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The idea of transplanting a sheet of laboratory-grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial tissue engineering. LCs were isolated from donor eyes, stored at -80 °C, and decellularized with water and trypsin-EDTA. The decellularization was investigated by nuclear staining and counting and the capsule thickness was determined by optical coherence tomography and compared with Descemet's membrane (DM). Transparency was examined by spectrometry, and collagenase degradation was performed to evaluate its resistance to degradation. Cell-scaffold interaction was assessed by measuring focal adhesions surface area on LC and plastic. Finally, primary corneal endothelial cells were grown on LCs to validate the phenotype. Trypsin-EDTA decellularized most effectively, removing 99% of cells. The mean LC thickness was 35.76 ± 0.43 μm, whereas DM measured 25.93 ± 0.26 μm (p < .0001). Light transmission was 90% for both LC and DM. On a collagenase challenge, LC and amniotic membrane were digested after 13 hr, whereas DM was digested after 17 hr. The surface area of focal adhesions for cells grown on coated LCs was at least double that compared with other conditions, whereas tight junctions, ion pumps, and hexagonal morphology were well maintained when endothelial cells were cultured on LCs. In conclusion, LCs demonstrate excellent scaffolding properties for tissue engineering and sustain the cell phenotype and can be considered a suitable substrate for ocular tissue engineering or as a template for future scaffolds.
The current study reports comparing the postoperative mechanical properties of the anterior capsule between femtosecond laser capsulotomy (FLC) and continuous curvilinear capsulorhexis (CCC) of variable size and shape in porcine eyes. All CCCs were created using capsule forceps. Irregular or eccentric CCCs were also created to simulate real cataract surgery. For FLC, capsulotomies 5.3 mm in diameter were created using the LenSx® (Alcon) platform. Fresh porcine eyes were used in all experiments. The edges of the capsule openings were pulled at a constant speed using two L-shaped jigs. Stretch force and distance were recorded over time, and the maximum values in this regard were defined as those that were recorded when the capsule broke. There was no difference in maximum stretch force between CCC and FLC. There were no differences in circularity between FLC and same-sized CCC. However, same-sized CCC did show significantly higher maximum stretch forces than FLC. Teardrop-shaped CCC showed lower maximum stretch forces than same-sized CCC and FLC. Heart-shaped CCC showed lower maximum stretch forces than same-sized CCC. Conclusively, while capsule edge strength after CCC varied depending on size or irregularities, FLC had the advantage of stable maximum stretch forces.
For more than a century there has been debate concerning the mechanism of accommodation--whether the lens capsule or lens material itself determines the functional relationship between ciliary muscle contractility and lens deformation during refractive adaptation. This morphological study in monkey eyes investigates the composition and distribution of several connective tissue components in the accommodative apparatus relaying muscle force to lens organization. Elastin distributes on the marginal surface of the ciliary process. A zonule is composed of fibrillin produced by epithelial cells of the process. In the progress of extension over the posterior chamber, fibrils unite into strands and possess longitudinal plasticity. By induction of the elastin network, strands extend in a concentric direction covering the equatorial region of the capsule. Upon tethering to the lens, the strand ramifies into fibrils, penetrating deeply close to the epithelial layer of the lens and binding with the collagen of the intercellular spaces. Tight linkage of the zonule with the capsule transmits precise contractility. Inside the lens, the cortical layer's elastic connective tissue network forms widely spaced lamellae of crystalline fibers. In contrast, the central nuclear lamellae are tightly opposed. The accumulation of lamellae is greater in the anterior cortex than in the posterior, yielding a more variable anterior chamber depth in the visual axis. The plasticity of the zonule and connective tissue distribution inside the lens produces an adjustable configuration. Thus, tight linkage between the dynamism of the capsule with interaction of the lenticular flexibility provides a novel understanding of accommodation.
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