Choi Bogyu, Cui Zhong-Kai, Kim Soyon, Fan Jiabing, Wu Benjamin M, Lee Min
Division of Advanced Prosthodontics, University of California Los Angeles, 10833 Le Conte Avenue, Los Angeles, California 90095, USA.
Department of Bioengineering, University of California Los Angeles, 410 Westwood Plaza, Los Angeles, California 90095, USA.
J Mater Chem B. 2015 Aug 21;3(31):6448-6455. doi: 10.1039/C5TB00843C.
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.
基于小干扰RNA（siRNA）的RNA干扰（RNAi）疗法在治疗疾病方面具有巨大潜力。尽管基于磷酸钙（CaP）的系统因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。在本研究中，我们通过添加高度阳离子化的谷氨酰胺共轭低聚壳聚糖（Gln-OChi）开发了一种具有增强细胞内摄取能力的稳定CaP纳米载体系统。涂有Gln-OChi的CaP纳米颗粒（CaP/Gln-OChi）在永生化细胞系（HeLa）和原代间充质干细胞（MSC）中均显著提高了基因转染和敲低效率，且细胞毒性最小。通过使用包裹MSC和负载靶向骨形态发生蛋白拮抗剂头蛋白的siRNA颗粒的光交联壳聚糖水凝胶，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一种高效且安全的基因递送系统。
