Cataract Lens Epithelial Cells Research Articles

In vitro assessment of the severity of deoxyribonucleic acid damage in different types of cataracts directly in lens epithelial cells.

This study aimed to assess the severity of deoxyribonucleic acid (DNA) damage in lens epithelial cells (LECs) of senile cortical, nuclear, and posterior subcapsular cataracts. LECs were obtained from senile cortical, nuclear, and subcapsular types of cataracts after surgery. DNA damage in the cells was immediately assessed quantitatively using the CometScore™ software. Comets were found in cataractous LECs. The formation of "comets" in the DNA of LECs can be visualized using single-cell gel electrophoresis and indicates DNA strand breaks because the damaged DNA migrates at a different rate than the nondamaged DNA. Maximal damage was observed in Grade 3 cortical, nuclear, and subcapsular forms of cataracts. Statistically significant DNA damage was seen between grades 1 and 3 of cortical type of cataract, grades 1 and 3 of nuclear type of cataract, and grades 2 and 3 and grades 1 and 3 of posterior subcapsular type of cataract. In patients with senile cataract, DNA of LECs was randomly damaged, and this type of damage was possibly caused by reactive oxygen species (ROS). Maximum DNA damage was found in patients with Grade 3 senile cortical, nuclear, and subcapsular type cataracts. The pathogenesis of senile cataracts is multifactorial and includes continuous molecular stress resulting from photooxidative stress, UV irradiation, and oxidative reactions.

Immunoreactivity of ICAM-1, MMP-2, and Nesfatin-1 in lens epithelial cells of patients with diabetes mellitus with or without diabetic retinopathy

The aim of this study was to evaluate the immunoreactivity of matrix metalloproteinases-2 (MMP-2), intercellular adhesion molecule-1 (ICAM-1), and nesfatin-1 in cataract lens epithelial cells (LECs) of patients with diabetes mellitus (DM) and to investigate the relationship of these markers with DM cataract and diabetic retinopathy (DR). Ninety patients were included in the study. The patients were divided into three groups (n = 30): Group 1 (control; patients without DM or DR); Group 2 (patients with DM only), and Group 3 (patients with both DM and DR). Lens capsule samples were collected during intraoperative cataract surgery. Samples were immunohistochemically stained for MMP-2, ICAM-1, and nesfatin-1 and their immunoreactivity was evaluated. The number of immunoreactive cells was determined with a microscope at ×400 magnification. Increased MMP-2 and ICAM-1 immunoreactivity was detected in the LECs of patients with DM, and especially in patients with DR (p < 0.001, p < 0.001, respectively). Nesfatin-1 immunoreactivity was significantly lower in LECs of diabetic patients (p < 0.001). The mean of MMP-2 immunoreactive cells were 7.47 ± 8.18, 22.80 ± 15.70, and 34.80 ± 20.85 in Groups 1, 2, and 3, respectively. The mean of ICAM-1 immunoreactive cells were 17.10 ± 9.83, 38.50 ± 23.55, and 56.93 ± 20.94 in Groups 1, 2, and 3, respectively. Nesfatin-1, MMP-2, and ICAM-1 and could potentially play important roles in the pathogenesis of cataracts in patients with DM.

Synchrotron-based FTIR microspectroscopy of protein aggregation and lipids peroxidation changes in human cataractous lens epithelial cells

Cataract is the leading cause of blindness worldwide but the mechanisms involved in the process of cataractogenesis are not yet fully understood. Two most prevalent types of age-related cataracts are nuclear (N) and cortical (C) cataracts. A common environmental factor in most age-related cataracts is believed to be oxidative stress. The lens epithelium, the first physical and biological barrier in the lens, is build from lens epithelial cells (LECs). LECs are important for the maintenance of lens transparency as they control energy production, antioxidative mechanisms and biochemical transport for the whole lens. The purpose of this study is to characterize compounds in LECs originated from N and C cataracts, by using the synchrotron radiation-based Fourier Transform Infrared (SR-FTIR) microspectroscopy, in order to understand the functional importance of their different bio-macromolecules in cataractogenesis. We used the SR-FTIR microspectroscopy setup installed on the beamline MIRAS at the Spanish synchrotron light source ALBA, where measurements were set to achieve single cell resolution, with high spectral stability and high photon flux. The results showed that protein aggregation in form of fibrils was notably pronounced in LECs of N cataracts, while oxidative stress and the lipids peroxidation were more pronounced in LECs of C cataracts.

