transforming-growth-factor-beta and Corneal-Opacity

To compare the incidence and degree of corneal haze formation following laser subepithelial keratomileusis (LASEK) and epithelial laser in situ keratomileusis (epi-LASIK), and examine its correlation with tear film transforming growth factor-beta1 (TGF-beta1) levels.. This prospective, interventional, clinical trial included 20 eyes (20 patients) randomly assigned to undergo LASEK or epi-LASIK. The level of TGF-beta1 in tear fluid was measured preoperatively and 1, 3, and 5 days postoperatively. Corneal haze was graded at 1 and 3 months after surgery, and the relationship with TGF-beta1 levels was determined.. Mean preoperative spherical equivalent refraction was -4.50 +/- 1.44 diopters (D) (range: -1.50 to -6.00 D) for LASEK eyes and -4.90 +/- 1.26 D (range: -1.75 to -6.00 D) for epi-LASIK eyes. Although mean corneal haze scores at 1 month were significantly higher in LASEK-treated eyes than in epi-LASIK treated eyes (P=.031), these scores were similar at 3 months (P=.608). Tear fluid TGF-beta1 levels were similar in LASEK and epi-LASIK eyes before surgery (P=.458) and significantly higher in the LASEK group at 1, 3, and 5 days postoperatively (P=.015, P=.023, and P=.039, respectively). A positive correlation was noted between tear TGF-beta1 levels on the first postoperative day and the degree of corneal haze at 1 month (r=0.501, P=.016).. Less corneal haze was noted after epi-LASIK than LASEK. A positive correlation between corneal haze and tear fluid TGF-beta1 levels on the first postoperative day suggest a possible mechanism for the observed difference.

Topics: Adult; Corneal Opacity; Female; Humans; Incidence; Keratectomy, Subepithelial, Laser-Assisted; Keratomileusis, Laser In Situ; Male; Myopia; Postoperative Complications; Prospective Studies; Tears; Transforming Growth Factor beta; Transforming Growth Factor beta1

Granular corneal dystrophy type 2 (GCD2) is an autosomal dominant corneal stromal dystrophy that is caused by p.Arg124His mutation of transforming growth factor β induced (TGFBI) gene. It is characterized by well demarcated granular shaped opacities in central anterior stroma and as the disease progresses, extrusion of the deposits results in ocular pain due to corneal epithelial erosion. Also, diffuse corneal haze which appears late, causes decrease in visual acuity. The prevalence of GCD2 is high in East Asia including Korea. Homozygous patients show a severe phenotype from an early age, and the heterozygote phenotype varies among patients, depending on several types of compound heterozygous TGFBI mutations. In the initial stage, conservative treatments such as artificial tears, antibiotic eye drops, and bandage contact lenses are used to treat corneal erosion. Different surgical methods are used depending on the depth and extent of the stromal deposits. Phototherapeutic keratectomy removes anterior opacities and is advantageous in terms of its applicability and repeatability. For deeper lesions, deep anterior lamellar keratoplasty can be used as the endothelial layer is not always affected. Recurrence following these treatments are reported within a wide range of rates in different studies due to varying definition of recurrence and follow-up period. In patients who have undergone corneal laser vision-correction surgeries such as photorefractive keratectomy, LASEK, or LASIK including SMILE surgery, corneal opacity exacerbates rapidly with severe deterioration of visual acuity. Further investigations on new treatments of GCD2 are necessary.

Topics: Cornea; Corneal Dystrophies, Hereditary; Corneal Opacity; Corneal Ulcer; Humans; Keratomileusis, Laser In Situ; Photorefractive Keratectomy; Transforming Growth Factor beta

To evaluate the effect of topical suberanilohydroxamic acid (SAHA) and 5-methyl-1-phenyl-2[1H]-pyridone (pirfenidone) on the degree of corneal haze in the stromal wounded ex vivo canine cornea.. Twenty-four corneoscleral rims from normal dogs were uniformly wounded with an excimer laser and placed into culture medium with an air-liquid interface. The control group (n = 8) contained placebo-treated corneas. Treatment group 1 (n = 8) received SAHA topically every 6 hours. Treatment group 2 (n = 8) received pirfenidone topically every 6 hours. Each cornea was fluorescein stained and macrophotographed every 6 hours to assess epithelialization rate. All corneas were also macrophotographed weekly to assess optical clarity (haze). Images were analyzed for differences in pixel intensity between wounded (haze) and unwounded (nonhaze) regions, and haze surface area for each cornea was calculated.. The mean epithelialization time was 47.25 hours in the control group, 45.00 hours in the SAHA group, and 43.50 hours in the pirfenidone group, revealing no significant difference (P = 0.368). The median difference in pixel intensity between haze and nonhaze areas was 21.5 in the control group, 8.0 in the SAHA group, and 8.0 in the pirfenidone group, which is significant (P < 0.01). The median haze surface area was 12.96 mm2 in the control group, 5.70 mm2 in the SAHA group, and 5.92 mm2 in the pirfenidone group, which is significant (P < 0.01).. Stromal-wounded ex vivo canine corneas exhibited greater optical clarity when treated with SAHA and pirfenidone than when placebo treated at 21 days. There was no significant difference in epithelialization rate between groups. Corneal contour was correlated with geographic haze distribution.

Topics: Actins; Administration, Ophthalmic; Animals; Anti-Inflammatory Agents, Non-Steroidal; Connective Tissue Growth Factor; Cornea; Corneal Injuries; Corneal Opacity; Corneal Stroma; Dogs; Epithelium, Corneal; Histone Deacetylase Inhibitors; Immunohistochemistry; Lasers, Excimer; Models, Animal; Organ Culture Techniques; Pyridones; Re-Epithelialization; Real-Time Polymerase Chain Reaction; Transforming Growth Factor beta; Visual Acuity; Vorinostat; Wound Healing

To describe 2 unrelated families with multiple members demonstrating a less commonly recognized vortex pattern of corneal deposits confirmed to be granular corneal dystrophy type 1 (GCD1) after identification of the p.(Arg555Trp) mutation in the transforming growth factor β-induced gene (TGFBI).. A slit-lamp examination was performed on individuals from 2 families, one of Mexican descent and a second of Italian descent. After DNA extraction from affected individuals and their unaffected relatives, TGFBI screening was performed.. Eight of 20 individuals in the Mexican family and 20 of 55 in the Italian family demonstrated corneal stromal opacities. Seven of the 8 affected individuals in the Mexican family and 4 of the 20 affected individuals in the Italian family demonstrated a phenotype characterized by a "sea fan" or vortex pattern of superficial stromal corneal deposits originating from the inferior aspect of the cornea. Screening of TGFBI in both families revealed a heterozygous missense mutation [p.(Arg555Trp)] in exon 12, confirming the diagnosis of GCD1.. Our findings demonstrate that GCD1 may present with a vortex pattern of anterior stromal deposits. Although this pattern of dystrophic deposits is not recognized by clinicians as a typical phenotype of GCD1, it is consistent with the production of the majority of the TGFBI protein by the corneal epithelium.

Topics: Adolescent; Aged, 80 and over; Child; Corneal Dystrophies, Hereditary; Corneal Opacity; Corneal Stroma; DNA Mutational Analysis; Extracellular Matrix Proteins; Female; Gene Frequency; Heterozygote; Humans; Italy; Male; Mexico; Middle Aged; Mutation, Missense; Pedigree; Polymerase Chain Reaction; Slit Lamp; Transforming Growth Factor beta; Young Adult

The aim of this study was to examine whether mesenchymal stem cells (MSCs) and/or corneal limbal epithelial stem cells (LSCs) influence restoration of an antioxidant protective mechanism in the corneal epithelium and renewal of corneal optical properties changed after alkali burns. The injured rabbit corneas (with 0.25 N NaOH) were untreated or treated with nanofiber scaffolds free of stem cells, with nanofiber scaffolds seeded with bone marrow MSCs (BM-MSCs), with adipose tissue MSCs (Ad-MSCs), or with LSCs. On day 15 following the injury, after BM-MSCs or LSCs nanofiber treatment (less after Ad-MSCs treatment) the expression of antioxidant enzymes was restored in the regenerated corneal epithelium and the expressions of matrix metalloproteinase 9 (MMP9), inducible nitric oxide synthase (iNOS), α-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), and vascular endothelial factor (VEGF) were low. The central corneal thickness (taken as an index of corneal hydration) increased after the injury and returned to levels before the injury. In injured untreated corneas the epithelium was absent and numerous cells revealed the expressions of iNOS, MMP9, α-SMA, TGF-β1, and VEGF. In conclusion, stem cell treatment accelerated regeneration of the corneal epithelium, restored the antioxidant protective mechanism, and renewed corneal optical properties.

