Inhibition of lens epithelial cell migration using different square-edge intraocular lenses in an in vitro capsular bag model / 中华实验眼科杂志

Objective:To establish an in vitro capsular bag model and compare the inhibitory effects of different 360° square-edge intraocular lens (IOL) on lens epithelial cells (LECs) migration. Methods:In vitro capsular bag model with posterior capsule opacification (PCO) was established using Transwell compartment, cell climbing slices, human collagen type Ⅳ, and IOL.The models were divided into Plate-loop HydroSmart group, C-loop HydroSmart group, and C-compensation-loop Hydrophobic group according to the different square-edge IOL implanted.A blank control group was set using the Transwell compartment without IOL.The early PCO pathological manifestations in lens epithelial cell line SRA01/04 cultured in the Transwell compartment were observed with an inverted microscope.The cell morphology in different groups was observed by hematoxylin and eosin staining.The cell counting and cell migration inhibition rate of anterior capsule and posterior capsule were calculated by Transwell assay and cell-exclusion zone assay, respectively. Results:The early pathological characteristics of PCO, such as early Soemmering ring and small Elschnig pearl, could be found in cells in the in vitro capsular bag model after 48-hour culture.The migrating cells in model groups were fibrous.No changes mentioned above were found in blank control group.The number of migrating cells in the anterior capsule of Plate-loop HydroSmart group, C-loop HydroSmart group, C-compensation-loop Hydrophobic group was 18.80±5.53, 24.67±9.80, and 34.47±10.80, respectively, and the number of migrating cells in the optical area of the posterior capsule of the three groups was 56.43±9.00, 162.20±16.38, and 121.30±12.01, respectively.The cell migration inhibition rate in the anterior capsule of Plate-loop HydroSmart group, C-loop HydroSmart group, C-compensation-loop Hydrophobic group was (92.02±1.94)%, (89.76±3.10)%, (86.27±4.54)%, respectively, and the cell migration inhibition rate in optical area of the posterior capsule of the three groups was (91.60±3.65)%, (70.14±5.35)%, (78.43±3.48)%, respectively.The number of migrating cells in the anterior capsule was lower and the cell migration rate inhibition was higher in Plate-loop HydroSmart group than C-compensation-loop Hydrophobic group, with significant differences (both at P<0.05). The number of migrating cells in the optical area of the posterior capsule and the cell migration inhibition rate was greater than those of C-loop HydroSmart group and C-compensation-loop Hydrophobic group, showing statistically significant differences (all at P<0.001). Conclusions:The in vitro capsular bag model can be used in PCO research.Compared with C-loop HydroSmart IOL and C-compensation-loop Hydrophobic IOL, Plate-loop HydroSmart IOL can more effectively inhibit the migration of LECs to the optical area of the posterior capsule.

Applications of capsular bag model in posterior capsular opacification / 中华实验眼科杂志

Posterior capsular opacification (PCO),also known as after-cataract,is the most frequent complication and the primary cause for visual decrease after extracapsular cataract surgery.At present,there is no effective way to treat PCO,so more attentions are focused on preventive reseaching of PCO and treatment methods.Although a variety of studies have increased our understanding of the pathogenesis of PCO,the cellular mechanisms responsible for PCO are still unclear.Cultured capsular bag model in vitro could effectively simulate lens capsular membrane and cells survival environment after cataract extraction and IOL implantation.However,lens capsular bag cultivation with different methods have their own characreristics.The material source,preparation methods of capsular bag model,characreristics of materials which maintain capsular bag contours and its application in PCO were reviewed.