MicroRNA let-7b induces lens epithelial cell apoptosis by targeting leucine-rich repeat containing G protein-coupled receptor 4 (Lgr4) in age-related cataract

Owing to a rapidly aging population, vision impairment caused by age-related cataract has become very common. Age-related cataract has also become one of the principal causes of blindness, and apoptosis of lens epithelial cells contributes to non-congenital cataract development. Previous studies have reported that microRNA let-7b (let-7b) is upregulated in cataractous lens epithelial cells, and the expression level of let-7b is positively associated with N, C and P cataract scores. However, the role of let-7b in the development of age-related cataract remains unclear. Here, we observed that the expression level of let-7b in the anterior lens capsules of age-related cataract was significantly higher than that in the normal anterior lens capsules. We performed ultraviolet (UV) irradiation to induce lens epithelial cell apoptosis. The results showed that the expression level of let-7b in lens epithelial cells which were treated by UV irradiation was significantly higher than that in the control, and let-7b promoted UV irradiation-induced apoptosis. Furthermore, we showed that leucine-rich repeat containing G protein-coupled receptor 4 (Lgr4) was a direct target of let-7b, and let-7b modulated lens epithelial cell apoptosis by directly targeting Lgr4. These findings will offer new insights into our understanding of the molecular mechanisms underlying the pathogenesis of cataract.

Effects of lentiviral RNA interference-mediated downregulation of integrin-linked kinase on biological behaviors of human lens epithelial cells.

To investigate the effects of lentivirus (LV) mediated integrin-linked kinase (ILK) RNA interference (RNAi) on biological behaviors of human lens epithelial cells (LECs). Human cataract LECs and immortalized human LEC line, human lens epithelial (HLE) B-3 cells were transfected by lentiviral vector expressing ILK-specific short hairpin RNA (shRNA) and then stimulated by transforming growth factor-β (TGF-β), the silencing of ILK gene and protein was identified by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot methods; biological behaviors including cell cycle and apoptosis, cell morphology, α-smooth muscle actin (SMA) stress fiber formation and cell migration were examined. Remarkable decreases of ILK protein expression were detected in LECs carrying lentiviral ILK-shRNA vector; flow cytometry revealed arresting of cell cycle progression through the G1/S transition and higher apoptosis rate in ILK-RNAi-LV transfected cells. Less α-SMA stress fiber formation and migration was observed in ILK-RNAi-LV transfected LECs. The present study demonstrated that ILK was an important regulator for LECs proliferation and migration. LV mediated ILK RNAi is an effective way to decrease ILK-regulated cell growth by arresting cell cycle progression and increasing cell apoptosis, as well as, to prevent cell migration by inhibiting TGF-β induced α-SMA stress fiber formation. Thus, LV mediated ILK RNAi might be useful to prevent posterior capsular opacification.

Estradiol Biosynthesis in Canine Lens Epithelial Cells

ABSTRACTPurpose: To confirm that lens epithelial cells (LEC) synthesize 17β-estradiol, active estrogen, and to identify the pathway(s) by which normal and cataractous LEC synthesize 17β-estradiol.Methods: ELISA was used to measure estradiol in aqueous humor; immunohistochemical staining was used to localize estradiol, testosterone and sulfatase; tritiated water release assay was used to measure aromatase activity; and qRT-PCR was used to quantify expression of aromatase and sulfatase in normal and cataractous canine and human LEC.Results: Canine eyes with and without cataracts had no differences in aqueous humor estradiol levels; however, cataractous LEC had more intense immunoreactivity for estradiol than normal LEC. There were little to no differences in canine sulfatase protein and mRNA expression when normal and cataractous LEC were compared. qRT-PCR demonstrated that canine cataractous LEC had significantly higher expression of aromatase; this was confirmed with the tritiated water release assay. Similar to dogs, human cataracts had both sulfatase and aromatase mRNA expression.Conclusions: Normal and cataractous LEC can synthesize estradiol by the sulfatase pathway; however, cataractous LEC appear to use the aromatase pathway as well. Because no differences in aqueous humor estradiol levels were detected, we suspect that estradiol synthesized by the sulfatase pathway is secreted into the aqueous humor; whereas, estradiol synthesized by the aromatase pathway is used for, as yet unknown, intracrine purposes.

