and treated with retinoic acid (RA) to further investigate the expression of ALDHs.. In healthy corneas, ALDH1A1 and ALDH1A3 were markedly expressed in basal cells of corneal epithelium. In LSCD diseased corneas, ALDH1A1 and ALDH1A3 were markedly expressed in the conjunctivalized apical epithelial cells, the goblet cells, and the stroma. CAOMECS grafted corneas showed a decreased expression of ALDHs as compared to LSCD diseased corneas. Western blot analysis confirmed the up regulation of ALDH1A1 and ALDH1A3 expression in LSCD-diseased corneal epithelial cells. CAOMECS expressed low levels of ALDH1A1 and ALDH1A3, as compared to diseased CECs (D-CEC). When ALDH1A3 was up regulated by retinoic acid treatment in OMECS, Pax-6 expression was down regulated, suggesting a decrease in regenerative capacity when ALDH enzymes are up regulated.. These findings report for the first time the up regulation of ALDH1A1 and ALDH1A3 in rabbit corneas with LSCD and document that CAOMECS grafting used to reconstruct corneal epithelium may reduce the expression levels of ALDH enzymes.

Topics: Aldehydes; Animals; Corneal Diseases; Epithelial Cells; Limbus Corneae; Oxidoreductases; Rabbits; Stem Cells; Tretinoin

Congenital PAX6-aniridia is a rare panocular disease resulting from limbal stem cell deficiency. In PAX6-aniridia, the downregulation of the retinol-metabolizing enzymes ADH7 (All-trans-retinol dehydrogenase 7) and ALDH1A1/A3 (Retinal dehydrogenase 1, Aldehyde dehydrogenase family 1 member A3) have been described in limbal epithelial cells (LECs) and conjunctival epithelial cells. The aim of this study was to identify the role of retinol derivates in the differentiation of human LEC and its potential impact on aniridia-associated keratopathy development. Human LEC were isolated from healthy donor corneas and were cultured with retinol, retinoic acid, or pan-retinoic acid receptor antagonist (AGN 193109) acting on RARα, β, γ (NR1B1, NR1B2 NR1B3) or were cultured with pan-retinoid X receptor antagonist (UVI 3003) acting on RXR α, β, γ (retinoid X receptor, NR2B1, NR2B2, BR2B3). Using qPCR, differentiation marker and retinoid-/fatty acid metabolism-related mRNA expression was analysed. DSG1 (Desmoglein 1), KRT3 (Keratin 3), and SPINK7 (Serine Peptidase Inhibitor Kazal Type 7) mRNA expression was downregulated when retinoid derivates were used. AGN 193109 treatment led to the upregulation of ADH7, KRT3, and DSG1 mRNA expression and to the downregulation of KRT12 (Keratin 12) and KRT19 (Keratin 19) mRNA expression. Retinol and all-trans retinoic acid affect some transcripts of corneal LEC in a similar way to what has been observed in the LEC of PAX6-aniridia patients with the altered expression of differentiation markers. An elevated concentration of retinol derivatives in LEC or an altered response to retinoids may contribute to this pattern. These initial findings help to explain ocular surface epithelia differentiation disorders in PAX6-aniridia and should be investigated in patient cells or in cell models in the future in more detail.

Topics: Aniridia; Corneal Diseases; Down-Regulation; Tretinoin

To evaluate the long-term follow-up of recurrent conjunctival and corneal intraepithelial neoplasia (CCIN) treated with combination topical interferon alfa-2b and retinoic acid (I/RA).. Our study represents a retrospective observational interventional series of 82 eyes from 82 patients from a single institution, reviewed for CCIN. All were administered topical interferon alfa-2b 1 million IU/mL QID and retinoic acid 0.01% every other day. Patients had been diagnosed by biopsy. A Kaplan-Meier survival analysis, Wilcoxon signed-rank test and a multivariate logistic regression were statistical tests used to correlate recurrence with patient and tumor variables.. 79 eyes assessed for CCIN diagnoses and treated with I/RA achieved tumor resolution. The median tumor-free follow-up was ~109.1 months with a median time to resolution being ~2.8 months. Our median treatment duration was ~11.3 months. The greatest difference in the mean total residual tumor size was identified between Months 0-1 [-7.63 mm. Combination I/RA was effective in treating CCIN lesions with few transient side effects. The combination of retinoids and interferons may represent a viable topical therapeutic agent with an extended tumor-free follow-up and a large proportion of our study's patients achieving >10 year's tumor-free follow-up. Our treatment duration is long, but by cost-comparing surgical against medical interventions, topical I/RA may serve as a safe and effective alternative.

