Soluble lumican glycoprotein purified from human amniotic membrane promotes corneal epithelial wound healing

Lung Kun Yeh, Wei Li Chen, Wei Li, Edgar M. Espana, Jie Ouyang, Tetsuya Kawakita, Winston W Y Kao, Scheffer C G Tseng, Chia Yang Liu

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

PURPOSE. To purify and characterize the glycoprotein lumican, isolated from human amniotic membrane (AM), and to examine its efficacy in treating corneal epithelium debridement. METHODS. An affinity-purified, anti-human lumican antibody-conjugated protein A Sepharose column was used to purify soluble lumican protein from human AM. The purified AM lumican was characterized by two-dimensional and SDS gel electrophoresis, plus Western blot analysis with anti-lumican antibody. The effects of lumican on corneal epithelial wound healing were examined in an organ culture mouse eye model. RESULTS. Lumican was found to be abundantly present in the stroma of human AM. It was extracted from the AM by isotonic, 1 M NaCl, and 4 M guanidine HCl solutions, suggesting that it is present in both the soluble and matrix-bound states. In two-dimensional gel electrophoresis, the 50-kDa human amniotic lumican purified by antibody-conjugated affinity chromatography migrated in a smear between pH 3.0 and 6.0. After endo-β-galactosidase digestion, it existed as a single core protein at pH 6.0, suggesting that native human amniotic lumican is a glycoprotein with short sugar moiety. Addition of purified human AM lumican to cultured medium promoted re-epithelialization and enhanced cell proliferation of wild-type mouse corneal epithelial cells in an organ culture. In lumican-knockout (lum-/-) mice, the effect of human lumican on promoting corneal epithelial wound healing was even more dramatic than in wild-type mice. CONCLUSIONS. The diversified functions of lumican include modulation of epithelial cells in wound healing and serving as an extracellular matrix component. Administration of lumican may be beneficial for treating epithelial defects in the cornea and other tissues.

Original languageEnglish
Pages (from-to)479-486
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume46
Issue number2
DOIs
Publication statusPublished - 2005 Feb
Externally publishedYes

Fingerprint

Amnion
Wound Healing
Glycoproteins
Organ Culture Techniques
Electrophoresis, Gel, Two-Dimensional
Lumican
Epithelial Cells
Galactosidases
Re-Epithelialization
Corneal Epithelium
Antibody Affinity
Guanidine
Debridement
Affinity Chromatography
Population Groups
Knockout Mice
Cornea
Extracellular Matrix
Digestion
Anti-Idiotypic Antibodies

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Soluble lumican glycoprotein purified from human amniotic membrane promotes corneal epithelial wound healing. / Yeh, Lung Kun; Chen, Wei Li; Li, Wei; Espana, Edgar M.; Ouyang, Jie; Kawakita, Tetsuya; Kao, Winston W Y; Tseng, Scheffer C G; Liu, Chia Yang.

In: Investigative Ophthalmology and Visual Science, Vol. 46, No. 2, 02.2005, p. 479-486.

Research output: Contribution to journalArticle

Yeh, LK, Chen, WL, Li, W, Espana, EM, Ouyang, J, Kawakita, T, Kao, WWY, Tseng, SCG & Liu, CY 2005, 'Soluble lumican glycoprotein purified from human amniotic membrane promotes corneal epithelial wound healing', Investigative Ophthalmology and Visual Science, vol. 46, no. 2, pp. 479-486. https://doi.org/10.1167/iovs.04-1014
Yeh, Lung Kun ; Chen, Wei Li ; Li, Wei ; Espana, Edgar M. ; Ouyang, Jie ; Kawakita, Tetsuya ; Kao, Winston W Y ; Tseng, Scheffer C G ; Liu, Chia Yang. / Soluble lumican glycoprotein purified from human amniotic membrane promotes corneal epithelial wound healing. In: Investigative Ophthalmology and Visual Science. 2005 ; Vol. 46, No. 2. pp. 479-486.
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abstract = "PURPOSE. To purify and characterize the glycoprotein lumican, isolated from human amniotic membrane (AM), and to examine its efficacy in treating corneal epithelium debridement. METHODS. An affinity-purified, anti-human lumican antibody-conjugated protein A Sepharose column was used to purify soluble lumican protein from human AM. The purified AM lumican was characterized by two-dimensional and SDS gel electrophoresis, plus Western blot analysis with anti-lumican antibody. The effects of lumican on corneal epithelial wound healing were examined in an organ culture mouse eye model. RESULTS. Lumican was found to be abundantly present in the stroma of human AM. It was extracted from the AM by isotonic, 1 M NaCl, and 4 M guanidine HCl solutions, suggesting that it is present in both the soluble and matrix-bound states. In two-dimensional gel electrophoresis, the 50-kDa human amniotic lumican purified by antibody-conjugated affinity chromatography migrated in a smear between pH 3.0 and 6.0. After endo-β-galactosidase digestion, it existed as a single core protein at pH 6.0, suggesting that native human amniotic lumican is a glycoprotein with short sugar moiety. Addition of purified human AM lumican to cultured medium promoted re-epithelialization and enhanced cell proliferation of wild-type mouse corneal epithelial cells in an organ culture. In lumican-knockout (lum-/-) mice, the effect of human lumican on promoting corneal epithelial wound healing was even more dramatic than in wild-type mice. CONCLUSIONS. The diversified functions of lumican include modulation of epithelial cells in wound healing and serving as an extracellular matrix component. Administration of lumican may be beneficial for treating epithelial defects in the cornea and other tissues.",
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AU - Yeh, Lung Kun

