epidermal-growth-factor and Cataract

To determine the effect of sodium hyaluronate combined with recombinant human epidermal growth factor (rhEGF) on clinical symptoms and inflammation in patients with newly diagnosed xerophthalmia after cataract surgery.. A total of 106 patients who underwent cataract surgery and were newly diagnosed with xerophthalmia in our hospital between June 2018 and August 2019 were enrolled. Of these, 50 patients who were treated with sodium hyaluronate (0.1%) were assigned to the monotherapy group (MG) and the remaining 56 patients who were treated with sodium hyaluronate (0.1%) combined with rhEGF (20 μg/ml) were assigned to the combination group (CG). The 2 groups were compared based on ocular surface disease index (OSDI) score, break-up time (BUT), fluorescein corneal staining level, Schirmer I test (SI) level, clinical efficacy (disappearance of typical symptoms, including eyes drying, burning sensation, foreign body sensation, etc), and interleukin (IL)-1, IL-6, and tumor necrosis factor-α (TNF-α) levels. Spearman correlation analysis was conducted to analyze the relationship between IL-1, IL-6, TNF-α and clinical efficacy. In addition, receiver operating characteristic curves were drawn to analyze the predictive value of IL-1, IL-6, and TNF-α in efficacy on xerophthalmia.. After treatment, the CG showed reduced OSDI score compared with the MG. The CG showed increased BUT (s) and SI (mm) levels compared with MG. After treatment, the CG exhibited decreased levels of IL-1(ng/mL), IL-6 (ng/mL), and TNF-α (ng/mL) compared with the MG. Spearman correlation analysis revealed that IL-1, IL-6, and TNF-α were negatively correlated with clinical efficacy. The areas under the curves of IL-1, IL-6, and TNF-α were 0.801, 0.800, and 0.736 respectively.. Sodium hyaluronate combined with rhEGF is helpful to alleviate clinical symptoms and inflammation in patients with xerophthalmia undergoing cataract surgery.

Topics: Cataract; Cataract Extraction; Epidermal Growth Factor; Humans; Hyaluronic Acid; Inflammation; Ophthalmic Solutions; Recombinant Proteins; Xerophthalmia

To determine the factors that influence the ability of dexamethasone (dex) to inhibit or stimulate the growth of lens epithelial cells.. Different growth factors with or without dex (10. Dex was found to both stimulate and inhibit mitogen-induced. The study demonstrated that dex reduces the overall growth and activity of lens epithelial cells in vitro. This result provides insight into the risk of developing posterior subcapsular cataracts (PSC) in patients on oral glucocorticoid therapy. Understanding the basic mechanisms by which steroids mediate lens cell growth may provide the ability to more accurately predict who will develop PSC. The present studies show the difference in the effect of dex from lens cell to lens cell, but, more importantly, suggest a pattern of dependent variables that might prove useful in such predictions.

Topics: Cataract; Cells, Cultured; Dexamethasone; DNA; Epidermal Growth Factor; Epithelial Cells; Humans; Insulin; Progesterone; Thymidine

Cataractogenesis begins from the dynamic lens epithelial cells (LECs) and adjacent fiber cells. LECs derived from cell lines cannot maintain the crystalline expression as the primary LECs. The current study aimed to efficiently generate large numbers of human LECs from patient-specific induced pluripotent stem cells (iPSCs). Anterior lens capsules were collected from cataract surgery and were used to culture primary hLECs. iPSCs were induced from these primary hLECs by lentiviral transduction of Oct4, Sox2, Klf4, and c-Myc. Then, the generated iPSCs were re-differentiated into hLECs by the 3-step addition of defined factor combinations (Noggin, BMP4/7, bFGF, and EGF) modified from an established method. During the re-differentiation process, colonies of interest were isolated using a glass picking tool and cloning cylinders based on the colony morphology. After two steps of isolation, populations of LEC-like cells (LLCs) were generated and identified by the expression of lens marker genes by qPCR, western blot and immunofluorescence staining. The study introduced a modified protocol to isolate LLCs from iPSCs by defined factors in a short time frame. This technique could be useful for mechanistic studies of lens-related diseases. J. Cell. Physiol. 231: 2555-2562, 2016. © 2016 Wiley Periodicals, Inc.

