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Park, Park, Lee, and Lee: Effects of Human Serum on Human Corneal Epithelial Cells in Vitro
Original Article

Journal of the Korean Ophthalmological Society 2017; 58(12): 1333-1340.

Published online: 14 December 2017

DOI: https://doi.org/10.3341/jkos.2017.58.12.1333

Effects of Human Serum on Human Corneal Epithelial Cells in Vitro

Young Min Park, MD1, 2, Jae Sung Park, MD3, In Ho Lee, MD4, Jong Soo Lee, MD, PhD4

1Department of Ophthalmology, Gyeongsang National University School of Medicine, Jinju, Korea.

2Department of Ophthalmology, Gyeongsang National University Changwon Hospital, Changwon, Korea.

3Danggam Choi Eye Clinic, Busan, Korea.

4Department of Ophthalmology, Pusan National University School of Medicine, Yangsan, Korea.

Address reprint requests to Jong Soo Lee, MD, PhD. Department of Ophthalmology, Pusan National University Hospital, #179 Gudeok-ro, Seo-gu, Busan 49241, Korea. Tel: 82-51-240-7321, Fax: 82-51-240-7341, jongsool@pusan.ac.kr

Received 16 March 2017       Revised 27 June 2017       Accepted 29 November 2017

©2017 The Korean Ophthalmological Society

The Korean Ophthalmological Society

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

To investigate the effect of human serum on corneal epithelial cells.

Methods

Changes of corneal epithelial cells were evaluated after 1, 4, 12, and 24 hours (hrs) of exposure to various concentrations of human serum (3, 5, 8, and 16%). Cellular metabolic activity and the extent of cellular damage were measured. Effect of human serum on cell migration was also examined. Concentration of procollagen type-I COOH-terminal peptide (PIP), epidermal growth factor (EGF), and laminin after exposure to human serum was further observed.

Results

In every concentration of human serum, metabolic activity of the corneal epithelial cells temporarily decreased at 4 hrs of exposure and recovered to baseline levels afterward. With the same exposure time, there was no statistically significant difference in metabolic activity between the human serum-exposed group and the control group. Cellular toxicity of human serum exhibited a time- and dose-dependent relationship. Cellular migration was observed after 24 hrs of exposure to 5% concentration of human serum and after 12 hrs of exposure to 8% and 16% concentration of human serum. The PIP, EGF, and laminin titers increased in time- and dose-dependent manners.

Conclusions

Human serum does not decrease the metabolic activity of corneal epithelial cells as the concentration and exposure time increase, but it can induce cytotoxicity. Considering cellular migration, a serum concentration of 5% or higher should be used.

Keywords: Corneal epithelium, Metabolic activity, Scratch wound assay, Serum, Toxicity

Figures and Tables

Figure 1

The absorption rate of the water-insoluble formazan dye in corneal epithelial cells. Metabolic activity of the corneal epithelial cells temporarily decreased at 4 hrs of exposure and recovered to baseline levels afterward. With the same exposure time, there was no statistically significant difference in metabolic activity between the human serum-exposed group and the control group. *p < 0.05 (Kruskal-Walis test, intra-group comparison). hr = hour(s).

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Figure 2

Lactate dehydrogenase (LDH) assay results. LDH titers of cultured corneal epithelial cells exposed to human serum. ¥: Significant changes (p < 0.05, Kruskal-Walis test) compared with the control group. hr = hour(s).

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Figure 3

Procollagen type I COOH-terminal peptide (PIP), Laminin, and epidermal growth factor (EGF) titers of cultured corneal epithelial cells exposed to human serum. The PIP (A), EGF (B), and laminin (C) titers increased in timeand dose-dependent manners. hr = hour(s).

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Figure 4

Scratch assay of corneal epithelial cells after exposure to human serum. Cellular migration was observed after 24 hrs of exposure to 5% concentration of human serum and after 12 hrs of exposure to 8% and 16% concentration of human serum. hr = hour(s).

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Table 1

Comparison of the cell migration assay after exposure to human serum

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Acellular Zone: values are mean ± standard deviation. p-value: Kruskal-Wallis test (1 h vs. tested h).

h = hour(s).

Notes

Conflicts of Interest The authors have no conflicts to disclose.

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