Biomarkers of oxidative stress and cataract. Novel drug delivery therapeutic strategies targeting telomere reduction and the expression of telomerase activity in the lens epithelial cells with N-acetylcarnosine lubricant eye drops: anti-cataract which helps to prevent and treat cataracts in the eyes of dogs and other animals.

Cataracts in small animals are shown to be at least partially caused by oxidative damage to lens epithelial cells (LECs) and the internal lens; biomarkers of oxidative stress in the lens are considered as general biomarkers for life expectancy in the canine and other animals. Telomeres lengths and expressed telomerase activity in canine LECs may serve as important monitors of oxidative damage in normal LECs with documented higher levels of telomerase activity in cataractous LECs during cells' lifespan. Loss of functional telomere length below a critical threshold in LECs of canines during the effect of UV and chronic oxidative stress or metabolic failure, can activate programs leading to LEC senescence or death. Telomerase is induced in LECs of canines at critical stages of cataractogenesis initiation and exposure to oxidative stress through the involvement of catalytically active prooxidant transition metal (iron) ions. This work documents that transition metal ions (such as, ferrous ions- catalytic oxidants) might induce premature senescence in LECs of canines, telomere shortening with increased telomerase activity as adaptive response to UV light, oxidative and metabolic stresses. The therapeutic treatment with 1% N-acetylcarnosine (NAC) prodrug delivery is beneficial for prevention and dissolution of ripe cataracts in canines. This biological activity is based on the findings of ferroxidase activity pertinent to the dipeptide carnosine released ophthalmically from NAC prodrug of L-carnosine, stabilizing properties of carnosine on biological membranes based on the ability of the imidazole-containing dipeptides to interact with lipid peroxidation products and reactive oxygen species (ROS), to prevent membrane damage and delute the associated with membrane fragements protein aggregates. The advent of therapeutic treatment of cataracts in canines with N-acetylcarnosine lubricant eye drops through targeting the prevention of loss of functional telomere length below a critical threshold and "flirting" with an indirect effect with telomerase expression in LECs of canines during the effects of UV, chronic oxidative stress increases the successful rate of cataract management challenges in home veterinary care.

Dynamic and differential regulation in the microRNA expression in the developing and mature cataractous rat lens

Recent evidence supports a role for microRNAs (miRNAs) in regulating gene expression, and alterations in gene expression are known to affect cells involved in the development of ageing disorders. Using developing rat lens epithelial cells (LECs), we profiled the expression of miRNAs by a microarray-based approach. Few gene expression changes known to be involved in pathogenesis or cytoprotection were uniquely influenced by miRNA expression. Most miRNAs increased or decreased in abundance (let 7b, let 7c, miR29a, miR29c, miR126 and miR551b) in LECs/lenses during late embryonic and post-natal development and in cataract. Among them, miR29a, miR29c and miR126 were dramatically decreased in cataractous LECs from Shumiya Cataract Rats (SCRs). Specifically, the cytoskeleton remodelling genes tropomyosin (Tm) 1α and 2β, which have been implicated in the initiation of pathophysiology, were targets of miR29c and were over-stimulated as demonstrated by inhibitor experiments. In transfection experiments, increasing the level of miR29c caused a corresponding decrease in the expression of Tm1α and 2β, suggesting that miR29c may regulate the translation of Tm1α and 2β. 3′UTR luciferase activity of Tm1α, not 2β, was significantly decreased in miR29c-transfected mouse LECs. These findings demonstrate changes in miRNAs expression, and target molecules have potential as diagnostic indicators of ageing and as a foundation of miR-based therapeutics for age-related diseases.

