Electron microscopy analysis of femtosecond laser-assisted capsulotomy before and after lens fragmentation.

Studying anterior lens capsule cutting edge profiles from femtosecond laser-assisted capsulotomy procedures performed before and after lens fragmentation. Twenty eyes (10 patients) with age-related cataract underwent femtosecond laser-assisted surgery (FLACS) using the Ziemer Z8 platform. First step of laser surgery was either capsulotomy (group first) or fragmentation (group second). One eye of each patient was assigned randomly, the second eye treated with the different sequence of procedures. After anterior capsule removal, tissue was fixed in cacodylate-buffered solution and cutting-edge profiles were analysed using scanning electron microscopy (SEM). All cases had cataract grade 2 and 3 based on LOCS III grading. SEM analysis showed more smooth edges in the first group, especially in cases with pseudoexfoliation (P = 0.037); more tags and bridges and a significant number of staggered cutting patterns (7 out of 10 cases) in the second group. All cases evolved the same microgroves with "valleys and mountains " as signs of the photodisruption process. Femtosecond laser capsulotomy should be performed before lens fragmentation minimizing the rate of cutting errors. Especially in eyes with advanced cataract, as intracapsular pressure may increase due to lens fragmentation without anterior capsular opening.

MicroRNA Let-7c-5p-Mediated Regulation of ERCC6 Disrupts Autophagic Flux in Age-Related Cataract via the Binding to VCP.

Purpose: DNA damage contributes to the pathogenesis of age-related cataract (ARC) and is repaired through the nucleotide excision repair (NER) pathway, which includes ERCC6. Evidence has demonstrated that defective autophagy leads to lens organelle degradation and cataract. This study aimed to investigate the effects of ERCC6 on autophagy and determine its mechanisms in ARC.Methods: The clinical case-control study comprised 30 patients with ARC and 30 age-matched controls who received transparent lens extraction. Transmission electron microscopy was used to assess the ultrastructure of autophagic vesicles in lens anterior capsule tissues and lens epithelial cell line (SRA01/04). Real-time polymerase chain reaction and western blot analyses were performed to measure relative gene expression levels. Gene expression levels and localization were assessed by immunofluorescence. A coimmunoprecipitation assay was used to investigate the relationship between CSB which encoded by ERCC6 and VCP. ERCC6-siRNA and let-7 c-5p mimic were used to alter the expression of ERCC6 and let-7 c-5p.Results: Autophagy induction occurred in lens anterior capsule tissues of patients with ARC and in UVB-induced SRA01/04 cells, where the number of LC3B puncta was increased. Consistent with this result, the expression of beclin1 (BECN1) and LC3B, in addition to that of p62, was increased. Additionally, ERCC6 expression decreased, and silencing ERCC6 induced increases in the expression of BECN1, LC3B and p62. Moreover, CSB interacted with VCP, and let-7 c-5p induced dysregulation of autophagy by targeting ERCC6.Conclusion: In ARC, Let-7 c-5p-mediated downregulation of ERCC6 might prevent the degradation of autophagic vacuoles. CSB binds to VCP, inducing autophagosomes to combine with lysosomes and be degraded.

Lens Epithelial Surface Disorders in Exfoliation Syndrome: A Scanning and Transmission Electron Microscopy Study.

BACKGROUND: Hydrodissection was recently reported to occur more easily in patients with exfoliation syndrome (XFS). Transmission electron microscopy (TEM) studies have already revealed alterations of the lens epithelial cells (LECs) and their apical membrane towards the lens fibers. OBJECTIVE: The aim of this work was to examine the three-dimensional appearance of the lens epithelium in patients with XFS. METHODS: Fourteen patients with senile cataract, 7 of whom had XFS, were included. Anterior lens capsules (aLCs) were obtained with continuous curvilinear capsulorrhexis (CCC) during phacoemulsification and were examined with scanning electron microscopy (SEM) and TEM. RESULTS: Exfoliation samples exhibited an overall more irregular apical surface of the lens epithelium compared to control aLCs. The height of LECs varied extensively. On the apical surface of LECs, amorphous, crystalline-like, or microgranular extracellular material and membranous, oval-shaped structures were documented with SEM. All findings were connected to corresponding observations with TEM and were not correlated to the type of cataract. CONCLUSIONS: In XFS patients, the lens epithelial surface exhibited a highly irregular margin, with extracellular material covering the apical membrane of LECs. We suggest that XFS probably causes both epithelial and lens fiber degeneration which, during CCC and mechanical extraction of the aLC from the lens cortex, result in diverse alterations.

