트리판 블루가 배양된 섬유주세포의 생존에 미치는 영향
Effect of Trypan Blue on the Survival of Cultured Trabecular Meshwork Cells
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목적: 백내장 수술에 사용하는 트리판 블루가 배양된 섬유주세포의 생존에 미치는 영향을 알아보고자 하였다. 대상과 방법: 일차배양한 인체의 섬유주세포에 0.05%, 0.10%, 0.50%의 트리판 블루에 1분, 5분, 30분간 노출시켰으며, 상용의 0.06% 트리판 블루에도 노출시켰다. 세포의 생존은 MTT assay로 알아보았고, 세포고사의 유발을 알아보기 위하여 Annexin-propidium iodide 이중염색을 이용하여 유세포분석을 시행하였다. 결과: 트리판 블루는 농도와 노출시간에 비례하여 유의하게 세포의 생존을 감소시켰으며, 0.50%의 농도에 30분간 노출시킨 경우 유의하게 세포고사를 유발하였다(p<0.05). 상용의 트리판 블루는 5분 이상 노출시킨 경우 세포의 생존을 저하시켰으며 30분간 노출시킨 경우 세포고사를 유발하였다(p<0.05). 결론: 트리판 블루는 저농도에서도 섬유주세포에 장시간 노출시킬 경우 세포손상을 유발할 수 있으며 그 기전에는 세포고사가 관여한다. 따라서 백내장 수술 시 트리판 블루를 사용할 경우 가능한 한 단시간 사용하고 완전히 제거해야 할 것으로 생각된다
Abstract
Purpose: To evaluate the effects of trypan blue (TB) on the survival of cultured human trabecular meshwork cells (HTMCs). Methods: Primarily cultured HTMCs were exposed to 0.05, 0.10 or 0.50% TB for 1, 5 or 30 min. Cellular survival was assessed using the MTT assay and degree of apoptosis was analyzed with flow cytometry using annexin-V/propidium iodide double staining. Results: Long-term exposure or high concentration of TB decreased the survival of HTMCs (p < 0.05). In flow cytometric analysis, exposure to 0.50% TB for 30 min increased the degree of apoptosis (p < 0.05). Commercial TB decreased cell survival after exposure for 5 min and increased the degree of apoptosis after exposure for 30 min (p < 0.05). Conclusions: TB may cause cellular damage of cultured HTMCs and apoptosis could be the underlying mechanism. In TB-assisted cataract surgery, TB should be used for the shortest time possible and removed completely.
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