J Korean Ophthalmol Soc.  2017 Mar;58(3):327-332. 10.3341/jkos.2017.58.3.327.

Effect of Trypan Blue on the Survival of Cultured Trabecular Meshwork Cells

• Kang HR
• Hwang SJ
• Kim JW

• KMID: 2371914
• DOI: http://doi.org/10.3341/jkos.2017.58.3.327

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.

Keyword

Apoptosis; Flow cytometry; Survival; Trabecular meshwork cell; Trypan blue

MeSH Terms

Apoptosis
Cataract
Cell Survival
Flow Cytometry
Humans
Trabecular Meshwork*
Trypan Blue*
Trypan Blue

Figure

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  Figure 1 Effects of trypan blue on the survival of human trabecular meshwork cells. Exposure to trypan blue for 1 minute decreased cellular survival significantly at the concentration of 0.50%. *p < 0.05.

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  Figure 2 Effects of trypan blue on the survival of human trabecular meshwork cells. Exposure to trypan blue for 5 minutes decreased cellular survival significantly at the concentration of 0.50%. *p < 0.05.

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  Figure 3 Effects of trypan blue on the survival of human trabecular meshwork cells. Exposure to trypan blue for 30 minutes decreased cellular survival significantly at the concentration of 0.10% and 0.50%. *p < 0.05.

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  Figure 4 Effects of 0.06% commercial trypan blue on the survival of human trabecular meshwork cells. Exposure to trypan blue for 5 minutes or 30 minutes decreased cellular survival. *p < 0.05.

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  Figure 5 Flow cytometric analysis of apoptosis using annexin-PI double staining. Exposure to 0.50% trypan blue for 30 minutes increased the degree of apoptosis significantly compared to non-exposed control. *p < 0.05.

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  Figure 6 Flow cytometric analysis of apoptosis using annexin-propidium iodide double staining. 0.06% commercial trypan blue exposed for 30 minutes increased the degree of apoptosis significantly compared to non-exposed control. *p < 0.05.

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