epidermal-growth-factor and Pterygium

To observe the effect of recombinant human epidermal growth factor (rhEGF) on ocular surface re-epithelization and the efficacy of amniotic membrane transplantation in patients with pterygium.. Amniotic membrane transplantation was performed on 40 eyes of 36 patients with pterygium after removal of pterygium. All the eyes were randomly divided into 2 groups: 20 eyes received rhEGF ophthalmic solution and the other 20 drug vehicle. The healing rates of the corneal and conjunctival epithelium were observed respectively. Follow-up study lasting for 3 to 12 months was conducted in all the cases postoperatively.. The healing rate of both the corneal and conjunctival epithelium in rhEGF-treated group (74.983+/-18.998 micrometer/h and 36.584+/-7.888 micrometer/h respectively) was significantly faster than the control (59.372+/-17.197 +/-m/h and 29.18+/-5.450 micrometer/h respectively, P<0.01 for both comparisons). Follow-up witnessed no recurrence of the disease.. rhEGF effectively promotes the healing of ocular surface epithelium defect, and amniotic membrane transplantation is a good surgical procedure for treating pterygium.

Topics: Adult; Aged; Amnion; Conjunctiva; Cornea; Epidermal Growth Factor; Epithelium; Eye; Female; Humans; Male; Middle Aged; Pterygium; Recombinant Proteins; Time Factors; Treatment Outcome

Recently, it has been reported that miRNA is involved in pterygium, however the exact underlying mechanism in pterygium is unrevealed and require further investigation.. The differential expression of miRNA in pterygium was profiled using microarray and validated with quantitative real-time polymerase chain reaction (qRT-PCR). Human conjunctival epithelial cells (HCEs) were cultured and treated with transforming growth factor β (TGF-β) and epidermal growth factor (EGF) and transfected with miR-199a-3p/5p mimic and inhibitor. Markers of epithelial-mesenchymal transition (EMT) in HCEs were detected using western blot and immunohistochemistry. Cell migration ability was determined using wound healing and transwell assay, while apoptosis was determined by flow cytometry. The target genes of miR-199a were confirmed by the dual-luciferase reporter assay.. TGF-β and EGF could induced EMT in HCEs and increase miR-199a-3p/5p but suppress target genes, DUSP5 and MAP3K11. With the occurrence of EMT, cell migration ability was enhanced, and apoptosis was impeded. Promoting miR-199a-3p/5p expression could induce EMT in HCEs without TGF-β and EGF, while suppressing miR-199a-3p/5p could inhibit EMT in TGF-β and EGF induced HCEs. In a word, TGF-β and EGF induced EMT could be regulated with miR-199a-3p/5p-DUSP5/MAP3K11 axes. The validated results in tissues showed that, compared with control conjunctival tissues, miR-199a-3p/5p were more overexpressed in pterygium, while DUSP5/MAP3K11 were lower expressed. In addition, bioinformatics analysis indicated the miR-199a-3p/5p-DUSP5/MAP3K11 was belong to MAPK signalling pathway.. TGF-β and EGF induce EMT of HCEs through miR-199a-3p/5p-DUSP5/MAP3K11 axes, which explains the pathogenesis of EMT in pterygium and may provide new targets for pterygium prevention and therapy.

Topics: Dual-Specificity Phosphatases; Epidermal Growth Factor; Epithelial-Mesenchymal Transition; Humans; MicroRNAs; Pterygium; Transforming Growth Factor beta