Topics: Adipocytes; Alkalies; Animals; Antioxidants; Burns, Chemical; Cell Differentiation; Corneal Opacity; Corneal Pachymetry; Epithelium, Corneal; Female; Gene Expression Regulation; Immunohistochemistry; Limbus Corneae; Matrix Metalloproteinase 9; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Nitric Oxide Synthase Type II; Protective Agents; Rabbits; Superoxide Dismutase; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

In humans suffering from pulmonary disease and a mouse model, transient receptor potential vanilloid 4 (TRPV4) channel activation contributes to fibrosis. As a corneal alkali burn induces the same response, we determined if such an effect is also attributable to TRPV4 activation in mice. Accordingly, we determined if the alkali burn wound healing responses in wild-type (WT) mice are different than those in their TRPV4-null (KO) counterpart. Stromal opacification due to fibrosis in KO (n = 128) mice was markedly reduced after 20 days relative to that in WT (n = 157) mice. Immunohistochemistry revealed that increases in polymorphonuclear leukocytes and macrophage infiltration declined in KO mice. Semi-quantitative real time RT-PCR of ocular KO fibroblast cultures identified increases in proinflammatory and monocyte chemoattractant protein-1 chemoattractant gene expression after injury. Biomarker gene expression of fibrosis, collagen1a1 and α-smooth muscle actin were attenuated along with macrophage release of interleukin-6 whereas transforming growth factor β, release was unchanged. Tail vein reciprocal bone marrow transplantation between WT and KO chimera mouse models mice showed that reduced scarring and inflammation in KO mice are due to loss of TRPV4 expression on both corneal resident immune cells, fibroblasts and infiltrating polymorphonuclear leukocytes and macrophages. Intraperitoneal TRPV4 receptor antagonist injection of HC-067047 (10 mg/kg, daily) into WT mice reproduced the KO-phenotype. Taken together, alkali-induced TRPV4 activation contributes to inducing fibrosis and inflammation since corneal transparency recovery was markedly improved in KO mice.

Topics: Actins; Alkalies; Animals; Cornea; Corneal Opacity; Eye Burns; Fibrosis; Gene Expression Regulation; Gene Knockout Techniques; Inflammation; Interleukin-6; Mice; Transforming Growth Factor beta; TRPV Cation Channels; Vascular Endothelial Growth Factor A

Injuring mouse corneas with alkali causes myofibroblast expression leading to tissue opacification. However, in transient receptor potential vanilloid 1 channel (TRPV1-/-) knockout mice healing results in transparency restoration. Since TGFβ is the primary inducer of the myofibroblast phenotype, we examined the mechanism by which TRPV1 affects TGFβ-induced myofibroblast development. Experiments were performed in pig corneas and human corneal fibroblasts (HCFs). Immunohistochemical staining of α-smooth muscle actin (α-SMA) stress fibers was used to visualize myofibroblasts. Protein and phosphoprotein were determined by Western blotting. siRNA transfection silenced TRPV1 gene expression. Flow cytometry with a reactive oxygen species (ROS) reporting dye analyzed intracellular ROS. [Ca2+]I was measured by loading HCF with fura2. In organ cultured corneas, the TRPV1 antagonist capsazepine drastically reduced by 75% wound-induced myofibroblast development. In HCF cell culture, TGF-β1 elicited rapid increases in Ca2+ influx, phosphorylation of SMAD2 and MAPKs (ERK1/2, JNK1/2 and p38), ROS generation and, after 72 hrs myofibroblast development. SMAD2 and p38 activation continued for more than 16 h, whereas p-ERK1/2 and p-JNK1/2 waned within 90 min. The long-lived SMAD2 activation was dependent on activated p38 and vice versa, and it was essential to generate a > 13-fold increase in α-SMA protein and a fully developed myofibroblast phenotype. These later changes were markedly reduced by inhibition of TRPV1 or reduction of the ROS generation rate. Taken together our results indicate that in corneal derived fibroblasts, TGFβ- induced myofibroblast development is highly dependent on a positive feedback loop where p-SMAD2-induced ROS activates TRPV1, TRPV1 causes activation of p38, the latter in turn further enhances the activation of SMAD2 to establish a recurrent loop that greatly extends the residency of the activated state of SMAD2 that drives myofibroblast development.

Topics: Actins; Alkalies; Animals; Calcium; Cornea; Corneal Injuries; Corneal Opacity; Feedback, Physiological; Gene Expression Regulation; Humans; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase 8; Mitogen-Activated Protein Kinase 9; Myofibroblasts; p38 Mitogen-Activated Protein Kinases; Reactive Oxygen Species; Signal Transduction; Smad2 Protein; Swine; Tissue Culture Techniques; Transforming Growth Factor beta; TRPV Cation Channels

Corneal stromal scarring partly involves the production of corneal myofibroblasts. The purpose of this study was to examine the effects of rapamycin (an inhibitor of the mammalian target of rapamycin [mTOR] pathway) on myofibroblast formation in vitro and in-vivo.. Human corneal fibroblasts were grown in culture and transformed into myofibroblasts using TGF-β (2 ng/mL). The phosphorylation (activation) of the mTOR pathway was examined by immunoblotting. Cell proliferation with and without rapamycin was examined by thiazolyl blue tetrazolium bromide (MTT) assay and Ki67 staining. The expression of the myofibroblast differentiation marker smooth muscle actin (SMA) was examined by immunostaining and immunoblotting. The functional effects of rapamycin were measured using a gel contraction assay. For in vivo studies, 140 μm laser ablation was performed on rabbit corneas followed by subconjunctival rapamycin or vehicle. Corneal haze development was graded at 4 weeks, while the expression of myofibroblast markers was examined by immunostaining and immunoblotting.. The TGF-β activated the mTOR pathway with peak phosphorylation at 2 to 4 hours. Treatment of corneal fibroblasts with rapamycin reduced their proliferation by 46% compared to control. Rapamycin significantly inhibited TGF-β-induced expression of myofibroblast markers (17.2% SMA positive cells with rapamycin compared to 69.0% in control). Rapamycin also significantly inhibited TGF-β-induced collagen gel contraction. In the rabbit eyes treated with rapamycin, corneal haze development was significantly less compared to controls (0.75 ± 0.4 vs. 2.17 ± 0.7).. Rapamycin appears to inhibit proliferation and differentiation of corneal myofibroblasts and, thus, may provide an effective therapeutic measure for preventing corneal scarring.

