Role of calcifying nanoparticles in the calcification process of mesenchymal stem cells in vitro

The role of calcifying nanoparticles (CNPs) in the calcification process of the outer cyst wall of hepatic cystic echinococcosis is still unknown. CNPs were isolated from the tissues of patients with cystic hepatic echinococcosis. Western blotting, alkaline phosphatase (ALP) staining, and Alizarin-Red S staining were performed to detect cellular calcium ion deposition induced by CNPs. CCK-8 and flow cytometry assays were conducted to determine the effect of CNPs on the apoptosis of mesenchymal stem cells (MSCs). Western blot experiments were performed to examine the expression level of apoptosis-related factors and TGF-β1/Smad signaling pathway activation. CNPs treatment induced the MSCs to differentiate into osteoblast-like cells. Calcification-related proteins including OPN, BMP-2, and RUNX2 were upregulated under CNPs treatment. Similarly, the CNPs exposure also increased cellular calcium ion deposition in MSCs. Furthermore, the expression of Bax and Caspase-8 was elevated by CNPs in MSCs. Treatment of CNPs promoted MSC apoptosis and inhibited the growth of MSCs. The TGF-β1/Smad signaling pathway was also activated after CNPs treatment. This study demonstrated that CNPs may play a critical role in initiating the calcification of the outer cyst wall of cystic hepatic echinococcosis and promoting the decay of the echinococcosis, providing a new strategy for the treatment of hepatic echinococcosis.

肝囊型棘球蚴病外囊壁钙化过程中，钙化纳米颗粒（CNPs）的作用尚不明确。CNPs是从肝囊型棘球蚴病患者组织中分离得到的。本研究通过Western blotting、碱性磷酸酶（ALP）染色以及阿尔扎林红S染色技术，检测了CNPs诱导的细胞钙离子沉积。CCK-8和流式细胞术实验用于确定CNPs对间充质干细胞（MSCs）凋亡的影响。Western blot实验用于检测凋亡相关因子及TGF-β1/Smad信号通路活化的表达水平。CNPs处理诱导MSCs向成骨细胞样细胞分化。在CNPs处理下，与钙化相关的蛋白如OPN、BMP-2和RUNX2表达上调。同样，CNPs暴露也增加了MSCs中的细胞钙离子沉积。此外，CNPs在MSCs中上调了Bax和Caspase-8的表达。CNPs处理促进了MSCs的凋亡并抑制了MSCs的生长。CNPs处理后，TGF-β1/Smad信号通路也被激活。本研究证实，CNPs可能在启动肝囊型棘球蚴病外囊壁的钙化过程中发挥关键作用，并促进棘球蚴的退化，为肝棘球蚴病的治疗提供了一种新的策略。