Observation on the ultrastructure of cataract lens epithelial cells

Objective To study the relationship of cataract with lens epithelial cell apoptosis and autophagy by observing the ultrastructure of age-related and diabetic cataract lens epithelial cells.Methods Transmission electron microscope was used to observe the apoptotic and autophagic morphology of the lens epithelial cells in 5 cases with age-related and diabetic cataract respectively,and in 2 cases with normal lens.Results The apoptosis and autophagy of lens epithelial cells was found in cases with age-related and diabetic cataract,while no expression was observed in control lens epithelial cells.Conclusion The apoptosis and autophagy of lens epithelial cells are closely related to age-related and diabetic cataract formation. Key words: Cataract; Lens epithelial cells; Apoptosis; Autophagy

Elevated tropomyosin expression is associated with epithelial–mesenchymal transition of lens epithelial cells

Injury to lens epithelial cells (LECs) leads to epithelial–mesenchymal transition (EMT) with resultant fibrosis. The tropomyosin (Tpm) family of cytoskeleton proteins is involved in regulating and stabilizing actin microfilaments. Aberrant expression of Tpms leads to abnormal morphological changes with disintegration of epithelial integrity. The EMT of LECs has been proposed as a major cause of posterior capsule opacification (PCO) after cataract surgery. Using in vivo rodent PCO and human cataractous LECs, we demonstrated that the aberrant expression of rat Tpm and human Tpm1α/2β suggested their association in remodelling of the actin cytoskeleton during EMT of LECs. Expression analysis from abnormally growing LECs after lens extraction revealed elevated expression of α-smooth muscle actin (α-SMA), a marker for EMT. Importantly, these cells displayed increased expression of Tpm1α/2β following EMT/PCO formation. Expression of Tpm1α/2β was up-regulated in LECs isolated from cataractous lenses of Shumiya Cataract Rats (SCRs), compared with non-cataractous lenses. Also, LECs from human patients with nuclear cataract and anterior subcapsular fibrosis (ASF) displayed significantly increased expression of Tpm2β mRNA, suggesting that similar signalling invokes the expression of these molecules in LECs of cataractous SCR and human lenses. EMT was observed in LECs overexpressed with Tpm1α/2β, as evidenced by increased expression of α-SMA. These conditions were correlated with remodelling of actin filaments, possibly leading to EMT/PCO and ASF. The present findings may help clarify the condition of the actin cytoskeleton during morphogenetic EMT, and may contribute to development of Tpm-based inhibitors for postponing PCO and cataractogenesis.

Differences of bFGF gene expression in lens epithelial cells between fetuses and cataract patients.

To study the differences of basic fibroblast growth factor (bFGF) gene expression in lens epithelial cells (LECs) between fetuses and cataract patients. In situ hybridization was used to detect bFGF mRNA in the LECs that were cultured and in tissue sections from fetuses and in the LECs from the anterior capsule of cataract patients. Image analysis was used for the relative quantitative analysis of bFGF mRNA. bFGF gene existed in the LECs that were cultured and in tissue sections from fetuses and in the LECs from the anterior capsule of cataract patients. The integral absorbance for the fetal cultured cells, the fetal tissue sections and the capsule membrane cells of cataract patients were 627.1±268.7, 131.5±42.8 and 79.2±26.3 respectively. The integral absorbance of fetal cultured LECs was significantly higher than that of fetal section LECs (P<0.01). The integral absorbance of cataract LECs was significantly lower than that of fetal LECs (P<0.01). The in vitro culture of LECs can improve bFGF gene expression. The bFGF gene expression in fetal LECs is significantly higher than that in cataract LECs.