Topics: Animals; Blotting, Western; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cicatrix; Corneal Opacity; Corneal Stroma; Disease Models, Animal; Female; Humans; Immunosuppressive Agents; Myofibroblasts; Photorefractive Keratectomy; Rabbits; Sirolimus; Transforming Growth Factor beta

To describe new affected individuals of Franceschetti's original pedigree of hereditary recurrent erosion and to classify a unique entity called Franceschetti corneal dystrophy.. Observational case series.. Slit-lamp examination of 10 affected individuals was conducted. Biomicroscopic examinations were supplemented by peripheral corneal biopsy in 1 affected patient with corneal haze. Tissue was processed for light and electron microscopy and immunohistochemistry was performed. DNA analysis was carried out in 12 affected and 3 nonaffected family members.. All affected individuals suffered from severe ocular pain in the first decade of life, attributable to recurrent corneal erosions. Six adult patients developed bilateral diffuse subepithelial opacifications in the central and paracentral cornea. The remaining 4 affected individuals had clear corneas in the pain-free stage of the disorder. Histologic and immunohistochemical examination of the peripheral cornea in a single patient showed a subepithelial, avascular pannus. There was negative staining with Congo red. DNA analysis excluded mutations in the transforming growth factor beta-induced (TGFBI) gene and in the tumor-associated calcium signal transducer 2 (TACSTD2) gene.. We have extended the pedigree of Franceschetti corneal dystrophy and elaborated its natural history on the basis of clinical examinations. A distinctive feature is the appearance of subepithelial opacities in adult life, accompanied by a decreased frequency of recurrent erosion attacks. Its clinical features appear to distinguish it from most other forms of dominantly inherited recurrent corneal erosion reported in the literature.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers; Biopsy; Cadherins; Cell Adhesion Molecules; Child; Chondroitin; Claudins; Corneal Dystrophies, Hereditary; Corneal Opacity; Decorin; Dermatan Sulfate; DNA Mutational Analysis; Extracellular Matrix Proteins; Eye Pain; Female; Humans; Immunohistochemistry; Male; Pedigree; Recurrence; Transforming Growth Factor beta

To develop ex vivo organ culture models of human corneal scarring suitable for pharmacological testing and the study of the molecular mechanisms leading to corneal haze after laser surgery or wounding.. Corneas from human donors were cultured ex vivo for 30 days, either at the air-liquid interface (AL) or immersed (IM) in the culture medium. Histological features and immunofluorescence for fibronectin, tenascin C, thrombospondin-1, and α-smooth muscle actin were graded from 0 to 3 for control corneas and for corneas wounded with an excimer laser. The effects of adding 10 ng/ml transforming growth factor-β1 (TGF-β1) to the culture medium and of prior complete removal of the epithelium and limbus, thus preventing reepithelialization, were also analyzed on wounded corneas. Collagen III expression was detected with real-time PCR.. Wounding alone was sufficient to induce keratocyte activation and stromal disorganization, but it was only in the presence of added TGF-β1 that intense staining for fibronectin and tenascin C was found in the AL and IM models (as well as thrombospondin-1 in the AL model) and that α-smooth muscle actin became detectable. The scar-like appearance of the corneas was exacerbated when TGF-β1 was added and reepithelialization was prevented, resulting in the majority of corneas becoming opaque and marked upregulation of collagen III.. THE MAIN FEATURES OF CORNEAL SCARRING WERE REPRODUCED IN THESE TWO COMPLEMENTARY MODELS: the AL model preserved differentiation of the epithelium and permits the topical application of active molecules, while the IM model ensures better perfusion by soluble compounds.

Topics: Actins; Biomarkers; Cicatrix; Collagen Type III; Cornea; Corneal Injuries; Corneal Keratocytes; Corneal Opacity; Culture Media; Fibronectins; Gene Expression Regulation; Humans; Lasers, Excimer; Organ Culture Techniques; Re-Epithelialization; Surface Properties; Tenascin; Thrombospondin 1; Transforming Growth Factor beta

Granular corneal dystrophy type 2 (GCD2) causes the formation of corneal deposits having 3 different morphological types. We used Fourier domain optical coherence tomography to assess the depths of each type according to the morphology.. A prospective study was performed in 54 eyes of 54 heterozygous patients with GCD2. Corneal deposits of 54 patients with GCD2 were classified into 3 morphological types: type 1, diffuse haze; type 2, granular shape (2 subgroups: type 2a, round granulated and type 2b, round spiculated); and type 3, linear shape (2 subgroups: type 3a, short side branched and type 3b, long side branched). Using Fourier domain optical coherence tomography, we measured the distances from the Bowman layer to the upper surface of the deposits (USBL), to the lower surface of the deposits (LSBL), and the thickness of the deposits (TD). The deposits formed along the flap interface were also examined among 19 patients who had LASIK.. Types 1 and 2 deposits were always adjacent to the Bowman layer; thus the USBLs for each were 0.0 ± 0.0 μm, whereas that of type 3 deposits was 65.4 ± 48.0 μm (P < 0.0001). The LSBL and TD of linear deposits with long side branches (type 3) (313.3 ± 71.4 and 246.2 ± 71.9 μm) were greater than those of type 1 (47.7 ± 10.2 and 47.7 ± 10.2 μm) and type 2 (91.3 ± 39.5 and 91.3 ± 39.5 μm) (P < 0.0001). There were no differences in the measurements between the subgroups type 2a and type 2b or between types 3a and 3b. USBL of the laser in situ keratomileusis group was 54.5 ± 29.8 μm.. The depths of corneal deposits in patients with GCD2 were associated with the morphology of the deposits. The linear deposits were located most deeply in the cornea, followed by granular deposits and diffuse haze moving anteriorly. Several deposits have distinct depths according to the morphological types.

Topics: Adult; Aged; Aged, 80 and over; Cornea; Corneal Dystrophies, Hereditary; Corneal Opacity; Extracellular Matrix Proteins; Female; Fourier Analysis; Humans; Keratomileusis, Laser In Situ; Male; Middle Aged; Point Mutation; Prospective Studies; Reproducibility of Results; Tomography, Optical Coherence; Transforming Growth Factor beta

The purpose of this study was to investigate the role of transforming growth factor beta (TGFβ) and/or platelet-derived growth factor-B (PDGF-B) blockade on the differentiation of vimentin and alpha-smooth muscle actin (αSMA)-expressing myofibroblasts associated with haze in mice. Mouse corneas had haze-generating irregular PTK (phototherapeutic keratectomy) and topical treatment with the vectors. Six study groups of PTK treated corneas, with four corneas per group in each experiment, were Group 1) treated with TGFβ-KDEL vector interfering with TGFβ signaling through anomalous sorting of cytokine bound to the expressed altered receptor; Group 2) treated with PDGF-B-KDEL vector interfering with PDGF signaling through anomalous sorting of cytokine bound to the expressed altered receptor; Group 3) treated with both TGFβ-KDEL vector and PDGF-B-KDEL vector to interfere with signaling of both cytokines; Group 4) empty pGFPC1 vector; Group 5) empty pCMV vector; and Group 6) no vector treatment control. At one month after surgery, the corneas were analyzed by immunocytochemistry (IHC) for central stromal cells expressing myofibroblast markers vimentin and αSMA. The stroma of corneas treated with the TGFβ-KDEL vector alone (p < 0.05) or both the TGFβ-KDEL and PDGF-B-KDEL vectors (P < 0.05) had significantly lower density of vimentin-positive cells compared to the corresponding control group. The central stroma of corneas treated with the TGFβ-KDEL vector (p < 0.05) or the PDGF-B-KDEL vector (p < 0.05) had lower density of αSMA-positive cells compared to the corresponding control group. The density of αSMA-positive stromal cells was also significantly lower (p < 0.05) when both the TGFβ-KDEL and PDGF-B-KDEL and vectors were applied together compared to the corresponding control groups. This study provides in situ evidence that TGFβ and PDGF-B have important roles in modulating myofibroblast generation in the mouse cornea after haze-associated injury.

