June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Preparation of decellularized porcine cornea for corneal tissue engineering by supercritical carbon dioxide extraction technology
Author Affiliations & Notes
  • Yi-Hsun Huang
    Ophthalmology, National Cheng Kung University Hospital, Tainan, Taiwan
  • Ming-Lung YEH
    Biomedical Engineering, National Cheng Kung University, Tainan, Taiwan
  • Dar-Jen Hsieh
    ACRO Biomedical Co., Ltd., Kaohsiung, Taiwan
  • Fang-Wei Tseng
    ACRO Biomedical Co., Ltd., Kaohsiung, Taiwan
  • I-Chen Peng
    Ophthalmology, National Cheng Kung University Hospital, Tainan, Taiwan
  • Wen-Shin Chang
    Biomedical Engineering, National Cheng Kung University, Tainan, Taiwan
  • Footnotes
    Commercial Relationships   Yi-Hsun Huang, None; Ming-Lung YEH, None; Dar-Jen Hsieh, ACRO Biomedical Co., Ltd. (P); Fang-Wei Tseng, ACRO Biomedical Co., Ltd. (P); I-Chen Peng, None; Wen-Shin Chang, None
  • Footnotes
    Support  MOST 105-2314-B-006–020
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4341. doi:
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      Yi-Hsun Huang, Ming-Lung YEH, Dar-Jen Hsieh, Fang-Wei Tseng, I-Chen Peng, Wen-Shin Chang; Preparation of decellularized porcine cornea for corneal tissue engineering by supercritical carbon dioxide extraction technology. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4341.

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

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Abstract

Purpose : To evaluate the stability and biocompatibility of decellularized porcine cornea that was prepared by supercritical carbon dioxide extraction (scCO2) technology.

Methods : Cornea was harvested from porcine eye ball and incubated in reverse osmosis water and 2M NaCl solution in sequential fashion for three times. The scCO2 extraction of porcine cornea was carried out using a high-pressure reaction apparatus. The decellularization of cornea was confirmed by histological study and quantitative assay of the residual DNA and glycosaminoglycans (GAGs). For in vivo-testing, the decellularized corneal matrix of 300 μm thickness and 6.0 mm diameter was transplanted onto a 6.0 mm diameter keratectomy wound in New Zealand White rabbits. Triton® X-100 treated corneal matrix was used as control. After eight weeks, the rabbit eyes were enucleated to examine the tissue compatibility of the implanted matrix.

Results : Histology results showed complete removal of cells. The processed corneas showed excellent biocompatibility in cell cultures. SDS-PAGE and western blot revealed the absence of cell marker proteins. The scCO2 processed porcine cornea showed good appearance in lamellar keratoplasty performed on rabbits. There were no epithelial defect, neovascularization, immunological reaction and rejection 8 weeks post transplantation.

Conclusions : The scCO2 extraction technology successfully produce an ideal scaffold that exhibited good biocompatibility both in vitro and in vivo for corneal tissue engineering.

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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