Morphological observations of rat corneal endothelial cells after exposure to ozonated solution

Suzuki, Hajime; Satō, Shigeru; Murano, N.; Matsui, Hironori; Oharazawa, Hideaki; Takahashi, Hiroshi

doi:10.1007/s10384-008-0629-4

Cited by 6 publications

(5 citation statements)

References 30 publications

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“…These activities of toxicant-induced damage related to enzymes and DNA fragmentations can be detected by electrophoresis or enzyme-linked immunoassay techniques. 15 The major goal of this study was to attain and develop a new bioanalytical method capable of continuous real-time monitoring without any reagent addition. The present model was used to reveal an unconventional method for monitoring photodamaging effects in retinal cells by assessing light responses in vitro.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Other existing methods to monitor toxicant-induced stress and cells undergoing changes were established by radioactive labeling, which is assessed by microscopy and biochemical assays. These activities of toxicant-induced damage related to enzymes and DNA fragmentations can be detected by electrophoresis or enzyme-linked immunoassay techniques . The major goal of this study was to attain and develop a new bioanalytical method capable of continuous real-time monitoring without any reagent addition.…”

Section: Results and Discussionmentioning

confidence: 99%

“…However, the potential role of these disorders is neither well understood nor explored. Several studies have been conducted to obtain qualitative results of light-induced stress-related changes. − Toxicity, cell viability, cell proliferation, and apoptosis analysis methods are frequently used in high-throughput screening, ,,− although they are not suitable for continuous or real-time analysis.…”

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confidence: 99%

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Impedance-Based Cell Culture Platform To Assess Light-Induced Stress Changes with Antagonist Drugs Using Retinal Cells

Bennet

Kim

2013

Anal. Chem.

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This Article describes an unprecedented, simple, and real-time in vitro analytical tool to measure the luminous effect on the time responses function of retinal ganglion cells (RGC-5) by electric cell substrate impedance sensing (ECIS) system. The ECIS system was used for the continuous measurement of different color light-induced effects on the response of cells that exposed to protective drugs. The measurement suggests that the association of photo-oxidative stress was mediated by reactive oxygen species (ROS), which plays a critical role that leads to cell stress, damages, and retinopathy, resulting in eye degenerative diseases. Continuous light radiation caused time-dependent decline of RGC-5 response and resulted in photodamage within 10 h due to adenosine 5'-triphosphate depletion and increased ROS level, which is similar to in vivo photodamage. The ECIS results were correlated with standard cell viability assay. ECIS is very helpful to determine the protective effects of analyzed drugs such as β-carotene, quercetin, agmatine, and glutathione in RGC-5 cells, and the maximum drug activity of nontoxic safer drug concentrations was found to be 0.25, 0.25, 0.25, and 1.0 mM, respectively. All drugs show protection against light radiation toxicity in a dose-dependent manner; the most effective drug was found to be glutathione. The proposed system identifies the phototoxic effects in RGC-5 and provides high throughput drug screening for photo-oxidative stress during early stages of drug discovery. This study is convenient and potential enough for the direct measurements of the photoprotective effect in vitro and would be of broad interest in the field of therapeutics.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Results and Discussionmentioning

confidence: 99%

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confidence: 99%

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Impedance-Based Cell Culture Platform To Assess Light-Induced Stress Changes with Antagonist Drugs Using Retinal Cells

Bennet

Kim

2013

Anal. Chem.

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“…ADSCs revealed polygonal morphology from spindle shape after our sequential non-genetic reprogramming treatment, co-culture with R-CECs and seeded onto decellularized corneal stroma. At the same time, induced cells converted to weakly express CD31, AQP-1 and ZO-1, which were endothelial markers [55] , [56] . Wu’s laboratory demonstrated that adult rat CEC-derived conditioned medium could induce rat neural crest cells (NCCs) to differentiate into polygonal CEC-like cells.…”