To assess the efficacy of combined 5FU and Avastin injections in the treatment of primary pterygium METHODS: Sixteen eyes with primary pterygium received intralesional 5 fluorouracil and Avastin (2.5-5 mg) injections every 2 weeks for a maximum of five injections. Fourteen eyes of 14 patients received five injections, one eye received three injections and one eye received two injections. All eyes were followed at monthly intervals for 3 months after last injection. Tissue was obtained by surgical excision of primary pterygium from four eyes who received injections and three eyes with primary pterygium who did not receive injections (control) and subjected to immunohistological examination for beta fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), von-Willebrand factor (vWF), lymphatic vessel endothelial hyaluronan receptor (LYVE-1) and collagen-I.. Pterygium progression was arrested in all patients. Sixty-two percent of patients had improvement of redness while 89% had reduced thickness of the lesion. VEGF, bFGF, EGF, vWF, LYVE-1 and collagen-I were all reduced in the injected samples.. The injection of 5 fluorouracil and Avastin act synergistically to arrest progression and induce atrophy in primary pterygium. This is related to the effect of agents on fibroblasts, collagen, and vascular tissues. Such medical intervention is a safe and viable option in the management of primary pterygium though excision of residual tissue is still required in some cases. Longer follow up is needed to ascertain whether this will reduce the recurrence rate following excision.

Topics: Adult; Aged; Angiogenesis Inhibitors; Antimetabolites; Bevacizumab; Biomarkers; Collagen Type I; Drug Therapy, Combination; Epidermal Growth Factor; Female; Fibroblast Growth Factors; Fluorescent Antibody Technique, Indirect; Fluorouracil; Humans; Injections, Intralesional; Male; Middle Aged; Pterygium; Vascular Endothelial Growth Factor A; Vesicular Transport Proteins; von Willebrand Factor; Young Adult

The heat-sensitive transient receptor potential vanilloid type-1 (TRPV1) channel (i.e., capsaicin [CAP] receptor) is upregulated in numerous cancers. This study determined if this response occurs in fresh and cultured hyperplastic human pterygial epithelial tissues.. Reverse transcriptase PCR and quantitative real-time PCR, along with immunohistochemistry and Western blotting, characterized TRPV1 expression patterns in pterygial and healthy conjunctival tissue, primary and immortalized pterygial cells (hPtEC), and primary and immortalized conjunctival epithelial cells (HCjEC). Imaging of Ca2+ and planar whole-cell patch-clamping evaluated TRP channel activity. An MTS assay measured cell metabolic activity and a cell growth assay monitored proliferation.. Capsaicin (20 μM) and elevating bath temperature above 43°C activated Ca2+ transients more in hPtEC than HCjEC. Capsaicin induced corresponding changes in inward currents that were inhibited by 20 μM capsazepine (CPZ). Vascular endothelial growth factor (VEGF) also increased Ca2+-influx and induced corresponding inward currents more in hPtEC than in HCjEC, whereas CPZ (20 μM), BCTC (20 μM), or La3+ (500 μM) reduced these responses, respectively. Whereas epidermal growth factor (EGF) increased proliferation more in hPtEC than in HCjEC, VEGF had no effect on this response. Capsazepine suppressed hPtEC proliferation induced by EGF and VEGF, whereas it was cytotoxic to HCjEC.. Mitogenic responses to EGF and VEGF are mediated through TRPV1 transactivation. Only in hPtEC do the increases in proliferation induced by EGF exceed those in HCjEC. Therefore, TRPV1 is a potential drug target whose clinical relevance in treating pterygium warrants further assessment.

Topics: Blotting, Western; Calcium; Capsaicin; Cell Proliferation; Cells, Cultured; Conjunctiva; DNA; Epidermal Growth Factor; Gene Expression Regulation; Humans; Immunohistochemistry; Patch-Clamp Techniques; Pterygium; Reverse Transcriptase Polymerase Chain Reaction; Transcriptional Activation; TRPV Cation Channels; Up-Regulation; Vascular Endothelial Growth Factor A

The expression of growth factors in ophthalmic pterygium and phenotypically normal conjunctiva was examined and correlated with the clinical findings. Fifteen specimens of ophthalmic pterygia and 8 specimens of phenotypically normal conjunctiva were examined. Total RNA was extracted from all specimens and mRNA levels for transforming growth factor (TGFB1), vascular endothelial growth factor (VEGFA), basic fibroblast growth factor (FGF2), epidermal growth factor (EGF) and insulin-like growth factor (IGF1) were measured using real-time reverse-transcription PCR (qRT-PCR). Differences in the expression of these factors between pterygium and conjunctival specimens were examined, as well as correlations between RNA levels and clinical parameters. mRNA levels for VEGFA and FGF2 were significantly higher in pterygium, compared with conjunctival specimens, whereas the respective differences for other factors examined were statistically not significant. The mRNA levels for VEGFA and FGF2 were also significantly higher in recurrent compared with primary pterygium. Correlations between the mRNA levels and other clinical parameters were statistically not significant for the growth factors examined. The higher levels of FGF2 or VEGFA mRNA in pterygium imply that these factors may be involved in the pathogenesis or clinical behavior of the pterygium, including postoperative recurrence. VEGFA expression may have potential for use in the selective treatment of pterygium with anti-VEGFA agents.