Topics: Administration, Topical; Antineoplastic Agents; Carcinoma in Situ; Conjunctival Neoplasms; Corneal Diseases; Follow-Up Studies; Humans; Interferon alpha-2; Neoplasm Recurrence, Local; Retrospective Studies; Treatment Outcome; Tretinoin

PAX6-related Aniridia is a sight-threatening disease involving progression of secondary glaucoma and aniridia related keratopathy (ARK). Change or loss of limbal epithelial progenitors causes epithelial surface defects. We analyzed the effect of PAX6 on mRNA expression changes with a two-step approach, as follows. First, we sequenced mRNA from limbal epithelial cells isolated from controls and aniridia patients. Second, we confirmed the bioinformatics and literature-based result list for a small interfering RNA (siRNA)-based primary aniridia cell model (PAX6 knockdown). With this approach, we expected that the genes directly influenced by PAX6 would be distinguishable from those affected secondarily by the ARK disease state. Therefore, epithelial cells were isolated from the limbus region of two patients with aniridia and cultured in keratinocyte serum-free medium. Normal control cells were obtained from the limbus region of corneal donors. For the siRNA-based aniridia cell model, cells were transfected with Lipofectamine and 5 nM siRNA against PAX6 or control treatment. All cells were lysed to yield DNA, RNA, and protein. Reduction of PAX6 protein was assessed by western blot. Aniridia and control Poly-A-enriched RNA libraries were subjected to next-generation sequencing. The differential analysis was a combination of quantification with RSEM and differential tests with edgeR. Gene lists were filtered by comparison to NCBI GEO datasets, annotated with DAVID, and manually annotated using a literature search. Based on the resulting filtered gene list, qPCR primers were purchased, and candidate genes (TP63, ABCG2, ADH7, ALDH1A1, PITX1, DKK1, DSG1, KRT12, KRT3, KRT13, SPINK6, SPINK7, CTSV, SERPINB1) were verified by qPCR on the siRNA-based aniridia cell model. We identified genes that might be regulated by PAX6 and showed that SPINK7 mRNA, which codes for a protease inhibitor, is downregulated in patients as well as in our primary aniridia cell model. ALDH1A1 and AHD7 mRNA levels were reduced in limbal epithelial cells of aniridia patients, and both transcripts were downregulated by PAX6 knockdown in our cell model. This siRNA-based aniridia cell model is a valuable tool for confirming identified PAX6-affected genes that might promote ARK pathogenesis. The model recapitulated expression changes for SPINK7, ADH7, and ALDH1A1 that were also observed in patient samples. These results provide evidence that PAX6 might drive corneal epithelial differentiation by d

Topics: Alcohol Dehydrogenase; Aldehyde Dehydrogenase; Aldehyde Dehydrogenase 1 Family; Aniridia; Blotting, Western; Cell Differentiation; Cells, Cultured; Corneal Diseases; Epithelium, Corneal; Gene Expression Regulation; High-Throughput Nucleotide Sequencing; Humans; Limbus Corneae; Models, Biological; PAX6 Transcription Factor; Real-Time Polymerase Chain Reaction; Retinal Dehydrogenase; RNA, Messenger; RNA, Small Interfering; Serine Peptidase Inhibitors, Kazal Type; Signal Transduction; Transfection; Tretinoin

Topics: Administration, Topical; Adult; Aged; Aged, 80 and over; Corneal Diseases; Epithelium, Corneal; Female; Humans; Keratolytic Agents; Limbus Corneae; Male; Middle Aged; Retrospective Studies; Stem Cells; Tretinoin

