磺化壳聚糖的制备及其对生长因子活性的保护作用

以壳聚糖为基材,分别以氯磺酸、三甲胺.三氧化硫和丙磺酸内酯为磺化试剂,制备了3,6-O-磺化壳聚糖(OCS)、2-N-磺化壳聚糖(NCS)和2-N-磺丙基壳聚糖(PCS)3种磺化壳聚糖.采用红外光谱、核磁共振谱证明了磺化壳聚糖的结构,元素分析测定了磺化壳聚糖的磺化率.以组织修复过程中一种重要的活性因子——碱性成纤维细胞生长因子(bFGF)为目标因子,通过体外成纤维细胞培养实验,评价了不同磺化壳聚糖对bFGF活性的保护能力.结果显示,相对于未改性壳聚糖(CS),磺化壳聚糖对成纤维细胞的形态无显著影响.细胞活性检测结果显示,OCS和PCS对bFGF活性无明显的保护能力;NCS能有效提高bFGF促进成纤维细胞活性的能力,其对bFGF活性的保护能力可达肝素70%左右.磺化壳聚糖的bFGF活性保护能力不随磺化率的改变而变化.     

Preparation of mannan modified anionic PCL-PEG-PCL nanoparticles at one-step for bFGF antigen delivery to improve humoral immunity

In this article, blank anionic poly(-caprolactone)-poly(ethylene glycol)-poly(-caprolactone) (PCEC) and anionic mannan modified PCEC (MPCEC) nanoparticles with nearly the same particle size and zeta potential were prepared by emulsion solvent evaporation method. Human basic fibroblast growth factor (bFGF) was absorbed onto anionic nanoparticles surface due to electrostatic interaction. The obtained bFGF-nanoparticles complexes were injected subcutaneously into C57BL/6 mice at 20 μg of bFGF/dose on week 0, 1, 2 and 3. The mice serum was collected on week 4, and bFGF-specific autoantibody total IgG, IgG1 and IgG2a titer in serum was determined by ELISA. The results indicated that the autoantibody IgG, IgG1 and IgG2a titer of the mice immunized by bFGF–MPCEC complexes were higher than that immunized by either bFGF–PCEC or bFGF–Alum. This phenomenon might be due to that mannan functionalized MPCEC nanoparticles could be targeted to dendritic cells (DCs) to improve humoral immunity. The prepared anionic mannan modified PCEC nanoparticles (MPCEC) might have great potential application in vaccine delivery systems.