Lee Sohyun, Park Seoyeon, Kim Tae-Il
Department of Agriculture, Forestry and Bioresources, College of Agriculture and Life Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
Research Institute of Agriculture and Life Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
Pharmaceutics. 2024 Dec 22;16(12):1623. doi: 10.3390/pharmaceutics16121623.
BACKGROUND/OBJECTIVES: In this study, HECP2k polymer, polyethylenimine2k (PEI2k)-modified hydroxyethyl cellulose (HEC) was utilized to form the nanocomplexes with receptor activator of nuclear factor k-B (RANK) siRNA and zoledronate (Zol) for osteoclast inhibition. HECP2k/(RANK siRNA + Zol) nanocomplexes prepared by simple mixing were anticipated to overcome the low transfection efficiency of siRNA and the low bioavailability of Zol.
The characterization of both HECP2k/(pDNA + Zol) nanocomplexes and HECP2k/(RANK siRNA + Zol) nanocomplexes was performed.
The nanocomplexes were successfully formed even in the presence of Zol, showing about 200 nm sizes and about 20 mV of positive zeta potential values suitable for efficient cellular uptake. They also possessed high endosome buffering ability by PEI and Zol, suggesting the potential for efficient endosomal escape. It was found that the low cytotoxic nanocomplexes (>90% cell viability) displayed greater transfection efficiency than PEI25k and even HECP2k polyplexes. Finally, it was found by tartrate-resistant acid phosphatase (TRAP) assay and qPCR analysis that HECP2k/(RANK siRNA + Zol) nanocomplexes could inhibit the TRAP to about 50% value and another characteristic osteoclastic gene expression, increasing FAS gene expression to about 16 times higher than control and more efficiently (about 3 times and 5 times higher, respectively) than HECP2k/siRNA polyplexes and Zol only.
HECP2k/(RANK siRNA + Zol) nanocomplexes formed by simple mixing showed great potential for inhibiting osteoclast differentiation and osteoclast activity, inducing the apoptosis via combinatorial effects of RANK siRNA and Zol.
背景/目的：在本研究中，使用了己二胺环氧化合物-2k聚合物、聚乙烯亚胺-2k（PEI2k）修饰的羟乙基纤维素（HEC）与核因子κB受体激活剂（RANK）小干扰RNA（siRNA）和唑来膦酸（Zol）形成纳米复合物以抑制破骨细胞。通过简单混合制备的HECP2k/（RANK siRNA + Zol）纳米复合物有望克服siRNA转染效率低和Zol生物利用度低的问题。
对HECP2k/（质粒DNA + Zol）纳米复合物和HECP2k/（RANK siRNA + Zol）纳米复合物进行了表征。
即使在存在Zol的情况下也成功形成了纳米复合物，其尺寸约为200 nm，zeta电位正值约为20 mV，适合细胞有效摄取。它们还通过PEI和Zol具有较高的内体缓冲能力，表明具有有效内体逃逸的潜力。发现低细胞毒性的纳米复合物（细胞活力>90%）显示出比PEI25k甚至HECP2k多聚体更高的转染效率。最后，通过抗酒石酸酸性磷酸酶（TRAP）测定和定量聚合酶链反应（qPCR）分析发现，HECP2k/（RANK siRNA + Zol）纳米复合物可将TRAP抑制至约50%，并增加另一个破骨细胞特征性基因的表达，使FAS基因表达比对照高约16倍，且比HECP2k/siRNA多聚体和单独的Zol更有效（分别高约3倍和5倍）。
通过简单混合形成的HECP2k/（RANK siRNA + Zol）纳米复合物在抑制破骨细胞分化和破骨细胞活性、通过RANK siRNA和Zol的联合作用诱导细胞凋亡方面显示出巨大潜力。
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