Abstract: Hydrogel microneedles (MNs) have demonstrated promising application potential in the field of wound healing due to their excellent biocompatibility, high drug-loading capacity, and controllable drug release characteristics. However, most current hydrogel microneedles rely on photoinitiator-induced crosslinking, which may pose biosafety risks. Therefore, in this study, a photothermal-responsive hydrogel microneedle was constructed using o-nitrobenzyl alcohol-modified gelatin (GelNB) as the matrix and incorporating cuttlefish ink nanoparticles (CINP) as the photothermal conversion material. Under ultraviolet light irradiation, GelNB can generate aldehyde groups in situ, which subsequently undergo self-crosslinking with amino groups on its molecular chains, thus obviating the requirement for photoinitiators. The fabricated GelNB/CINP MNs exhibited excellent mechanical properties (0.99 N/needle) and could effectively penetrate the stratum corneum of the skin. Meanwhile, the incorporation of CINP endowed the MNs with favorable photothermal performance. Drug release experiments using rhodamine B as a model drug demonstrated that GelNB/CINP MNs possessed superior near-infrared light (NIR)-triggered controlled drug release capability. In vitro cell experiments revealed that GelNB/CINP MNs exhibited good biocompatibility and cell proliferation-promoting functions. Furthermore, under NIR irradiation, GelNB/CINP MNs displayed significant antibacterial activity against Escherichia coli (antibacterial rate >90%). Therefore, the GelNB/CINP MNs developed in this study provide a promising novel drug delivery platform for safe and efficient wound treatment.