Inhibitory effect of propylene glycol mannate sulfate on growth of rabbit lens epithelial cells in vitro / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the inhibitory effect of rabbit lens epithelial cell(RLEC)survival and growth by propylene glycol mannate sulfate(PGMS)on the rabbit capsular bag in vitro.METHODS;Capsular bags were prepared from rabbit eyes after extracapsular cataract extraction(ECCE)and incubated in 0.2,0.4,0.8g/L PGMS in 2,5,10 minutes incubation periods.After treatment,the capsular bags were cultured for 7 days in Dulbecco minimum essential medium(DMEM)supplemented with 50mL/L fetal calf serum(FCS).The specimens were examined with light microscopy and transmission electron microscopy(TEM).Capsular bags without receiving PGMS only served as controls.RESULTS:PGMS inhibited the proliferation of RLEC in the manner of concentration and time dependentment.At the threshold protocol of incubation in PGMS at 0.8g/L for 5 or 10 minutes,proliferative activity of cells were largely arrested and nearly no RLEC was seen on the posterior capsule(P＜0.05).Control group had no effect on structure and proliferative activity of RLEC,and the growth proceeded rapidly so that the posterior capsule were totally covered by a confluent monolayer of cell by the end of 7 days.Under TEM,the cells in the control group were tightly arrayed with clearly defined cellular boundary and structure;while cellular deformity and undefined intracellular structure could be seen in the 0.4g/L and 0.8g/L experimental groups.CONCLUSION:PGMS can effectively inhibit the proliferation of RLEC.

Inhibitory Effects of Paclitaxel and Cisplatin on Transdifferentiation of Lens Epithelial Cells into Fibroblast

PURPOSE: This study investigated the inhibitory effects of Paclitaxel by altering tubulin assembly and cisplatin exposure by binding DNA of the lens epithelial cells (LECs) during epithelial cell cultures in the capsular bag model. METHODS: In the capsular bag model, the LECs were cultured with exposure to Paclitaxel (1, 10, 100 nM) and Cisplatin (1, 10, 100 micro M) for 3 min. The effect of Paclitaxel and Cisplatin was analyzed by observing the cell number of fibroblasts per field, Western blots for type IV collagen, TUNEL assay and the Proliferating Cell Nuclear Antigen (PCNA) and Bromodeoxyuridine (BrdU) incorporated proliferating cells. RESULTS: An increase in concentration of Paclitaxel and Cisplatin resulted in a decrease in the number of fibroblasts and spindle-shaped cells. The number of proliferating cells showing PCNA positivity and BrdU incorporation in the nuclei was decreased in a dose dependent manner by treatments of Paclitaxel and Cisplatin. Expression of type IV collagen also decreased after treatment with these two agents. Results of the TUNEL assay showed no change in the apoptosis of cells with regard to an increase in concentration of Paclitaxel and Cisplatin. CONCLUSIONS: This study showed inhibitory effects of Paclitaxel and Cisplatin on the proliferation and transdifferentiation of LECs into fibroblasts using the capsular bag model.

Inhibitory Effects of Imatinib Mesylate on Transdifferentiation of Lens Epithelial Cells into Fibroblast

PURPOSE: To investigate whether Imatinib mesylate (IM) could inhibit the transdifferentiation of the lens epithelial cells (LECs) into fibroblasts using the capsular bag model. METHODS: In the capsular bag model, LECs were cultured by exposure to IM at various concentrations for 3 min. The effect of IM was analysed by observing the covering area, numbers of alpha-SMA positive cell, and BrdU incorporated proliferating cells. The same analysis was performed in the culture of LECs with TGF-beta. RESULTS: The covering area was significantly decreased by the treatment of 30 micro M IM, and the positive cells for alpha-SMA and BrdU were also decreased by IM treatment in a dose dependent manner. In addition, increasing TGF-beta concentration accelerated transdifferentiation, but suppressed the acceleration of the transdifferentiation induced by TGF-beta. CONCLUSIONS: In the capsular bag model, IM effectively inhibited not only the transdifferentiation of LECs into fibroblasts but also the TGF-beta induced acceleration of the transdifferentiation.

Differentiation and Transdifferentiation of Lens Epithelial Cells in Capsular Bag Model Culture

