Characterization of rabies pDNA nanoparticulate vaccine in poloxamer 407 gel
Characterization of rabies pDNA nanoparticulate vaccine in poloxamer 407 gel
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DOI：
10.1016/j.ijpharm.2018.05.018
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发表时间：
2018-07-10
影响因子：
5.8
通讯作者：
D'Souza, Martin J.
中科院分区：
文献类型：
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作者：
Bansal, Amit;Wu, Xianfu;D'Souza, Martin J.
质粒DNA(PDNA)疫苗具有预防包括狂犬病在内的多种疾病的潜力，但降解速度快，抗原提呈细胞摄取能力差。为了克服这些限制，我们制造了一种pDNA纳米颗粒疫苗。将带负电荷的多聚脱氧核糖核酸吸附在阳离子聚(D，L-丙交酯-乙交酯)-壳聚糖纳米粒表面，作为载体。为了创造一种可持续释放疫苗的水凝胶，我们将PDNA纳米颗粒分散在泊洛沙姆407凝胶中，该凝胶在4摄氏度时是液体，在37℃时变成软凝胶，提供了易于给药和防止PDNA突然释放的能力。当PDNA与纳米粒子的比例(P/N)为1/50时，可将PDNA完全固定到阳离子纳米粒子上。细胞对纳米粒的摄取具有时间和浓度依赖性，并遵循饱和动力学，K-m和V-max分别为139.48和11.389微克/毫升·h。体外释放研究表明，纳米颗粒疫苗具有长达24天的持续释放。综上所述，PDNA PLGA-壳聚糖纳米粒是无毒的，其缓冲能力和细胞摄取能力增强，并保持了PDNA的释放。我们预计我们的PDNA疫苗的效力将在动物实验中得到极大的提高。
Plasmid DNA (pDNA) vaccines have the potential for protection against a wide range of diseases including rabies but are rapid in degradation and poor in uptake by antigen-presenting cells. To overcome the limitations, we fabricated a pDNA nanoparticulate vaccine. The negatively charged pDNA was adsorbed onto the surface of cationic PLGA (poly (D, L-lactide-co-glycolide))-chitosan nanoparticles and were used as a delivery vehicle. To create a hydrogel for sustainable vaccine release, we dispersed the pDNA nanoparticles in poloxamer 407 gel which is liquid at 4 degrees C and turns into soft gels at 37 degrees C, providing ease of administration and preventing burst release of pDNA. Complete immobilization of pDNA to cationic nanoparticles was achieved at a pDNA to nanoparticles ratio (P/N) of 1/50. Cellular uptake of nanoparticles was both time and concentration dependent and followed a saturation kinetics with V-max of 11.389 mu g/mL h and K-m of 139.48 mu g/mL. The in vitro release studies showed the nanoparticulate vaccine has a sustained release for up to 24 days. In summary, pDNA PLGA-chitosan nanoparticles were non-cytotoxic, their buffering capacity and cell uptake were enhanced, and sustained the release of pDNA. We expect our pDNA vaccine's potency will be greatly improved in the animal studies.