Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Shatin, Hong Kong.
Acta Orthop. 2009 Jun;80(3):363-7. doi: 10.3109/17453670902988386.
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.
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.
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).
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.
先前有关人肌腱细胞培养的研究表明,地塞米松和曲安奈德会降低细胞活力、抑制细胞增殖并减少胶原合成。然而,这样的细胞培养缺乏正常肌腱的细胞外基质和三维结构,从而影响它们对刺激的反应。我们建立了人肌腱组织块培养体系,并检测了地塞米松和曲安奈德对细胞活力的影响。
从健康的腘绳肌腱中制备原代人肌腱组织块培养物。从腘绳肌腱中采集肌腱条,并在添加 2%胎牛血清的 DMEM 中的 24 孔板中进行培养。将肌腱组织块用 0µM(对照)、10µM 或 100µM 地塞米松磷酸钠或 0µM(对照)、10µM 或 100µM 曲安奈德处理 96 小时。在糖皮质激素处理前后通过 Alamar Blue 检测评估细胞活力。
与对照组相比,10µM 和 100µM 地塞米松孵育分别使肌腱组织块的细胞活力降低 35%和 45%(p=0.01,混合效应方差分析)。10µM 和 100µM 曲安奈德孵育分别使细胞活力降低 33%和 36%(p=0.07,混合效应方差分析)。
人肌腱组织块培养物可用于研究糖皮质激素对人肌腱的影响。地塞米松和曲安奈德在具有基质锚定的天然 3 维环境中抑制人肌腱的细胞活力。人肌腱组织块培养物为进一步研究糖皮质激素对人肌腱细胞外基质代谢及其机械性能的影响提供了种特异性模型。