Section: Discussionmentioning

confidence: 94%

Non-Genetic Direct Reprogramming and Biomimetic Platforms in a Preliminary Study for Adipose-Derived Stem Cells into Corneal Endothelia-Like Cells

et al. 2014

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Cell fate and function can be regulated and reprogrammed by intrinsic genetic program, extrinsic factors and niche microenvironment. Direct reprogramming has shown many advantages in the field of cellular reprogramming. Here we tried the possibility to generate corneal endothelia (CE) -like cells from human adipose-derived stem cells (ADSCs) by the non-genetic direct reprogramming of recombinant cell-penetrating proteins Oct4/Klf4/Sox2 (PTD-OKS) and small molecules (purmorphamine, RG108 and other reprogramming chemical reagents), as well as biomimetic platforms of simulate microgravity (SMG) bioreactor. Co-cultured with corneal cells and decellularized corneal ECM, Reprogrammed ADSCs revealed spherical growth and positively expressing Nanog for RT-PCR analysis and CD34 for immunofluorescence staining after 7 days-treatment of both purmorphamine and PTD-OKS (P-OKS) and in SMG culture. ADSCs changed to CEC polygonal morphology from spindle shape after the sequential non-genetic direct reprogramming and biomimetic platforms. At the same time, induced cells converted to weakly express CD31, AQP-1 and ZO-1. These findings demonstrated that the treatments were able to promote the stem-cell reprogramming for human ADSCs. Our study also indicates for the first time that SMG rotary cell culture system can be used as a non-genetic means to promote direct reprogramming. Our methods of reprogramming provide an alternative strategy for engineering patient-specific multipotent cells for cellular plasticity research and future autologous CEC replacement therapy that avoids complications associated with the use of human pluripotent stem cells.

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“…Ethanol is used to disinfect the skin but is not suitable for disinfection of the cornea [44]. Ozonated solution (4 ppm) is effective for disinfecting the eye surface [45,46], but has several issues including short half-life, need for ozone generator and damaging to corneal endothelial cells after exposure for over 10 s [47].…”

Section: Povidone-iodine As a "Border Control" Measurementioning

confidence: 99%

Cataract Surgery by Intraoperative Surface Irrigation with 0.25% Povidone–Iodine

Shimada

Nakashizuka

2021

JCM

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Postoperative endophthalmitis after cataract surgery is typically caused by the patient’s own conjunctival normal bacterial flora. A three-step approach is recommended to prevent endophthalmitis: (1) “border control” to prevent microorganisms from entering the eye by disinfecting the ocular surface is the most important measure; (2) bacteria that have gained access into the anterior chamber are reduced by irrigation; (3) bacteria remaining in the anterior chamber and vitreous at the end of surgery are controlled by antibacterial drugs. We have devised a method, “the Shimada technique”, for irrigating the ocular surface with povidone-iodine, a disinfectant with potent microbicidal effect and established effective and safe concentrations for eye tissues. Povidone-iodine exhibits a bactericidal effect for a wide concentration range of 0.005–10%, but 0.1% povidone-iodine has the highest activity and requires the shortest time of only 15 s to achieve microbicidal effect. When used to irrigate the ocular surface every 20–30 s during cataract surgery, 0.25% povidone-iodine is conceivably diluted to around 0.1%. Irrigation with 0.25% povidone-iodine during cataract surgery significantly reduced bacteria contamination rate in the anterior chamber compared with saline (p = 0.0017) without causing corneal endothelial damage.

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Morphological observations of rat corneal endothelial cells after exposure to ozonated solution

Abstract: A short period of irrigation of the anterior chamber with ozonated solution does not harm the corneal endothelium even when used in combination with ascorbic acid.

Cited by 6 publications

References 30 publications

Impedance-Based Cell Culture Platform To Assess Light-Induced Stress Changes with Antagonist Drugs Using Retinal Cells

Impedance-Based Cell Culture Platform To Assess Light-Induced Stress Changes with Antagonist Drugs Using Retinal Cells

Non-Genetic Direct Reprogramming and Biomimetic Platforms in a Preliminary Study for Adipose-Derived Stem Cells into Corneal Endothelia-Like Cells

Cataract Surgery by Intraoperative Surface Irrigation with 0.25% Povidone–Iodine

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