Comparison of anterior capsulotomy techniques: continuous curvilinear capsulorhexis, femtosecond laser-assisted capsulotomy and selective laser capsulotomy.

PURPOSE: To compare the anterior capsulotomy edge tear strength created by manual continuous curvilinear capsulorhexis (CCC), femtosecond laser-assisted capsulotomy (FLACS), and selective laser capsulotomy (SLC). SETTING: Singapore National Eye Centre, Singapore and Excel-Lens, Livermore, California, USA. DESIGN: Three armed study in paired human eyes. METHODS: Capsulotomies were performed in 60 cadaver eyes of 30 donors using CCC, Victus Femtosecond Laser, (Bausch & Lomb, Rochester, New York, USA) or CAPSULaser, (Excel-Lens, Los Gatos, California, USA). Three pairwise study groups each involved 10 pairs of eyes. Study group 1: SLC eyes compared with fellow eyes with CCC. Study group 2: CCC eyes compared with fellow eyes with FLACS. Study group 3: FLACS eyes compared with fellow eyes with SLC.A shoe-tree method was used to apply load to the capsulotomy edge, and Instron tensile stress instrument measured distension and threshold load applied to initiate capsule fracture. Relative fracture strengths and distension of CCC, FLACS and SLC were determined. Scanning electron microscopy (SEM) of capsule edges were reviewed RESULTS: Anterior capsulotomies behave as non-linear elastic (elastomeric) systems when exposed to an external load. The pairwise study demonstrated that the SLC fracture strength was superior to that of CCC by a factor of 1.46-fold with SLC 277±38 mN versus CCC with 190±37 mN. Furthermore, CCC fracture strength was superior to that of FLACS by a factor of 1.28-fold with CCC 186 + 37 mN versus FLACS 145 ± 35 mN (p < 0.001). This was determined by statistical analysis utilising the Wilcoxon matched-pairs signed-ranks test and in accordance with the Consolidated Standards of Reporting Trials guidelines. The capsule edge of SLC on SEM demonstrated a rolled over edge anteriorly and an alteration of collagen. CONCLUSIONS: The strength of the capsulotomy edge for SLC was significantly stronger than that of CCC which and both were significantly stronger than FLACS. The relative strengths can be explained by SEM of each type of capsulotomy.

Exfoliation Fibrils within the Basement Membrane of Anterior Lens Capsule: A Transmission Electron Microscopy Study.

Purpose: The aim of the present study was to demonstrate novel findings recognized within the basement membrane of the anterior lens capsules (ALCs) in exfoliation syndrome, in a transmission electron microscopy (TEM) study. Materials and methods: Twenty-one patients with age-related cataract were included in the study. Eleven out of them suffered from exfoliation syndrome (XFS). Anterior lens capsules were obtained from patients during phacoemulsification, applying continuous curvilinear capsulorhexis and then were examined by transmission electron microscope. Results: Six out of 11 examined basement membranes with XFS had a granular appearance with dotted deposits of electron-dense material. Fibrils were clearly detected, longitudinally, within the basement membrane, usually located in the outer quarter of the basement membrane, towards the anterior chamber. Conclusion: New data about the exfoliation material and its location within the basement membrane of the anterior lens capsules of XFS patients are presented. These findings constitute new evidence for XFS's histopathogenesis and might help clarify the lenticular exfoliation material's (XFM) mechanisms of origin.

Intraoperative performance and ultrastructural integrity of human capsulotomies created by the improved precision pulse capsulotomy device.

PURPOSE: To investigate the intraoperative performance and the ultrastructural features of anterior capsulotomy performed with the improved Zepto precision pulse capsulotomy (PPC) device. SETTING: Launceston Eye Institute, Launceston, Tasmania, Australia. DESIGN: Prospective, consecutive case series. METHODS: Intraoperative observation of performance and complications were made on consecutive eyes having capsulotomies with the improved PPC device. The capsulotomy specimens were examined under scanning electron microscope and compared with manual continuous curvilinear capsulorhexis and femtosecond laser-assisted cataract surgery specimens. RESULTS: The study comprised 52 eyes. Intended capsulotomy occurred in 50 eyes with 2 device failures. Complete free-floating capsulotomies were achieved in 48 eyes (96%). Anterior capsule tears occurred in 2 eyes (4%). CONCLUSIONS: The ability of the PPC device to create complete free-floating capsulotomies improved; however, the radial tear rate remained high. The ultrastructural features in ex vivo human capsulotomy specimens still showed areas of irregular capsule margin with frayed edges.

