Wagenbrenner Mike, Heinz Tizian, Horas Konstantin, Jakuscheit Axel, Arnholdt Joerg, Mayer-Wagner Susanne, Rudert Maximilian, Holzapfel Boris M, Weißenberger Manuel
Department of Orthopaedic Surgery, University of Wuerzburg, Koenig-Ludwig-Haus, Brettreichstr. 11, 97074 Wuerzburg, Germany.
Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistraße 15, 81377 Munich, Germany.
J Clin Med. 2020 Nov 29;9(12):3880. doi: 10.3390/jcm9123880.
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.
氨甲环酸(TXA)局部应用有助于预防全关节置换术后失血。尽管有这些发现,但对关节及关节周围组织的影响仍不清楚。因此,本体外研究考察了不同暴露时间和浓度的TXA对软骨细胞和人骨髓间充质干细胞(hMSC)增殖率、基因表达及分化能力的影响,其中hMSC进行了成骨分化。软骨细胞和hMSC在单层细胞培养中分离并增殖。使用含有特定生长因子的分化培养基诱导hMSC成骨分化21天。使用ATP检测分析细胞增殖情况。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。用50mg/mL的TXA处理后,观察到细胞增殖率下降。此外,用20mg/mL的TXA处理至少48小时导致软骨细胞明显脱离。用50mg/mL的TXA处理至少24小时导致软骨细胞和成骨分化的hMSC中特定标记基因的表达下降。对于浓度超过20mg/mL的TXA与暴露时间少于24小时的情况,未观察到显著影响。因此,这可能代表体内局部应用的安全限度。关于体内条件以及对hMSC功能影响的进一步研究对于全面确定TXA对关节及关节周围组织的影响是必要的。