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The effects of commercially available preservative-free FDA-approved triamcinolone (Triesence®) on retinal cells in culture
- Source :
- Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics. 27(2)
- Publication Year :
- 2011
-
Abstract
- To evaluate the effects of Triesence® (TRI), a new preservative-free triamcinolone approved by the U.S. Food and Drug Administration (FDA) for intraocular use, on human retina pigment epithelial (ARPE-19) and rat neurosensory (R28) cells in culture.ARPE-19 and R28 cell cultures were treated 24 h with 1,000, 500, 200, or 100 μg/mL of crystalline (cTRI) or 1,000, 500, or 200 μg/mL of solubilized (sTRI). TRI was solubilized by centrifuging the drug, discarding the supernatant containing the vehicle and then resuspending the drug pellet in an equivalent amount of Dimethyl sulfoxide to achieve the same concentration as the commercial preparation. Percentage of cell viability (CV) was evaluated by a trypan blue dye-exclusion assay. The mitochondrial membrane potential (ΔΨm) was analyzed with the JC-1 assay. The caspase-3/7 activity was measured by a fluorochrome assay.In the ARPE-19 cultures, the cTRI caused a decrease in CV at 1,000 μg/mL (13.03±6.51; P0.001), 500 μg/mL (28.87±9.3; P0.001), 200 μg/mL (54.93±5.61; P0.001), and 100 μg/mL (82.53±0.65; P0.005) compared with the untreated controls (96.98±0.16). In R28 cultures, the cTRI treatment also reduced CV values significantly (P0.001) for the 1,000 μg/mL (22.73±2.44), 500 μg/mL (34.63±1.91), 200 μg/mL (58.70±1.39), and 100 μg/m (75.33±2.47) compared with the untreated controls (86.08±3.54). Once the TRI was solubilized (sTRI), the CV and ΔΨm remained similar to the untreated controls for both ARPE-19 and R28 cells. The sTRI treatment with 1,000, 500, and 200 μg/mL increased in caspase-3/7 activity in ARPE-19 cells (P0.01) and in R28 cells (P0.05) compared with dimethyl sulfoxide equivalent controls.The crystalline form of TRI (cTRI) can cause a significant decrease in CV to cultured retinal cells. Once the TRI is solubilized (sTRI), at the same concentrations, the cells remain viable with no decrease in CV or ΔΨm. The sTRI can, however, increase caspase-3/7 activity, thus suggesting some degree of apoptosis.
- Subjects :
- Preservative
medicine.medical_specialty
Triamcinolone acetonide
medicine.drug_class
Cell Survival
Apoptosis
Triamcinolone
Retina
chemistry.chemical_compound
Internal medicine
medicine
Animals
Humans
Pharmacology (medical)
Viability assay
Cells, Cultured
Pharmacology
Caspase 7
Chromatography
Dose-Response Relationship, Drug
Dimethyl sulfoxide
business.industry
Caspase 3
Retinal
Rats
Ophthalmology
Endocrinology
chemistry
Cell culture
Corticosteroid
Trypan blue
business
medicine.drug
Subjects
Details
- ISSN :
- 15577732
- Volume :
- 27
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics
- Accession number :
- edsair.doi.dedup.....e6ad4d76146953206cab14a7c656c178