Reconstructing protein conformations to activate follicular enzyme-nutrient reservoirs for reversing follicle aging.

Hair regeneration depends on a reservoir of follicular enzymes that regulate the microenvironment and provide metabolic support. However, in degenerative alopecia, this enzymatic reservoir is disrupted, impairing microenvironmental homeostasis. Here, through bioinformatics and molecular dynamics analyses, we revealed that several Zn²⁺/Cu²⁺-dependent proteases (<i>e.g.</i>, LOX and SOD1) were downregulated and exhibit aberrant conformations and activities in degenerative alopecia, further weakening microenvironmental support. To address this, we developed a Zn²⁺/Cu²⁺-doped decellularized extracellular matrix microneedle patch (dECMMA^Zn/Cu) to rapidly restore the follicular microenvironment and enzymatic reservoir function. <i>In vitro</i>, microneedle delivery of dECMMA^Zn/Cu significantly upregulated key Zn²⁺/Cu²⁺-dependent proteases. Restoration of the enzymatic reservoir reduced p16 expression in dermal papilla stem cells, inhibited cellular senescence(definition), and promoted stem cell proliferation. In a classical degenerative alopecia animal model, dECMMA^Zn/Cu microneedle treatment resulted in denser and more continuous dermal collagen, a significant increase in hair follicle numbers, and promoted hair regeneration. In summary, this study establishes an innovative microneedle material that restores follicular function by remodeling protein conformation and activating the enzymatic reservoir.