AU - Chen, Wei Li

AU - Li, Wei

AU - Espana, Edgar M.

AU - Ouyang, Jie

AU - Kawakita, Tetsuya

AU - Kao, Winston W Y

AU - Tseng, Scheffer C G

AU - Liu, Chia Yang

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N2 - PURPOSE. To purify and characterize the glycoprotein lumican, isolated from human amniotic membrane (AM), and to examine its efficacy in treating corneal epithelium debridement. METHODS. An affinity-purified, anti-human lumican antibody-conjugated protein A Sepharose column was used to purify soluble lumican protein from human AM. The purified AM lumican was characterized by two-dimensional and SDS gel electrophoresis, plus Western blot analysis with anti-lumican antibody. The effects of lumican on corneal epithelial wound healing were examined in an organ culture mouse eye model. RESULTS. Lumican was found to be abundantly present in the stroma of human AM. It was extracted from the AM by isotonic, 1 M NaCl, and 4 M guanidine HCl solutions, suggesting that it is present in both the soluble and matrix-bound states. In two-dimensional gel electrophoresis, the 50-kDa human amniotic lumican purified by antibody-conjugated affinity chromatography migrated in a smear between pH 3.0 and 6.0. After endo-β-galactosidase digestion, it existed as a single core protein at pH 6.0, suggesting that native human amniotic lumican is a glycoprotein with short sugar moiety. Addition of purified human AM lumican to cultured medium promoted re-epithelialization and enhanced cell proliferation of wild-type mouse corneal epithelial cells in an organ culture. In lumican-knockout (lum-/-) mice, the effect of human lumican on promoting corneal epithelial wound healing was even more dramatic than in wild-type mice. CONCLUSIONS. The diversified functions of lumican include modulation of epithelial cells in wound healing and serving as an extracellular matrix component. Administration of lumican may be beneficial for treating epithelial defects in the cornea and other tissues.

AB - PURPOSE. To purify and characterize the glycoprotein lumican, isolated from human amniotic membrane (AM), and to examine its efficacy in treating corneal epithelium debridement. METHODS. An affinity-purified, anti-human lumican antibody-conjugated protein A Sepharose column was used to purify soluble lumican protein from human AM. The purified AM lumican was characterized by two-dimensional and SDS gel electrophoresis, plus Western blot analysis with anti-lumican antibody. The effects of lumican on corneal epithelial wound healing were examined in an organ culture mouse eye model. RESULTS. Lumican was found to be abundantly present in the stroma of human AM. It was extracted from the AM by isotonic, 1 M NaCl, and 4 M guanidine HCl solutions, suggesting that it is present in both the soluble and matrix-bound states. In two-dimensional gel electrophoresis, the 50-kDa human amniotic lumican purified by antibody-conjugated affinity chromatography migrated in a smear between pH 3.0 and 6.0. After endo-β-galactosidase digestion, it existed as a single core protein at pH 6.0, suggesting that native human amniotic lumican is a glycoprotein with short sugar moiety. Addition of purified human AM lumican to cultured medium promoted re-epithelialization and enhanced cell proliferation of wild-type mouse corneal epithelial cells in an organ culture. In lumican-knockout (lum-/-) mice, the effect of human lumican on promoting corneal epithelial wound healing was even more dramatic than in wild-type mice. CONCLUSIONS. The diversified functions of lumican include modulation of epithelial cells in wound healing and serving as an extracellular matrix component. Administration of lumican may be beneficial for treating epithelial defects in the cornea and other tissues.

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