Topics: Actins; Animals; Cells, Cultured; Cornea; Corneal Opacity; Corneal Surgery, Laser; Disease Models, Animal; Down-Regulation; Female; Genetic Therapy; Immunohistochemistry; Mice; Mice, Inbred C57BL; Myofibroblasts; Proto-Oncogene Proteins c-sis; Time Factors; Transfection; Transforming Growth Factor beta; Vimentin

Topics: Aged; Cataract; Corneal Dystrophies, Hereditary; Corneal Opacity; Diabetic Retinopathy; Exons; Extracellular Matrix Proteins; Humans; Male; Point Mutation; Polymerase Chain Reaction; Transforming Growth Factor beta

The purpose of this study was to report the association of phenotypic features characteristic of lattice corneal dystrophy (LCD) with a monoclonal gammopathy of undetermined significance (MGUS) after exclusion of a coding region mutation in transforming growth factor beta-induced (TGFBI) gene.. Case report.. Slit-lamp examination and collection of DNA for TGFBI screening were performed. A systemic evaluation was also performed to evaluate for conditions associated with systemic amyloidosis.. A 65-year-old man demonstrated bilateral linear branching corneal stromal opacities characteristic of classic LCD. No mutations were found in any of the 17 exons of TGFBI or in the intron-exon boundary regions. Four previously described single nucleotide polymorphisms were identified: c.698C>G (p.Leu217Leu; rs1442), c.1028A>G (p.Val327Val; rs1054124), c.1416C>T (p.Leu472Leu; rs1133170), and c.1667T>C (p.Phe540Phe; rs4669). Serum protein electrophoresis revealed the presence of a monoclonal spike, and based on the results of additional investigations, the patient was diagnosed with MGUS.. Although the presence of bilateral thin branching lattice lines in the corneal stroma is characteristic of classic LCD, this distinctive phenotype may not be associated with a TGFBI coding region mutation but instead with a myeloproliferative disorder such as MGUS. Therefore, appropriate genetic and serologic testing should be performed in patients with a late-onset LCD phenotype in the absence of a positive family history.

Topics: Aged; Blood Proteins; Corneal Dystrophies, Hereditary; Corneal Opacity; Corneal Stroma; Electrophoresis; Exons; Extracellular Matrix Proteins; Genetic Testing; Humans; Male; Mutation; Paraproteinemias; Phenotype; Polymorphism, Single Nucleotide; Transforming Growth Factor beta

Trichostatin A (TSA), a histone deacetylase inhibitor, has been shown to suppress TGF-beta-induced fibrogenesis in many nonocular tissues. The authors evaluated TSA cytotoxicity and its antifibrogenic activity on TGF-beta-driven fibrosis in the cornea with the use of in vitro and in vivo models.. Human corneal fibroblasts (HSFs) were used for in vitro studies, and New Zealand White rabbits were used for in vivo studies. Haze in the rabbit cornea was produced with photorefractive keratectomy (PRK) using excimer laser. Trypan blue exclusion and MTT assays evaluated TSA cytotoxicity to the cornea. Density of haze in the rabbit eye was graded with slit lamp biomicroscopy. Real-time PCR, immunoblotting, or immunocytochemistry was used to measure alpha-smooth muscle actin (SMA), fibronectin, and collagen type IV mRNA or protein levels. TUNEL assay was used to detect cell death.. TSA concentrations of 250 nM or less were noncytotoxic and did not alter normal HSF morphology or proliferation. TGF-beta1 treatment of HSF significantly increased mRNA and protein levels of SMA (9-fold), fibronectin (2.5-fold), and collagen type IV (2-fold). TSA treatment showed 60% to 75% decreases in TGF-beta1-induced SMA and fibronectin mRNA levels and 1.5- to 3.0-fold decreases in protein levels but had no effect on collagen type IV mRNA or protein levels in vitro. Two-minute topical treatment of TSA on rabbit corneas subjected to -9 D PRK significantly decreased corneal haze in vivo.. TSA inhibits TGF-beta1-induced accumulation of extracellular matrix and myofibroblast formation in the human cornea in vitro and markedly decreases haze in rabbit cornea in vivo.

Topics: Actins; Animals; Cell Proliferation; Cell Survival; Cells, Cultured; Collagen Type IV; Corneal Opacity; Corneal Stroma; Disease Models, Animal; Enzyme Inhibitors; Fibroblasts; Fibronectins; Fibrosis; Histone Deacetylase Inhibitors; Humans; Hydroxamic Acids; Immunoblotting; Immunohistochemistry; In Situ Nick-End Labeling; Photorefractive Keratectomy; Rabbits; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta

To report a photorefractive keratectomy (PRK)-treated patient who first developed stromal opacities with a typical granular pattern 9 years after surgery, which was confirmed as heterozygous Avellino dystrophy by DNA analysis.. A 37-year-old woman, who had undergone PRK 17 years ago, has been followed for glaucoma and corneal dystrophy. She had preoperative clear corneas in both eyes.. Nine years after PRK, at the age of 29 years, 2-3 small, white, anterior stromal corneal opacities were first detected in both eyes. Seventeen years postoperatively, multiple anterior stromal corneal deposits with a dot, crumb, or snowflake appearance were noted to the same degree in both eyes. The intervening cornea was relatively clear. Her DNA was heterozygous for the R124H mutation.. Slowly developing, mild corneal deposits occurred after PRK in this patient with heterozygote Avellino corneal dystrophy. Various clinical manifestations can occur after excimer laser refractive surgery in patients with Avellino corneal dystrophy.

Topics: Adult; Corneal Dystrophies, Hereditary; Corneal Opacity; Corneal Stroma; DNA Mutational Analysis; Extracellular Matrix Proteins; Female; Humans; Lasers, Excimer; Myopia; Photorefractive Keratectomy; Postoperative Complications; Transforming Growth Factor beta

To compare corneal haze and transforming growth factor (TGF)-beta expression in rat eyes with mechanical debridement of corneal epithelium or a chemically induced epithelial flap.. Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.. Sixty corneas from 60 Sprague-Dawley rats were treated in 1 of 4 ways using a 2.0 mm trephine: corneal epithelium mechanically removed (Group 1), central cornea exposed to 20% ethanol for 30 seconds (Group 2), corneal epithelial flap made by applying 20% ethanol for 30 seconds and flap amputated (Group 3), corneal epithelial flap repositioned after ethanol-assisted detachment of epithelium as in Group 3 (Group 4). Corneal haze was graded. The TGF-beta expression was measured in corneas and lacrimal glands using reverse transcription polymerase chain reaction and immunohistochemistry until day 30. Semiquantitative analysis was done in a density ratio of TGF-beta/beta-actin with image analyzer and densitometry.. Corneal haze was more severe in Groups 1 and 3 than in Groups 2 and 4. By day 7, mRNA expression of TGF-beta2 and type II receptor in corneas had increased more in Groups 1 and 3 than in Groups 2 and 4. In lacrimal glands, only TGF-beta1 in Group 3 increased until day 7. In corneas, staining of both TGF-beta1 and beta2 increased, more prominently in Groups 1 and 3. Lacrimal gland staining was more intense in Groups 1 and 3.. Well-positioned corneal epithelial flaps may decrease corneal haze by reducing expression of TGF-beta; inadvertent removal of an epithelial flap made by ethanol seems to exacerbate haze by increasing TGF-beta.

Topics: Animals; Cornea; Corneal Opacity; Debridement; Epithelium, Corneal; Ethanol; Gene Expression; Immunoenzyme Techniques; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Surgical Flaps; Transforming Growth Factor beta

To report a possible case of phenotypic non-penetrance in granular corneal dystrophy type II (GCD-II).. DNA analysis was performed on 11 patients with white granular corneal opacities and 50 normal controls after informed consent was obtained. The TGFBI gene was analyzed by sequencing DNA from epidermal keratinocytes obtained using adhesive tape.. The heterozygous R124H mutation of TGFBI gene was found in all 11 patients. Although 49 normal controls had no mutation in the TGFBI gene, one normal control, a 26-year-old man, had the heterozygous R124H mutation of TGFBI gene. His 55-year-old father had the same mutation, but no corneal opacities.. As not all mutations are expressed in the phenotype, GCD-II gene mutation may have non-penetrance. This report documents a possible case of phenotypic non-penetrance in GCD-II.

