Ultrastructure of equine endothelial cells exposed to endotoxin and flunixin meglumine and equine neutrophils.
Abstract: An in vitro system of cultured equine endothelial cells was evaluated as a model for endotoxin (ET) exposure in the horse. Primary cell lines from pulmonary vessels and aortas were cultured from tissues of 6 horses. Effects of ET alone with and without serum and in combination with the cyclo-oxygenase inhibitor flunixin meglumine and isolated equine neutrophils were evaluated by transmission electron microscopy. Cells plus serum were incubated with 10, 25, 50, or 100 micrograms of ET/ml of incubation medium for 1, 3, 8, or 24 hours. Cells without serum were cultured for 1 and 3 hours. Flunixin meglumine was used at a concentration of 20 micrograms/ml. Cells also were incubated in the presence of 1,000, 5,000, or 20,000 neutrophils/ml plus ET and in the presence of a combination of ET and flunixin meglumine for 1 or 3 hours. Endotoxin alone did not cause cell damage, and the only evidence of an effect was an increased number of secondary lysosomes at incubation hour 8. At incubation hour 24, cells appeared normal. Endotoxin plus neutrophils caused cells to become round and detach from the growth substrate. Cell pathologic changes included swollen and distorted mitochondria and cytoplasmic vacuolization. Response to the ET plus neutrophil combination was variable and ranged from 5% to 50% of the cells being affected. The variability appeared to have some correlation with cell age, as well as individual preparation of neutrophils.
Publication Date: 1987-09-01 PubMed ID: 3310767
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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This research study explores the impact of endotoxin and flunixin meglumine exposure on equine endothelial cells, using an in vitro system. The interaction with neutrophils is also examined. Researchers observed responses through transmission electron microscopy on primary cell lines from horses’ pulmonary vessels and aortas, noting changes in cell shape and internal structures.
Research Methodology
- Endothelial cells from equine pulmonary vessels and aortas were cultured, using samples from six horses to set up an in vitro model for the experimentation.
- The cells were subjected to endotoxin (ET) at different concentrations and time intervals, both alone and with serum included. Effects of these conditions were evaluated using transmission electron microscopy.
- A cyclo-oxygenase inhibitor called flunixin meglumine was also introduced at a specific concentration to study how the cells would respond.
- Isolated equine neutrophils were added in defined quantities, combined with endotoxin, and the observed effects were noted.
Findings
- Endotoxin alone did not result in any substantial damage to the endothelial cells. However, there was a noticeable increase in lysosomes (cellular units used to digest foreign particles) at the eight-hour mark.
- By the 24-hour mark, the endothelial cells returned to their normal state, showing no detectable effects of endotoxin exposure.
- When the endotoxin was combined with neutrophils, the cells exhibited significant change in morphology, becoming rounder and detaching from the growing substrate. Additionally, changes within the cells were also noticed, including swelling of mitochondria and the formation of compartments filled with fluid (vacuolization) in the cytoplasm.
- Cell responses to the combination of endotoxin and neutrophils varied considerably (from 5% to 50% of the cells affected), suggesting cell age and the way neutrophils were prepared affected the outcomes.
How this study advances existing knowledge:
The findings from this study can serve as a foundation for further explorations into the effects of endotoxin, flunixin meglumine, and neutrophils on equine endothelial cell health and functionality. It introduces a viable in vitro model using horse endothelial cells for future research.
Cite This Article
APA
Turek JJ, Lamar CH, Fessler JF, Bottoms GD.
(1987).
Ultrastructure of equine endothelial cells exposed to endotoxin and flunixin meglumine and equine neutrophils.
Am J Vet Res, 48(9), 1363-1366.
Publication
Researcher Affiliations
- Department of Veterinary Anatomy, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.
MeSH Terms
- Animals
- Aorta / cytology
- Cells, Cultured
- Clonixin / analogs & derivatives
- Clonixin / pharmacology
- Endothelium, Vascular / cytology
- Endothelium, Vascular / drug effects
- Endothelium, Vascular / ultrastructure
- Endotoxins / pharmacology
- Escherichia coli
- Horses / physiology
- Neutrophils / physiology
- Nicotinic Acids / pharmacology
- Pulmonary Veins / cytology
Citations
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