Transmission electron microscopy study of undescribed material at the anterior lens capsule in exfoliation syndrome.

PURPOSE: In the present study, we aimed to examine the anterior lens capsule using transmission electron microscopy (TEM) and compare the findings in patients with and without exfoliation syndrome (XFS). METHODS: Eighteen patients with senile cataract, including 10 with XFS, were included. Anterior capsule specimens were obtained from patients during phacoemulsification through continuous curvilinear capsulorrhexis, and were examined via TEM. RESULTS: In the XFS group, in addition to the typical fibrillar material above the basement membrane of the lens capsule, another unknown, microgranular, electron-dense, unbound material was observed beneath the lens epithelium. Larger formations appeared to detach from the surface of the epithelial cells, and oval or crescent-shaped structures of an electron-denser material were less commonly observed on the apical side of the epithelium. Membranous structures were occasionally attached to epithelial cells that often exhibited thicker or ruptured cell membranes on their apical-free side, along with deposits of electron-dense material. Degenerative lesions of various severities were observed at the epithelium in both groups. CONCLUSION: The anterior lens capsule epithelium in patients with XFS exhibits a highly irregular and rough margin on its free side, with unbound material over its membrane, which probably contributes to loose contact with the underlying lens and leads to different clinical behaviors in XFS eyes during cataract surgery.

Ultrastructural effects of silicone oil on the clear crystalline lens of the human eye.

PURPOSE: To evaluate light and electron microscopic changes of the anterior capsule and its epithelium after clear lens extraction of vitrectomized myopic eyes with silicone oil tamponade. METHODS: This prospective, controlled, non-randomized, interventional study included 20 anterior lens capsular specimens that were excised during combined clear lens extraction and silicone oil removal from previously vitrectomized highly myopic patients with silicone oil tamponade for previous retinal detachment surgeries. The specimens were examined via light microscopy and electron microscopy and compared with 20 anterior capsule specimens removed during clear lens extraction of non-vitrectomized highly myopic eyes. RESULTS: Light microscopic examination of clear lens anterior capsule specimens of vitrectomized myopic eyes filled with silicone oil showed relatively more flat cells with irregular outline of lens' epithelial cells with wide intercellular spaces, deeply stained nuclei, and multiple intracytoplasmic vacuoles. Scanning electron microscopy revealed collagenous surfaces filled with multiple pits, depressions, and abnormal deposits. Transmission electron microscopy revealed lens epithelial cells with apoptotic changes, many cytoplasmic vacuoles, and filopodia-like protrusions between lens epithelial cells and the capsule. Epithelial proliferation and multilayering were also observed. CONCLUSION: silicone oil may play a role in the development of apoptotic and histopathological changes in clear lens epithelial cells. Clarity of the lens at the time of silicone oil removal does not indicate an absence of cataractous changes. We found justification of combined clear lens extraction and silicone oil removal or combined phacovitrectomy when silicone oil injection is planned, but further long-term studies with larger patient groups are required.

Ideal parameters for femto-second laser-assisted anterior capsulotomy: Animal studies.

In femtosecond laser-assisted cataract surgery, the parameter such as horizontal spot spacing and energy level can be adjusted. Although there have been several studies reported on various laser systems, showing the effects of varying energy levels and horizontal spot spacing on lens capsulotomy cut edges, none have been reported on the Catalys laser system (Abbott Medical Optics, Inc., Santa Ana, CA). The aim of this study is to evaluate, using scanning electron microscopy (SEM), the quality of the cut edges of the laser lens capsulotomy obtained using the Catalys Laser System, using different horizontal spot spacing and energy levels, and to determine the ideal parameters based on SEM results. Fifty rabbit capsulorhexis specimens from a femtosecond laser with different spot spacing and energy settings were divided into five groups randomly. Spot spacing was 3 um and laser pulse energy was 4 uJ in group 1. The respective values were 5 um and 2 uJ in group 2, 5 um and 4 uJ in group 3, 5 um and 6 uJ in group 4, and 7 um and 4 uJ in group 5. All samples were evaluated using SEM to compare the number of tags per capsulotomy and the laser emission time. Group 1 had a significantly lower tag formation than groups 3 and 5 (P = 0.042 and 0.021, respectively). Although the laser emission time increased about 1.5 sec as the spot spacing increased from 3 to 7 um, the quality of the cut was smoother in group 1 because of overlapping effect of photodisruption cavities. There was no significant difference between groups 2, 3 and 4 at different laser energy settings. In an ex-vivo study, samples from an energy setting of 10 uJ showed increased irregularity and damage. The degree of irregularity was higher at increasing spot spacing and laser energy settings, with abundant tag formation. Dense spot spacing with low-energy settings provide a better cut quality, which is probably correlated with the reduction in anterior capsular tear complications.

