Ciprofloxacin Decreases Collagen in Mouse Tympanic Membrane Fibroblasts. (1st March 2016)

• Record Type: Journal Article
• Authors: Orobello, Nicklas C.; Dirain, Carolyn O.; Schultz, Gregory; Milne‐Davies, Bailey A.; Ng, Maria R. A.; Antonelli, Patrick J.
• Is Part Of: Otolaryngology--head and neck surgery. Volume 155:Number 1(2016:Jul.)
• Page Start: 127
• Page End: 132
• Publication Date: 2016-03-01
• Subjects: ciprofloxacin -- collagen -- tubulin -- fibroblast -- eardrop; Head -- Surgery -- Periodicals; Neck -- Surgery -- Periodicals; Otolaryngology -- Periodicals; 617.51
• DOI: 10.1177/0194599816633671
• Languages: English
• ISSNs: 0194-5998

Abstract

Abstract : Objectives: To determine how collagen production by tympanic membrane fibroblasts is affected by ciprofloxacin at levels found in eardrops. Study Design: Prospective, controlled, and blinded cell culture study. Setting: Academic tertiary medical center. Subjects: Cell culture of mouse fibroblasts. Methods: A primary fibroblast culture was established from mouse tympanic membranes. Fibroblasts were cultured until they were 75% confluent, then treated with dilute hydrochloric acid (control) or ciprofloxacin (0.01% or 0.3%) for 24 or 72 hours for Western blotting and for 24 or 48 hours for cytotoxicity assay. Cells were observed with phase‐contrast microscope. Western blotting was performed for collagen type 1 α1 (collagen 1A1) and α‐tubulin. Results: Fibroblasts treated with 0.01% and 0.3% ciprofloxacin for 24 hours had lower levels of collagen 1A1 ( P =. 0005 and P <. 0001, respectively) and α‐tubulin (both P <. 0001) than control fibroblasts. Collagen 1A1 and α‐tubulin levels were lower in fibroblasts treated with 0.3% than with 0.01% ciprofloxacin ( P =. 02 and P =. 014). After 72 hours, 0.3% ciprofloxacin completely eliminated collagen 1A1 and α‐tubulin ( P <. 001). Cells treated with 0.01% ciprofloxacin for 72 hours also had lower collagen 1A1 ( P <. 0001) and α‐tubulin ( P =. 005) as compared with the control. Seventy‐two‐hour incubation in 0.01% or 0.3% ciprofloxacin resulted in lower levels of collagen 1A1 ( P =. 009 and P <. 0001, respectively) and α‐tubulin ( P =. 007 and P <. 0001, respectively) than 24‐hour incubation. Cytotoxicity assay and phase‐contrast microscopy mirrored these findings. Conclusions: Treatment of tympanic membrane fibroblasts with 0.3% ciprofloxacin, as found in eardrops, reduces fibroblast viability and collagen and α‐tubulin protein levels. These findings could explain tympanic membrane healing problems associated with quinolone eardrops.