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Impact of Tranexamic Acid on Chondrocytes and Osteogenically Differentiated Human Mesenchymal Stromal Cells (hMSCs) In Vitro

机译:Tranexamic酸对软骨细胞和骨骺分化的人间充质基质细胞(HMSCs)的影响

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摘要

The topical application of tranexamic acid (TXA) helps to prevent post-operative blood loss in total joint replacements. Despite these findings, the effects on articular and periarticular tissues remain unclear. Therefore, this in vitro study examined the effects of varying exposure times and concentrations of TXA on proliferation rates, gene expression and differentiation capacity of chondrocytes and human mesenchymal stromal cells (hMSCs), which underwent osteogenic differentiation. Chondrocytes and hMSCs were isolated and multiplied in monolayer cell cultures. Osteogenic differentiation of hMSCs was induced for 21 days using a differentiation medium containing specific growth factors. Cell proliferation was analyzed using ATP assays. Effects of TXA on cell morphology were examined via light microscopy and histological staining, while expression levels of tissue-specific genes were measured using semiquantitative RT-PCR. After treatment with 50 mg/mL of TXA, a decrease in cell proliferation rates was observed. Furthermore, treatment with concentrations of 20 mg/mL of TXA for at least 48 h led to a visible detachment of chondrocytes. TXA treatment with 50 mg/mL for at least 24 h led to a decrease in the expression of specific marker genes in chondrocytes and osteogenically differentiated hMSCs. No significant effects were observed for concentrations beyond 20 mg/mL of TXA combined with exposure times of less than 24 h. This might therefore represent a safe limit for topical application in vivo. Further research regarding in vivo conditions and effects on hMSC functionality are necessary to fully determine the effects of TXA on articular and periarticular tissues.
机译:Tranexamic酸(TXA)的局部施用有助于防止总关节置换术后失血。尽管存在这些发现,但对关节和面膜组织的影响仍然尚不清楚。因此,这种体外研究检测了不同暴露时间和TXA浓度对软骨细胞和人间充质基质细胞(HMSCs)的增殖率,基因表达和分化能力的影响,该细胞和人间充质细胞(HMSCs)的效果。将软骨细胞和HMSCs分离并乘以单层细胞培养物。使用含有特异性生长因子的分化培养基诱导HMSCs的骨质植物分化21天。使用ATP测定分析细胞增殖。通过光学显微镜和组织学染色检查TXA对细胞形态的影响,同时使用半定量RT-PCR测量组织特异性基因的表达水平。在用50mg / ml TXA处理后,观察到细胞增殖率降低。此外,浓度为20mg / ml TXA的处理至少48小时导致软骨细胞的可见脱离。用50mg / ml的TXA处理至少24小时导致软骨细胞中特定标记基因表达的减少,并且骨开口分化的HMSCs。对于超过20mg / ml TXA的浓度没有观察到显着效果,其暴露时间小于24小时。因此,这可能代表体内局部应用的安全限制。关于体内病症和对HMSC功能的影响的进一步研究是必要的,以完全确定TXA对关节和围绕组织的影响。

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