Therapeutic efficiency of tissue-engineered human corneal endothelium transplants on rabbit primary corneal endotheliopathy

Fan, Ting-Jun; Zhao, Jian; Hu, Xiuzhong; Ma, Xiya; Zhang, Wenbo; Yang, Cui

doi:10.1631/jzus.b1000199

Cited by 8 publications

(14 citation statements)

References 12 publications

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“…Our results of DiI fluorescence observations indicated that the endothelial cells in the TE-HCE-transplanted rabbit corneas were all from the TE-HCE, not from autogenous proliferated rabbit corneal endothelial cells. This finding was also supported by our previous reports (Fan et al, , 2013Fan TJ et al, 2011). ECD and integrity of corneal endothelium are important indicators of endothelial keratoplasty (Schierhölter and Honegger, 1975;Srinivas, 2012), and substantial loss of postoperative HCE cells might result in visual acuity impairment and even graft failure (Terry et al, 2008;Lass et al, 2010).…”

Section: Discussionsupporting

confidence: 86%

“…Based on our previous achievements (Fan et al, 2010(Fan et al, , 2013Fan TJ et al, 2011), the present study was designed to construct a high density TE-HCE using nontransfected HCE cells at low passage as seeder cells and modified denuded amniotic membrane (mdAM) as a scaffold, and to evaluate its biological functions by corneal transplantation in rabbit models.…”

Section: Introductionmentioning

confidence: 99%

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Construction of a tissue-engineered human corneal endothelium and its transplantation in rabbit models

Zhao¹,

Ma²,

Fan³

2016

Turk J Biol

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Human corneal endothelium (HCE) is vital in maintaining corneal transparency and thickness, and damages to it may result in endotheliopathy and even corneal blindness, which can only be cured by keratoplasty. Due to the shortage of donor corneas, tissueengineered HCE (TE-HCE) has become an equivalent to HCE. This study constructs a high endothelial cell density (ECD) TE-HCE and evaluates its functions by corneal transplantation in rabbits. A TE-HCE was constructed by culturing DiI-labeled nontransfected HCE cells on modified denuded amniotic membrane (mdAM) using collagen IV, fibronectin, and laminin, and rabbit corneas were characterized both in vivo and ex vivo after TE-HCE transplantation. Our results indicated that the constructed TE-HCE with a high ECD of 3611 ± 56.66 cells/mm2 had similar morphology and structure to a native HCE. In vivo postsurgery detections showed that the TE-HCE-transplanted cornea gained transparency and recovered its thickness gradually during a monitoring period of 358 days, while the mdAM-transplanted cornea remained opaque and edematous. Ex vivo examinations revealed that a native-like corneal endothelium with an ECD of 2703 ± 70.37 cells/mm2 was reconstructed by the transplanted TE-HCE. Our findings suggested that the TE-HCE might be used as a HCE equivalent and has promising applications in therapy of corneal endotheliopathy.

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Section: Discussionsupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Construction of a tissue-engineered human corneal endothelium and its transplantation in rabbit models

Zhao¹,

Ma²,

Fan³

2016

Turk J Biol

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“…Non‐transfected HCE cells, especially their monoclonal strains, may be considered favorable for HCE equivalent construction and future clinical use . However, cell density of previously constructed TE‐HCE is not high enough and there have only been transplantation experiments on rabbit and cat models . Since the species of monkey, whose visual system resembles human’s, represents the major nonhuman primate resource for biomedical research, it is of great importance to reconstruct a high cell density TE‐HCE and examine its functions after transplantation into monkey models .…”

Section: Discussionmentioning

confidence: 99%

“…22,23 However, cell density of previously constructed TE-HCE is not high enough and there have only been transplantation experiments on rabbit and cat models. 25,26 Since (Table S1), equivalent to that of a normal HCE in vivo of a 3-year-old child, 29 which is "young" enough to be used for clinical transplantation.…”

Section: Discussionmentioning

confidence: 99%

Construction of a high cell density human corneal endothelial equivalent and its transplantation in primate models

