Arnedo A, Irache J M, Merodio M, Espuelas Millán M S
Centro Galénico, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de Navarra, Irunlarrea 1, Ap. 177, 31080 Pamplona, Spain.
J Control Release. 2004 Jan 8;94(1):217-27. doi: 10.1016/j.jconrel.2003.10.009.
The goal of this study was to evaluate the potential of albumin nanoparticles as a delivery system for antisense oligonucleotides. Nanoparticles were prepared by a coacervation process and cross-linkage with glutaraldehyde. Phosphodiester (PO) and phosphorotioate (PS) oligonucleotides were either adsorbed on the surface of nanoparticles (PO-NPA and PS-NPA) or incorporated in the nanoparticle matrix (PO-NPB and PS-NPB). When PO-loaded nanoparticles were incubated with phosphodiesterase, only NPB was able to keep the oligonucleotide hybridization capability for at least 60 min. The antiviral activity was evaluated in MRC-5 fibroblasts infected with human cytomegalovirus at a MOI of 0.0035. Both PO nanoparticle formulations significantly increased the antiviral activity of free PO (P<0.001) and NPB showed slightly higher efficacies than NPA (P<0.05). On the other hand, PS exhibited significant higher activity than free PO (P<0.001), however, no significant differences were found between PS-nanoparticle and PO-nanoparticle formulations. These findings were well correlated with the intracellular distribution observed for fluorescent oligonucleotide-loaded albumin nanoparticles. Even these carriers delayed and decreased the uptake of PO by MRC-5 cells, they finally induced a diffused cytoplasmic distribution and major nuclear accumulation. In summary, albumin nanoparticles partially protected a PO against enzymatic degradation and improved their presence in the nucleus and thus, increased its efficiency.
本研究的目的是评估白蛋白纳米颗粒作为反义寡核苷酸递送系统的潜力。纳米颗粒通过凝聚过程和与戊二醛交联制备。磷酸二酯（PO）和硫代磷酸酯（PS）寡核苷酸要么吸附在纳米颗粒表面（PO-NPA和PS-NPA），要么掺入纳米颗粒基质中（PO-NPB和PS-NPB）。当将负载PO的纳米颗粒与磷酸二酯酶一起孵育时，只有NPB能够在至少60分钟内保持寡核苷酸杂交能力。在感染人巨细胞病毒且感染复数为0.0035的MRC-5成纤维细胞中评估抗病毒活性。两种PO纳米颗粒制剂均显著提高了游离PO的抗病毒活性（P<0.001），且NPB的疗效略高于NPA（P<0.05）。另一方面，PS表现出比游离PO显著更高的活性（P<0.001），然而，PS纳米颗粒制剂和PO纳米颗粒制剂之间未发现显著差异。这些发现与负载荧光寡核苷酸的白蛋白纳米颗粒的细胞内分布密切相关。即使这些载体延迟并减少了MRC-5细胞对PO的摄取，但它们最终诱导了弥散的细胞质分布和主要的核积累。总之，白蛋白纳米颗粒部分保护PO免受酶降解，并改善了其在细胞核中的存在，从而提高了其效率。