Topics: Adult; Arginine; Base Sequence; Corneal Dystrophies, Hereditary; Corneal Opacity; Epidermis; Extracellular Matrix Proteins; Heterozygote; Histidine; Humans; Keratinocytes; Male; Middle Aged; Mutation; Mutation, Missense; Penetrance; Phenotype; Transforming Growth Factor beta

Beta igh3 is a transforming growth factor-beta-inducible cell adhesion molecule and its mutations are responsible for human autosomal dominant corneal dystrophies. Previously, we have studied the molecular properties of beta igh3 in vitro and reported that beta igh3 polymerizes to form a fibrillar structure and interacts with several extracellular matrix proteins including type I collagen. This study aimed to understand the role of elevated circulating levels of normal beta igh3 in eye development and corneal diseases.. We generated Alb-hss igh3 transgenic mice that have liver-specific expression of human beta igh3 (hss igh3) under the control of the albumin (Alb) enhancer/promoter and investigated the influence of beta igh3 overexpression in mouse eye. Polymerase chain reaction (PCR) genotyping, western blotting, and ELISA were performed to generate Alb-hss igh3 transgenic mouse lines. To identify the ocular pathology, electron microscopy and histological staining were employed in Alb-hss igh3 transgenic mice and wild-type mice.. Normal hss igh3 was ectopically overexpressed in the liver, secreted into blood stream, and reached the cornea of Alb-hss igh3 transgenic mice. Among transgenic mice, some mice had anterior segment defects including corneal opacity, disorganization of the collagen layers in the corneal stroma, and corneolenticular adhesion.. These results suggest that beta igh3 may be involved in anterior segment morphogenesis and eye development in mice. In addition, this indicates that the level of normal beta igh3 expression must be properly maintained during ocular development. The phenotype observed in Alb-hss igh3 transgenic mice is similar to human eye disorders such as anterior segment dysgenesis and Peters' anomaly. Thus, this model provides a very useful tool to study human eye diseases and the control of proliferation and differentiation of neural crest-originated cells.

Topics: Animals; Anterior Eye Segment; Collagen; Cornea; Corneal Diseases; Corneal Opacity; Corneal Stroma; Extracellular Matrix Proteins; Eye; Humans; Lens Diseases; Liver; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microscopy, Electron; Phenotype; Tissue Distribution; Transforming Growth Factor beta

To report the clinical and molecular study of a family with an autosomal dominant stromal granular dystrophy of the cornea caused by a novel and unusual TGFBI gene mutation.. A complete ophthalmological examination, corneal dystrophy phenotype characterization, PCR amplification, and automated nucleotidic sequencing of exons 4, 11,12, 13, and 14 of the TGFBI gene was carried out on the family. DNA from 40 unrelated ethnically matched healthy individuals were analyzed as controls.. Corneal dystrophy in two sisters was characterized by multiple grayish-white lesions located in the anterior and mid-stroma. Numerous small sized non-coalescent opacities were observed in the peripheral cornea while fewer larger lesions were apparent towards the central part of the cornea. A heterozygous missense mutation, consisting of a G to A transition at nucleotide position 384 in TGFBI exon 4 that predicts a valine (GTT) to isoleucine (ATT) replacement in residue 113 (Val113Ile) of the TGFBI protein was identified.. This is the most 5' located mutation detected so far in subjects with TGFBI-linked corneal dystrophy. Valine 113 is strictly conserved in TGFBI from several species and we suggest that the phenotype observed in these patients is related to the unusual location of the mutation. Our results expand the mutational spectrum in the group of TGFBI-linked corneal dystrophies.

Topics: Adenine; Adult; Base Sequence; Corneal Dystrophies, Hereditary; Corneal Opacity; Corneal Stroma; Extracellular Matrix Proteins; Female; Guanine; Heterozygote; Humans; Isoleucine; Mutation, Missense; Transforming Growth Factor beta; Valine

To report the first case of Avellino corneal dystrophy exacerbation after LASIK in a white or North American patient.. Case report and literature review.. A 25-year-old white female developed progressive corneal opacities after LASIK. Preoperative examination had revealed only subtle white corneal opacities in each eye. The patient's mother had similar corneal opacities. DNA analysis of the patient revealed a heterozygous mutation at the R124H location in the BIGH3 gene.. LASIK can exacerbate Avellino corneal dystrophy and should be avoided in patients with this condition. A careful history and genetic analysis can identify affected patients and those at risk.

Topics: Adult; Corneal Dystrophies, Hereditary; Corneal Opacity; Disease Progression; Extracellular Matrix Proteins; Female; Georgia; Humans; Keratomileusis, Laser In Situ; Mutation; Myopia; Transforming Growth Factor beta

To determine whether topical tranilast might reduce corneal haze through suppression of transforming growth factor (TGF)-beta1 synthesis in keratocyte after photorefractive keratectomy.. Department of Ophthalmology, Korea University College of Medicine, Seoul, Republic of Korea.. Photorefractive keratectomy was performed on 48 eyes of 28 white rabbits and 24 eyes in a tranilast group were treated with tranilast solution, and the other 24 eyes in control group were treated with saline after laser ablation. The grades of corneal haze at 1, 2, 4, and 8 weeks after surgery were evaluated in 10 eyes of each group for comparison. Immunohistochemistry was performed on 10 eyes of each group, and Western blot analysis was done on 4 eyes of each group for studying TGF-beta1 expression at postoperative day 7.. There was no statistically significant difference in corneal haze between 2 groups from week 1 to week 4 after surgery, but a significant difference was found at week 8 after photorefractive keratectomy (P=.02). The mean number of keratocytes that expressed TGF-beta1 in the tranilast group was 58.3 (+/-17.2), which showed significant difference, compared with that of the control group, 104.5 (+/-23.0) (P<.01). Western blot analysis also revealed that the amount of TGF-beta1 in tranilast group was slightly less than the control group.. Topical tranilast could reduce corneal haze by suppressing TGF-beta1 expression in keratocytes after photorefractive keratectomy.

Topics: Administration, Topical; Animals; Anti-Allergic Agents; Blotting, Western; Cornea; Corneal Opacity; Fibroblasts; Immunoenzyme Techniques; Lasers, Excimer; Ophthalmic Solutions; ortho-Aminobenzoates; Photorefractive Keratectomy; Rabbits; Transforming Growth Factor beta; Transforming Growth Factor beta1

To quantify central corneal regrowth and haze development after LASIK in rabbits.. New Zealand White rabbits received an 89 microm (-8 diopters) myopic LASIK and were evaluated during 4 months using slit-lamp and in vivo confocal microscopy to monitor changes in central corneal morphology, epithelial and stromal thickness, flap and bed thickness, and corneal light backscattering (haze). At various time-points, corneas were processed for histology.. Using in vivo confocal microscopy, LASIK induced no detectable morphological changes besides a slightly elevated light backscattering at the interface. Correspondingly, all corneas remained clear with no haze development by slit-lamp biomicroscopy. Corneal thickness was stable by 8 weeks after an increase of 17 +/- 4 microm that consisted of a 13 +/- 3 microm stromal regrowth and a 4 +/- 2 microm epithelial hyperplasia. At the LASIK interface, less than 4 microm new extracellular matrix was deposited. Accordingly, all LASIK flaps were easily pulled off by 6 months.. LASIK induces a minimal wound healing response in rabbit corneas with no haze development and a regrowth (regression) of only 17 microm of an 89-microm photoablation. Three main factors contributed to the observed regrowth: epithelial hyperplasia (approximately 4 microm), matrix deposition at the LASIK interface (approximately 4 microm), and stromal growth outside the interface within the flap and wound bed (approximately 9 microm).