Explant culture of human traumatic cataract lens epithelial cells

目的 建立人外伤性白内障晶状体上皮细胞体外培养的简单有效方法,观察晶状体上皮细胞的体外生长规律和特点.方法 应用改良组织块贴附培养法对儿童及成人外伤性自内障的晶状体上皮细胞进行体外培养,倒置显微镜下观察其生长规律.结果 儿童和成人外伤性白内障晶状体上皮细胞都具有增生能力,晶状体上皮细胞均可传3代,晶状体上皮细胞多次传代后生长缓慢.结论 儿童和成人外伤性白内障晶状体上皮细胞体外培养增生能力有限,儿童晶状体上皮细胞增生能力较强.改良组织块贴附培养法简单易行,重复性好,是晶状体上皮细胞体外培养的较好方法。

Ultraviolet irradiation up-regulates telomerase transcription and activity in lens epithelial cells

Ultraviolet irradiation (UVR) increases telomerase activity in various cell types including skin, a sun-exposed organ. The lens is also continually exposed to UVR and we hypothesized that lenses exposed to UVR would have increased telomerase activity, with up-regulated TERT and TR, the two main components of the telomerase holoenzyme. To evaluate whether the cornea would protect lenses from such changes, whole globes, as well as isolated lenses, were exposed to UVR, and lenses were evaluated for changes in telomerase activity. There were three parts to this project. The first part of this experiment evaluated freshly harvested normal adult canine lenses exposed to 0, 300, 600, or 1200 J/m(2) UVR, and then allowed to recover for 1, 2, 3 and 4 h. Since only 600 J/m(2) UVR increased telomerase activity, four more postexposure recovery time-points for this UVR dose were evaluated: 10 min, 30 min, 8 h and 24 h. The second part of this experiment used freshly enucleated whole canine globes exposed to 0, 50, 100, 150, 300, 600 or 1200 J/m(2) and incubated overnight; lens epithelial cells (LEC) were evaluated for telomerase activity. The third part evaluated lenses that were exposed to 0 or 600 J/m(2) UVR, and then allowed to recover for 8 and 24 h, before TERT and TR mRNA levels were measured. Isolated lenses exposed to 600 J/m(2) UVR had significantly higher telomerase activity than unexposed controls and other UVR doses, at all time-points except 24 h postexposure. Lenses from whole globes exposed to UVR showed a dose-dependent increase in telomerase activity except at 50 J/m(2) and 1200 J/m(2). Isolated lenses exposed to 600 J/m(2) UVR and then allowed to recover for 8 and 24 h significantly up-regulated TERT and TR mRNAs compared to unexposed control lenses. Telomerase activity is regulated at both the transcriptional and post-translational levels in canine LEC. Previous work in our laboratory showed dose, time, and age-dependent changes in telomerase activity in the lens. The present study showed that TERT and TR mRNA transcription was increased for up to 24 h following an acute dose of UVR. Both telomerase activity and TERT and TR mRNA levels were elevated until 24 h post-UVR exposure, TERT in combination with TR functions in proliferation-related telomerase activity, but TERT alone has an anti-apoptotic function and its up-regulation may protect LEC from the acute effects of UVR. We are continuing to evaluate the mechanisms by which telomerase is regulated in normal and cataractous LEC.

Evaluation of advanced glycation end-products in diabetic and inherited canine cataracts

The receptor for advanced glycation end-products (RAGE) increases in the human cataract and should correlate with increased DNA damage and proliferation of lens epithelial cells (LECs). The purpose of this study was to measure and immunolocalize RAGE in normal and cataractous canine LECs, and to determine whether there was a correlation between RAGE and DNA damage (gadd45), cell-cycle regulation (p21), and LEC proliferation (proliferating cell nuclear antigen, PCNA). Thirty-two anterior lens capsules from 22 dogs that underwent cataract surgery and 10 lenses from dogs with normal eyes were evaluated. Eleven of the cataractous lenses were from diabetic patients (n=16), and eleven were from patients with inherited cataracts (n=16). Standard immunohistochemical staining was performed using antibodies against RAGE, gadd45, p21, PCNA, alpha-smooth muscle actin, and TGF-beta. Immunostaining intensity for each antibody was given a score of 0-4+. Standard Western blot analysis on normal and cataractous lens capsules was performed using the same antibodies as in the immunohistochemical staining. Comparisons were also made based on age and sex. Real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) was performed for RAGE. There was an increase in RAGE expression with age in normal LECs, but no significant difference was seen when normal adult LECs were compared to cataractous LECs. The stage of the cataract and the presence of LIU were not associated with a significant increase in RAGE expression. There was no age-dependent difference in the normal lenses for gadd45, p21, or PCNA. Significant up-regulation of p21 (P < 0.05) and PCNA (P < 0.05) was seen in diabetic cataracts compared to inherited cataracts. RAGE and PCNA expression did not increase with cataractogenesis, possibly due to overexpression associated with normal aging and constant exposure to oxidative stress from sunlight-related ultraviolet irradiation, respectively. However, p21 and PCNA increased in diabetic cataractogenesis suggesting cell cycle and proliferation dysregulation. This may be related to the rapid onset in this type of cataract compared with the more chronic and slower-to-develop inherited cataracts.

