June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Effect of the cell density of cultivated corneal endothelial cells on tissue engineering for the treatment of corneal endothelial dysfunction
Author Affiliations & Notes
  • Naoki Okumura
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Shinichiro Nakano
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Ayaka Kusakabe
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Ryota Inoue
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Yugo Okazaki
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Kazuya Kakutani
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Shigeru Kinoshita
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Noriko Koizumi
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Footnotes
    Commercial Relationships Naoki Okumura, JCR Pharmaceuticals Co.,Ltd. (P), Senju Pharmaceutical Co.,Ltd. (P); Shinichiro Nakano, None; Ayaka Kusakabe, None; Ryota Inoue, None; Yugo Okazaki, None; Kazuya Kakutani, None; Shigeru Kinoshita, Alcon (C), AMO (C), HOYA (C), JCR Pharmaceuticals Co.,Ltd. (P), Otsuka Pharmaceutical Co. (C), Senju Pharmaceutical Co.,Ltd. (P); Noriko Koizumi, JCR Pharmaceuticals Co.,Ltd. (P), Senju Pharmaceutical Co.,Ltd. (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1174. doi:
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      Naoki Okumura, Shinichiro Nakano, Ayaka Kusakabe, Ryota Inoue, Yugo Okazaki, Kazuya Kakutani, Shigeru Kinoshita, Noriko Koizumi; Effect of the cell density of cultivated corneal endothelial cells on tissue engineering for the treatment of corneal endothelial dysfunction. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1174.

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

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Abstract

Purpose: We have initiated clinical research of a cultured corneal endothelial cell (CEC) injection therapy for the treatment of corneal endothelial dysfunction. CEC density (CD) is an important index for evaluating the health of corneal endothelium in the clinical setting. Here we investigated the effect of cultivated CEC CD on the reconstruction of corneal endothelium by cell-injection in an animal model.

Methods: Corneal endothelium was mechanically removed in 6 eyes of 6 rabbits. A 5x105 amount of high-CD CECs (1834.7±149.3 cells/mm2) with Rho-associated protein kinase-inhibitor Y-27632 was then injected into the anterior chamber of 3 eyes (high-CD group), while low-CD CECs (503.0±133.7 cells/mm2) were injected into 3 eyes (low-CD group). The anterior segment in each eye was then evaluated by slit-lamp microscopy for 2 weeks. Corneal thickness and volume were evaluated by ultrasound pachymetry and a Scheimpflug camera system (Pentacam® HR). At 2-weeks post treatment, the phenotype of the reconstructed corneal endothelium was evaluated by immunohistochemical analysis of ZO-1, N-cadherin, and Na+/K+-ATPase.

Results: Corneal clarity was recovered in all eyes of both groups. However, corneal clarity and corneal thickness recovered faster in the high-CD group than in the low-CD group. Mean corneal thickness in the high-CD group was significantly lower than that in the low-CD group at 2-weeks post treatment (391.7±7.0μm and 598.3±11.0μm, respectively). Pentacam® HR examinations revealed that the mean corneal volume in the high-CD group was significantly less than that in the low-CD group (42.7±2.2mm3 and 68.4±5.1mm3, respectively). Immunohistochemical analysis showed regenerated corneal endothelium in the form of a monolayer that expressed ZO-1, N-cadherin, and Na+/K+-ATPase. Of interest, the mean regenerated CEC CD in the high-CD group was 3312.0±429.0 cells/mm2, while that in the low-CD group was 1642.7±279.2 cells/mm2 (p<0.05).

Conclusions: Our findings suggest that an injection-therapy of high-CD cultured CECs can regenerate high-CD corneal endothelium. Further study is needed to elucidate the optimal criteria of cultured CEC CD to regenerate high-CD and enable a good prognosis post corneal endothelial regenerative medicine treatment for corneal endothelial dysfunction.

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