Effects of Long-Chain Non-Coding Ribonucleic Acid Hox Tran Antisense RNA on Proliferation and Migration of Epithelial Cells of Cataract Lens

In order to explore effects of long-chain non-coding ribonucleic acid (RNA) HOTAIR on proliferation and migration of human lens epithelial cells, SRA01/04 cells were selected as the research strain in this study and divided into S1 group (no HOTAIR transfection), S2 group (siHOTAIR transfection), S3 group (siHOTAIR+10 ng/mL TGF-β2), and S4 group (no HOTAIR transfection+10 ng/mL TGF-β2) according to the presence or absence of transforming growth factor (TGF)-β2 and silent HOTAIR treatment. 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) colorimetric method was applied to detect cell proliferation.Western blot was used for detection of E-cadherin, zonula occluden-1 (ZO-1), Vimentin, α-smooth muscle actin (SMA), Snail, Slug, zinc finger E-box binding homeobox 1 (ZEB1), and Smad-2 expressions. Results showed that the number of transmembrane cells in S4 group was higher markedly than that of the other groups, but that of S2 group dropped steeply compared with the other groups (P <0.05); E-cadherin (2.59±0.58) and ZO-1 (1.95±0.56) of S2 group increased hugely compared with the other groups, while Vimentin (0.57±0.14) and α-SMA (0.64±0.28) decreased sharply compared with the other groups (P < 0.05); Snail (2.51±0.59), Slug (2.11±0.47), and ZEB1 (2.83±0.53) of S4 group rose obviously compared with the other groups, but the above of S2 group reduced hugely compared with the other groups (P < 0.05); pSmad-2 and pSmad-3 of S4 group elevated greatly compared with the other groups, and those of S2 group reduced hugely compared with the other groups (P < 0.05). In conclusion, HOTAIR high expression could promote TGF-β2-induced SRA01/04 cell proliferation, migration, invasion, and epithelial-mesenchymal trans-differentiation, which was related to TGF-β/Smad signaling pathway.

In Vivo Quasi-Elastic Light Scattering Eye Scanner Detects Molecular Aging in Humans and Mice

The absence of clinical tools to evaluate individual variation in the pace of aging represents a major impediment to understanding aging and maximizing health throughout life. The lens is an ideal tissue for quantitative assessment of molecular aging in vivo. Long-lived proteins in lens fiber cells are expressed during fetal life, do not undergo turnover, accumulate molecular alterations throughout life, and are optically accessible in vivo. We used quasi-elastic light scattering (QLS) to measure age-dependent signals in lenses of both healthy human subjects and wild-type C57BL/6 mice. Age-dependent QLS signal changes detected in vivo in humans and mice recapitulated time-dependent changes in hydrodynamic radius, protein polydispersity, and supramolecular order of human lens proteins during long-term incubation (~1 year) and in response to sustained oxidation (~2.5 months) in vitro. Our findings demonstrate that QLS analysis of lens proteins provides a practical technique for noninvasive assessment of molecular aging in vivo.

Characterisation and Modelling of an Artificial Lens Capsule Mimicking Accommodation of Human Eyes

A synthetic material of silicone rubber was used to construct an artificial lens capsule (ALC) in order to replicate the biomechanical behaviour of human lens capsule. The silicone rubber was characterised by monotonic and cyclic mechanical tests to reveal its hyper-elastic behaviour under uniaxial tension and simple shear as well as the rate independence. A hyper-elastic constitutive model was calibrated by the testing data and incorporated into finite element analysis (FEA). An experimental setup to simulate eye focusing (accommodation) of ALC was performed to validate the FEA model by evaluating the shape change and reaction force. The characterisation and modelling approach provided an insight into the intrinsic behaviour of materials, addressing the inflating pressure and effective stretch of ALC under the focusing process. The proposed methodology offers a virtual testing environment mimicking human capsules for the variability of dimension and stiffness, which will facilitate the verification of new ophthalmic prototype such as accommodating intraocular lenses (AIOLs).

Exosomal miR-29b found in aqueous humour mediates calcium signaling in diabetic patients with cataract

AIM: To investigate the role of exosomal miR-29b and Ca2+ in regulating the function of human lens epithelial cells (HLECs). METHODS: Exosomes were isolated from human aqueous humour (AH) by ultracentrifugation, and visualized by nanoparticle tracking and transmission electron microscopy. Exosomal miRNA sequencing was performed to identify differentially expressed miRNAs between diabetes with cataracts (DMC) group and age-related cataracts (ARC) group. TargetScan was used to predict potential target of certain miRNA. The expression of CACNA1C mRNA was determined by quantitative real-time polymerase chain reaction and CACNA1C protein was determined by Western blotting. Concentration of Ca2+ in human AH and the culture supernatant of cells were detected by the calcium assay kit. Cell counting kit-8 was used to determine cell viability. RESULTS: Exosomes were isolated from human AH, which had a typical cup-shaped phenotype and a particle size distribution in accordance with micro extracellular vesicles. Exosomal miRNA sequencing revealed that miR-29b was significantly downregulated in DMC group compared with ARC. Ca2+ concentration of human AH in DMC was higher than that in ARC. The culture supernatant of cells transfected with miR-29b inhibitors had a higher concentration of Ca2+ than that transfected with miR-29b mimics. miR-29b reduced the viability of HLECs by upregulating CACNA1C expression. CONCLUSION: Exosomes isolated from human AH contains abundant miRNAs. A significantly expressed miRNA, miR-29b, can affect the concentration of Ca2+ and regulate HLEC processes by upregulating CACNA1C.

Quantitative Evaluation of Human Lens Epithelial Cell Viability and Cytolysis by Distilled Water Ex Vivo

Purpose: To investigate the role of distilled water (DW) in isolated human lens epithelial cells (LECs) viability and lysis ex vivo.Methods: After immersion in DW or balanced salt solution (BSS) for 1-, 2-, and 3-minutes, respectively, the cell viability of LECs was quantitatively evaluated. In addition, the capsule samples soaked in DW or BSS for 1-, 2-, and 3-minutes were combined with rinse for 1 minute to analyze the difference of LECs shedding percentage in each subgroup. The histopathological changes of the samples after treating were observed.Results: The percentage of LECs shed in DW immersion combined with rinse was significantly higher than in DW immersion alone (p all <0.001). In the subgroup soaked in DW for 3 minutes, the death number, mortality, and the percentage of cell shedding of LECs was the most (p all <0.001). The histopathological changes showed that the cell destruction in the DW subgroup for 1-, 2-, and 3-minutes, and the transmission electron microscope results showed that the cells were partially detached from the capsule in the DW 3 minutes subgroup.Conclusions: Soaking in the DW can cause LECs death, and DW immersion combined with rinse was an effective method to remove LECs. The histopathology changes of treated DW suggested cellular necrosis was one type of LECs death mechanism.