PURPOSE: To investigate the differentiation of lens epithelial cells (LECs) to lens fiber, and the transdifferentiation of LECs to fibroblast in capsular bag culture. METHODS: After observing the changes of LECs by using phase-contrast microscopy, we observed a cross section of capsular bag by using light microscope (LM) and electron microscope (EM). In addition, the expressions of alpha A-crystallin, a marker of differentiation of LEC to lens fiber, and of alpha-smooth muscle actin, a marker of LEC to fibroblast, were examined during the culture period by western blot. RESULTS: On phase-contrast microscopy, 7 to 14 days after culture, the portion of LECs was gradually elongated and cytoplasm became transparent, so that the differentiation resembled lens fiber. One to 7 days after culture, the portion of LECs changed to spindle shape and the transdifferentiation resembled fibroblast. LM and EM observations indicated that changes of each LEC were lens fiber, and fibroblast. According to Western blot, the expression of alpha A-crystallin was increased by 10 days after culture. The alpha-smooth muscle actin showed an increased expression 10 to 30 days after culture. CONCLUSIONS: From the capsular bag model, we observed the resemblances of the differentiation and transdifferentiation of LECs with lens fiber and fibroblast.

Inhibitory Effects of Mitomycin-C and 5-Fluorouracil on the Proliferation of Lens Epithelial Cells using Capsular Bag Model

PURPOSE: We investigated the inhibitory effects of mitomycin-C (MMC) and 5-fluorouracil (5-FU) on the proliferation of the lens epithelial cells (LECs) depending on time and concentration during the epithelial cell culture using capsular bag model. METHODS: We cultured LECs using capsular bag model after exposure to various time and concentration of MMC and 5-FU. We compared the half coverage time and appearance of posterior capsule by LECs. We examined the proliferative and inhibitory effect on LECs using BrdU immune staining. We observed the morphologic change of LECs on histologic section. RESULTS: Half coverage time of posterior capsule by LEC was 18.2 +/- 3.5 days in control group, whereas 27.5 +/- 3.8 days and 26.8 +/- 4.2 days when treated with 0.2 mg/ml MMC, 50 mg/ml 5-FU for 3 minutes, repectively. The increase of concentration and the exposure time of MMC or 5-FU resulted in the delay of coverage time of posterior capsule by LECs and reduction of BrdU incorporation in the nucleus of proliferating cells. On histologic section, reduction of LECs' multilayering and few cytoplasmic organells were observed. CONCLUSIONS: Using capsular bag model, we found the inhibitory effect of MMC and 5-FU on LECs proliferation depending on the concentration and exposure time. Capsular bag model would be contribute to the study about after-cataract.

The Effect of the Remnants after Cataract Extraction on the Lens Epithelial Cell Culture

PURPOSE: To assess the effect of the remnants after lens extraction on posterior capsular opacification with lens epithelial cell culture through in vitro capsular bag model. METHODS: After isolating porcine lens capsules, sterile non-toxic PMMA (polymethyl- mathacrylate) tension ring was inserted into the capsule. These were placed in organ culture medium up to 6 weeks. The grade of cell coverage of the posterior lens capsule was recorded to check the proliferative activity. RESULTS: In the process of cell culture, outgrowth of the epithelial cells was observed across the posterior capsule after a lag period. The rate of cell coverage was dependent upon the added factors. The proliferative activity was the greatest in the group where lens cortical and nuclear materials were added, and other groups showed no difference from a control group. CONCLUSIONS: To reduce posterior capsular opacification, it is important that we should not leave the lens cortical material behind during cataract surgery.

The Effect of Lens Nucleus and Cortex Material on Lens Epithelial Cell Culture through In vitro Capsular Bag Model

PURPOSE: This study attempts to evaluate the effect of lens cortex and nucleus remnants on posterior capsular opacification with method of the cell culture according to in vitro capsular bag model. METHODS: After bovine lens were isolated, we performed continuous curvilinear capsulorhexis and hydrodissectioin of the lens fiber mass. At this stage a tension ring was implanted and then the preparations placed in organ culture for up to 6 weeks. Lens cortex and nucleus material was added at the culture media in group 2, 3, 4, 5, 6 with amount of 1/16, 1/32, 1/64, 1/96, 1/128 of one lens volume. Group 1 was control group that was not added lens materials. Cell coverage of the posterior lens capsule was recorded and the capsules were examined, both pre-and post-coverage, for proliferative activity. RESULTS: After a lag period outgrowth was observed across the posterior capsule. The proliferative activity was greater at the groups that were added more amount of the lens cortex and nucleus material. CONCLUSIONS: it is important that we should not remain any lens cortex material remnant at cataract surgery.