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Home / Equine Research Bank / Equine Vet J / Amikacin induces rapid dose-dependent apoptotic cell death i...
Equine veterinary journal2020; 52(5); 715-724; doi: 10.1111/evj.13243

Amikacin induces rapid dose-dependent apoptotic cell death in equine chondrocytes and synovial cells in vitro.

Abstract: Equine veterinarians frequently inject aminoglycoside antibiotics intra-articularly, either to treat septic arthritis or for prophylaxis with other medications when injecting joints for osteoarthritis. Although aminoglycosides have been demonstrated to be toxic to equine mesenchymal stem cells (MSC), their effects on resident joint cells have not been previously investigated. Moreover, safe and effective intra-articular doses have not been defined. Objective: To determine effects of concentration, duration of exposure, pH and the presence of synovial fluid on the cytotoxic effects of amikacin on equine chondrocytes, synoviocytes and bone marrow- and adipose-derived MSC. Methods: In vitro experimental study. Methods: Four cell types were harvested from three donor horses and plated in triplicate wells for 48 hours prior to the addition of amikacin. The effects of amikacin on cell viability were assessed for different exposure times, concentrations and with pH buffered or unbuffered in media, as well as in the presence of synovial fluid. Cell metabolism/viability was assessed by colorimetric MTT assay. Cell proliferation was assessed by live cell imaging. Cell viability was assessed using trypan blue and dimeric cyanine nucleic acid stain (yoyo-1). To determine the mechanism of cell death, apoptosis was evaluated using Annexin V and 7AAD staining with flow cytometric quantification. Induction of apoptotic cell death pathways was assessed using caspase-3 expression. Results: Amikacin is cytotoxic to equine joint cells and MSC in a rapid, dose-dependent, pH-independent manner, which occurs primarily by apoptosis. Amikacin cytotoxicity was not mitigated by the addition of synovial fluid in vitro. Conclusions: Further studies are necessary to determine whether these in vitro results predict joint injury in live animal models. Conclusions: Amikacin at clinically applied doses induces rapid, pronounced cell death of equine joint cells. These findings suggest that amikacin doses currently used intra-articularly should be reconsidered pending in vivo joint titration studies.
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Publication Date: 2020-02-21 PubMed ID: 31997397DOI: 10.1111/evj.13243Google Scholar: Lookup
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  • Journal Article

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.

This research investigates how antibiotic amikacin affects equine joint cells and stem cells. The study found that amikacin, commonly used in treating joint issues in horses, leads to rapid and dose-dependent cell death, suggesting a potential reevaluation of its clinical use.

Background and Objectives

  • Veterinarians typically use the aminoglycoside antibiotic, amikacin, in intra-articular injections to treat septic arthritis or for prophylaxis mixed with other medications when treating osteoarthritis in horses.
  • Although previous studies have shown aminoglycosides can be toxic to equine mesenchymal stem cells (MSC), the effects of these antibiotics on joint cells present in the horse (chondrocytes, synoviocytes) have not been extensively examined.
  • The primary objective of this study was to understand the effects of amikacin on equine chondrocytes, synovial cells, and MSC derived from bone marrow and fat tissues. The researchers also aimed to determine how factors like concentration, exposure duration, pH levels, and synovial fluid presence influenced the observed cytotoxic impacts.

Methods

  • The researchers conducted an in vitro experimental study using cells from three donor horses.
  • Cell viability under varying exposure times, concentrations, and media conditions (buffered/unbuffered pH, synovial fluid presence) was assessed using MTT assay, trypan blue staining, yoyo-1 dimeric cyanine nucleic acid stain, and live cell imaging.
  • The researchers also evaluated the occurrence and intensity of apoptosis (or programmed cell death) via Annexin V and 7AAD staining along with flow cytometric quantification.
  • Finally, the researchers evaluated caspase-3 expression to determine the activation of apoptotic cell death pathway.

Results

  • The study reproducibly found that amikacin is cytotoxic to equine joint cells and MSC, inducing cell death in a rapid, dose-dependent, pH-independent manner.
  • The cell death mainly occurs through apoptosis, not toxic reactions.
  • Synovial fluid, which naturally occurs in joints, does not mitigate amikacin’s cytotoxic effects.

Conclusions

  • The findings recommend a reconsideration of the intra-articular doses of amikacin currently used in equine treatment.
  • More research is necessary to understand if the adverse effects observed in the in vitro study will also occur in live animals.

Cite This Article

APA
Pezzanite L, Chow L, Soontararak S, Phillips J, Goodrich L, Dow S. (2020). Amikacin induces rapid dose-dependent apoptotic cell death in equine chondrocytes and synovial cells in vitro. Equine Vet J, 52(5), 715-724. https://doi.org/10.1111/evj.13243

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 52
Issue: 5
Pages: 715-724

Researcher Affiliations

Pezzanite, Lynn
  • Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
Chow, Lyndah
  • Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
Soontararak, Sirikul
  • Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
Phillips, Jennifer
  • Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
Goodrich, Laurie
  • Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
Dow, Steven
  • Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.

MeSH Terms

  • Amikacin
  • Animals
  • Apoptosis
  • Chondrocytes
  • Horses
  • Synovial Fluid
  • Synoviocytes

Grant Funding

  • USDA National Institute of Food and Agriculture
  • Verdad Foundation
  • Shipley Foundation
  • NI18AHDR1019785G010 / Animal Health and Disease
  • Colorado State University CVMBS Veterinary Summer Scholars Program
  • Young Investigator Award in Companion Animal Studies

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