van der Maaden Koen, Sekerdag Emine, Schipper Pim, Kersten Gideon, Jiskoot Wim, Bouwstra Joke
†Division of Drug Delivery Technology, Leiden Academic Centre for Drug Research (LACDR), Leiden University, P.O. Box 2300 RA, 2311 EZ Leiden, The Netherlands.
‡Institute for Translational Vaccinology (Intravacc), P.O. Box 450, 3720 AL Bilthoven, The Netherlands.
Langmuir. 2015 Aug 11;31(31):8654-60. doi: 10.1021/acs.langmuir.5b01262. Epub 2015 Jul 27.
The aim of this work was to coat pH-sensitive microneedle arrays with inactivated polio vaccine (IPV) particles and N-trimethyl chitosan chloride (TMC) via electrostatic interactions, and assess the immunogenicity of the vaccine after topical application of the coated microneedles in rats. The surface of 200 μm long microneedles was first chemically modified with pH-sensitive (pyridine) groups and then coated with negatively charged IPV and a positively charged polymer (TMC). To obtain a sufficient high antigen dose, 10 layers of IPV were alternately coated with TMC. The binding of IPV and TMC onto pH-sensitive microneedles was quantified and visualized by using fluorescently labeled TMC and IPV. The release of IPV and TMC from the microneedles was evaluated in ex vivo human skin by fluorescence and the immunogenicity of (unlabeled) IPV was assessed after topical application of the coated microneedles in rats. pH-sensitive microneedles were homogeneously coated with 10 layers of both IPV and TMC, resulting in 45 D antigen units IPV and 700 ng TMC per microneedle array. Fluorescence microscopy imaging revealed that both IPV and TMC were released into ex vivo human skin upon application of the coated microneedles. Finally, in vivo application of IPV-TMC-coated pH-sensitive microneedles in rats led to the induction of IPV specific antibody responses, illustrating that they are practically applicable. Topical administration of pH-sensitive microneedles coated with polyelectrolyte multinanolayers of antigens and oppositely charged polymers may be a useful approach for microneedle-based vaccination.
本研究旨在通过静电相互作用，用灭活脊髓灰质炎疫苗（IPV）颗粒和氯化N-三甲基壳聚糖（TMC）包覆pH敏感微针阵列，并评估在大鼠局部应用包覆微针后疫苗的免疫原性。首先用pH敏感（吡啶）基团对200μm长的微针表面进行化学修饰，然后包覆带负电荷的IPV和带正电荷的聚合物（TMC）。为了获得足够高的抗原剂量，将10层IPV与TMC交替包覆。通过使用荧光标记的TMC和IPV对IPV和TMC与pH敏感微针的结合进行定量和可视化。通过荧光在离体人皮肤中评估IPV和TMC从微针中的释放，并在大鼠局部应用包覆微针后评估（未标记）IPV的免疫原性。pH敏感微针均匀包覆了10层IPV和TMC，每微针阵列产生45 D抗原单位的IPV和700 ng TMC。荧光显微镜成像显示，在应用包覆微针后，IPV和TMC均释放到离体人皮肤中。最后，在大鼠体内应用IPV-TMC包覆的pH敏感微针导致诱导出IPV特异性抗体反应，说明它们具有实际应用价值。局部施用包覆有抗原和带相反电荷聚合物的聚电解质多层膜的pH敏感微针可能是基于微针的疫苗接种的一种有用方法。
