The effect of glucocorticoids on tendon cell viability in human tendon explants.
OBJECTIVE
Previous studies on the culture of human tenocytes have shown that dexamethasone and triamcinolone reduce cell viability, suppress cell proliferation, and reduce collagen synthesis. However, such cell cultures lack the extracellular matrix and three-dimensional structure of normal tendons, which affects their response to stimuli. We established a human tendon explant culture system and tested the effects of dexamethasone and triamcinolone on cell viability.


METHODS
Primary human tendon explant cultures were prepared from healthy hamstring tendons. Tendon strips were harvested from hamstring tendons and cultured in 24-well plates in Dulbecco's modification of Eagle's Medium (DMEM) supplemented with 2% fetal calf serum. The tendon explants were treated with 0 microM (control), 10 microM, or 100 microM dexamethasone sodium phosphate or 0 microM (control), 10 microM, or 100 microM triamcinolone acetonide in DMEM for 96 h. Cell viability was measured by Alamar blue assay before and after glucocorticoid treatment.


RESULTS
Incubation with 10 microM and 100 microM dexamethasone reduced cell viability in human tendon explants by 35% and 45%, respectively, as compared to a 6% increase in the controls (p = 0.01, mixed-effects ANOVA). Triamcinolone at 10 microM and 100 microM reduced cell viability by 33% and 36%, respectively, as compared to a 9% increase in the controls (p = 0.07, mixed-effects ANOVA).


CONCLUSIONS
Human tendon explant cultures can be used to study the effects of glucocorticoids on human tendon. Dexamethasone and triamcinolone suppress the cell viability of human tendon in its natural 3-dimensional environment with matrix anchorage. Human tendon explant cultures provide a species-specific model for further investigation of the effects of glucocorticoids on the metabolism of the extracellular matrix of human tendon, and on its mechanical properties.
Wong
Margaret Wan Nar
MW
Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Shatin, Hong Kong. mwnwong@cuhk.edu.hk
Lui
Wai Ting
WT
Fu
Sai Chuen
SC
Lee
Kwong Man
KM
D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D013710:Tendons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014221:Triamcinolone / Q000494:pharmacology
D005938:Glucocorticoids; D014221:Triamcinolone; D003907:Dexamethasone