In the near future oral polio vaccine (OPV) will be replaced by inactivated polio vaccine (IPV) as part of the eradication program of polio. For that reason, there is a need for substantial amount of safe and more affordable IPV for low-income countries. Bioneedles, which are biodegradable mini-implants, have the potential to deliver vaccines outside the cold-chain and administer them without the use of needles and syringes. In the current study, Bioneedles were filled with IPV, subsequently lyophilized, and antigenic recoveries were determined both directly after IPV-Bioneedle preparation as well as after elevated stability testing. Further, we assessed the immunogenicity of IPV-Bioneedles in rats and the residence time at the site of administration. Trivalent IPV was formulated in Bioneedles with recoveries of 101±10%, 113±18%, and 92±15%, respectively for serotypes 1, 2 and 3. IPV in Bioneedles is more resistant to elevated temperatures than liquid IPV: liquid IPV retained less than half of its antigenicity after 1 day at 45°C and IPV in Bioneedles showed remaining recoveries of 80±10%, 85±4% and 63±4% for the three serotypes. In vivo imaging revealed that IPV administered via Bioneedles as well as subcutaneously injected liquid IPV showed a retention time of 3 days at the site of administration. Finally, an immunogenicity study showed that IPV-filled Bioneedles are able to induce virus-neutralizing antibody titers similar to those obtained by liquid intramuscular injection when administered in a booster regime. The addition of LPS-derivate PagL in IPV-filled Bioneedles did not increase immunogenicity compared to IPV-Bioneedles without adjuvant. The current study demonstrates the pre-clinical proof of concept of IPV-filled Bioneedles as a syringe-free alternative delivery system. Further pre-clinical and clinical studies will be required to assess the feasibility whether IPV-Bioneedles show sufficient safety and efficacy, and may contribute to the efforts to eradicate and prevent polio in the future.