The combination of topical interferon α-2b (IFN α-2b) and all-trans retinoic acid (ATRA) 0.01% has previously been shown to be effective in conjunctival and corneal intraepithelial neoplasia. This combination was incidentally found to be effective in a patient with partial limbal stem cell deficiency (LSCD), a condition which can be challenging to treat if conservative measures fail. This retrospective study evaluates the combination of topical IFN α-2b and ATRA 0.01% in the treatment of partial LSCD.. Five patients from one institution with a clinical and/or histopathological diagnosis of LSCD had failed a period of conservative treatment with cessation of toxic stimuli and use of lubricating eye-drops. These patients were treated with a combination regimen of topical IFN α-2b and ATRA 0.01%.. All five patients had partial LSCD, but limbal involvement was significantly worse in one patient who later progressed to total LSCD. Complete clinical resolution of signs of LSCD was achieved in the four patients with partial LSCD after a mean of 9 months of treatment. The one patient who progressed to total LSCD did not respond to treatment. Duration of follow-up after clinical resolution in the four patients with partial LSCD was at least 18 months, with no signs of recurrence seen. Aside from the complaint of ocular irritation in one patient, no other side effects were observed.. Combination treatment of topical IFN α-2b and ATRA 0.01% can be considered in partial LSCD, where adjacent parts of the limbus remain intact. This treatment is associated with minimal side effects and no recurrence of signs of LSCD at least 18 months after clinical resolution.

Topics: Administration, Topical; Aged; Aged, 80 and over; Antineoplastic Agents; Corneal Diseases; Drug Therapy, Combination; Female; Follow-Up Studies; Humans; Interferon alpha-2; Interferon-alpha; Limbus Corneae; Male; Middle Aged; Ophthalmic Solutions; Recombinant Proteins; Retrospective Studies; Risk Factors; Stem Cells; Treatment Outcome; Tretinoin; Visual Acuity

Topics: Antineoplastic Agents; Carcinoma in Situ; Conjunctival Neoplasms; Corneal Diseases; Eye Neoplasms; Female; Humans; Interferon-alpha; Male; Tretinoin

Topics: Antineoplastic Agents; Carcinoma in Situ; Conjunctival Neoplasms; Corneal Diseases; Eye Neoplasms; Female; Humans; Interferon-alpha; Male; Tretinoin

Retinoic acid (RA) is a metabolite of vitamin A that plays a fundamental role in the development and function of the human eye. The purpose of this study was to investigate the effects of RA on the phenotype of corneal stromal keratocytes maintained in vitro for extended periods under serum-free conditions.. Keratocytes isolated from human corneas were cultured up to 21 days in serum-free media supplemented with RA or DMSO vehicle. The effects of RA and of its removal after treatment on cell proliferation and morphology were evaluated. In addition, the expression of keratocyte markers was quantified at the transcriptional and protein levels by quantitative PCR and immunoblotting or ELISA, respectively. Furthermore, the effects of RA on keratocyte migration were tested using scratch assays.. Keratocytes cultured with RA up to 10 × 10(-6) M showed enhanced proliferation and stratification, and reduced mobility. RA also promoted the expression of keratocyte-characteristic proteoglycans, such as keratocan, lumican, and decorin, and increased the amounts of collagen type-I in culture while significantly reducing the expression of matrix metalloproteases 1, 3, and 9. RA effects were reversible, and cell phenotype reverted to that of control after removal of RA from media.. Retinoic acid was shown to control the phenotype of human corneal keratocytes cultured in vitro by regulating cell behavior and extracellular matrix composition. These findings contribute to our understanding of corneal stromal biology in health and disease, and may prove useful in optimizing keratocyte cultures for applications in tissue engineering, cell biology, and medicine.

