Uptake and transfection with polymeric nanoparticles are dependent on polymer end-group structure, but largely independent of nanoparticle physical and chemical properties.
Uptake and transfection with polymeric nanoparticles are dependent on polymer end-group structure, but largely independent of nanoparticle physical and chemical properties.
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DOI：
10.1021/mp3004176
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发表时间：
2012-11-05
影响因子：
4.9
通讯作者：
Green JJ
中科院分区：
文献类型：
--
作者：
Sunshine JC;Peng DY;Green JJ
开发用于基因递送的非病毒颗粒需要更好地理解使基因递送颗粒能够克服细胞内DNA递送的众多障碍的性质。线性聚（β-氨基）酯（PBAE）已显示出基因递送的实质性前景，但其有效性背后的机制并没有很好地量化这些障碍。在这项研究中，我们合成，表征，并评估基因传递的线性PBAE阵列，不同的小变化沿着的骨干，侧链，和端基的聚合物。我们研究了粒径和表面电荷，聚合物分子量，聚合物降解速率，缓冲能力，细胞摄取，转染，和细胞毒性的纳米粒子与这些聚合物配制。值得注意的是，这是第一项研究，量化了这类聚合物的小差异结构变化如何调节缓冲能力和聚合物降解速率，并将这些发现与基因递送功效联系起来。所有聚合物均形成带正电荷（ζ电位21-29 mV）的纳米级制品（~ 150 nm）。聚合物在生理条件下水解降解迅速，根据聚合物结构，半衰期为90分钟至6小时。PBAE在相关pH范围（pH 5.1 - 7.4）内的缓冲容量从34%变化到95%可质子化胺，并且基于单位质量，PBAE缓冲1.4-4.6 mmol H+/g。当与25 kDa支链聚乙烯亚胺（PEI）相比时，PBAE缓冲显著更少的质子/质量，因为PEI在相同范围内缓冲6.2mmol H+/g。然而，由于PBAE的细胞毒性相对较低，因此可以使用比PEI更高的聚合物质量来形成颗粒，并且基于PBAE的颗粒的总缓冲能力显著超过PEI的总缓冲能力。COS-7细胞的摄取范围为0%至95%的细胞，转染范围为0%至93%的细胞，这取决于基础聚合物结构和所检查的末端修饰。五种聚合物实现了比支链PEI对照更高的摄取和转染功效，具有更低的毒性。令人惊讶的是，丙烯酸酯封端的基础聚合物在吸收方面都比它们的封端版本显著地更不有效。（丙烯酸酯为1-3%，封端为75-94%）和转染效率（0-1%对20-89%），尽管丙烯酸酯和封端聚合物在凝胶电泳中的DNA阻滞、粒度、ζ电位方面存在最小差异，和细胞毒性。这些研究进一步阐明了聚合物结构在基因递送中的作用，并强调了线性聚合物的小分子端基修饰对于细胞摄取可能是至关重要的，其方式在很大程度上不依赖于聚合物/DNA结合、粒径和颗粒表面电荷。
Development of non-viral particles for gene delivery requires a greater understanding of the properties that enable gene delivery particles to overcome the numerous barriers to intracellular DNA delivery. Linear poly(beta-amino) esters (PBAE) have shown substantial promise for gene delivery, but the mechanism behind their effectiveness is not well quantified with respect to these barriers. In this study, we synthesized, characterized, and evaluated for gene delivery an array of linear PBAEs that differed by small changes along the backbone, side chain, and end-group of the polymers. We examined particle size and surface charge, polymer molecular weight, polymer degradation rate, buffering capacity, cellular uptake, transfection, and cytotoxicity of nanoparticles formulated with these polymers. Significantly, this is the first study that has quantified how small differential structural changes to polymers of this class modulate buffering capacity and polymer degradation rate and relates these findings to gene delivery efficacy. All polymers formed positively charged (zeta potential 21–29 mV) nanosized articles (~ 150 nm). The polymers hydrolytically degraded quickly in physiological conditions, with half-lives ranging from 90 minutes to 6 hours depending on polymer structure. The PBAE buffering capacities in the relevant pH range (pH 5.1 – 7.4) varied from 34% to 95% protonable amines, and on a per mass basis, PBAEs buffered 1.4–4.6 mmol H+/g. When compared to 25 kDa branched polyethyleneimine (PEI), PBAEs buffer significantly fewer protons/mass, as PEI buffers 6.2 mmol H+/g over the same range. However, due to the relatively low cytotoxicity of PBAEs, higher polymer mass can be used to form particles than with PEI and total buffering capacity of PBAE-based particles significantly exceeds that of PEI. Uptake into COS-7 cells ranged from 0% to 95% of cells and transfection ranged from 0% to 93% of cells, depending on the base polymer structure and the end-modifications examined. Five polymers achieved higher uptake and transfection efficacy with less toxicity than branched-PEI control. Surprisingly, acrylate-terminated base polymers were dramatically less efficacious than their end-capped versions, both in terms of uptake (1–3% for acrylate, 75–94% for end-capped) and transfection efficacy (0–1% vs. 20–89%), even though there are minimal differences between acrylate and end-capped polymers in terms of DNA retardation in gel electrophoresis, particle size, zeta potential, and cytotoxicity. These studies further elucidate the role of polymer structure for gene delivery and highlight that small molecule end-group modification of a linear polymer can be critical for cellular uptake in a manner that is largely independent of polymer/DNA binding, particle size, and particle surface charge.
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影响因子：
14
作者：
Eltoukhy, Ahmed A.;Siegwart, Daniel J.;Alabi, Christopher A.;Rajan, Jay S.;Langer, Robert;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.
DOI：
10.1016/j.jconrel.2011.07.009
发表时间：
2011-10-30
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Johnson RN;Chu DS;Shi J;Schellinger JG;Carlson PM;Pun SH
通讯作者：
Pun SH
影响因子：
14
作者：
Bhise, Nupura S.;Gray, Ryan S.;Sunshine, Joel C.;Htet, Soe;Ewald, Andrew J.;Green, Jordan J.
通讯作者：
Green, Jordan J.
影响因子：
4.7
作者：
Akinc, A;Anderson, DG;Langer, R
通讯作者：
Langer, R
影响因子：
29.4
作者：
Green, Jordan J.;Zugates, Gregory T.;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.