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Equine veterinary journal2011; 44(3); 355-360; doi: 10.1111/j.2042-3306.2011.00437.x

The in vitro effects of antibiotics on cell viability and gene expression of equine bone marrow-derived mesenchymal stromal cells.

Abstract: To investigate the effects of commonly used antibiotics on cell viability and gene expression of equine bone marrow-derived mesenchymal stromal cells (MSC) in vitro. Methods: Bone marrow-derived MSC were cultured in media containing gentamicin, amikacin, penicillin, enrofloxacin or ceftiofur at concentrations of 50, 100, 200 and 500 µg/ml. The alamarBlue fluorescence assay was used to assess cell viability over 48 h. After 5 days the cells were released and lysed prior to RNA extraction and reverse transcription. RNA levels were assessed using spectrophotometry and quantitative PCR was used to analyse gene expression of COL1A2, COL5A1, TNC, TNFα, CASP3, BCl2 and TGFβR2 relative to the reference gene GAPDH. Results: Enrofloxacin produced a significant concentration-dependent reduction in cell viability at 200 µg/ml and higher concentrations (P = 0.009). Amikacin significantly reduced cell viability at 500 µg/ml (P = 0.002). Penicillin had no effect on cell viability at the concentrations tested (P = 0.18). Gentamicin and ceftiofur showed some interaction with the assay but had no overall effect on cell viability. At 500 µg/ml gentamicin (P<0.001), amikacin (P = 0.03), enrofloxacin (P<0.001) and ceftiofur (P<0.001) caused significant reductions in RNA levels. At 50 µg/ml gentamicin (P<0.001) and amikacin (P = 0.019) reduced BCl2 expression. Enrofloxacin produced a significant increase in COL1A2 expression (P<0.001). Conclusions: Enrofloxacin reduced MSC viability in vitro and may require cautious use in clinical situations. Penicillin has minimal detrimental effects on MSC in vitro and its use in conjunction with MSC at implantation appears safe. Further work is needed to fully investigate the effects of gentamicin, amikacin and ceftiofur. Conclusions: Clinicians using local antibiotic administration should consider the potential for local toxicity as well as the need for effective concentrations of the antibiotic.
© 2011 EVJ Ltd.
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Publication Date: 2011-09-01 PubMed ID: 21883415DOI: 10.1111/j.2042-3306.2011.00437.xGoogle Scholar: Lookup
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The researchers have carried out an in vitro study to understand how certain antibiotics impact the viability and gene expression of bone marrow-derived mesenchyme stromal cells (MSC) in horses.

Research Methodology

  • This study used equine bone marrow-derived mesenchymal stromal cells (MSC) and treated them with gentamicin, amikacin, penicillin, enrofloxacin, or ceftiofur at varying concentrations.
  • The AlamarBlue fluorescence assay, a test that measures cell viability, was applied over a period of 48 hours to track the impact of these antibiotics on MSC.
  • The cells were lysed and RNA extracted and transcribed to assess the RNA levels and determine gene expression of COL1A2, COL5A1, TNC, TNFα, CASP3, BCl2, and TGFβR2 relative to the common reference gene GAPDH.

Research Findings

  • Enrofloxacin had a concentration-dependent effect, significantly reducing cell viability at concentrations of 200µg/ml and above.
  • Amikacin notably reduced cell viability at a high concentration of 500µg/ml.
  • No significant impact on cell viability was noted with penicillin at the tested concentrations, indicating that it seems safe for use with MSC.
  • While gentamicin and ceftiofur interacted slightly with the viability assay, they didn’t affect cell viability overall.
  • On testing gene expression, a significant reduction in RNA levels was observed with concentrations of 500µg/ml of gentamicin, amikacin, enrofloxacin, and ceftiofur.
  • Significant reduction in the expression of gene BCl2 was observed with gentamicin and amikacin at 50µg/ml.
  • Enrofloxacin led to a significant increase in COL1A2 expression, indicating that it may impact the cellular functions governed by this gene.

Conclusion of the Study

  • The study concludes that enrofloxacin, in particular, can reduce MSC viability in vitro and hence needs to be used cautiously.
  • Penicillin displayed minimal detrimental effects on MSC in vitro, indicating its safety for use in conjunction with MSC during implantation.
  • There’s a need for further research on the effects of gentamicin, amikacin, and ceftiofur.
  • Lastly, clinicians using local antibiotic administration should consider potential local toxicity alongside the need for adequate concentrations of the antibiotic.

Cite This Article

APA
Parker RA, Clegg PD, Taylor SE. (2011). The in vitro effects of antibiotics on cell viability and gene expression of equine bone marrow-derived mesenchymal stromal cells. Equine Vet J, 44(3), 355-360. https://doi.org/10.1111/j.2042-3306.2011.00437.x

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 44
Issue: 3
Pages: 355-360

Researcher Affiliations

Parker, R A
  • University of Edinburgh, Large Animal Hospital, Easter Bush Veterinary Centre, Midlothian, UK. Russell.Parker@ed.ac.uk
Clegg, P D
    Taylor, S E

      MeSH Terms

      • Animals
      • Anti-Bacterial Agents / pharmacology
      • Bone Marrow Cells / drug effects
      • Bone Marrow Cells / metabolism
      • Cell Survival / drug effects
      • Cells, Cultured
      • Gene Expression Regulation / drug effects
      • Horses / genetics
      • Horses / metabolism
      • Mesenchymal Stem Cells / drug effects
      • Mesenchymal Stem Cells / metabolism

      Citations

      This article has been cited 11 times.
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        doi: 10.1111/evj.13314pubmed: 32544273google scholar: lookup
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        doi: 10.1161/STROKEAHA.115.008663pubmed: 26138122google scholar: lookup
      9. Radtke CL, Nino-Fong R, Rodriguez-Lecompte JC, Esparza Gonzalez BP, Stryhn H, McD￮ LA. Osteogenic potential of sorted equine mesenchymal stem cell subpopulations. Can J Vet Res 2015 Apr;79(2):101-8.
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      10. Gittel C, Brehm W, Burk J, Juelke H, Staszyk C, Ribitsch I. Isolation of equine multipotent mesenchymal stromal cells by enzymatic tissue digestion or explant technique: comparison of cellular properties. BMC Vet Res 2013 Oct 29;9:221.
        doi: 10.1186/1746-6148-9-221pubmed: 24168625google scholar: lookup
      11. Tan H, Ma R, Lin C, Liu Z, Tang T. Quaternized chitosan as an antimicrobial agent: antimicrobial activity, mechanism of action and biomedical applications in orthopedics. Int J Mol Sci 2013 Jan 16;14(1):1854-69.
        doi: 10.3390/ijms14011854pubmed: 23325051google scholar: lookup
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