Topics: Blotting, Western; Cataract; Cell Differentiation; Epidermal Growth Factor; Epithelial Cells; Fluorescent Antibody Technique; Humans; Induced Pluripotent Stem Cells; Kruppel-Like Factor 4; Lens, Crystalline; Male; Middle Aged; Real-Time Polymerase Chain Reaction

In this report we present a patient with unilateral membranous cataract and describe the histological and biochemical findings accompanying this rare condition.. The patient underwent an uneventful cataract extraction. Aqueous humor (20 μl) was aspirated from the anterior chamber intraoperatively and processed for fibroblast growth factor (FGF) and epidermal growth factor (EGF) using an immunoassay method (ELISA). The lens material was subjected to histological examination.. The patient had increased levels of FGF and EGF in the aqueous humor, as measured by ELISA. Histological examination of the lens material showed a marked fibrous metaplasia and thickening of the anterior lens capsule, while the lens epithelial cells were transformed to active myofibroblasts which generated a fibrous matrix of collagen lamellae. Unfortunately, visual function was not restored postoperatively due to underlying amblyopia.. Our histological and biochemical findings suggest that FGF and EGF may play a key role in the formation of membranous cataract, and therefore their impact on lens physiology should be further investigated.

Topics: Adult; Aqueous Humor; Cataract; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Female; Fibroblast Growth Factors; Humans; Lens, Crystalline; Risk Factors

Phase-contrast and transmission electron microscopic studies were performed to evaluate the effect of epidermal growth factor (EGF) upon the morphology of lens epithelial cells (LECs) obtained from human cataractous lenses, cultured on human anterior lens capsules, and divided into the following three groups: group 1 receiving no EGF supplement, group 2 supplemented with 1 ng/ml EGF, and group 3 supplemented with 10 ng/ml EGE Phase-contrast microscopy revealed that the cells in the group supplemented with 1 ng/ml EGF concentration had elongated, and those supplemented with 10 ng/ml EGF disclosed changes indicating fibroblast-like cells. Under transmission electron microscopy, the cells in the group supplemented with 1 ng/ml EGF concentration had become multilayered and a few cells had a ball-and-socket junction structure and nucleolar chromatin condensation, while those supplemented with 10 ng/ml EGF showed changes not only in the cells but also in the anterior lens capsules to present numerous microfibers. This is the first study to confirm the effect of EGF upon LECs cultured on anterior lens capsules, a condition closely resembling the intraocular environment, and these findings are significant.

Topics: Aged; Cataract; Cells, Cultured; Culture Media; Epidermal Growth Factor; Epithelial Cells; Follow-Up Studies; Humans; Lens, Crystalline; Microscopy, Electron; Microscopy, Phase-Contrast; Recombinant Proteins

It has been assumed that lens epithelial cells (LECs) existing at the capsulotomy edge have been traumatized through anterior capsulotomy in cataract extraction. In this study, the correlation between traumatized LECs remaining at the anterior capsulotomy edge and epidermal growth factor (EGF) found in the aqueous humor, a cell growth factor though to affect cell morphology, was determined. Anterior lens capsules with adhering LECs were obtained following anterior capsulotomy performed during cataract surgery to first confirm the presence of EGF receptors on LECs, which are needed for EGF to be biologically active. Besides, to identify any EGF receptors on traumatized LECs, I next intentionally traumatized the cells by pressing them with a forceps from the anterior capsular side. It has been found that the LECs containing EGF receptors were always those existing at the edge of the anterior capsular opening and LECs containing EGF receptors existed along the pressed region too. The present results indicate that traumatized LECs along the capsulotomy edge have undergone changes to manifest EGF receptors, thus allowing EGF from the aqueous humor to become more active. The physiological effect of EGF upon these LECs may therefore be one of the causative factors of fibrous opacification of the anterior capsulotomy edge after cataract extraction.

Topics: Aged; Aged, 80 and over; Aqueous Humor; Cataract; Cataract Extraction; Coloring Agents; Epidermal Growth Factor; Epithelial Cells; ErbB Receptors; Fibrosis; Humans; Lens Capsule, Crystalline; Middle Aged; Postoperative Complications; Trypan Blue; Wounds, Nonpenetrating

