Division of Hand Surgery, Plastic Surgery and Aesthetic Surgery, University Hospital, LMU Munich, Ziemssenstraße 5, 80336 Munich, Germany.
Int J Mol Sci. 2024 Jul 13;25(14):7690. doi: 10.3390/ijms25147690.
This study investigates the impact of acetylsalicylic acid (ASA), also known as aspirin, on adipose tissue-derived stem cells (ASCs), aiming to elucidate its dose-dependent effects on morphology, viability, proliferation, and osteogenic differentiation. Isolated and characterized human ASCs were exposed to 0 µM, 100 µM, 200 µM, 400 µM, 800 µM, 1000 µM, 10,000 µM, and 16,000 µM of ASA in vitro. Cell morphology, viability, and proliferation were evaluated with fluorescent live/dead staining, alamarBlue viability reagent, and CyQUANT cell proliferation assay, respectively. Osteogenic differentiation under stimulation with 400 µM or 1000 µM of ASA was assessed with alizarin red staining and qPCR of selected osteogenic differentiation markers (RUNX2, SPP1, ALPL, BGLAP) over a 3- and 21-day-period. ASA doses ≤ 1000 µM showed no significant impact on cell viability and proliferation. Live/dead staining revealed a visible reduction in viable cell confluency for ASA concentrations ≥ 1000 µM. Doses of 10,000 µM and 16,000 µM of ASA exhibited a strong cytotoxic and anti-proliferative effect in ASCs. Alizarin red staining revealed enhanced calcium accretion under the influence of ASA, which was macro- and microscopically visible and significant for 1000 µM of ASA ( = 0.0092) in quantification if compared to osteogenic differentiation without ASA addition over a 21-day-period. This enhancement correlated with a more pronounced upregulation of osteogenic markers under ASA exposure (ns). Our results indicate a stimulatory effect of 1000 µM of ASA on the osteogenic differentiation of ASCs. Further research is needed to elucidate the precise molecular mechanisms underlying this effect; however, this discovery suggests promising opportunities for enhancing bone tissue engineering with ASCs as cell source.
本研究旨在探讨乙酰水杨酸（ASA），又称阿司匹林，对脂肪组织来源的干细胞（ASCs）的影响，阐明其对形态、活力、增殖和成骨分化的剂量依赖性作用。分离并鉴定的人 ASCs 体外暴露于 0µM、100µM、200µM、400µM、800µM、1000µM、10000µM 和 16000µM 的 ASA 中。通过荧光死活染色、alamarBlue 活力试剂和 CyQUANT 细胞增殖测定分别评估细胞形态、活力和增殖。通过茜素红染色和选定的成骨分化标志物（RUNX2、SPP1、ALPL、BGLAP）的 qPCR 在 3 天和 21 天的时间内评估刺激下 400µM 或 1000µM 的 ASA 诱导的成骨分化。≤1000µM 的 ASA 剂量对细胞活力和增殖没有显著影响。活/死染色显示，1000µM 以上 ASA 浓度的活细胞融合度明显降低。10000µM 和 16000µM 的 ASA 剂量对 ASCs 表现出强烈的细胞毒性和抗增殖作用。ASA 影响下的茜素红染色显示钙积累增强，在宏观和微观上是可见的，在 21 天的时间内与没有添加 ASA 的成骨分化相比，1000µM 的 ASA（=0.0092）具有统计学意义。这种增强与 ASA 暴露下成骨标志物的更明显上调相关（无统计学意义）。我们的结果表明 1000µM 的 ASA 对 ASCs 的成骨分化具有刺激作用。需要进一步研究阐明这种作用的确切分子机制；然而，这一发现为使用 ASCs 作为细胞来源增强骨组织工程提供了有前景的机会。
