Journal List > J Korean Ophthalmol Soc > v.49(4) > 1008245

Lee, Lee, and Kim: Effect of Nitric Oxide on the Migration and Fibroblast-mediated Contraction of Collagen Gels

Abstract

Purpose

To investigate the role of nitric oxide (NO) on the migration of cultured human Tenon's capsule fibroblasts (HTCF) and contraction of collagen gel.

Methods

After artificial wounding, the primarily cultured HTCF were exposed to an NO donor such as sodium nitroprusside (SNP), S-Nitroso-N-acetylpenicillamine (SNAP), or dexamethasone at various concentrations. The cellular migration was measured up to five days. After embedding the cells in the collagen gels, the amount of contraction by the gels was also measured. Cellular survival and NO production were measured with MTT assay and Griess assay, respectively.

Results

Cellular survival was decreased by both NO donors but not by dexamethasone. SNP inhibited migration of HTCF in a dose-dependent manner and enhanced contraction of collagen gels. However, SNAP had no effect on the cellular migration or gel contraction. Dexamethasone inhibited cellular migration but did not affect the contraction of collagen gels.

Conclusions

Among the NO donors, only SNP inhibited migration of HTCF and enhanced contraction of collagen gels in vitro. Thus, the effects between the two NO donors on fibroblast‐ induced wound healing differ.

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Figure 1.
Effect of NO donors and dexamethasone on the survival of fibroblasts. NO donors decreased cellular survival in a dose-dependent manner compared to non-exposed control. (*; p<0.05)
jkos-49-661f1.tif
Figure 2.
Effect of NO donors on the production of nitric oxide. Both NO donor increased NO in a dose-dependent manner. (*; p<0.05)
jkos-49-661f2.tif
Figure 3.
Effect of sodium nitroprusside on the migration of fibroblasts. Sodium nitroprusside inhibited migration of fibroblasts in a dose-dependent manner from 10 µM of concentration after 3 days. (*; p<0.05)
jkos-49-661f3.tif
Figure 4.
Effect of SNAP on the migration of fibroblsts. SNAP did not affect on the migration of fibroblasts up to 5 days compared to non-exposed controls.
jkos-49-661f4.tif
Figure 5.
Effect of dexamethasone on the migration of fibroblasts. Dexamethasone inhibited the migration of fibroblasts after 4 days at 10 µM and after 3 days at 100 µM, compared to non-exposed controls. (*; p<0.05)
jkos-49-661f5.tif
Figure 6.
Effect of sodium nitroprusside on the speed of migration in cultured Tenon fibroblasts. Sodium nitroprusside decreased the speed of migration significantly compared to non-exposed control up to 5 days. (*; p<0.05)
jkos-49-661f6.tif
Figure 7.
Effect of SNAP on the speed of migration in cultured Tenon capsule fibroblasts. SNAP did not affect on the speed of migration significantly compared to non-exposed control.
jkos-49-661f7.tif
Figure 8.
Effect of dexamethasone on the speed of migration in cultured human Tenon capsule fibroblasts. Dexamethasone decreased the speed of migration at high concentration. (*; p<0.05)
jkos-49-661f8.tif
Figure 9.
Effect of NO donor on the contraction of collagen gels after 5 days. Sodium nitroprusside augmented gel contraction significantly compared to non-exposed control. (*; p<0.05)
jkos-49-661f9.tif
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