Topics: Adult; Aged; Aged, 80 and over; Conjunctiva; Epidermal Growth Factor; Female; Fibroblast Growth Factor 2; Gene Expression Regulation; Humans; Insulin-Like Growth Factor I; Intercellular Signaling Peptides and Proteins; Male; Middle Aged; Pterygium; RNA, Messenger; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

To investigate the growth promoting and chemotactic effects of heparin binding epidermal growth factor-like growth factor (HB-EGF), recently shown to be upregulated by ultraviolet irradiation in pterygium-derived epithelium cells (PECs) and pterygium fibroblasts (PFs).. PECs and PFs were incubated with various concentrations of HB-EGF. Cell proliferation was evaluated by measurement of [3H]thymidine incorporation. The potential chemotactic effect of HB-EGF on these two cell lines was assessed with migration assays, using modified Boyden chambers and checkerboard analysis.. Incubation of PECs and PFs with HB-EGF resulted in a significant increase in [3H]thymidine incorporation compared with that of control cells. HB-EGF stimulated chemotaxis of both PECs and PFs. Maximum stimulation occurred at 1 ng/mL for PFs and 10 ng/mL for PECs. These effects were abolished by the addition of a neutralizing antibody to HB-EGF.. The findings demonstrate the potential proliferative and chemotactic effects of HB-EGF on both PECs and PFs. This is the first study to illustrate the positive effect of a specific growth or chemotactic factor on the cellular elements of a pterygium.

Topics: Cell Division; Cell Line; Chemotaxis; Epidermal Growth Factor; Epithelial Cells; Fibroblasts; Flow Cytometry; Heparin-binding EGF-like Growth Factor; Humans; Intercellular Signaling Peptides and Proteins; Pterygium

Ultraviolet (UV) light is one of the major factors implicated in the pathogenesis of pterygium. The mechanism by which UV light induces this disease remains elusive. The aim of this study was to evaluate the effects of UVB irradiation on the expression of growth factors in cultured pterygium epithelial cells and to demonstrate their distribution within pterygium. We cultured pterygial epithelial cells from pterygium explants and these cells were exposed to 20 mJ/cm(2) of UVB. Total RNA was extracted at 0, 6, and 12 hours after irradiation. (32)P-labeled cDNA was synthesized and analyzed using microarray technology to determine the differential expression of 268 growth factor and cytokine related genes. Semiquantitative reverse transcriptase-polymerase chain reaction was used to corroborate this data. Conditioned media derived from cells exposed to UVB irradiation was analyzed for protein expression by enzyme-linked immunosorbent assay. Immunohistochemistry was used to evaluate the distribution of heparin-binding epidermal growth factor-like growth factor (HB-EGF) in pterygium tissue. Analysis of the hybridization signals revealed that the genes encoding HB-EGF, fibroblast growth factor 3, and cytotoxic trail ligand receptor were consistently elevated at 6 and 12 hours after UVB treatment. HB-EGF mRNA was elevated 6.8-fold at 6 hours after irradiation and was augmented in culture supernatants after the same treatment. Furthermore, HB-EGF reactivity was identified in the epithelium and vasculature of pterygium by immunohistochemistry. HB-EGF was present in normal limbal epithelium, although it was not induced in cultured limbal epithelial cells by UV irradiation. HB-EGF is a potent mitogen, localized in pterygium tissue, and significantly induced by UVB in pterygium-derived epithelial cells. We postulate that this growth factor is a major driving force in the development of pterygia and a means by which UV irradiation causes the pathogenesis of pterygium.

