Rheumatoid arthritis (RA) is an angiogenic and chronic inflammatory disease. One of the most extensively used first-line drugs against RA is methotrexate (MTX), but it shows poor solubility, short in vivo circulation, and off-target binding, leading to strong toxicity. To overcome these shortcomings, the present study loaded MTX into nanoparticles of human serum albumin modified with mannose (MTX-M-NPs) to target the drug to neutrophils. MTX-M-NPs were prepared, and their uptake by neutrophils was studied using laser confocal microscopy and flow cytometry. A chick chorioallantoic membrane assay was used to assess their ability to inhibit angiogenesis. The pharmacokinetics and tissue distribution of MTX-M-NPs were investigated using fluorescence microscopy and high-performance liquid chromatography. Their pharmacodynamics was evaluated in a rat model with arthritis induced by collagen. Neutrophils took up MTX-M-NPs significantly better than the same nanoparticles (NPs) without mannose. MTX-M-NPs markedly suppressed angiogenesis in chick embryos, and the MTX circulation was significantly longer when it was delivered as MTX-M-NPs than as a free drug. MTX-M-NPs accumulated mainly in arthritic joints. The retention of NPs was promoted by mannose-derived coating in arthritic joints. Serum levels of inflammatory cytokines, joint swelling, and bone erosion were significantly decreased by MTX-M-NPs. In conclusion, these NPs can prolong the in vivo circulation of MTX and target it to the sites of inflammation in RA, reducing drug toxicity. MTX-M-NPs allow the drug to exert its intrinsic anti-inflammatory, antiangiogenic, and analgesic properties, making it a useful drug delivery system in RA.

中文翻译：

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甘露糖包被靶向中性粒细胞的血清白蛋白纳米颗粒治疗类风湿关节炎

类风湿关节炎（RA）是一种血管生成性慢性炎症性疾病。甲氨蝶呤（MTX）是抗RA用途最广泛的一线药物，但它显示出较差的溶解度，体内循环时间短和脱靶结合，导致强烈的毒性。为了克服这些缺点，本研究将MTX加载到经甘露糖修饰的人血清白蛋白纳米颗粒（MTX-M-NPs）中，以将药物靶向中性粒细胞。制备MTX-M-NP，并使用激光共聚焦显微镜和流式细胞术研究嗜中性粒细胞的摄取。鸡绒膜尿囊膜测定用于评估其抑制血管生成的能力。使用荧光显微镜和高效液相色谱法研究了MTX-M-NPs的药代动力学和组织分布。在具有胶原蛋白诱导的关节炎的大鼠模型中评估了它们的药效学。中性粒细胞吸收MTX-M-NP明显优于不含甘露糖的相同纳米颗粒（NP）。MTX-M-NPs显着抑制了鸡胚的血管生成，当以MTX-M-NPs的形式传递时，MTX循环的时间明显长于游离药物。MTX-M-NP主要在关节炎关节中积累。NPs的保留是由关节炎关节中甘露糖衍生的涂层促进的。MTX-M-NP可显着降低血清炎症细胞因子，关节肿胀和骨侵蚀的水平。总之，这些NP可以延长MTX的体内循环并将其靶向RA的炎症部位，从而降低药物毒性。MTX-M-NP可让药物发挥其固有的抗炎，抗血管生成和止痛特性，