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Original research
Potential adverse effects of ciprofloxacin and tetracycline on ARPE-19 cell lines
  1. Nasim Salimiaghdam1,
  2. Lata Singh1,
  3. Kevin Schneider1,
  4. Angele Nalbandian1,
  5. Marilyn Chwa1,
  6. Shari R Atilano1,
  7. Andrea Bao1,
  8. M Cristina Kenney2,3
  1. 1Ophthalmology, University of California, Irvine, California, USA
  2. 2Ophthalmology, University of California School of Medicine, Irvine, California, USA
  3. 3Department of Pathology and Laboratory Medicine, University of California Irvine School of Medicine, Irvine, California, USA
  1. Correspondence to Dr M Cristina Kenney; mkenney{at}hs.uci.edu

Abstract

Background We aim to determine the possible adverse effects of ciprofloxacin (CPFX) and tetracycline (TETRA), as examples of bactericidal and bacteriostatic agents, respectively, on cultured human retinal pigment epithelial cells (ARPE-19).

Methods Cells were treated with 30, 60 and 120 µg/mL of CPFX and TETRA. Cell metabolism was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. JC-1 dye (5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide) assay was conducted to measure the mitochondrial membrane potential (MMP). The level of reactive oxygen species (ROS) was measured using the -2’,7’-dichlorodihydrofluorescein diacetate assay (H2DCFDA). Quantitative real-time PCR was performed to analyse the gene expression levels associated with apoptosis (BAX, BCL2-L13, BCL2, Caspase 3, Caspase 7 and Caspase 9), inflammatory (interleukin-1β (IL-1β), IL-6, IL-33, transforming growth factor-α (TGF-α), TGF-β1 and TGF-β2) and antioxidant pathways (SOD2, SOD3, GPX3 and NOX4), along with the mitochondrial DNA (mtDNA) copy numbers.

Results Results illustrated that while all three concentrations of CPFX decreased cellular viability of ARPE-19 during all incubation periods, the 120 µg/mL TETRA resulted in increased cellular viability. At 48 and 72 hours, levels of MMP and ROS decreased significantly with each antibiotic. BAX, BCL2-L13, CASP-7, CASP-9, SOD2 and GPX3 genes overexpressed by either antibiotics. There was higher expression of IL-6 and IL-1B with TETRA treatment. The level of mtDNA decreased using both treatments.

Conclusions Clinically relevant concentrations of CPFX and TETRA have detrimental impacts on ARPE-19 cell lines in vitro, including upregulation of genes related to apoptosis, inflammation and antioxidant pathways. Additional studies are warranted to investigate if these harmful effects might be seen in retinal degeneration models in vivo.

  • retina
  • apotosis
  • drugs
http://creativecommons.org/licenses/by-nc/4.0/

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.

https://doi.org/10.1136/bmjophth-2020-000458

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    Footnotes

    • Contributors NS planned the study, designed experiments, wrote the manuscript, and analysed data. LS and AN edited the manuscript. MCK, the PI, developed the concepts, edited the manuscript, provided resources for the study. KS, SA, MC and AB helped with the experimental designs and performed some of the experiments. NS is an Arnold and Mabel Beckman Retinal Degeneration Fellow. KS is a Genentech AMD Research Fellow.

    • Funding This work was supported by the Discovery Eye Foundation, Polly and Michael Smith, Edith and Roy Carver, Iris and B. Gerald Cantor Foundation, Max Factor Family Foundation, and NEI R01 EY0127363 (MCK). Supported in part by an Unrestricted Departmental Grant from Research to Prevent Blindness. We acknowledge the support of the Institute for Clinical and Translational Science (ICTS) at University of California Irvine.

    • Competing interests None declared.

    • Patient and public involvement Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.

    • Patient consent for publication Not required.

    • Provenance and peer review Not commissioned; externally peer reviewed.

    • Data availability statement All data relevant to the study are included in the article.