Alendronate Alters the Release of EVs by Raw 264.7 Osteoclasts.

IF 1.5 4区 生物学 Q4 CELL BIOLOGY
Lorraine Perciliano de Faria, Victor E Arana-Chavez, Lexie Shannon Holliday
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Abstract

Alendronate (ALN), a nitrogen-containing bisphosphonate, is widely used to treat bone disorders. While its inhibitory effect on osteoclast activity is well-established, its impact on the release of extracellular vesicles (EVs) is less understood. This study investigated the effect of ALN on the quantity and size distribution of EVs released by osteoclasts cultured on bovine bone slices pretreated with 10-µM ALN, 100-µM ALN, or vehicle. Raw 264.7 cells were differentiated into osteoclasts using RANK-ligand, and EVs were isolated from conditioned media. Tartrate-resistant acid phosphatase (TRAP) staining, phalloidin staining for actin rings, and nanoparticle tracking analysis (NTA) were performed. TRAP staining showed a significant reduction in the number of TRAP-positive multinucleated cells in the 100-µM ALN group, confirming that high-concentration ALN also impairs osteoclast formation. Phalloidin staining showed a significant decrease in actin ring formation in the 100-µM ALN group, confirming ALN's inhibitory effect on osteoclast activity. NTA revealed a lower total EV concentration in the 100-µM ALN group, with a distinct peak of smaller EVs (<100 nm), suggestive of exosomes. These findings indicate that ALN, especially at higher concentrations, alters the release profile of osteoclast-derived EVs, potentially affecting intercellular communication and bone remodeling beyond its direct inhibition of resorption.

阿仑膦酸钠改变破骨细胞对ev的释放。
阿仑膦酸钠(ALN)是一种含氮双膦酸盐,被广泛用于治疗骨疾病。虽然其对破骨细胞活性的抑制作用是公认的,但其对细胞外囊泡(ev)释放的影响尚不清楚。本研究考察了ALN对破骨细胞在10µM ALN、100µM ALN和载药预处理的牛骨片上释放的ev数量和大小分布的影响。利用rank配体将264.7细胞分化为破骨细胞,并在条件培养基中分离ev。进行抗酒石酸酸性磷酸酶(TRAP)染色、肌动蛋白环的phalloidin染色和纳米颗粒跟踪分析(NTA)。TRAP染色显示100µM ALN组TRAP阳性的多核细胞数量显著减少,证实高浓度ALN也会损害破骨细胞的形成。Phalloidin染色显示100µM ALN组肌动蛋白环形成明显减少,证实了ALN对破骨细胞活性的抑制作用。NTA显示,100µM ALN组的总EV浓度较低,且较小的EV有明显的峰值(
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来源期刊
CiteScore
6.00
自引率
0.00%
发文量
32
审稿时长
1 months
期刊介绍: Journal of Histochemistry & Cytochemistry (JHC) has been a pre-eminent cell biology journal for over 50 years. Published monthly, JHC offers primary research articles, timely reviews, editorials, and perspectives on the structure and function of cells, tissues, and organs, as well as mechanisms of development, differentiation, and disease. JHC also publishes new developments in microscopy and imaging, especially where imaging techniques complement current genetic, molecular and biochemical investigations of cell and tissue function. JHC offers generous space for articles and recognizing the value of images that reveal molecular, cellular and tissue organization, offers free color to all authors.
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