Topics: Cell Culture Techniques; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Epithelium, Corneal; Heparin; Heparin-binding EGF-like Growth Factor; Humans; Intercellular Signaling Peptides and Proteins; Oligonucleotide Array Sequence Analysis; Pigment Epithelium of Eye; Pterygium; Reverse Transcriptase Polymerase Chain Reaction; Ultraviolet Rays

Pterygia are characterised by a fleshy outgrowth of altered conjunctival tissue over the cornea and are most common in tropical regions. Pterygial fibroblasts are characteristically distinct from normal conjunctival fibroblasts, and therefore the aim of this study was to determine the presence and functional significance of histamine and epidermal growth factor (EGF) receptors in these cells. Pterygial specimens were cultured in vitro and cellular outgrowths were phenotypically characterised as fibroblasts using vimentin and cytokeratin staining. Intracellular calcium mobilization was used to characterise the functional activity of histamine receptors on these cells. Maximal response was obtained with 100 microM histamine. However, lower concentrations of histamine also caused mobilization of calcium that were totally abolished by pre-incubation with H1 but not H2 or H3 receptor antagonists. EGF receptor was diffusely expressed over the cell surfaces. EGF stimulated receptor internalization, ERK protein phosphorylation and intracellular calcium mobilization. Therefore, fibroblasts derived from human pterygia express functionally active histamine and epidermal growth factor receptors. Controlled modification of either the receptors or the appropriate ligands could have beneficial effects in pterygia treatment.

Topics: Calcium; Cells, Cultured; Epidermal Growth Factor; ErbB Receptors; Fibroblasts; Histamine; Humans; Immunohistochemistry; Intracellular Fluid; Mitogen-Activated Protein Kinase 1; Pterygium; Receptors, Histamine

To investigate the effects of recombinant epidermal growth factor on the wounded corneal healing after excision of pterygium.. Simple and unrecurrent pterygium was selected, excised and sewed up conjunctival flap under local anaesthesia and microscope. After operation two drops of EGF eyedrop were used, and five minutes later 0.3% Tarivid Eye Oint was used again, and then the operated eye was enswathed. Everyday the wound cornea was observed and the EGF eyedrop was used.. The wound cornea healing time of control group was seven days and that of the EGF group was five days and gave a remarkable difference (P < 0.05). The same results were obtained after comparing the effective group and ineffective group, P < 0.05.. EGF eyedrop can accelerate proliferation and recovery of wound corneal epithelial cells. In the clinical trial, every body felt well and no side-effect was observed.

Topics: Adolescent; Adult; Aged; Epidermal Growth Factor; Epithelium, Corneal; Female; Humans; Male; Middle Aged; Pterygium; Recombinant Proteins; Wound Healing

Pterygium is a degenerative corneal limbal process and UV irradiation has been suggested as being a major environmental predisposing factor. The invasive nature of the fibroblasts associated with pterygia raises the question as to whether these cells are transformed. To test this hypothesis, we established fibroblast strains from autologous and heterologous pterygial and conjunctival specimens, respectively, from subjects between 40 to 50 yr of age, and compared their growth characteristics in culture. All pterygial fibroblast strains exhibited a reduced dependence on serum and exogenous growth factors for growth and reached a saturation population density that was threefold higher than conjunctival fibroblasts cultured under the same conditions. In addition, all pterygial fibroblast strains were able to form colonies in soft agar in 5% fetal bovine serum at a 6.0 to 7.5% efficiency. Under the same experimental conditions, none of the conjunctival fibroblast strains were able to grow. The results presented support the conclusion that pterygial fibroblasts have acquired many of the properties of the transformed phenotype.

Topics: Adult; Cell Adhesion; Cell Division; Cell Line, Transformed; Conjunctiva; Epidermal Growth Factor; Fibroblast Growth Factor 1; Fibroblasts; Humans; Male; Middle Aged; Phenotype; Pterygium; Time Factors; Transforming Growth Factor beta