Ultrastructural Integrity of Human Capsulotomies Created by a Thermal Device.

PURPOSE: To investigate the ultrastructural features of anterior capsulotomy performed with a thermal device, the precision pulse capsulotomy (PPC). DESIGN: Prospective, multicenter case series. PARTICIPANTS: Consecutive patients undergoing capsulotomy with the PPC device. METHODS: Prospective study of patients undergoing capsulotomy with the PPC by 2 surgeons, followed up by routine phacoemulsification cataract surgery, was undertaken. All capsulotomy specimens were collected for scanning electron microscopy (SEM). Observations were made regarding uniformity of the capsular edge and the presence of irregularities that may compromise integrity. Comparisons were made with manual continuous curvilinear capsulorrhexis. MAIN OUTCOME MEASURES: Ultrastructural features of PPC and presence of irregularities. RESULTS: Frayed appearance of the anterior capsule edge was noted in postoperative visits under slit-lamp examination. Scanning electron microscopy sampling showed a generally uniform rolled capsular edge, but interspersed with areas of irregularity with frayed appearance at the capsule margin. CONCLUSIONS: The PPC device is capable of creating reproducible, central, and precise circular capsulotomy. The ultrastructural features in ex vivo human capsulotomy specimens generally show eversion of the capsulotomy edge, but in some cases, this was accompanied by areas of irregular capsule margin with frayed edges, likely caused by dissipated thermal energy. The postoperative appearance and SEM features warrant further assessment of the PPC integrity and clinical correlation.

Effects of Fuchs uveitis syndrome on the ultrastructure of the anterior lens epithelium: A transmission electron microscopic study.

PURPOSE: The purpose of the study was to investigate the electron microscopic findings of the lens epithelial cells (LECs) in patients with Fuchs uveitis syndrome (FUS) who suffered from cataracts and to compare those with age-matched controls. METHODS: This study was a prospective, comparative case series. The anterior lens capsules (ALC: basement membrane and associated LECs) were taken from 12 eyes of 12 cases of FUS and ten eyes of ten control patients. The ALCs were obtained from cataract surgery and prepared for transmission electron microscopy (TEM). RESULTS: There were no statistically significant differences regarding the age or gender between the FUS group and the control group (P > 0.05). In the TEM examinations of the ALCs, all of the FUS cases revealed similar significant ultrastructural changes when compared to the control patients. In the FUS group, the LECs showed homogeneous thickening and irregularity which included some small vacuoles in different areas of the epithelial tissue. Moreover, in some areas of the LECs, widespread, oval-shaped, pigment clusters were detected. Conversely, in the control group, the LECs and all of their elements were in normal ultrastructural patterns, with the exception of some small intraepithelial vacuoles which were fewer and smaller than those in the FUS group. CONCLUSION: Ultrastructural analysis of the ALC of the patients with FUS disclosed some significant alterations which may be related to the summation of oxidative stress, intraocular inflammation, and iris atrophy.

Femtosecond Laser-Assisted Capsulotomy: Histological Comparison of Four Different Laser Platforms.

