A Novel PorcineEx VivoRetina Culture Model for Oxidative Stress Induced by H2O2

Hurst, José; Kuehn, Sandra; Jashari, Adelina; Tsai, Teresa; Bartz‐Schmidt, Karl Ulrich; Schnichels, Sven; Joachim, Stephanie C.

doi:10.1177/026119291704500105

Cited by 39 publications

(71 citation statements)

References 67 publications

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“…39 During research studies on RGCs, ex vivo cultures have been shown to remain viable for 7-10 days, which has facilitated the study of molecular mechanisms involved in disease. [83][84][85][86] A better awareness of the interactions between the retinal pigment epithelium and neural retina could provide a basis for investigating the role of signalling molecules or for testing the effects of potential new therapies. 39,[87][88][89] The adult porcine retina can be used as a suitable model for investigating photoreceptor-specific degeneration processes and enhancing the understanding of the mechanisms playing a role in degenerative neuronal cell death.…”

Section: Rabbitsmentioning

confidence: 99%

Retinal Organ Cultures as Alternative Research Models

et al. 2019

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Ex vivo organ cultures represent unique research models, as they combine the advantages of cell cultures with those of animal models. Being able to mimic in vivo situations through the use of organ cultures provides an excellent opportunity to investigate cellular processes, molecular pathways and cell–cell interactions, as well as structural and synaptic organisation. Human and animal organ cultures are now well established and comprise sensitive, easy-to-manipulate experimental systems that raise minimal ethical concerns. The eye, in particular, is a very complex organ that is not easy to reproduce in vitro. However, a lot of research has been dedicated to the development of suitable ocular organ cultures. This review covers the various ex vivo retinal organ culture systems available for use in ophthalmology research and compares them with commonly used animal models. In particular, bovine and porcine retinal organ culture systems are described, because the size, anatomy, physiology and vessel morphology of bovine and porcine eyes are similar to the human eye in an undisputed way, thus making them good models. In addition, these animals are widely used by the food industry and the eyes are considered surplus material. A short overview of murine, rat, rabbit, cat, canine and simian retinal organ cultures is also provided.

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Section: Rabbitsmentioning

confidence: 99%

Retinal Organ Cultures as Alternative Research Models

et al. 2019

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“…In the last years, we developed retinal organ culture models, to test new therapies without euthanasia of laboratory animals . These models are based on porcine retinae, which can be obtained as a waste product from the food industry.…”

Section: Introductionmentioning

confidence: 99%

iNOS‐inhibitor driven neuroprotection in a porcine retina organ culture model

Hurst

Mueller‐Buehl

Hofmann

et al. 2020

J Cellular Molecular Medi

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Since 1995, 11 million animals per year were used for research purposes in the EU, an average of 2.5 million in Germany alone, with an upward trend. 1 Ophthalmological research also frequently uses laboratory animals. An important field of research is the understanding of retinal degeneration, as its causes are not yet fully understood preventing successful treatment.In the last years, we developed retinal organ culture models, to test new therapies without euthanasia of laboratory animals. [2][3][4][5] These models are based on porcine retinae, which can be obtained as a waste product from the food industry. The morphology and AbstractNitrite oxide plays an important role in the pathogenesis of various retinal diseases, especially when hypoxic processes are involved. This degeneration can be simulated by incubating porcine retinal explants with CoCl 2 . Here, the therapeutic potential of iNOS-inhibitor 1400W was evaluated. Degeneration through CoCl 2 and treatment with the 1400W were applied simultaneously to porcine retinae explants.Three groups were compared: control, CoCl 2 , and CoCl 2 + iNOS-inhibitor (1400W).At days 4 and 8, retinal ganglion cells (RGCs), bipolar, and amacrine cells were analysed. Furthermore, the influence on the glia cells and different stress markers were evaluated. Treatment with CoCl 2 resulted in a significant loss of RGCs already after 4 days, which was counteracted by the iNOS-inhibitor. Expression of HIF-1α and its downstream targets confirmed the effective treatment with 1400W. After 8 days, the CoCl 2 group displayed a significant loss in amacrine cells and also a drastic reduction in bipolar cells was observed, which was prevented by 1400W. The decrease in microglia could not be prevented by the inhibitor. CoCl 2 induces strong degeneration in porcine retinae by mimicking hypoxia, damaging certain retinal cell types.Treatment with the iNOS-inhibitor counteracted these effects to some extent, by preventing loss of retinal ganglion and bipolar cells. Hence, this inhibitor seems to be a very promising treatment for retinal diseases. K E Y W O R D S hypoxia, iNOS-inhibitor 1400W, organ culture, retina, retinal ganglion cells | 4313 HURST eT al. How to cite this article: Hurst J, Mueller-Buehl AM, Hofmann L, et al. iNOS-inhibitor driven neuroprotection in a porcine retina organ culture model. J Cell Mol Med. 2020;24:4312-4323. https://doi.

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“…Lipid peroxidation is regarded as a vital marker for the toxicity induced by oxidative stress. It has been demonstrated that exogenous treatment with hydrogen peroxide increases the levels of LPO, even at low doses the treatments can cause necrosis and apoptosis of brain cells [16,17] suggesting the cellular injury by the action of the free radicals. In our study treatment of brain cultures with hydrogen peroxide resulted in significant high levels of the LPO when compared to the control.…”

Section: Discussionmentioning

confidence: 99%

Neuroprotective effect of quercetin in murine cortical brain tissue cultures

Haq

Alamro

2019

Clinical Nutrition Experimental

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Purpose: Quercetin (QR) is bioflavonoids known for its antioxidants property and its ability to alleviate oxidative stress and promote cellular survival. The aim of this study was to explore the neuroprotective potential of QR against the induced oxidative stress. Methods: Cortical brain tissue cultures from one week old Wister rats were set up in four groups. Group 1: the control group without any treatments; Group 2: cortical cultures treated with 1 mM H 2 O 2 for one hour; Group 3: Cortical cultures pretreated with varying doses of QR for 24 h followed by treatment with 1 mM H 2 O 2 for 1 h; Group 4: Cortical cultures treated with the vehicle alone (DMSO). The cortical Tissues from all four groups were homogenized, both the homogenized cortical tissues and conditioned medium was used for the biochemical assay. For histology studies cortical brain tissue were fixed in 10% formalin and stained with H&E. Results: Pretreatment with 100 mg/ml showed the optimum concentration, which completely ameliorates the effect of induced oxidative stress by H 2 O 2. All of the biochemical markers of oxidative stress such as lipid peroxidation, GST enzyme assay, DNA damage and fragmentation were completely reversed with the pretreatments of QR. Histology of the cortical tissues further confirmed the biochemical assays as it showed the pretreatments with QR resulted in the neuronal survival and viability. Conclusions: This study further reiterated the neuroprotective role of QR against oxidative stress-related neurodegenerative disorders.

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A Novel PorcineEx VivoRetina Culture Model for Oxidative Stress Induced by H₂O₂

Cited by 39 publications

References 67 publications

Retinal Organ Cultures as Alternative Research Models

Retinal Organ Cultures as Alternative Research Models

iNOS‐inhibitor driven neuroprotection in a porcine retina organ culture model

Neuroprotective effect of quercetin in murine cortical brain tissue cultures

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