The study of posterior capsular opacification after cataract surgery Senile cataractous lens epithelial cells culture

目的 观察老年性白内障晶状体上皮细胞体外培养的生长特点,为研究老年性白内障及术后后囊浑浊的发生机制及防治奠定基础.方法应用改良组织块培养法对超声乳化术中老年性白内障晶状体前囊上皮细胞进行体外培养,在倒置显微镜下观察其生长和分化的规律.结果前囊接种3～5天,有新生上皮细胞自囊片的边缘长出并向四周延伸,第3～4周部分细胞内出现空泡和颗粒等结构改变,生长近于停止;传代培养细胞不能增生.结论老年性白内障晶状体前囊上皮细胞体外增生能力有限,改良组织块培养法培养晶状体上皮细胞简单有效。

Sex hormones and their receptors in patients with age-related cataract

Purpose To determine the sex hormone levels in the serum and aqueous humor in patients with age-related cataract and the corresponding hormone receptors in cataract lens epithelial cells (LECs). Setting Research Laboratory, International Intraocular Implant Training Center, Tianjin Medical University, Tianjin, China. Methods Serum and aqueous humor were drawn from patients with cataract and a control group. Enzyme-linked immunosorbent assay was used to determine the levels of estradiol, progesterone, and testosterone in the serum and the aqueous. The anterior lens capsules with attached LECs were obtained during cataract surgery, and the expressions of estrogen receptor, progesterone receptor, or androgen receptor were examined by immunohistochemistry. Results The testosterone level in the serum was significantly higher in the control group than in the cataract group. In the cataract group, there was no difference between sexes in the serum levels of estradiol or progesterone; however, the testosterone level in men was significantly higher than in women. The aqueous level of each hormone was lower than in serum; however, there was no difference between sexes and no association with corresponding serum levels. No estrogen, progesterone, or androgen receptor was detected in the LECs of patients with age-related cataract. Conclusions There was no statistical difference between men and women or between the cataract and control groups in the levels of estradiol or progesterone in serum. There was no between-sex difference in the aqueous levels. It appears that sex hormone levels can be regarded only as a risk factor for cataractogenesis, not as a key factor.

Inhibition of lens epithelial cells by Fas-specific antibody activating Fas-Fas ligand system

Purpose. To detect cell specific apoptosis factors, Fas and Fas ligand, and the common intracellular apoptosis modulators, interleukin-1ß converting enzyme (ICE)-like protease (caspase 1), Bcl-2, Bcl-xL and Bax in lens epithelial cells (LEC) of human cataracts. To study the effects of Fas-stimulating monoclonal antibody on inhibition of LEC proliferation. Methods. Reverse-transcriptase-polymerase chain reaction (RT-PCR) was used to detect Fas, Fas ligand, caspase 1, Bcl-2, Bcl-xL and Bax, after cDNA was synthesized from the total RNA isolated from human cataractous LEC obtained by capsulotomy during cataract surgery. Fas-stimulating monoclonal antibody was added at the concentrations of 10, 30, 100, 300 and 1000 ng/ml to the incubation medium of human cataractous LEC; and the specimens were incubated for 24 h at 37°C with 5% CO 2 circulation and 100% humidity. The specimens were then stained with Hoechst 33342, and the number of apoptotic cells was counted. Results. Fas, caspase 1, Bcl-2, Bcl-xL and Bax mRNA were detected by RT-PCR. Fas ligand mRNA was not detected by RT-PCR. At each concentration, Fas-stimulating monoclonal antibody significantly inhibited LEC proliferation. Conclusions. Human cataractous LEC expressed mRNA of Fas and various modulators of apoptosis pathways. Fas-stimulating monoclonal antibody may have the potential to prevent posterior capsule opacification after cataract surgery by inhibiting LEC proliferation.

