Histopathology

One of the reasons why there are currently few reports describing intranasal vaccination using oil-based systems may pertain to concerns relating to potentially adverse effects on the nasal mucosa. To assess whether S/O nanodispersions could cause structural changes in the mucosal membrane, nasal toxicity was evaluated via H&E staining. Large areas of the nasal epithelium and cilia were damaged in the noses of mice treated with IPA, suggesting high toxicity of IPA to the nasal mucosa (Fig. 4-8). Conversely, in the group of mice administered S/O nanodispersions the nasal cilia appeared bushy and regular, and the epithelium was intact.

No removal of mucosa or cells of the epithelium was observed, which was similar to the group of mice administered PBS alone. Therefore, S/O nanodispersions may be safe for intranasal vaccination and appropriate for use as antigenic carriers in this context.

Fig. 4-8 Morphology of nasal mucosa in mice after intranasal administration of ovalbumin in solid-in-oil nanodispersions (S/O sol.), phosphate-buffered saline solution (PBS sol.), or isopropanol solution (IPA sol.). The images were captured using a fluorescence microscope with×4, ×20, and×40 objective lenses. Cilia are indicated by arrows.

To the best of our knowledge this is the first report describing intranasal vaccination using oil-based nanotechnology. Notably however, the mechanisms involved in the effects of intranasal immunization via S/O nanodispersions remain unclear. The main mechanism may be similar to that of water-based particle preparations in that antigens in the vaccine are transported to the immune system by M cells in NALT [6–8]. S/O nanodispersions maybe stable after contact with mucin on the surface of nasal mucosa, which has an amphiphilic structure, and first form a solid-in-oil-in-water emulsion and then be taken up by M cells. To further improve the immunogenicity of S/O nanodispersions, the incorporation of other adjuvants such as CpG oligoribodeoxynucleotides or resiquimod R848 will be investigated in the future [37,50].

4 Conclusion

Oil-based formulations based on S/O technology exhibit potential as intranasal vaccine carriers. In the present study, in in vivo nasal absorption experiments S/O formulations were associated with comparatively longer retention of OVA in the nasal cavity, and in vivo immunization induced strong mucosal and systemic immune responses. Intranasal vaccination using S/O nanodispersions may constitute a relatively simpler method of effective en masse immunization against infectious pathogens in the future.

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