인체 혈청이 배양된 인체 각막상피세포에 미치는 영향
Effects of Human Serum on Human Corneal Epithelial Cells in Vitro
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목적: 인체 각막상피세포에 대한 인체 혈청의 영향을 알아보고자 하였다. 대상과 방법: 인체 각막상피세포를 배양하여 3, 5, 8, 그리고 16% 농도의 인체 혈청에 1, 4, 12, 24시간 동안 접촉시킨 후, 세포 대사활성도 변화와 세포 독성을 확인하였다. 혈청 노출 후 Procollagen type-I COOH-terminal peptide (PIP), epidermal growth factor (EGF), 그리고 laminin 수치 변화를 측정하였으며, 창상 회복력을 확인하였다. 결과: 각막상피세포 대사 활성도는 인체 혈청의 모든 농도에서 노출 4시간째 일시적으로 감소한 후, 노출 시간이 증가함에 따라 지속적으로 증가하는 경향을 나타내었다. 노출 시간이 동일할 경우, 대조군과 혈청 노출군 사이 유의한 세포 대사 활성도 차이는 관찰되지 않았다. 세포 독성은 인체 혈청의 농도와 노출 시간이 증가함에 따라 증가하는 양상을 보였다. 5% 농도 인체 혈청의 경우 24시간째, 8%와 16% 농도의 경우 12시간째부터 유의한 각막 창상 회복이 관찰되었다. 높은 농도의 인체 혈청에 노출된 각막상피세포일수록 많은 양의 PIP, EGF, 그리고 laminin을 발현하였다. 결론: 인체 혈청은 농도가 증가하고 노출 시간이 증가함에 따라 각막상피세포의 대사 활성도 감소를 초래하지는 않지만 세포 독성을 초래할 수 있다. 또한 각막 창상 회복을 위해서는 최소 5% 이상 농도의 인체 혈청에 12시간 이상 노출이 필요할 것으로 생각된다.
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.
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