Cell viability effects of triamcinolone acetonide and preservative vehicle formulations

Abstract

Aim: To assess the effect of triamcinolone acetonide and preservative vehicle formulations on human retinal pigment epithelium (ARPE19) cells over a range of concentrations. Methods: Triamcinolone acetonide, in its trade and preservative free formulations, along with the preservative vehicle were added to ARPE19 cell cultures in various concentrations (0.01-1.0 mg/ml). Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay at day 5 after exposure. Functionality of the cultured ARPE19 cell line was confirmed by exposure to a previously characterised toxic agent, tamoxifen. Results: The ARPE19 cell line behaved as predicted with exposure to tamoxifen. All formulations caused significant reductions in ARPE19 cell viability at the highest concentrations (1.0 mg/ml for triamcinolone preparations and undiluted vehicle). Cell viability was reduced to the greatest degree in trade formulation triamcinolone, less so by the vehicle, and least by preservative free triamcinolone. At lower concentrations no significant effect on cell viability was observed, although cell viability was found to be inversely proportional to increasing concentration of all tested reagents Conclusions: Both the trade and preservative free formulations of triamcinolone acetonide as well as the vehicle result in cell loss at in vitro concentrations of 1 mg/ml. Although this represents theoretical vitreous concentrations achieved with current widespread therapeutic use it probably does not indicate the actual exposure of cells in their biological milieu. That cell viability was reduced most in the trade formulation suggests a possible potentiated inhibitory toxic effect of triamcinolone acetonide and vehicle at higher concentrations.

Different Ipsi- and Contralateral Glial Responses to Anti-VEGF and Triamcinolone Intravitreal Injections in Rats

“…Because these areas were positive for GFAP and vimentin, we interpreted them as areas of Müller end-foot cell hypertrophy. Although we removed most of the preservative present in the commercial preparation of triamcinolone as previously described, 88 the substance or the remaining preservative may have cellular toxic effects, 88,89 and the retina may react to stress with the initiation of glial blooms or preretinal membranes as has been observed in degenerating retinas. 87,90,91 On the other hand, treatment with the anti-VEGF aflibercept and ranibizumab has been shown to activate neonatal rat Müller cells in culture, and it has been proposed that it may be responsible for the therapeutic effects of these drugs.…”

Dystrophic Calcification in the Epidural and Extraforaminal Space Caused by Repetitive Triamcinolone Acetonide Injections

“…3) also showed that a benzyl alcohol concentration of 9 mg/mL caused extensive lysis of rabbit corneal endothelial cells within the first minute of exposure. Recently, Shaikh et al 17) showed that both the preserved TA formulation and its vehicle caused significant reductions in the viability of cultured retinal epithelial cells. Similarly, Narayanan et al 15) concluded that the vehicle was not totally responsible for toxicity but may initiate TA-dependent toxicity.…”

The Retinal Biocompatibility of Dyes in the ex vivo Model of the Isolated Superfused Vertebrate Retina

“…We could further determine a safe concentration for the intravitreal injection of TA. This is of particular importance in view of recent publications regarding a cytotoxic effect of TA in vitro [56,57]. However, cytotoxicity in vitro is contradicted by good clinical biocompatibility experiences.…”