PURPOSE: To compare histological analyses of capsulotomies from different laser systems with regard to tissue alteration, laser spot formation, and energy settings. METHODS: Four femtosecond laser platforms approved for cataract surgery were evaluated: LenSx (Alcon Laboratories, Inc., Fort Worth, TX), Catalys (Abbott Medical Optics, Santa Ana, CA), Victus (Tecnolas/Bausch & Lomb, Munich, Germany), and LensAR (Topcon, Gamagori, Japan). Ten human anterior lens capsulotomy disks from each laser system were analyzed: five by means of light microscopy and five by transmission electron microscopy (TEM). TEM was used especially to examine the cellular and stromal changes at the cutting edges. The cellular demarcation line was measured at 15 points along the rim and statistically compared with the findings from all laser platforms. Three capsules of each laser platform were prepared for TEM to evaluate the cellular and tissue edges of the laser cuttings. RESULTS: All capsulotomies revealed a circular demarcation zone of different sizes along the cutting edges. Light microscopy showed a radial tissue damage of 36.0 ± 12.3 µm after Victus laser capsulotomy, 8.9 ± 2.9 µm after Catalys laser capsulotomy, 25.2 ± 5.6 µm after LensAR laser capsulotomy, and 39.8 ± 5.1 µm after LenSx laser capsulotomy. Differences between the damaged tissue zones correlated with the laser systems and their energy settings. CONCLUSIONS: The use of less energy and larger spot separation led to smaller collateral damaged tissue areas along the cutting edges. The aberrant laser spots along the cutting edge may result in weakening of the remaining tissue, thereby enhancing the risk for capsular ruptures during surgery. [J Refract Surg. 2017;33(10):670-675.].

Performing Reliable Lens Capsulotomy in the Presence of Corneal Edema With a Femtosecond Laser.

Purpose: To determine the effects of the Ziemer LDV Z8 liquid interface femtosecond laser platform during capsulotomy under different energy settings in the presence of corneal edema. Methods: Cadaveric porcine eyes (n = 36) employed at less than 6 and greater than 24 post enucleation hours to simulate clear/edematous corneas, underwent capsulotomy with the Ziemer LDV Z8 femtosecond laser (5-mm diameter, energy 90%, 130%, or 150%). Lens capsules were removed for evaluation by scanning electron microscopy and rupture strengths determined by the single column universal testing system. Following ethical approval, 23 patients had lens capsules removed during routine cataract surgery following manual or Z8 capsulotomy and subjected to TUNEL assay. Results: There was no difference in edge morphology or rupture strength (120, 113, and 118 mN at increasing energy, P = 0.42) in the clear cornea. Only 50% of capsulotomies succeeded at 90% energy in an edematous cornea, improving with increased energy (75% completion at 130%, 100% at 150%). Rupture strength in edematous corneas was not significantly different at 112, 133, and 114 mN for 90%, 130%, and 150%, respectively (P = 0.3). In human samples, increased TUNEL-positive cells were seen at 130% energy, but not at 150% (0.0 manual vs. 0.2 [90%] vs. 2.1 [130%] vs. 0.6 [150%], P < 0.05). Conclusions: Because of the low energy delivered by a femtosecond nanojoule platform, even incremental increases in energy appeared to have minimal effect on lens capsule morphology and strength and negligible influence on cell death. Furthermore, increasing energy appeared to enhance consistency and the ability to complete a capsulotomy in an edematous cornea.

Anterior lens epithelium in cataract patients with retinitis pigmentosa - scanning and transmission electron microscopy study.

PURPOSE: In retinitis pigmentosa (RP) patients, relatively minor lens opacity in central part of posterior pole of the lens may cause disproportionate functional symptoms requiring cataract operation. To investigate the possible structural reasons for this opacity development, we studied the structure of the lens epithelium of patients with RP. METHODS: The anterior lens capsule (aLC: basement membrane and associated lens epithelial cells, LECs) was obtained from cataract surgery and prepared for scanning and transmission electron microscopy (SEM and TEM). RESULTS: Both SEM and TEM show a number of abnormal features in the anterior lens epithelium of cataract patients with RP. The abnormalities appear mainly as holes, thinning and degradation of the epithelium, with the dimensions from <1 µm to more than 50 µm. Other types of holes in size up to 20 µm were seen that may be formed by gradual stretching of the lens epithelium. Another type of abnormalities was cracks that were seen between adjacent LECs, with dimensions 0.1-2 µm × up to 10 µm. CONCLUSIONS: Abnormal structural features were observed in the anterior lens epithelium that may cause water influx into the lens. This may lead to clouding along the water clefts leading towards the posterior pole in the RP cataractous lens. We suggest that the lens epithelium has a role in the development of the cataract in patients with RP.

Superior Rim Stability of the Lens Capsule Following Manual Over Femtosecond Laser Capsulotomy.

