Glutamine-chitosan modified calcium phosphate nanoparticles for efficient siRNA delivery and osteogenic differentiation.
Glutamine-chitosan modified calcium phosphate nanoparticles for efficient siRNA delivery and osteogenic differentiation.
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DOI：
10.1039/c5tb00843c
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发表时间：
2015-08-21
期刊：
影响因子：
--
通讯作者：
Lee M
中科院分区：
文献类型：
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作者：
Choi B;Cui ZK;Kim S;Fan J;Wu BM;Lee M
利用小干扰RNA（siRNA）的基于RNA干扰（RNAi）的治疗显示出治疗疾病的巨大潜力。尽管基于磷酸钙（CaP）的系统由于其良好的生物相容性和与核酸的高亲和力而成为递送核酸的有吸引力的选择，但它们受到不可控的颗粒形成和不一致的转染效率的限制。在这项研究中，我们开发了一种稳定的钙磷纳米载体系统，通过添加高度阳离子，谷氨酰胺共轭低聚壳聚糖（Gln-OChi）增强细胞内摄取。用Gln-OChi（CaP/Gln-OChi）包被的CaP纳米颗粒显著增强永生化细胞系（HeLa）和原代间充质干细胞（MSC）中的基因转染和敲除效率，具有最小的细胞毒性。通过使用包封MSC的可光交联壳聚糖水凝胶和负载有靶向骨形态发生蛋白拮抗剂noggin的siRNA的颗粒，在三维环境中进一步证实了负载siRNA的CaP/Gln-OChi颗粒的成骨生物活性。这些发现表明我们的CaP/Gln-OChi纳米载体为治疗应用提供了有效且安全的基因递送系统。
RNA interference (RNAi)-based therapy using small interfering RNA (siRNA) exhibits great potential to treat diseases. Although calcium phosphate (CaP)-based systems are attractive options to deliver nucleic acids due to their good biocompatibility and high affinity with nucleic acids, they are limited by uncontrollable particle formation and inconsistent transfection efficiencies. In this study, we developed a stable CaP nanocarrier system with enhanced intracellular uptake by adding highly cationic, glutamine-conjugated oligochitosan (Gln-OChi). CaP nanoparticles coated with Gln-OChi (CaP/Gln-OChi) significantly enhanced gene transfection and knockdown efficiency in both immortalized cell line (HeLa) and primary mesenchymal stem cells (MSCs) with minimal cytotoxicity. The osteogenic bioactivity of siRNA-loaded CaP/Gln-OChi particles was further confirmed in three-dimensional environments by using photocrosslinkable chitosan hydrogels encapsulating MSCs and particles loaded with siRNA targeting noggin, a bone morphogenetic protein antagonist. These findings suggest that our CaP/Gln-OChi nanocarrier provides an efficient and safe gene delivery system for therapeutic applications.