This study presents the development of hydrogel microneedles (MN) with programmed mesophase transitions for controlled drug delivery. The MNs were fabricated from triblock amphiphiles with a hydrophilic poly(ethylene glycol) (PEG) middle block and two dendritic side-blocks with enzyme-cleavable hydrophobic end-groups. To enhance mechanical strength, a sodium alginate base layer and various polymeric excipients were incorporated. The MNs were successfully fabricated and exhibited favorable insertion efficiency and low height reduction when tested in a skin-simulant model. They demonstrated adequate mechanical strength under static forces up to 1000 g without fractures or broken segments. In buffer solution, the solid MNs swelled into a hydrogel within 30 seconds, followed by rapid disintegration into small hydrogel particles that could undergo slow enzymatic degradation into soluble polymers. In vitro release studies of dexamethasone (DEX) showed first-order release, with 90% released within 6 days. DEX-loaded MNs penetrated chicken skin completely, demonstrating their potential for self-administration, improved patient compliance, and sustained drug release. The study highlights the feasibility of programming hydrogel-forming MNs to undergo mesophase transitions and their potential as a delivery system for controlled drug release. The MNs can be used for targeted drug delivery, increasing the residence time of the drug at the target site, enabling better penetration and sustained delivery over an extended period while minimizing side effects. The results indicate that the mechanical properties of the MNs are significantly influenced by the molecular weight of PEG and the drug loading. The MNs showed good mechanical strength and were able to penetrate the skin, demonstrating their potential for transdermal drug delivery. The study also shows that the MNs can be used for controlled drug release, with the drug being released in a sustained manner. The results suggest that the MNs have the potential to be used for targeted drug delivery, increasing the residence time of the drug at the target site, enabling better penetration and sustained delivery over an extended period while minimizing side effects. The study demonstrates the feasibility of using triblock amphiphiles to fabricate microneedles that can be programmed to undergo sequential mesophase transitions. The MNs can transform from a solid structure into hydrogel-based microneedles upon absorbing water, and then undergo an in situ transition into hydrogel microparticles that can further undergo enzymatic degradation into soluble polymers. The study also showcases the potential of these MNs for controlled drug release by loading them with DEX as a model drug and monitoring the release kinetics. The programmable mesophase transitions can be applied to achieve local and more effective treatment by increasing the residence time of the drug at the target site, enabling better penetration and sustainable delivery for an extended period of time, while minimizing side effects.This study presents the development of hydrogel microneedles (MN) with programmed mesophase transitions for controlled drug delivery. The MNs were fabricated from triblock amphiphiles with a hydrophilic poly(ethylene glycol) (PEG) middle block and two dendritic side-blocks with enzyme-cleavable hydrophobic end-groups. To enhance mechanical strength, a sodium alginate base layer and various polymeric excipients were incorporated. The MNs were successfully fabricated and exhibited favorable insertion efficiency and low height reduction when tested in a skin-simulant model. They demonstrated adequate mechanical strength under static forces up to 1000 g without fractures or broken segments. In buffer solution, the solid MNs swelled into a hydrogel within 30 seconds, followed by rapid disintegration into small hydrogel particles that could undergo slow enzymatic degradation into soluble polymers. In vitro release studies of dexamethasone (DEX) showed first-order release, with 90% released within 6 days. DEX-loaded MNs penetrated chicken skin completely, demonstrating their potential for self-administration, improved patient compliance, and sustained drug release. The study highlights the feasibility of programming hydrogel-forming MNs to undergo mesophase transitions and their potential as a delivery system for controlled drug release. The MNs can be used for targeted drug delivery, increasing the residence time of the drug at the target site, enabling better penetration and sustained delivery over an extended period while minimizing side effects. The results indicate that the mechanical properties of the MNs are significantly influenced by the molecular weight of PEG and the drug loading. The MNs showed good mechanical strength and were able to penetrate the skin, demonstrating their potential for transdermal drug delivery. The study also shows that the MNs can be used for controlled drug release, with the drug being released in a sustained manner. The results suggest that the MNs have the potential to be used for targeted drug delivery, increasing the residence time of the drug at the target site, enabling better penetration and sustained delivery over an extended period while minimizing side effects. The study demonstrates the feasibility of using triblock amphiphiles to fabricate microneedles that can be programmed to undergo sequential mesophase transitions. The MNs can transform from a solid structure into hydrogel-based microneedles upon absorbing water, and then undergo an in situ transition into hydrogel microparticles that can further undergo enzymatic degradation into soluble polymers. The study also showcases the potential of these MNs for controlled drug release by loading them with DEX as a model drug and monitoring the release kinetics. The programmable mesophase transitions can be applied to achieve local and more effective treatment by increasing the residence time of the drug at the target site, enabling better penetration and sustainable delivery for an extended period of time, while minimizing side effects.