PURPOSE: Cataract surgery requires the removal of a circular segment of the anterior lens capsule (LC) by manual or femtosecond laser (FL) capsulotomy. Tears in the remaining anterior LC may compromise surgical outcome. We investigated whether biophysical differences in the rim properties of the LC remaining in the patient after manual or FL capsulotomy (FLC) lead to different risks with regard to anterior tear formation. METHODS: Lens capsule samples obtained by either continuous curvilinear capsulorhexis (CCC) or FLC were investigated by light microscopy, laser scanning confocal microscopy, and scanning electron microscopy; atomic force microscopy (AFM) was used to test the biomechanical properties of the LC. The mechanical stability of the LC following either of the two capsulotomy techniques was simulated by using finite-element modeling. RESULTS: Continuous curvilinear capsulorhexis produced wedge-shaped, uniform rims, while FLC resulted in nearly perpendicular, frayed rims with numerous notches. The LC is composed of two sublayers: a stiff epithelial layer that is abundant with laminin and a softer anterior chamber layer that is predominantly made from collagen IV. Computer models show that stress is uniformly distributed over the entire rim after CCC, while focal high stress concentrations are observed in the frayed profiles of LC after FLC, making the latter procedure more prone to anterior tear formation. CONCLUSIONS: Finite-element modeling based on three-dimensional AFM maps indicated that CCC leads to a capsulotomy rim with higher stress resistance, leading to a lower propensity for anterior radial tears than FLC.

Precision Pulse Capsulotomy: Preclinical Safety and Performance of a New Capsulotomy Technology.

PURPOSE: To assess the preclinical safety and performance of a new precision pulse capsulotomy (PPC) method. DESIGN: Human cadaver eye studies and surgical, slit-lamp, and histopathologic evaluation in a consecutive series of 20 live rabbits. PARTICIPANTS: Human cadaver eyes and New Zealand white rabbits. METHODS: Precision pulse capsulotomy uses a highly focused, fast, multipulse, low-energy discharge to produce a perfectly round anterior capsulotomy instantaneously and simultaneously along all 360°. Capsulotomies are performed using a disposable handpiece with a soft collapsible tip and circular nitinol cutting element. Miyake-Apple imaging and scanning electron microscopy (SEM) of PPC were conducted in human cadaver eyes. Surgical, postoperative slit-lamp, and histopathologic assessments of PPC were performed in 20 live rabbits and were compared with manual continuous curvilinear capsulorrhexis (CCC) in the fellow eye. Anterior chamber (AC) thermocouple temperature measurements were evaluated in a subset of rabbit eyes. MAIN OUTCOME MEASURES: Capsulotomy edge circularity, SEM morphologic features and zonular movement with PPC in human cadaver eyes. Anterior chamber temperature during PPC and grading of ocular inflammation, corneal endothelial damage, anterior capsular opacification (ACO), and posterior capsular opacification (PCO). RESULTS: Miyake-Apple imaging showed minimal zonular stress, and thermocouple measurements demonstrated negligible AC temperature changes during PPC. Precision pulse capsulotomy produced round, complete capsulotomies in all 20 rabbit eyes, leading to successful in-the-bag intraocular lens (IOL) implantation. Slit-lamp examinations at 3 days and 1, 2, and 4 weeks after surgery showed no significant differences between PPC and CCC in corneal edema, AC inflammatory reaction, capsular fibrosis, ACO, and PCO. Postmortem studies showed no difference in the corneal endothelium between PPC and CCC eyes. All IOLs were well centered in PPC eyes, and histopathologic analysis showed no greater inflammatory infiltrates. CONCLUSIONS: Precision pulse capsulotomy is a new method to automate consistent creation of a perfectly circular anterior capsulotomy with a disposable handheld instrument that can be used in the normal phacoemulsification surgical sequence. Compared with CCC in fellow rabbit eyes, PPC was equally safe and showed no greater zonular stress compared with CCC in human cadaver eyes. Human cadaver eye SEM showed a much smoother capsulotomy edge compared to those produced by femtosecond laser.

Anterior lens epithelium in intumescent white cataracts - scanning and transmission electron microscopy study.