Topics: Actins; Animals; Cornea; Corneal Opacity; Corneal Stroma; Epithelium, Corneal; Extracellular Matrix; Fibroblasts; Fibronectins; Keratomileusis, Laser In Situ; Light; Microscopy, Confocal; Microscopy, Fluorescence; Myopia; Rabbits; Scattering, Radiation; Surgical Flaps; Transforming Growth Factor beta; Wound Healing

To describe a Japanese patient with lattice corneal dystrophy type I (LCD I) who lacked the typical lattice lines.. Interventional case report.. A complete ophthalmologic examination was performed on a 54-year-old woman, and the TGFBI gene was analyzed by direct genomic sequencing.. The patient had diffuse opacification of the central corneal stroma but without lattice lines and corneal epithelial erosions bilaterally. Molecular genetic analysis identified a lattice corneal dystrophy I-associated heterozygous missense alteration (C417T) that changed arginine in codon 124 to cysteine (R124C) in the TGFBI gene.. The cornea of the patient appeared to represent late-stage lattice corneal dystrophy I, which suggests the existence of interactions of modifier genes, environmental factors during corneal aging, or both. The molecular genetic analysis of TGFBI can offer rapid, accurate diagnosis of patients with atypical corneal appearance.

Topics: Corneal Dystrophies, Hereditary; Corneal Opacity; Corneal Stroma; DNA Mutational Analysis; Extracellular Matrix Proteins; Female; Humans; Middle Aged; Mutation, Missense; Transforming Growth Factor beta; Visual Acuity

To report cases of Avellino corneal dystrophy (ACD) exacerbated by LASIK for myopia.. Retrospective, noncomparative, interventional case series and review of the literature.. Seven patients.. Six patients with exacerbation of granular corneal deposits after LASIK were examined for TGFBI mutations by polymerase chain reaction sequencing of DNA. One previously reported patient who was heterozygous for the ACD gene was followed up for 16 months after mechanical removal of granular deposits from the interface after LASIK.. Slit-lamp examination, visual acuity, manifest refraction, and DNA sequencing analysis.. All patients were heterozygous for the Avellino dystrophy gene. Corneal opacities appeared 12 months or more after LASIK. Best spectacle-corrected visual acuity decreased as the number and density of the opacities increased. One patient underwent mechanical removal of granules from the interface and had a severe recurrence within 16 months. Another patient had removal of the granules from the interface with PTK, followed by treatment with topical mitomycin C. In this patient, the cornea has remained relatively clear for 6 months.. Laser in situ keratomileusis increases the deposition of visually significant corneal opacities and is contraindicated in patients with ACD. Mechanical removal of the material from the interface does not prevent further visually significant deposits. Mitomycin C treatment, in conjunction with surgical removal of opacities, may be an effective treatment.

Topics: Adult; Corneal Dystrophies, Hereditary; Corneal Opacity; Extracellular Matrix Proteins; Female; Follow-Up Studies; Humans; Keratomileusis, Laser In Situ; Male; Mutation; Polymerase Chain Reaction; Refraction, Ocular; Retrospective Studies; Sequence Analysis, DNA; Transforming Growth Factor beta; Visual Acuity

To characterize the clinicopathologic phenotype as well as the molecular genetic basis of an autosomal dominant form of corneal amyloidosis.. Clinicopathologic and molecular genetic study of a family with a form of corneal amyloidosis.. Forty-nine individuals from one family were studied.. The medical records of affected family members were reviewed, and corneal tissue from those who had undergone penetrating keratoplasty (PK) was examined. Several family members were examined clinically, and corneas were photographed. Deoxyribonucleic acid from blood or buccal swabs was extracted from each consenting family member to determine the status of their transforming growth factor beta-induced (TGFBI) gene. The coding region of the TGFBI gene was analyzed for mutations in the proband's DNA, and compared with the nucleotide sequences of normal individuals. This was performed by amplifying and sequencing all exons of the TGFBI gene. In all other family members, only exons 4, 8, 11, and 12 of the gene were amplified, sequenced, and analyzed for mutations.. Clinicopathologic manifestations in relation to mutational status of the TGFBI gene.. Slit-lamp biomicroscopy revealed bilateral multiple polymorphic, polygonal, refractile, chipped ice-appearing gray and white opacities. There were also occasional deep filamentous lines that did not form a distinct lattice pattern. Corneal tissue of affected individuals who underwent PK contained widespread deposits of amyloid within the corneal stroma, particularly in the deep central stroma. Twelve members of the family were found to have a heterozygous single mutation in the TGFBI gene leading to a predicted amino acid substitution of aspartic acid for alanine (A546D). Nine of these individuals had ophthalmologist-documented corneal disease. The remaining 3, who were 11, 14, and 15 years old, were asymptomatic. In addition, 4 inconsequential polymorphisms with the nucleotide changes 387 G/A (R129R), 981 G/A (V327V), 1416 T/C (L472L), and 1620 C/T (F540F) were found.. A distinct, progressive form of corneal amyloidosis with an autosomal dominant mode of inheritance is characterized clinically by the presence of refractile polymorphic corneal opacities. We have designated this entity, which is caused by an A546D mutation in the TGFBI gene, polymorphic corneal amyloidosis.

Topics: Adolescent; Adult; Aged; Amyloidosis; Child; Corneal Diseases; Corneal Opacity; DNA Mutational Analysis; Extracellular Matrix Proteins; Female; Genes, Dominant; Humans; Keratoplasty, Penetrating; Male; Middle Aged; Mutation, Missense; Pedigree; Polymerase Chain Reaction; Reoperation; Transforming Growth Factor beta

The purpose of the study was to investigate the recurrence-free interval after phototherapeutic keratectomy (PTK) in patients with corneal dystrophies resulting from an Arg124His (R124H) mutation of the Big-h3 gene.. Patients with corneal dystrophy resulting from a genetically confirmed Big-h3 R124H mutation were examined with a slit lamp. The patients were divided into two groups on the basis of the mutation genotype, and the recurrence-free interval was analyzed.. In the 4 eyes of 3 homozygous patients, the mean (+/- standard deviation [SD]) recurrence-free interval was 9.5 +/- 3.1 months, whereas in the 7 eyes of 4 heterozygous patients it was 38.4 +/- 6.2 months. The former interval was statistically shorter than the latter (Kaplan-Meier survival analysis with log-rank test, p = 0.004).. These results strongly suggest that the mutation genotype of Big-h3 gene determined the recurrence-free interval as well as the clinical picture after PTK. Therefore, PTK should be considered for patients with Big-h3 R124H corneal dystrophy, on the basis of the expected recurrence-free interval deduced from molecular analysis of the zygosity of the Big-h3 R124H mutation.

Topics: Adult; Aged; Corneal Dystrophies, Hereditary; Corneal Opacity; Extracellular Matrix Proteins; Female; Heterozygote; Homozygote; Humans; Lasers, Excimer; Male; Middle Aged; Neoplasm Proteins; Photorefractive Keratectomy; Point Mutation; Recurrence; Time Factors; Transforming Growth Factor beta; Visual Acuity

To investigate the roles of transforming growth factor beta 1 (TGF-beta 1) during healing after photorefractive keratectomy (PRK).. Twenty of 24 rabbits underwent bilateral 193 nm excimer laser photorefractive keratectomy to correct 10 diopters of myopia, and the other 4 rabbits were taken as the normal group. The mRNA levels of TGF-beta 1 in the corneas of the rabbits were determined by in situ hybridization with a Digoxigenin labled probe.. 1. Corneal haze appeared in operated eyes from the 7th day, and peaked on the 28th day after PRK. The most severe haze reached Grade 3. 2. In the normal group, a low level of mRNA expression of TGF-beta 1 was observed in the epithelium, and no positive result in the stroma. On the 7th, 14th, and 28th days after the operation, the mRNA levels of TGF-beta 1 in the epithelium and stroma of the operation group were significantly higher than those in the normal group.. TGF-beta 1 may involve in corneal wound healing after PRK and may increase haze by promoting the synthesis of the extracellular matrix and the proliferation of keratocytes.

Topics: Animals; Cornea; Corneal Opacity; Female; Lasers, Excimer; Male; Photorefractive Keratectomy; Rabbits; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

To report a case of Avellino corneal dystrophy that increased in severity 1 year after uncomplicated laser in situ keratomileusis (LASIK) for myopia.. Avellino dystrophy was confirmed by polymerase chain reaction sequencing of DNA from the patient and her parents.. Best spectacle-corrected visual acuity decreased from 20/20 to 20/30 12 to 20 months after LASIK owing to opacities that appeared centrally in the corneal stroma and the LASIK flap and remaining posterior stroma interface.. LASIK is contraindicated in patients with Avellino corneal dystrophy because vision may be reduced by corneal opacities that appear in the interface of the flap and remaining posterior stroma postoperatively.