Topics: Blotting, Western; Cell Proliferation; Cells, Cultured; Corneal Diseases; Corneal Keratocytes; Corneal Stroma; Culture Media, Serum-Free; Enzyme-Linked Immunosorbent Assay; Eye Proteins; Fibroblasts; Gene Expression Regulation; Humans; Keratolytic Agents; Microscopy, Fluorescence; Phenotype; Polymerase Chain Reaction; RNA; Tretinoin

To evaluate the long-term recurrence rate of conjunctival and corneal intraepithelial neoplasia (CIN) treated with retinoic acid and topical interferon alfa-2b.. Retrospective, noncomparative, interventional case series.. A total of 89 eyes of 89 patients from 1 institution who were treated between September 2003 and February 2010 for CIN lesions used topical interferon alfa 1 million IU/ml drops 4 times daily and retinoic acid 0.01% once every second day.. Diagnosis was made by biopsy and impression cytology. Patients' notes and clinical photographs were reviewed, and data were analyzed. All eyes were monitored for the possibility of recurrence with a minimum of 1 year of follow-up from the time of documented clinical resolution.. All eyes were monitored for the possibility of recurrence with a minimum of 1 year of follow-up from the time of documented clinical resolution.. Complete clinical resolution of the CIN lesions was achieved in 87 of the 89 eyes treated (97.75%). Two of the 89 eyes treated (2.25%) had only a partial response to treatment; of these 2 patients, 1 was taking cyclosporine for keratitis sicca. For the 87 eyes with complete response, resolution occurred after a mean of 1.69 months (range, 19 days to 6.5 months). Mean follow-up after clinical resolution (tumor-free period) was 51.5 months (range, 11-84 months). Four of the 87 patients with complete response developed a mild allergic papillary conjunctivitis that settled on halving the interferon dose to 0.5 million IU drops and reducing the frequency to 3 times daily. Side effects were limited to 1 case of epithelial microcysts and 1 case of marginal keratitis.. In this group of patients observed with CIN lesions, combination treatment of topical retinoic acid and interferon alfa-2b was effective in treating lesions with minimal self-limited side effects with faster and greater resolution and a longer tumor-free period compared with studies using interferon alfa-2b alone. We hypothesize that topical all-trans retinoic acid and interferon alfa-2b may act synergistically. We believe that combination treatment of interferon alfa-2b and retinoic acid may offer a superior alternative to interferon alfa-2b alone in treating CIN.

Topics: Administration, Topical; Antineoplastic Agents; Carcinoma in Situ; Conjunctival Neoplasms; Corneal Diseases; Drug Therapy, Combination; Eye Neoplasms; Female; Follow-Up Studies; Humans; Interferon alpha-2; Interferon-alpha; Male; Neoplasm Recurrence, Local; Ophthalmic Solutions; Recombinant Proteins; Retrospective Studies; Treatment Outcome; Tretinoin

Topics: Administration, Topical; Antineoplastic Combined Chemotherapy Protocols; Carcinoma in Situ; Corneal Diseases; Epithelium, Corneal; Eye Neoplasms; Female; Humans; Interferon alpha-2; Interferon-alpha; Limbus Corneae; Middle Aged; Recombinant Proteins; Tretinoin

To determine the associations between corneal biomechanical parameters as measured by the Reichert Ocular Response Analyser (ORA) and disc morphology and retinal nerve fibre layer thickness (RNFL) measured by the Heidelberg Retinal Tomograph (HRT) II in Singaporean children.. This is a cross-sectional study conducted on a subset of children enrolled in the Singapore Cohort Study of the Risk Factors of Myopia (SCORM). Corneal hysteresis (CH), corneal resistance factor (CRF) and central corneal thickness (CCT) were measured with the ORA. Optic disc morphology and RNFL thickness were assessed by the HRT II. Cycloplegic refraction and ultrasound A-scans were also performed, and disc tilt was assayed from stereo photographs.. 102 subjects (mean age 12.01 (SD 0.57) years; range 11-14 years) were included in the study. The mean CH was 12.00 (1.40) mm Hg, the mean CRF was 11.99 (1.65) mm Hg, and the mean CCT was 581.12 (33.53) mum. Eyes with tilted discs had significantly longer axial lengths and more myopic refraction than eyes without tilted discs. There were no significant correlations between CH, CRF or CCT and the HRT II parameters, after the application of the Bonferroni correction. When stratified for disc tilt, however, the global disc area was significantly correlated with CCT (r = -0.49, p = 0.001).. Corneal biomechanical properties as measured with the ORA do not vary with optic disc parameters or RNFL. Central corneal thickness is correlated with disc area in Singaporean schoolchildren with tilted discs. This relationship may influence glaucoma risk in myopic subjects.

