Xiu Xueliang, Fu Hongyang, Zhang Ruipeng, Ma Shichao, Guo Panpan, Li Zhipeng, Zhu Yihan, Ma Fengsen
College of Pharmacy, Zhejiang University of Technology, Deqing, 313216, People's Republic of China.
Department of Dermatology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, 310006, People's Republic of China.
Int J Nanomedicine. 2025 Apr 30;20:5529-5549. doi: 10.2147/IJN.S502724. eCollection 2025.
AIM/BACKGROUND: Despite their superior safety and widespread use, split vaccines typically suffer from reduced immunogenicity due to the lack of an intact viral structure. Targeting the mannose receptors on antigen-presenting cells (APCs) with nanoparticles (NPs) and delivering them via microneedles (MNs) offers a promising solution. We designed and synthesized NPs that could form complexes with split H1N1 antigens, and evaluated the immunogenicity after loading them into dissolvable microneedle arrays (dMAs).
Man-N-HACC was synthesized by conjugating mannose moieties to N-2-hydroxypropyl trimethyl ammonium chloride chitosan (N-HACC), followed by cross-linking with tripolyphosphate to form Man-N-HACC NPs. The NPs were characterized in terms of morphology, size, zeta potential, spatial orientation, macrophage internalization, and stability. The microstructure, mechanical strength, skin penetration capability, and release behavior of dMAs loaded with Man-N-HACC NPs/H1N1 complexes were investigated. Finally, the efficacy of dMAs was assessed in a rat model using ELISA and hemagglutination inhibition (HAI) assay.
Characterization via Fourier transform infrared spectroscopy and nuclear magnetic resonance confirmed the synthesis of Man-N-HACC. The cross-linked generated Man-N-HACC NPs displayed uniform morphology and good stability over 28 days, along with confirmed spatial orientation of mannose ligands and macrophage internalization. The dMAs loaded with Man-N-HACC NPs/H1N1 exhibited mechanical robustness, capable of fully penetrating the skin and releasing nanovaccines. The increase in HAI titers and total IgG antibody levels in rat serum indicates the effectiveness of humoral immunity, and this effect only occurs after NPs formed post-crosslinking, rather than directly using raw nanomaterials, highlighting the critical role of the nanoparticle structure.
This study confirms that the delivery of Man-N-HACC NPs via dMAs provides a novel and promising approach for the administration of split influenza vaccines. Moreover, it underscores the great potential of nano-adjuvants in enhancing the efficacy of split vaccines.
目的/背景：尽管裂解疫苗具有更高的安全性且广泛应用，但由于缺乏完整的病毒结构，其免疫原性通常会降低。用纳米颗粒（NPs）靶向抗原呈递细胞（APCs）上的甘露糖受体并通过微针（MNs）进行递送提供了一种有前景的解决方案。我们设计并合成了能够与裂解H1N1抗原形成复合物的NPs，并将其加载到可溶解微针阵列（dMAs）中后评估其免疫原性。
通过将甘露糖部分与N-2-羟丙基三甲基氯化铵壳聚糖（N-HACC）共轭合成Man-N-HACC，然后与三聚磷酸交联形成Man-N-HACC NPs。对NPs的形态、大小、zeta电位、空间取向、巨噬细胞内化和稳定性进行了表征。研究了负载Man-N-HACC NPs/H1N1复合物的dMAs的微观结构、机械强度、皮肤穿透能力和释放行为。最后，使用酶联免疫吸附测定（ELISA）和血凝抑制（HAI）试验在大鼠模型中评估dMAs的效果。
通过傅里叶变换红外光谱和核磁共振表征证实了Man-N-HACC的合成。交联生成的Man-N-HACC NPs在28天内显示出均匀的形态和良好的稳定性，同时甘露糖配体的空间取向和巨噬细胞内化得到证实。负载Man-N-HACC NPs/H1N1的dMAs表现出机械稳健性，能够完全穿透皮肤并释放纳米疫苗。大鼠血清中HAI滴度和总IgG抗体水平的增加表明体液免疫有效，并且这种效果仅在交联后形成的NPs后出现，而不是直接使用原始纳米材料，突出了纳米颗粒结构的关键作用。
本研究证实通过dMAs递送Man-N-HACC NPs为裂解流感疫苗的给药提供了一种新颖且有前景的方法。此外，它强调了纳米佐剂在提高裂解疫苗效力方面的巨大潜力。