Zhao

Fan

et al. 2019

Xenotransplantation

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Background Recently, many patients with corneal blindness caused by endothelial dysfunction have no opportunity to receive keratoplasty therapy because of the extremely limited number of donor corneas. Corneal tissue engineering opens a new path for in vitro reconstruction of tissue‐engineered HCE which will cure the corneal endotheliopathy by clinical corneal transplantation. In this study, we construct a human corneal endothelium (HCE) equivalent with non‐transfected monoclonal HCE (mcHCE) cells and modified denuded amniotic membrane (mdAM), and evaluate its functions in monkey models. Methods Tissue‐engineered HCE (TE‐HCE) was constructed by culturing DiI‐labeled mcHCE cells on mdAMs in 20% fetal bovine serum‐containing DMEM/Ham’s Nutrient Mixture F12 (1:1) medium and 5% CO2 at 37°C on a 24‐well culture plate. The constructed TE‐HCE was transplanted into monkey corneas via penetrating keratoplasty with Descemet’s membrane and endothelium stripped. The corneal transparency, thickness, and intraocular pressure were monitored in vivo, and the corneal morphology and histological structure were examined ex vivo 181 days after surgery. Results The constructed TE‐HCE, with an average density of 3602.22 ± 45.22 cells/mm2, mimicked its natural counterpart both in morphology and histological structure. In vivo, corneal transparency was maintained, and the corneal thickness gradually decreased to 567.33 ± 72.77 μm at day 181 after TE‐HCE transplanted into monkey eyes, while intense corneal edema and turbid were found in mdAM‐transplanted eyes with their corneal thicknesses maintained over 1000 μm during the monitoring period. Ex vivo, a monolayer of corneal endothelium, consisting of mcHCE cells at a density of 2795.65 ± 156.83 cells/mm2, was reconstructed in transplanted monkey eyes. The cells in the transplanted area had the hexagonal or polygonal morphology and normal ultrastructure, and established plenty of cell‐cell and cell‐stromal matrix junctions. Besides, huge membrane‐bounded flat stacks with electric dense inclusions were found in mcHCE cells beneath the plasma membrane at the stromal side. Conclusions The constructed TE‐HCE has normal histological property and functions well in monkey models. The TE‐HCE could be used as a promising HCE equivalent in therapy of corneal endothelium dysfunction and corneal regenerative medicine.

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“…Thus, there is a clinical interest for engineering corneal endothelium for transplantation purposes. With increasing advances in regenerative medicine, several research groups have investigated on expansion of corneal endothelial cells and transplantation of tissue engineered corneal endothelium in experimental animal models [ 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ]. The present review focuses on emerging substrates for improved culturing of HCECs.…”

Section: Introductionmentioning

confidence: 99%

Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium

Navaratnam

Utheim

Rajasekhar

et al. 2015

JFB

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Corneal endothelium is a single layer of specialized cells that lines the posterior surface of cornea and maintains corneal hydration and corneal transparency essential for vision. Currently, transplantation is the only therapeutic option for diseases affecting the corneal endothelium. Transplantation of corneal endothelium, called endothelial keratoplasty, is widely used for corneal endothelial diseases. However, corneal transplantation is limited by global donor shortage. Therefore, there is a need to overcome the deficiency of sufficient donor corneal tissue. New approaches are being explored to engineer corneal tissues such that sufficient amount of corneal endothelium becomes available to offset the present shortage of functional cornea. Although human corneal endothelial cells have limited proliferative capacity in vivo, several laboratories have been successful in in vitro expansion of human corneal endothelial cells. Here we provide a comprehensive analysis of different substrates employed for in vitro cultivation of human corneal endothelial cells. Advances and emerging challenges with ex vivo cultured corneal endothelial layer for the ultimate goal of therapeutic replacement of dysfunctional corneal endothelium in humans with functional corneal endothelium are also presented.

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Therapeutic efficiency of tissue-engineered human corneal endothelium transplants on rabbit primary corneal endotheliopathy

Cited by 8 publications

References 12 publications

Construction of a tissue-engineered human corneal endothelium and its transplantation in rabbit models

Construction of a tissue-engineered human corneal endothelium and its transplantation in rabbit models

Construction of a high cell density human corneal endothelial equivalent and its transplantation in primate models

Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium

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