To determine whether the 12-lipoxygenase pathway of arachidonic acid metabolism is present in the human lens and whether 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) plays a role in regulating proto-oncogene expression and DNA synthesis in human lens epithelial cells (HLECs).. Second- and third-passage primary cultures of HLECs were used for analysis. Human cataract epithelia were obtained from surgery. 12-lipoxygenase mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the PCR product was sequenced. The 12-lipoxygenase protein was detected by immunoblotting. 12(S)-HETE was detected in HLEC-conditioned medium by radioimmunoassay. For studies of growth factor-induced mitogenesis, HLECs were serum starved, then stimulated with 15 ng/ml epidermal growth factor (EGF) and 1 microgram/ml insulin or with 0.3 microM 12(S)-HETE. The 12-lipoxygenase inhibitor, cinnamyl-3,4-dihydroxy-alpha-cyanocinnamate (CDC, 10 microM) was used to block endogenous 12-lipoxygenase activity. Expression of c-fos mRNA was determined by RT-PCR, and DNA synthesis was measured by 3H-thymidine incorporation.. 12-lipoxygenase mRNA and protein were detected in HLECs and in human cataract tissues. 12(S)-HETE was released into the medium by HLECs in the presence of EGF-insulin. Stimulation of c-fos mRNA expression and DNA synthesis by EGF-insulin was inhibited when the 12-lipoxygenase pathway was blocked by CDC. This inhibition was reversed completely by exogenously added 12(S)-HETE. However, exogenous 12(S)-HETE was unable to stimulate HLEC DNA synthesis in the absence of growth factors.. The 12-lipoxygenase pathway of arachidonic acid metabolism is present in human lens epithelial cells. 12(S)-HETE does not stimulate HLEC DNA synthesis in the absence of growth factors but enables the cellular response to EGF and insulin.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adult; Arachidonate 12-Lipoxygenase; Base Sequence; Caffeic Acids; Cataract; Cells, Cultured; DNA; DNA Primers; Epidermal Growth Factor; Epithelium; Gene Expression Regulation; Humans; Hydroxyeicosatetraenoic Acids; Hypoglycemic Agents; Infant; Insulin; Lens, Crystalline; Lipoxygenase Inhibitors; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; Proto-Oncogene Mas; Proto-Oncogene Proteins c-fos; Radioimmunoassay; RNA; RNA, Messenger; Transcription, Genetic

To assay the following cytokines in an incubation medium of cultured lens epithelial cells (LECs) derived from human cataracts: interleukin-1 alpha (IL-1), interleukin (IL-6), basic fibroblast growth factor (b-FGF), tumor necrosis factor-alpha (TNF-alpha), and epidermal growth factor (EGF).. Nishi Eye Hospital, Jinshikai Medical Foundation, Osaka, Japan.. The anterior lens capsule with attached LECs was obtained by capsulotomy during cataract surgery and cultured. The incubation medium was changed on days 1 and 2 of culture and thereafter weekly up to 7 weeks. The media collected from a specific number of cultures at each medium change were pooled and assayed for cytokines by enzyme-linked immunosorbent assay.. Interleukin-1 alpha was detected in one of the two pools of the 2 week cultures (207 pg/10(6) cells), in two of the three pools of the 3 week cultures (120 pg/10(6) and 139 pg/10(6) cells), and in one of the two pools of the 4 week cultures (111 pg/10(6) cells). Interleukin-6 was detected in one pool of the 1 week cultures (195 pg/10(5) cells) and in one pool of the 7 week cultures (81.6 pg/10(5) cells). Basic FGF was detected in the incubation media from three series of samples during the culture time course: 87 pg/ml in the 1 day cultures in the first series; 478, 310, and 269 pg/ml in the 1 day, 2 day, and 1 week cultures, respectively, in the second series; and 98 and 83 pg/ml in the 1 and 2 day cultures, respectively, in the third series. The TNF-alpha and EGF were not detected in any sample.. After cataract surgery, IL-1, IL-6, and b-FGF may be produced in vivo by residual LECs, causing postoperative inflammation and LEC proliferation.

Topics: Cataract; Cell Division; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Epithelium; Fibroblast Growth Factor 2; Follow-Up Studies; Humans; Interleukin-1; Interleukin-6; Lens, Crystalline; Tumor Necrosis Factor-alpha

The aqueous humor supplies the lens with nutrition. Recently, the existence of epidermal growth factor (EGF) and basic fibroblast growth factor (b-FGF) has been confirmed in the human aqueous humor. For this reason, it was thought that the function of these growth factors was related to the physiological condition of the lens epithelial cells (LEC). Therefore, we looked for the existence of receptors for EGF, FGF, and the internalization of EGF and b-FGF using LEC from human cataractous lenses in immunohistochemistry. We confirmed the existence of receptors for EGF and FGF as well as the internalization of EGF and b-FGF. From these results. We inferred that the function of EGF and b-FGF was strongly related to the physiological condition of LEC.