Topics: Adult; Contraindications; Corneal Dystrophies, Hereditary; Corneal Opacity; Disease Progression; DNA Mutational Analysis; Extracellular Matrix Proteins; Female; Humans; Keratomileusis, Laser In Situ; Myopia; Neoplasm Proteins; Polymerase Chain Reaction; Transforming Growth Factor beta; Vision Disorders; Visual Acuity

To investigate the opacity pattern in corneas with an Arg124His (R124H) homozygous mutation of the BIG-H3 gene.. Slit-lamp examination was performed on eight patients with corneal dystrophy resulting from a genetically confirmed BIG-H3 R124H homozygous mutation. The birthplace of each patient also was determined.. Slit-lamp examination disclosed two types of opacity patterns in corneas with the BIG-H3 R124H homozygous mutation. Type I (n = 4) is a spot-like opacity present in the anterior stroma in which the lesions are confluent. Type I is the same pattern that previous reports have shown to be caused by the BIG-H3 R124H homozygous mutation. The type II corneal opacity pattern (n = 4) is a reticular opacity in the anterior stroma with round translucent spaces. Type II opacity has not been reported previously in association with any corneal dystrophy. The patients with the type I opacity do not share a common birthplace; however, interestingly, the patients with the type II opacity traced their origin to Tottori prefecture in western Japan.. The BIG-H3 homozygous R124H mutation induces the development of two distinct patterns of corneal opacity, the recognition of which can establish an accurate diagnosis of corneal dystrophy caused by the homozygous BIG-H3 R124H mutation independent of genetic analysis. In addition, genetic factors or circumstantial influences other than the gene responsible for the corneal dystrophy may influence the pattern of corneal opacity.

Topics: Adult; Cornea; Corneal Dystrophies, Hereditary; Corneal Opacity; DNA Mutational Analysis; Extracellular Matrix Proteins; Female; Humans; Male; Middle Aged; Neoplasm Proteins; Point Mutation; Transforming Growth Factor beta; Visual Acuity

To report the recurrence pattern of corneal deposits after phototherapeutic keratectomy in patients with corneal dystrophies resulting from homozygous and heterozygous Arg124His (R124H) mutation of the BIG-H3 gene.. Slit-lamp examination was performed on patients with corneal dystrophy resulting from a genetically confirmed BIG-H3 R124H mutation.. The recurrence of corneal deposits after phototherapeutic keratectomy in patient with heterozygous R124H mutation was mild; the granular opacities occurred as spot lesions in the central cornea. The patient with a homozygous mutation had a more severe pattern, and the recurrent lesions were diffuse and occurred in the periphery between the corneal epithelium and laser-ablated stroma. The recurrence-free interval in homozygous patients was shorter.. The mutation genotype of BIG-H3 gene determines the recurrence pattern after phototherapeutic keratectomy.

Topics: Aged; Corneal Dystrophies, Hereditary; Corneal Opacity; Extracellular Matrix Proteins; Female; Humans; Lasers, Excimer; Male; Middle Aged; Neoplasm Proteins; Photorefractive Keratectomy; Point Mutation; Recurrence; Transforming Growth Factor beta; Visual Acuity

To report the chronic clinical course of two patients with homozygous R124H mutations in the betaig-h3 gene.. Case reports.. Two patients with homozygous R124H mutations in the betaig-h3 gene developed severe juvenile corneal opacities that required keratoplasty. After surgery, corneal opacities recurred and limited the recovery of visual acuity in the chronic follow-up.. In patients with homozygous R124H mutations in the betaig-h3 gene, recurrence of corneal opacities after keratoplasty limits the recovery of visual acuity in the chronic follow-up.

Topics: Child; Chronic Disease; Corneal Dystrophies, Hereditary; Corneal Opacity; Extracellular Matrix Proteins; Female; Humans; Keratoplasty, Penetrating; Middle Aged; Neoplasm Proteins; Point Mutation; Recurrence; Sequence Analysis, DNA; Transforming Growth Factor beta; Visual Acuity

To assess the role of the transforming growth factor (TGF)ss system in formation of corneal haze after excimer laser photorefractive keratectomy (PRK), levels of mRNAs for three TGFss isoforms (TGFss1, TGFss2, and TGFss3), the TGFss type II receptor (TssRII), and extracellular matrix (ECM) genes including fibronectin (FN), collagen I, collagen III, and collagen IV were measured in rat corneas.. Corneas were graded for corneal haze at 0, 1.5, 7, 21, 42, and 91 days after PRK. Total RNA was isolated from pooled corneas, and the levels of mRNAs were measured using competition-based quantitative reverse transcription-polymerase chain reaction (RT-PCR).. Severe corneal haze developed by day 42 and persisted to day 91. Levels of TGFss1 mRNA were high in rat corneas before PRK and remained relatively constant. In contrast, levels of TGFss2 and TGFss3 mRNAs were very low in normal corneas, increased 300-fold and 25-fold, respectively, on day 21, and remained elevated on day 91. Levels of mRNA for TssRII increased, with a peak elevation of 50-fold on day 42 after PRK. Levels of mRNAs for ECM proteins also increased. Fibronectin mRNA was nondetectable in normal corneas but rapidly increased to 675 copies/cell on day 7 and remained elevated to day 91. Collagen III mRNA levels peaked on day 21 with a 700-fold increase compared with a very low level of expression in normal cornea, and then decreased on day 91. Expression of collagen I mRNA lagged expression of collagen III mRNA and peaked at day 42 after PRK with a 1200-fold increase over normal cornea. In contrast, mRNA for collagen alpha(1)IV, a major component in basement membranes, remained relatively stable through day 21 and then increased slightly on days 42 and 91.. The synchronized increase in mRNA synthesis for both the TGFss system and key ECM genes supports the hypothesis that TGFss is a key growth factor promoting stromal haze formation in corneas after PRK and suggests that limiting TGFss system may reduce corneal scarring after excimer laser ablation.

Topics: Animals; Cornea; Corneal Edema; Corneal Opacity; DNA Primers; Extracellular Matrix Proteins; Eye Proteins; Gene Amplification; Gene Expression; Lasers, Excimer; Male; Photorefractive Keratectomy; Protein Isoforms; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Transforming Growth Factor beta; Wound Healing

Accumulating evidence suggests the involvement of TGF-beta in the process of corneal opacity, which is one of the serious causes of visual loss. However, whether TGF-beta is indeed critical for the pathogenesis remains unknown. We constructed an adenovirus expressing an entire ectodomain of the human type II TGF-beta receptor fused to Fc portion of human IgG (AdTbeta-ExR): this soluble receptor is secreted from AdTbeta-ExR-infected cells, binds to TGF-beta and inhibits TGF-beta signaling. When AdTbeta-ExR was injected into the femoral muscle of Balb/c mice, a high level of the soluble receptor protein (2.0-3.5 x 10(3) pM) was detectable in the serum and in the ocular fluid for at least 10 days. In the mice subjected to corneal injury with silver nitrate and to intramuscular injection with either saline or a control adenovirus expressing beta-galactosidase (AdLacZ), corneal opacification composed of extracellular matrix (ECM) accumulation, of infiltration of neutrophils and monocytes/macrophages, and of angiogenesis were all induced. In contrast, they were markedly reduced in the mice injected with AdTbeta-ExR. Immunohistochemical analysis revealed that TGF-beta, fibronectin, macrophage chemoattractant protein-1, and vascular endothelial growth factor were densely stained in the edge of wounded cornea, but they were scarcely present in the injured-cornea of AdTbeta-ExR-treated mice. Our results demonstrate that TGF-beta indeed plays a critical role in the process of cornea opacification, and that adenovirus-mediated expression of a soluble TGF-beta receptor can be therapeutically useful.

