Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter, Amarillo, Texas 79106, USA.
Mol Pharm. 2011 Apr 4;8(2):405-15. doi: 10.1021/mp100255c. Epub 2011 Jan 11.
Porous poly(L-lactic acid) (PLA) and poly(lactic-co-glycolic acid) (PLGA) nanoparticles were tested for pulmonary delivery of hepatitis B vaccine. In particular, the effects of particle size and hydrophobicity on mucosal and cell-mediated immune responses were investigated. Three formulations of PLA and PLGA nanoparticles containing a fixed amount of hepatitis B surface antigen (HBsAg) were prepared by a double-emulsion-solvent-evaporation method and characterized for surface morphology, charge, size, density and in vitro release. The immune responses were studied by measuring secretory IgA levels in mucosal fluids and quantitating cytokine levels in rat spleen homogenates. Particle uptake was studied in rat alveolar macrophages. Scanning electron microscopy revealed particles with smooth surfaces. Zeta potential measurements indicated that the particles carried negative surface charges. The antigen was continuously released for 42 days in phosphate buffer. Hydrophobic particles >500 nm elicited a more robust increase in secretary IgA, interleukin-2 and interferon-γ levels compared to hydrophilic particles <500 nm. Large hydrophobic particles were more efficiently internalized by rat alveolar macrophages compared to smaller hydrophilic particles. Calu-3 cell viability studies indicate that the viability of cells is not affected by nanoparticulate formulations. This study demonstrates that inhalable nanoparticles of HBsAg produce an enhancement of immune responses.
多孔聚（L-丙交酯）（PLA）和聚（丙交酯-乙交酯）（PLGA）纳米粒子被测试用于乙型肝炎疫苗的肺部递送。特别是，研究了粒径和疏水性对黏膜和细胞介导的免疫应答的影响。通过双乳液-溶剂蒸发法制备了三种含有固定量乙型肝炎表面抗原（HBsAg）的 PLA 和 PLGA 纳米粒子制剂，并对其表面形态、电荷、粒径、密度和体外释放进行了表征。通过测量黏膜液中分泌型 IgA 水平和定量大鼠脾匀浆中细胞因子水平来研究免疫反应。研究了颗粒摄取在大鼠肺泡巨噬细胞中的作用。扫描电子显微镜显示颗粒表面光滑。Zeta 电位测量表明颗粒带有负表面电荷。抗原在磷酸盐缓冲液中持续释放 42 天。与<500nm 的亲水性颗粒相比，>500nm 的疏水性颗粒引起的分泌型 IgA、白细胞介素-2 和干扰素-γ水平的增加更为显著。与较小的亲水性颗粒相比，大鼠肺泡巨噬细胞更有效地摄取大的疏水性颗粒。Calu-3 细胞活力研究表明，纳米颗粒制剂不会影响细胞活力。本研究表明，可吸入的 HBsAg 纳米颗粒可增强免疫应答。
