Nanobiofar Group, Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Santiago de Compostela, Campus Vida, E-15782, Santiago de Compostela, Spain.
Colloids Surf B Biointerfaces. 2013 Mar 1;103:615-23. doi: 10.1016/j.colsurfb.2012.11.009. Epub 2012 Nov 23.
Hyaluronic acid (HA) has been described as a biocompatibility enhancer for gene delivery systems; however, the mechanistic implications of its inclusion on the formation and activity of such systems and subsequent gene release are poorly understood. To better understand these issues, we describe herein the preparation and characterization of chitosan and chitosan-hyaluronic acid nanoparticles (CS and CS:HA NPs) for gene silencing. Different formulations were prepared by ionotropic gelation and evaluated for their physicochemical properties and biological activities in A549-Luc cells. Inclusion of HA to CS NPs resulted in a comparable silencing activity with Lipofectamine RNAiMAX (≈85% of luciferase knockdown) and significantly improved cell viability compared with CS NPs. As depicted by isothermal titration calorimetry, HA competed with siRNA for CS binding, lowering CS-siRNA binding strength by 25%. This suggests that besides improving cell biocompatibility of CS NPs, HA might also promote their gene release by loosening the CS-siRNA binding.
透明质酸 (HA) 已被描述为基因传递系统的生物相容性增强剂；然而，其对这些系统的形成和活性以及随后的基因释放的影响的机制尚不清楚。为了更好地理解这些问题，我们在此描述了壳聚糖和壳聚糖-透明质酸纳米粒子 (CS 和 CS:HA NPs) 的制备和表征，用于基因沉默。通过离子凝胶化制备了不同的配方，并在 A549-Luc 细胞中评估了它们的物理化学性质和生物活性。将 HA 包含到 CS NPs 中会导致与 Lipofectamine RNAiMAX 相当的沉默活性（≈85%的荧光素酶敲低），并且与 CS NPs 相比，细胞活力显著提高。如等温滴定量热法所示，HA 与 siRNA 竞争 CS 的结合，降低 CS-siRNA 结合强度 25%。这表明，除了提高 CS NPs 的细胞生物相容性外，HA 还可能通过松动 CS-siRNA 的结合来促进其基因释放。