Topics: Adenoviridae; Animals; Chemokine CCL2; Cornea; Corneal Edema; Corneal Opacity; Endothelial Growth Factors; Fibronectins; Flow Cytometry; Gene Expression; Genetic Therapy; Genetic Vectors; Humans; Immunohistochemistry; Injections, Intramuscular; Lymphokines; Macrophages; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; Neutrophils; Receptors, Transforming Growth Factor beta; Recombinant Fusion Proteins; Transfection; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

To develop a rabbit model of reproducible corneal haze after excimer laser keratectomy and to characterize expression of transforming growth factor beta (TGFbeta) and basic fibroblast growth factor (bFGF) in rabbit corneas during haze formation.. Seven rabbits underwent a 100 microm deep phototherapeutic keratectomy (PTK) in one eye and a 15-microm shallow PTK in the contralateral eye. Corneal haze was compared at 1-20 weeks after surgery. Subsequently, 16 rabbits underwent 100-microm PTK in one eye and 15-microm PTK in the contralateral eye. Four rabbits were killed at 1, 2, 3, and 4 weeks, respectively, after surgery. Immunohistochemistry was performed on the corneas to localize the expression of TGFbeta and bFGF. Control subjects were rabbits that underwent either epithelial debridement alone or no surgery.. A 100-microm PTK resulted in significantly more corneal haze than a 15-microm PTK at every postoperative examination (p < 0.05). Both TGFbeta and bFGF were expressed in the scars at 1-4 weeks after deep and shallow excimer ablations. bFGF was expressed in the keratocytes of both treated and control corneas. Minimal TGFbeta was detected in the keratocytes of the control corneas, whereas prominent TGFbeta expression was noted in the keratocyte-like cells adjacent to the postkeratectomy scars.. The 100-microm PTK ablation resulted in significantly more corneal scarring than the 15-microm PTK ablation. Even though there was no immunohistochemical difference in the pattern of TGFbeta and bFGF expression after deep and shallow ablations, there was an association between the expression of the growth factors and corneal scarring after excimer laser keratectomy.

Topics: Animals; Cornea; Corneal Opacity; Fibroblast Growth Factor 2; Immunoenzyme Techniques; Lasers, Excimer; Male; Photorefractive Keratectomy; Rabbits; Time Factors; Transforming Growth Factor beta; Wound Healing

Two patients were diagnosed with Reis-Bücklers corneal dystrophy (RBCD), although the pattern and severity of corneal opacification differed. To see whether there was a genetic basis for these phenotypic variations, we analyzed beta ig-h3, the gene that codes for kerato-epithelin and that contains a mutation (Arg555Gln) that causes RBCD.. A 30-year-old man with honeycomb-shaped subepithelial opacities in his central cornea and a 25-year-old man with progressive subepithelial geographic opacities were both considered to have RBCD. We isolated genomic DNA from leukocytes of the two patients and their family members and screened for an Arg555Gln kerato-epithelin mutation. Then we analyzed all exons of the gene using the single-strand conformation polymorphism (SSCP) technique to search for any other kerato-epithelin mutations.. The patient with honeycomb-shaped opacities had an Arg555Gln kerato-epithelin mutation that caused his RBCD, whereas the patient with geographic opacities did not; instead, he had a new kerato-epithelin mutation (Arg124Leu), which cosegregated with his family members.. The variant of RBCD characterized by honeycomb-shaped opacities is caused by an Arg555Gln kerato-epithelin mutation. On the other hand, a new kerato-epithelin mutation, Arg124Leu, was found to cause the RBCD variant characterized by recurrent epithelial erosions and progressive geographic subepithelial opacification. Codon 124 is a hot spot for kerato-epithelin mutations, where the mutations responsible for three autosomal dominant corneal dystrophies--lattice type I (Arg124Cys), Avellino (Arg124His), and the variant of RBCD with geographic rather than honeycomb opacities (Arg124Leu)--are located.

Topics: Adult; Cornea; Corneal Dystrophies, Hereditary; Corneal Opacity; DNA Mutational Analysis; DNA Primers; Exons; Extracellular Matrix Proteins; Female; Humans; Male; Neoplasm Proteins; Pedigree; Point Mutation; Polymorphism, Single-Stranded Conformational; Sequence Analysis, DNA; Transforming Growth Factor beta; Visual Acuity

Topics: Cicatrix; Collagen; Corneal Opacity; Corneal Ulcer; Epithelium, Corneal; Humans; Lasers, Excimer; Myopia; Photorefractive Keratectomy; Recurrence; Transforming Growth Factor beta

Tranilast (trade name Rizaben), an anti-allergic drug with anti-inflammatory effects, is thought to inhibit synthesis of extracellular matrix of fibroblasts through the suppression of TGF-beta. We evaluated the effect of topical tranilast on the subepithelial haze that developed after excimer laser keratectomy and its effect was compared with that of betamethasone eye drops. Excimer laser keratectomy (phototherapeutic keratectomy mode) was performed with the Nidek EC-5000 excimer laser on 16 rabbit corneas (eight rabbits). From the second postoperative day, topical 2% tranilast was instilled in the right eye and the control solution in the left eye, four times daily. Until the fourth week after the operation, we measured the densitometric values of scattered light intensity of the subepithelial haze with an anterior ocular analyzer, EAS-1000 (Nidek). At the fifth postoperative week, light and electron microscopy and immunohistochemistry with an antibody to TGF-beta were also performed.. Densitometric values of the subepithelial haze in the corneas treated with 2% tranilast were slightly less than those of the subepithelial haze in the control corneas. However, the values of the subepithelial haze in the betamethasone-treated corneas were significantly less than those in control corneas. Histochemical examinations revealed that topical tranilast had a small effect on the subepithelial haze after excimer laser keratectomy in rabbits.. Topical 0.1% betamethasone can limit the amount of subepithelial haze and tranilast may inhibit development of subepithelial haze by the suppression of TGF-beta.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Betamethasone; Cornea; Corneal Opacity; Epithelium; Immunohistochemistry; Lasers, Excimer; Ophthalmic Solutions; ortho-Aminobenzoates; Photorefractive Keratectomy; Rabbits; Transforming Growth Factor beta

To determine the relationship between anti-transforming growth factor-beta (anti-TGF-beta) antibodies and the amount of corneal stromal haze after excimer laser photorefractive keratectomy (PRK).. Wills Eye Hospital, Philadelphia, Pennsylvania, USA.. Nineteen rabbits had bilateral PRK. Dichlorotriazinyl fluorescein was used to stain the exposed stroma; all rabbits were then treated with antibiotic ointment for 4 days. Ten rabbits were randomized to treatment with topical anti-TGF-beta1, -beta2, and -beta3 antibody 50 microg three times a day for 4 days; the others received diluent three times a day for 4 days. Stromal haze was graded weekly for 8 weeks on a 0 to 4+ scale. At the end of the study, all corneas were examined histopathologically.. All treated eyes developed appreciable haze. Seven control rabbits and one antibody-treated rabbit had an epithelial erosion (P = .00001). Antibody-treated rabbits had significantly less haze at 3, 4, and 5 weeks (right eyes) and 3, 4, 5, 7 and 8 weeks (left eyes) (P < .05). Histopathology and fluorescence microscopy showed subepithelial collagen deposition consistent with clinical haze.. Topical anti-TGF-beta antibody reduced stromal haze after PRK in the rabbit model and may be clinically beneficial in humans.

Topics: Administration, Topical; Animals; Antibodies, Monoclonal; Collagen; Cornea; Corneal Opacity; Corneal Stroma; Disease Models, Animal; Lasers, Excimer; Male; Microscopy, Fluorescence; Ophthalmic Solutions; Photorefractive Keratectomy; Rabbits; Random Allocation; Recombinant Proteins; Transforming Growth Factor beta