PURPOSE: Our purpose was to study the structure of the lens epithelial cells (LECs) of intumescent white cataracts (IC) in comparison with nuclear cataracts (NC) in order to investigate possible structural reasons for development of IC. METHODS: The anterior lens capsule (aLC: basement membrane and associated LECs) were obtained from cataract surgery and prepared for scanning electron microscopy (SEM) and transmission electron microscopy (TEM). RESULTS: We observed by SEM that in IC, LEC swelling was pronounced with the clefts surrounding the groups of LECs. Another structural feature was spherical formations, that were observed on the apical side of LEC's, towards the fibre cell layer, both by SEM and TEM. Development of these structures, bulging out from the apical cell membrane of the LEC's and disrupting it, could be followed in steps towards the sphere formation. The degeneration of the lens epithelium and the structures of the aLC in IC similar to Morgagnian globules were also observed. None of these structural changes were observed in NC. CONCLUSIONS: We show by SEM and TEM that, in IC, LECs have pronounced structural features not observed in NC. This supports the hypothesis that the disturbed structure of LECs plays a role in water accumulation in the IC lens. We also suggest that, in IC, LECs produce bulging spheres that represent unique structures of degenerated material, extruded from the LEC.

Anterior lens epithelial cells attachment to the basal lamina.

PURPOSE: To study the structure of the anterior lens epithelial cells (aLECs) and the contacts of the aLECs with the basal lamina (BL) in order to understand their role in the lens epithelium's function. METHODS: The aLCs (BL and associated aLECs) were obtained from routine uneventful cataract surgery, prepared for and studied by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and confocal microscopy. RESULTS: SEM shows that the basal surface of the aLECs (~10-15 µm) is with aLECs foldings (~1-3 µm) and extensions (~0.5-3 µm) attached to the BL. Confocal microscopy images of the basal sections of the aLECs after membrane staining also suggest that the basal part of aLECs has foldings (~1-3 µm). TEM shows in the aLECs basal parts, towards BL, the structures that look like entanglement (~1-4 µm). In cases where there is a swelling of the cytoplasm and offset of the aLECs from the BL, individual extensions (~0.5-2 µm) that extend to the BL are visible by TEM. CONCLUSIONS: We provide detail evidence about the structural organization of the aLECs, in particular about their basal side which is in contact with the BL. This is supported by the complementary use of three techniques, SEM, TEM and confocal microscopy, each of them showing the same morphological features, the extensions and the entanglements of the aLECs cytoplasmic membrane at the border with the BL. The basal surface of the aLECs is increased. It suggests the functional importance of the contact between aLECs and BL.

Evaluation of the mechanical properties of the anterior lens capsule following femtosecond laser capsulotomy at different pulse energy settings.

PURPOSE: To evaluate and compare the mechanical properties of anterior capsule opening performed with femtosecond laser capsulotomy at different energy settings in ex vivo porcine anterior lens capsule specimens. METHODS: Twenty-five fresh porcine eyes per group were included in the study. Femtosecond laser capsulotomy was performed with three different pulse energy levels: 2 µJ (low energy group), 5 µJ (intermediate energy group), and 10 µJ (high energy group). The capsule openings were stretched with universal testing equipment until they ruptured. The morphologic profile of the cut capsule edges was evaluated using scanning electron microscopy. RESULTS: The high energy group had significantly lower rupture force (108 ± 14 mN) compared to the intermediate energy group (118 ± 10 mN) (P < .05) and low energy group (119 ± 11 mN) (P < .05), but the difference between the intermediate energy and low energy groups was not significant (P = .9479). The high energy group had significantly lower circumference stretching ratio (144% ± 3%) compared to the intermediate energy group (148% ± 3%) (P < .05) and low energy group (148% ± 3%) (P < .05), but the difference between the intermediate energy group and low energy group was not significant (P = .9985). Scanning electron microscopy images showed that the edge was only serrated with low and intermediate energy, but additional signs of collagen melting and denaturation were observed at high energy. CONCLUSIONS: Anterior capsule openings created at a high energy level were slightly weaker and less extensible than those created at low or intermediate levels, possibly due to the increased thermal effect of photo-disruption.

The use of Pentacam in the assessment of true exfoliation of the lens capsule.

PURPOSE: To highlight the use of Pentacam in the identification of true exfoliation of the lens capsule. METHODS: A 72-year-old woman presented for assessment of bilateral cataracts. An unusual appearance was noted in the right eye. A scrolled membrane was seen floating in the anterior chamber. RESULTS: Imaging with Pentacam showed its origin to be from the anterior lens capsule, suggesting the diagnosis of true exfoliation of the lens capsule. The patient went on to have uncomplicated cataract surgery and the lens capsule was sent for histopathology. CONCLUSIONS: This case highlights the use of anterior segment imaging, the classic histopathology findings, and the management of cataract surgery in these rare cases.