June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Construction of a bioengineered corneal substitute in animal study
Author Affiliations & Notes
  • Xinyi Wu
    Ophthal QiLu Hosp/Ophthal, Shandong University, Jinan, Shandong , China
  • Canwei Zhang
    Ophthal QiLu Hosp/Ophthal, Shandong University, Jinan, Shandong , China
  • Footnotes
    Commercial Relationships   Xinyi Wu, None; Canwei Zhang, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1395. doi:
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      Xinyi Wu, Canwei Zhang; Construction of a bioengineered corneal substitute in animal study. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1395.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose : The aim of this study were to fabricate a full-thickness artificial cornea substitute using limbal epithelial cell-like (LEC-like) cells and corneal endothelial cell-like (CEC-like) cells derived from human embryonic stem cells (hESCs) andacellular porcine corneal matrix (APCM).

Methods : LEC-like and CEC-like cells were differentiated from hESCs in vitro. Then, a 400 μm thickness, 11 mm diameter APCM lamella was prepared as the scaffold using trephine and a special apparatus made by ourselves. Cornea was simulated by an APCM seeded with LEC-like and CEC-like cells using a special insert of 24-well plates that enabled seeding both sides of the scaffold. Morphological characteristics and cell markers of the constructs were investigated by HE and immunofluorescence staining. Furthermore, the thickness, transmittance, and mechanical properties of the construct were also measured. Then, the biocompatibility and functionality of the construct were evaluated by penetrating keratoplasty in rabbits, and the animals were followed with slit lamp ophthalmic examination, optical coherent tomography and confocal laser microscopy.

Results : The differentiated LEC-like cells and CEC-like cells showed similar properties to native LECs and CECs and good survival on APCM. Three or four layers of epithelial cells were observed on the Bowman's membrane of APCM, and a uniform monolayer of CEC-like cells on the opposite side. Immunofluorescence showed expression of typical LEC and CEC markers on their corresponding sides. The thickness and mechanical properties of the construct were similar to native rabbit cornea. The transparency of the corneal substitute gradually increased after transplantation and almost completely restored on postoperative 8 weeks, although its transmittance (47%) was a little lower than that of native rabbit cornea (56%) in vitro.

Conclusions : The tissue-engineered full-thickness cornea substitute that we developed showed similar biological properties with the native cornea, and might be a suitable substitute of donor corneal graft for corneal transplantation.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.


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