Intracellular Delivery of DNA and Enzyme in Active Form Using Degradable Carbohydrate-Based Nanogels
Intracellular Delivery of DNA and Enzyme in Active Form Using Degradable Carbohydrate-Based Nanogels
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DOI：
10.1021/mp300255p
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发表时间：
2012-11-01
影响因子：
4.9
通讯作者：
Narain, Ravin
中科院分区：
文献类型：
--
作者：
Ahmed, Marya;Narain, Ravin
生物分子的简易封装沿着以及有效的配制和储存使得纳米凝胶成为药物和基因递送的理想候选物。到目前为止，纳米凝胶还没有用于质粒DNA和蛋白质的共递送，由于一些限制，包括类似电荷的生物分子的低包封效率和货物材料的大小。在这项研究中，温度和pH敏感的碳水化合物为基础的纳米凝胶合成通过可逆加成-断裂链转移（RAFT）聚合技术，并详细研究其封装和共递送质粒DNA和蛋白质的能力。纳米凝胶的温度敏感特性允许生物材料的容易封装，而其酸可降解特性允许生物分子在内体中的爆发释放。因此，这些材料有望作为有效的载体，单独或作为共递送系统递送所选的生物分子。所产生的纳米凝胶是相对单分散的，并且在37摄氏度下直径约为30-40 nm。纳米凝胶的DNA缩合效率取决于纳米凝胶核心的疏水性质。DNA-纳米凝胶复合物通过纳米凝胶的碳水化合物残基与DNA的相互作用形成，并且复合物进一步用线性阳离子糖共聚物稳定。还研究了DNA-纳米凝胶复合物的蛋白质负载能力。然后在体外研究纳米凝胶的降解以及DNA和蛋白质的控制释放。此外，发现向纳米凝胶-DNA复合物中加入无毒的阳离子糖共聚物可改善细胞摄取，从而改善基因表达。
The facile encapsulation of biomolecules along with efficient formulation and storage makes nanogels ideal candidates for drug and gene delivery. So far, nanogels have not been used for the codelivery of plasmid DNA and proteins due to several limitations, including low encapsulation efficacy of biomolecule of similar charges and the size of cargo materials. In this study, temperature and pH sensitive carbohydrate-based nanogels are synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization technique and are studied in detail for their capacity to encapsulate and codeliver plasmid DNA and proteins. The temperature sensitive property of nanogels allows the facile encapsulation of biomaterials, while its acid-degradable profile allows the burst release of biomolecules in endosomes. Hence these materials are expected to serve as efficient vectors to deliver biomolecules of choice either alone or as codelivery system. The nanogels produced are relatively monodisperse and are around 30-40 nm in diameter at 37 degrees C. DNA condensation efficacy of the nanogels is dependent on the hydrophobic property of the core of the nanogels. The DNA-nanogel complexes are formed by the interaction of carbohydrate residues of nanogels with the DNA, and complexes are further stabilized with linear cationic glycopolymers. The DNA-nanogels complexes are also studied for their protein loading capacity. The degradation of the nanogels and the controlled release of DNA and proteins are then studied in vitro. Furthermore, the addition of a nontoxic, cationic glycopolymer to the nanogel-DNA complexes is found to improve the cellular uptake and hence to improve gene expression.