# Dissolvable Microneedle Delivery of a Replication-Deficient Orthopoxvirus Vaccine: Formulation Screening and Immunogenicity Evaluation for Monkeypox Prevention

## TL;DR

Researchers developed a dissolvable microneedle vaccine for monkeypox that is stable at room temperature and induces strong immune responses in mice.

## Contribution

A thermostable dissolvable microneedle vaccine formulation for replication-deficient orthopoxvirus was developed and tested for monkeypox prevention.

## Key findings

- Formulation F2 retained potency with minimal loss after 2 months at 25 °C and 1 week at 37 °C.
- DMN patches dissolved within 5 minutes and induced robust MPXV-specific IgG and neutralizing antibodies in mice.
- The vaccine's immunogenicity was comparable to conventional intramuscular injection.

## Abstract

Background: The global spread of monkeypox virus (MPXV) highlights an urgent need for thermostable and easily administrable vaccines. Current orthopoxvirus vaccines are limited by cold-chain dependence and inconvenient injection-based delivery. Objectives: This study aimed to develop a dissolvable microneedle (DMN) vaccine against monkeypox based on a replication-deficient orthopoxvirus platform, through systematic formulation screening, stabilization mechanism exploration, and rigorous in vivo immunogenicity evaluation. Methods: A film-based approach was adopted for efficient, high-throughput formulation screening and thermostability assessment. NTV was mixed with excipients and dried into solid films. Stability was monitored via RT-qPCR after storage at 4 °C to 40 °C. The lead formulation was physically characterized, then used to fabricate MVA-BN-loaded DMN patches, which were further evaluated for in vivo immunogenicity via immunization in BALB/c mice. Results: The optimal formulation F2 (containing dextran, L-threonine, and BSA/HSA) showed a potency loss of only ~1 log10 after 2 months at 25 °C, and <1 log10 loss after 1 week at 37 °C. SEM revealed a porous virus-entrapment morphology, and FTIR indicated enhanced hydrogen bonding between the virus and the dextran matrix. The formulation was successfully manufactured into DMNs that dissolved within 5 min. In mice, these DMNs elicited robust MPXV-specific IgG and neutralizing antibody responses, with immunogenicity comparable to that induced by conventional intramuscular injection. Conclusions: This study successfully established a thermostable formulation and dissolvable microneedle delivery platform for replication-deficient orthopoxvirus vaccines against monkeypox. The optimized DMN vaccine induced robust MPXV-specific immune responses in mice with immunogenicity comparable to intramuscular injection, addressing the core limitations of current vaccines and providing a promising solution for monkeypox prevention.