Topics: Aged; Cataract; Epidermal Growth Factor; Epithelium; ErbB Receptors; Fibroblast Growth Factor 2; Humans; Immunohistochemistry; Lens, Crystalline; Middle Aged; Receptors, Fibroblast Growth Factor

Thirty-eight cataract eyes and 15 artificial aphakic or pseudophakic eyes were enrolled in this study to determine levels of human epidermal growth factor (hEGF) and basic fibroblast growth factor (bFGF) in human aqueous, using a radioimmunoassay. Neither hEGF nor bFGF were detected in either congenital cataract eyes or senile cataract eyes without any complication. In cases of senile cataract complicated with glaucoma and traumatic cataract, bFGF range from 0.4 to 0.8 ng/ml whereas hEGF was not detected. In cases of senile cataract with myocardial or brain infarction and anterior subcapsular cataract, bFGF was detected ranging from 0.4 to 1.0 ng/ml and in some of the cases hEGF was detected at a level of 1.0 ng/ml. hEGF was detected at 1.0 ng/ml in some cases of secondary cataract after uveitis, cataract complicated with retinal detachment, cataract after scleral buckling or vitrectomy, and aphakic or pseudophakic eyes after extracapsular cataract extraction or phacoemulsification. These results showed that hEGF and bFGF exist at a level of about 1 ng/ml in human aqueous in some pathological states and it seems possible that proliferation of lens epithelial cells is promoted by the growth factors, with the result that after cataract, anterior capsular opacities and shrinkage become severe.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aqueous Humor; Cataract; Child; Child, Preschool; Epidermal Growth Factor; Epithelium; Female; Fibroblast Growth Factor 2; Humans; Infant; Lens, Crystalline; Male; Middle Aged; Radioimmunoassay

By using a highly specific and sensitive homologous radioimmunoassay, we found that the content of epidermal growth factor (EGF) in the lateral one-third of whole cataractous human lenses (age range 45-85 yr) extracted at elective intracapsular lens surgery, varied from undetectable to 106.25 pg mg-1 water soluble protein (WSP) (mean +/- S.D. = 39.70 +/- 38.90). When the lenses were grouped according to the stage of the cataract, i.e. immature (n = 3), mature (n = 4), and hypermature (n = 3), the means +/- S.D. were 92.56 +/- 26.23, 23.89 +/- 7.71, and 7.92 +/- 2.00 pg mg-1 WSP, respectively. In ten age-matched whole 'normal' lenses that we removed within 2-12 hr after death, the values in EGF of the lateral one-third of the lenses ranged from 2.91 to 36.40 pg mg-1 WSP (19.39 +/- 13.65). No correlation between the age of the lenses and the content of EGF could be demonstrated at the 95% confidence interval for the cataractous and 'normal' lenses. The quantity of endogenous EGF correlated significantly (P less than 0.01) with the clinical stage of the cataract and is probably related to the mitotic activity of the equatorial proliferative zone. We discuss the importance of EGF in normal and cataractous lenses and postulate that EGF in the lens is endogenous in origin.

Topics: Aged; Aged, 80 and over; Cataract; Epidermal Growth Factor; Humans; Lens, Crystalline; Middle Aged; Radioimmunoassay

Epidermal growth factor (EGF) is a polypeptide that stimulates the growth of various tissues, including the cornea. The presence of EGF in tears from normal volunteers and in aqueous humor from cataract patients was investigated via human EGF (hEGF)-specific radioimmunoassay. Immunoreactive hEGF was found to be present at similar concentrations in both reflex (ranging from 0.7 to 8.1 ng/ml) and non-reflex tears (ranging from 1.9 to 9.7 ng/ml), but was undetectable in aqueous humor. Immunoreactive EGF in human tears was indistinguishable immunologically, biologically and biochemically from urine EGF and standard hEGF.

Topics: Adult; Aged; Aqueous Humor; Binding, Competitive; Cataract; Chromatography, Gel; Epidermal Growth Factor; Female; Humans; Male; Middle Aged; Radioimmunoassay; Radioligand Assay; Recombinant Proteins; Tears