Targeted delivery of complexes of biotin-PEG-polyethylenimine and NF-κB decoys to brain-derived endothelial cells in vitro
Targeted delivery of complexes of biotin-PEG-polyethylenimine and NF-κB decoys to brain-derived endothelial cells in vitro
复制标题
DOI：
10.1007/s11095-007-9389-y
复制
发表时间：
2008-03-01
影响因子：
3.7
通讯作者：
Bickel, Ulrich
中科院分区：
文献类型：
--
作者：
Bhattacharya, Raktima;Osburg, Berit;Bickel, Ulrich
目的。评估将含有寡脱氧核苷酸（ODN）诱饵的多聚体摄取机制从吸收介导转向受体介导的内吞作用对核因子κ B （nf - κ B）的影响。材料与方法。将odn与生物素-聚乙二醇和聚亚胺（BPP）共聚物结合抗体8D3和链霉亲和素（8D3SA）靶向脑源性内皮细胞。采用动态光散射法测定了ODN/BPP配合物的尺寸和稳定性。用共聚焦显微镜观察细胞摄取。研究了肿瘤坏死因子刺激小鼠bEnd5细胞的细胞活力和药理作用。ODN/BPP配合物的尺寸为116 +/- 2.3 nm，与8D3SA偶联后增大了40 nm，在生理体液中稳定。靶向复合物被完整地内化到内体腔室中。处理条件对bEnd5细胞VCAM-1、ICAM-1、I κ B α和iNOS mRNA表达有显著抑制作用，但不影响细胞活力。在0.5 μ M时，诱饵ODN作为8d3sa靶向复合物递送，可显著抑制单核细胞对bEnd5单层的粘附，而较高浓度的非靶向复合物则无效。nf - κ B诱骗物和BPP复合物可靶向转铁蛋白受体，是治疗影响血脑屏障的神经炎性疾病的有前途的候选药物。
Purpose. To evaluate the effect of re-directing the uptake mechanism of polyplexes containing oligodeoxynucleotide (ODN) decoys to nuclear factor kappa B (NF-kappa B) from absorptive-mediated to receptor-mediated endocytosis.Materials and Methods. Complexes of ODNs and a co-polymer of biotin-polyethylenglycol and polyethylenimine (BPP) were targeted to brain-derived endothelial cells with a conjugate of antibody 8D3 and streptavidin (8D3SA). Size and stability of ODN/BPP complexes was measured by dynamic light scattering. Cellular uptake was studied by confocal microscopy. Cell viability and pharmacological effects were investigated on murine bEnd5 cells stimulated with tumor necrosis factor.Results. ODN/BPP complexes showed sizes of 116 +/- 2.3 nm, which increased by 40 nm when coupled to 8D3SA, and were stable in physiological fluids. Targeted complexes were internalized intact into endosomal compartments. Treatment conditions, which yielded significant inhibitory effects on mRNA expression of VCAM-1, ICAM-1, I kappa B alpha and iNOS by bEnd5 cells, did not affect viability. At 0.5 mu M, decoy ODN significantly inhibited monocyte adhesion to bEnd5 monolayers when delivered as 8D3SA-targeted complex, while higher concentrations of untargeted complex were ineffective.Conclusions. The complex of NF-kappa B decoys and BPP, which can be targeted to transferrin receptors, is a promising drug candidate for neuroinflammatory diseases affecting the blood-brain barrier.