Topics: Adolescent; Biomechanical Phenomena; Child; Cornea; Corneal Diseases; Cross-Sectional Studies; Diagnostic Techniques, Ophthalmological; Female; Glaucoma; Humans; Intraocular Pressure; Male; Myopia; Nerve Fibers; Optic Disk; Refractive Errors; Singapore; Tretinoin

We report four cases of corneoconjunctival keratinization that were successfully treated with topical retinoic acid ointment. In two cases keratinization was due to squamous metaplasia and in two others it was secondary to intraepithelial corneoconjunctival neoplasia. Treatment reversed severe keratinization in a case of drug-induced pseudopemphigoid and stabilized the disease in one of the two affected eyes without additional treatment. In a case of ocular cicatricial pemphigoid, retinoic acid was useful as an adjuvant therapy to immunosuppression, by reversing keratinization of the conjunctiva. In two cases of corneoconjunctival neoplasia, lesions regressed markedly. Long-term treatment was well tolerated in three patients. Our findings suggest that retinoic acid ointment is effective in treating severe squamous metaplasia in cicatrizing diseases of the conjunctiva. Our findings indicate further that retinoic acid seems to inhibit growth of corneoconjunctival neoplasias and thus might be useful complementary therapy in this situation.

Topics: Administration, Topical; Aged; Aged, 80 and over; Conjunctiva; Conjunctival Diseases; Conjunctival Neoplasms; Conjunctivitis; Cornea; Corneal Diseases; Epithelium; Female; Humans; Keratins; Male; Metaplasia; Tretinoin

Topics: Aged; Calcinosis; Corneal Diseases; Dry Eye Syndromes; Female; Humans; Male; Tretinoin

Several reports have appeared on the efficacy of topically applied 0.01% or 0.1% all-trans retinoic acid (0.04-0.4 millimolar) for treatment of xerophthalmia, conjunctival squamous metaplasia, and corneal epithelial erosions in humans and animals. An observation common to many of these studies is the occurrence of an adverse reaction to retinoic acid in the form of lid margin hyperemia and blepharoconjunctivitis. Since retinoic acid is biologically active at micromolar to nanomolar concentrations, it may be possible to reduce side effects while maintaining therapeutic effectiveness by reducing the retinoic acid concentration in ophthalmic formulations. In the present study, topical 0.005% retinoic acid in petrolatum ointment reversed corneal keratinization in xerophthalmic, vitamin A-deficient rabbits in 3-4 days while 0.0005% (2 micromolar) retinoic acid ointment was effective in 4-6 days. Further clinical trials of topical retinoic acid for treatment of ocular surface disease should be conducted using micromolar concentrations of retinoic acid which are expected to maintain a therapeutic effect while reducing adverse reactions.

Topics: Administration, Topical; Animals; Corneal Diseases; Osmolar Concentration; Rabbits; Time Factors; Tretinoin; Wound Healing; Xerophthalmia

Retinoic acid 0.1% in arachis oil was applied to one eye and arachis oil alone to the other eye, of each of 19 patients with equivalent degrees of corneal xerophthalmia in the fellow eyes. Even with concomitant systemic vitamin A therapy, topical retinoic acid was associated with more rapid healing of corneal lesions in a substantial proportion of cases. Application of retinoic acid three times a day produced no significant side effects; application five times a day, however, resulted in moderate to severe conjunctival injection and increased corneal vascularization and scarring.

Topics: Administration, Topical; Corneal Diseases; Humans; Tretinoin; Vitamin A; Xerophthalmia

The entire corneal epithelium of each of 42 rabbits was removed bilaterally. Tretinoin or 0.5% or 0.1% ethylretinamide drops were applied topically five times a day to the eyes of one-half of the experimental animals, and peanut oil was applied to the eyes of the other half (the controls). The healing of the denuded corneas of the animals receiving tretinoin or 0.5% ethylretinamide was significantly more advanced than the healing of the corneas of the control animals. The 0.1% ethylretinamide solution did not enhance the healing process.

Topics: Animals; Cornea; Corneal Diseases; Epithelium; Female; Male; Rabbits; Time Factors; Tretinoin; Wound Healing