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首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >Preparation and Characterization of SDF-1α-Chitosan-Dextran Sulfate Nanoparticles
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Preparation and Characterization of SDF-1α-Chitosan-Dextran Sulfate Nanoparticles

机译：SDF-1α-壳聚糖-葡聚糖硫酸盐纳米粒子的制备与表征

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摘要

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under appropriate conditions. The glycan nanoparticles are useful carriers for protein factors, which facilitate the in vivo delivery of the proteins and sustain their retention in the targeted tissue. The glycan polyelectrolyte complexes are also ideal for protein delivery, as the incorporation is carried out in aqueous solution, which reduces the likelihood of inactivation of the proteins. Proteins with a heparin-binding site adhere to dextran sulfate readily, and are, in turn, stabilized by the binding. These particles are also less inflammatory and toxic when delivered in vivo. In the protocol described below, SDF-1α (Stromal cell-derived factor-1α), a stem cell homing factor, is first mixed and incubated with dextran sulfate. Chitosan is added to the mixture to form polyelectrolyte complexes, followed by zinc sulfate to stabilize the complexes with zinc bridges. The resultant SDF-1α-DS-CS particles are measured for size (diameter) and surface charge (zeta potential). The amount of the incorporated SDF-1α is determined, followed by measurements of its in vitro release rate and its chemotactic activity in a particle-bound form.
机译：壳聚糖（CS）和硫酸葡聚糖（DS）是带电荷的多糖（聚糖），在适当条件下混合后会形成基于聚电解质复合物的纳米颗粒。聚糖纳米颗粒是用于蛋白质因子的有用载体，其促进蛋白质的体内递送并维持其在靶组织中的保留。聚糖聚电解质复合物也是蛋白质递送的理想选择，因为掺入是在水溶液中进行的，这降低了蛋白质失活的可能性。具有肝素结合位点的蛋白质容易粘附到硫酸葡聚糖上，并通过结合而稳定。当在体内递送时，这些颗粒的炎症和毒性也较小。在下面描述的方案中，首先将干细胞归巢因子SDF-1α（Stromal cell-derivedfactor-1α，Stromal cell-derivedfactor-1α）（干细胞归巢因子）混合并与硫酸葡聚糖温育。将壳聚糖添加到混合物中以形成聚电解质复合物，然后添加硫酸锌以稳定具有锌桥的复合物。测量所得的SDF-1α-DS-CS颗粒的尺寸（直径）和表面电荷（ζ电势）。确定掺入的SDF-1α的量，然后以颗粒结合形式测量其体外释放速率和趋化活性。

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