Costa Lima Sofia A, Reis Salette
UCIBIO/REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
UCIBIO/REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
Colloids Surf B Biointerfaces. 2015 Sep 1;133:378-87. doi: 10.1016/j.colsurfb.2015.04.048. Epub 2015 Apr 30.
Inflammation plays a crucial role in rheumatoid arthritis progress. In the present work, a novel stealth polymeric nanospheres platform able to carry anti-inflammatory drugs and an imaging agent was developed. Incorporation of gold nanoparticles will allow photoacoustic imaging and near infra-red photothermal application. Through emulsion-diffusion evaporation technique methotrexate and gold nanoparticles were incorporated in the pegylated-poly(DL-lactic-co-glycolic acid) nanospheres. In vitro drug release assays revealed pH and temperature-dependence on gold nanoparticles. Blank nanospheres exhibited negligible in vitro cytotoxicity, while methotrexate-loaded nanospheres hampered monocytes and macrophages viability at a higher level than free methotrexate. Confocal fluorescent microscopy and flow cytometry revealed effective nanospheres internalization, and that their cellular uptake was energy dependent mediated by caveolae and clathrin-endocytosis mechanism. Finally, MTX-loaded multifunctional nanospheres containing gold lead to a significant reduction of IL-1β, IL-6 and TNF-α inflammatory cytokines produced by monocytes and macrophages upon in vitro inflammatory stimulation, suggesting a favorable anti-inflammatory activity. These results confirm that the multifunctional nanospheres represent a promising theranostic platform for RA diagnosis and intracellular treatment, by combining methotrexate and gold nanoparticles for a highly effective targeted chemo-photothermal therapy.
炎症在类风湿性关节炎的发展过程中起着关键作用。在本研究中，开发了一种新型的隐形聚合物纳米球平台，该平台能够携带抗炎药物和一种成像剂。金纳米颗粒的掺入将实现光声成像和近红外光热应用。通过乳液扩散蒸发技术，将甲氨蝶呤和金纳米颗粒掺入聚乙二醇化聚（DL-乳酸-乙醇酸）纳米球中。体外药物释放试验表明，药物释放对金纳米颗粒具有pH和温度依赖性。空白纳米球在体外表现出可忽略不计的细胞毒性，而载有甲氨蝶呤的纳米球比游离甲氨蝶呤更能抑制单核细胞和巨噬细胞的活力。共聚焦荧光显微镜和流式细胞术显示纳米球能有效内化，且其细胞摄取是能量依赖的，由小窝和网格蛋白介导的内吞作用机制介导。最后，载有甲氨蝶呤的含金多功能纳米球在体外炎症刺激后，能显著降低单核细胞和巨噬细胞产生的白细胞介素-1β、白细胞介素-6和肿瘤坏死因子-α等炎性细胞因子，表明其具有良好的抗炎活性。这些结果证实，通过将甲氨蝶呤和金纳米颗粒结合用于高效靶向化学光热疗法，多功能纳米球代表了一种用于类风湿性关节炎诊断和细胞内治疗的有前景的诊疗平台。
