Role of Oxidative Stress in the Disruption of the Endothelial Apical Junctional Complex During Corneal Cold Storage

Thanuja, M. Y.; Ranganath, Sudhir H.; Srinivas, Sangly P.

doi:10.1089/jop.2022.0082

Cited by 5 publications

(4 citation statements)

References 61 publications

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“…Both P38 MAPK inhibitors, SB203580 and SB202191, demonstrated excellent efficacy in increasing the ECD by 59% and 65%, respectively, while also maintaining the differentiated state of hCECs. SB203580 was reported for its protection of CECs’ barrier function in vitro and ex vivo [ 79 , 80 ] and its numerous beneficial effects on hCEC culture through anti-senescence mechanisms [ 38 ].…”

Section: Discussionmentioning

confidence: 99%

Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture

Aouimeur

Sagnial

Coulomb

et al. 2023

Cells

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Corneal endothelial diseases are the leading cause of corneal transplantation. The global shortage of donor corneas has resulted in the investigation of alternative methods, such as cell therapy and tissue-engineered endothelial keratoplasty (TEEK), using primary cultures of human corneal endothelial cells (hCECs). The main challenge is optimizing the hCEC culture process to increase the endothelial cell density (ECD) and overall yield while preventing endothelial–mesenchymal transition (EndMT). Fetal bovine serum (FBS) is necessary for hCEC expansion but contains TGF-βs, which have been shown to be detrimental to hCECs. Therefore, we investigated various TGF-β signaling pathways using inhibitors to improve hCEC culture. Initially, we confirmed that TGF-β1, 2, and 3 induced EndMT on confluent hCECs without FBS. Using this TGF-β-induced EndMT model, we validated NCAM as a reliable biomarker to assess EndMT. We then demonstrated that, in a culture medium containing 8% FBS for hCEC expansion, TGF-β1 and 3, but not 2, significantly reduced the ECD and caused EndMT. TGF-β receptor inhibition had an anti-EndMT effect. Inhibition of the ROCK pathway, notably that of the P38 MAPK pathway, increased the ECD, while inhibition of the ERK pathway decreased the ECD. In conclusion, the presence of TGF-β1 and 3 in 8% FBS leads to a reduction in ECD and induces EndMT. The use of SB431542 or LY2109761 may prevent EndMT, while Y27632 or Ripasudil, and SB203580 or SB202190, can increase the ECD.

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Section: Discussionmentioning

confidence: 99%

Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture

Aouimeur

Sagnial

Coulomb

et al. 2023

Cells

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“…Although cold storage has been shown to damage the junctional complex of the endothelium, the unavailability of corneas preserved at 30 °C from our research's tissue-supplying eye banks limits our experimental scope for investigating the effect of hypothermic storage in this study. Considering that the cornea is composed of five primary layers, interactions between these layers are crucial for vision focus . As we obtained only the stromal layer, limitations arise in investigating interlayer crosstalk.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

“…Considering that the cornea is composed of five primary layers, interactions between these layers are crucial for vision focus. 70 As we obtained only the stromal layer, limitations arise in investigating interlayer crosstalk. Future proteomic analysis of completed corneal tissue can deepen our understanding of the molecular interaction with corneal tissues.…”

Section: ■ Conclusion and Discussionmentioning

confidence: 99%

Comparative Proteomics Reveals Prolonged Corneal Preservation Impaired Ocular Surface Immunity Accompanied by Fibrosis in Human Stroma

Wu,

Liu,

Wong

et al. 2023

J. Proteome Res.

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Cornea transplantation is one of the most commonly performed allotransplantations worldwide. Prolonged storage of donor corneas leads to decreased endothelial cell viability, severe stromal edema, and opacification, significantly compromising the success rate of corneal transplantation. Corneal stroma, which constitutes the majority of the cornea, plays a crucial role in maintaining its shape and transparency. In this study, we conducted proteomic analysis of corneal stroma preserved in Optisol-GS medium at 4 °C for 7 or 14 days to investigate molecular changes during storage. Among 1923 identified proteins, 1634 were quantifiable and 387 were significantly regulated with longer preservation. Compared to stroma preserved for 7 days, proteins involved in ocular surface immunomodulation were largely downregulated while proteins associated with extracellular matrix reorganization and fibrosis were upregulated in those preserved for 14 days. The increase in extracellular matrix structural proteins together with upregulation of growth factor signaling implies the occurrence of stromal fibrosis, which may compromise tissue clarity and cause vision impairments. This study is the first to provide insights into how storage duration affects corneal stroma from a proteomic perspective. Our findings may contribute to future research efforts aimed at developing long-term preservation techniques and improving the quality of preserved corneas, thus maximizing their clinical application.

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“…Specically, we have shown that exposure to hypothermia induces microtubule disassembly and disruption of the perijunctional actomyosin ring (PAMR) and leads to a loss of barrier function in cultured corneal endothelial cells and ex vivo corneal endothelium. 9,14 In addition, TNFa (tumor necrosis factor-a), a pro-inammatory cytokine secreted during allogra rejection, induces disassembly of microtubules and loss of barrier function. 10 Pharmacological pre-treatment of the donor corneal endothelium with microtubule stabilizers, 11 Rho kinase inhibitors, 15 cold stress protectants, 16 anti-apoptotic genes 17 and their sustained intracellular delivery are imperative to preserve its functionality (barrier function), cell density and tackle potential adverse effects induced by hypothermia during storage and cytokine stress aer transplantation.…”

Section: Introductionmentioning

confidence: 99%

Fusogenic liposome-coated nanoparticles for rapid internalization into donor corneal endothelial tissue to enable prophylaxis before transplantation

M. Y.,

Tellakula,

Suryavanshi

et al. 2023

Nanoscale Adv.

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Role of Oxidative Stress in the Disruption of the Endothelial Apical Junctional Complex During Corneal Cold Storage

Cited by 5 publications

References 61 publications

Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture

Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture

Comparative Proteomics Reveals Prolonged Corneal Preservation Impaired Ocular Surface Immunity Accompanied by Fibrosis in Human Stroma

Fusogenic liposome-coated nanoparticles for rapid internalization into donor corneal endothelial tissue to enable prophylaxis before transplantation

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