3Dアストロサイト微小環境モデルによる酸化神経炎症の迅速なカルシウム分解能解析と治療調節の実現

Astrocytes orchestrate key neuroinflammatory processes, yet conventional two dimentional (2D) cultures distort Ca²⁺signaling and elevate inflammatory noise. Here, we introduce the Astrocytic Reactive-Calcium 3D microenvironment (ARC-3D), a tunable methacrylated gelatin (GelMA)-based 3D astrocyte microenvironment that enables stable long-term culture, improved transcriptional fidelity, and rapid Ca²⁺-resolved functional assessment under perfusion. The optimized 3.5 wt% matrix supported quiescent, structurally mature astrocytes and markedly reduced the inflammatory baseline observed in 2D systems. Upon oxidative stimulation with H₂O₂, ARC-3D recapitulated hallmark features of reactive astrogliosis, including elevated basal Ca²⁺ levels, impaired receptor-evoked Ca²⁺ transients, induction of GFAP and MAO-B, and secretion of IL-6 and IL-1β, captured at second-to-minute timescales. Treatment with KDS12025, an H₂O₂-scavenging MAO-B modulator, partially restored Ca²⁺ homeostasis, suppressed Ca²⁺-dependent cytokines, and selectively recovered gliogenic and metabolic signatures in transcriptomic analysis. In vivo intracerebroventricular H₂O₂ challenge validated the ARC-3D pathological trajectory and therapeutic modulation, highlighting strong in vitro-in vivo concordance. Collectively, ARC-3D offers a mechanistically precise, Ca²⁺-resolved, and translationally aligned platform for decoding oxidative neuroinflammation and for advancing astrocyte-targeted therapeutic strategies.