Detection of integrins in cataract lens epithelial cells

PurposeTo detect the expression of integrin subunits in lens epithelial cells (LECs) of human cataracts. SettingResearch Laboratory, International Intraocular Implant Training Centre, Tianjin Medical University, China. MethodsThe circular sections of the anterior capsules with attached LECs were obtained during cataract surgery from 100 patients. The LECs were stained with an avidin-biotin-complex immunohistochemical technique using 5 monoclonal antibodies specific for alpha subunits 2, 3, 5 and beta subunits 1, 2. ResultsAll integrin subunits studied were found to varying degrees in specimens. The positive percentages were 70%, 65%, 75%, 70%, and 80%, respectively. ConclusionIntegrin subunits were present in LECs of human cataracts. These molecules may serve in the adhesion of LECs to the lens capsule and play a role in cell–posterior capsule interaction after cataract surgery.

Degeneration and transdifferentiation of human lens epithelial cells in nuclear and anterior polar cataracts

Purpose To study the possible mechanisms of cataractogenesis by evaluating the characteristics of cataractous lens epithelial cells (LECs) in different types of human cataract. Setting Kangnam St. Mary’s Hospital, The Catholic University of Korea, Seoul, Korea. Methods Lens epithelial cells attached to the anterior capsules in eyes with nuclear or anterior subcapsular were analyzed for morphological characteristics by electron microscopy and for cellular characteristics by immunohistochemistry. Results Human LECs beneath the anterior capsule were degenerated in nuclear cataracts and were transdifferentiated in anterior polar cataracts. In senile nuclear cataractous lenses, LECs beneath the anterior capsule showed degenerative changes in morphology. In nuclear cataracts, LECs were immunohistochemically positive for cytokeratin and vimentin, while those in anterior polar cataracts were positive for vimentin only. The LECs of anterior subcapsular cataracts were transdifferentiated into spindle-shaped fibroblast-like cells without cellular junctions and embedded within a fibrillar meshwork mass. The extracellular matrixes in the anterior capsule of anterior subcapsular cataracts were immunohistochemically positive for fibronectin, laminin, collagen type I, and collagen type IV. Conclusions Lens epithelial cells in different types of cataracts have distinct cellular characteristics and may possess a bipotential nature with the ability to transdifferentiate into mesenchymal cells. This may be an underlying mechanism for the development of cataract and capsule opacification.

Effects of the cytokines on the proliferation of and collagen synthesis by human cataract lens epithelial cells.

To assess the effects of the cytokines, interleukin-1 (IL-1), IL-1 receptor antagonist (IL-1ra), transforming growth factor-beta 2 (TGF-beta 2) and basic fibroblast growth factor (b-FGF), on the mitosis of and collagen synthesis by lens epithelial cells (LECs) of human cataracts. The anterior lens capsule with attached LECs was obtained by capsulotomy during cataract surgery and cultured. The cultures at 2 to 3 weeks before confluency were used for the experiments. To quantify the mitosis and collagen synthesis, the incorporation of 3H-thymidine and 3H-proline, respectively, into the LECs was measured by a scintillation counter at 48 hours and 24 hours, respectively, after addition of the cytokine at various concentrations into the incubation medium. IL-1 and b-FGF increased the mitosis and collagen synthesis significantly, but IL-1ra significantly decreased the mitosis while leaving the collagen synthesis intact. TGF-beta 2 decreased the mitosis significantly, but increased the collagen synthesis significantly. These cytokines may play an important role in an autocrine or paracrine pathway in the proliferation of residual LECs after cataract surgery. Elucidation of the role of these cytokines may lead to the development of new therapies for the prevention of secondary cataract.