Effect of various solvents on the cytotoxic activity and stability of Bleomycin for in vitro cell culture based assays

다국어 초록 (Multilingual Abstract)

Bleomycin sulfate is a metal‐complexing glycopeptide that is commonly used as an experimental model for pulmonary fibrosis. Pulmonary fibrosis induced by Bleomycin sulfate is mediated via DNA damage and cytotoxic actions on the alveolar epithelial cells. Among various factors, the in vitro actions of Bleomycin sulfate depend considerably on its chemical stability and the solvent used to dissolve the drug. In this study, we evaluated the effects of various solvents [water, dimethyl sulfoxide (DMSO), saline and Dulbecco's Modified Eagle Medium (DMEM)] on the in vitro cytotoxic activity and stability of Bleomycin.
The cytotoxic effect of Bleomycin sulfate was evaluated on alveolar epithelial cells (A549) using various concentration (10, 15 and 20 mcg/mL) of the drug dissolved in different solvents. A549 cells were seeded at a concentration of 5 × 103 cells per well in a 96 well‐plate and incubated for 24 hours. These cells were then exposed to various concentrations of Bleomycin sulfate in the different solvents for the next 24 hours. The cell viability was determined using the MTT (3‐(4, 5‐dimethylthiazolyl‐2)‐2, 5‐diphenyltetrazolium bromide) reduction assay.
Significant cytotoxic response was observed with all the doses of Bleomycin sulfate (10, 15 and 20 mcg/mL) when it was dissolved in either water or DMSO. However, this was not the case with DMEM or saline since lower concentration of Bleomycin sulfate (10 mcg/mL) failed to induce cell death. We noticed that treatment of A549 cells with any of the solvents by themselves (vehicle control) did not induce cell death. With regards to DMSO, we used a final concentration of 0.025% v/v in the cell culture media.
We conclude that water or DMSO (at a final concentration of 0.025% v/v) may serve as suitable solvents to dissolve Bleomycin sulfate for carrying out in vitro cell culture assays. Also, it appears that higher concentrations of Bleomycin sulfate are required to show cytotoxic activity when DMEM or saline is used as a solvent. Further studies are warranted using high performance liquid chromatography (HPLC) to explore drug stability in the various solvents used.
Support or Funding Information
Seed Funding from California Health Sciences University
This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.