Single-cell transcriptome-guided biomimetic magnetothermal hydrogel microspheres for multimodal eradication of residual glioblastoma

Incomplete resection and rapid postoperative recurrence remain major challenges in glioblastoma (GBM) treatment. Inspired by the self-healing microduct architecture of pine resin, we developed an injectable magnetothermal-responsive hierarchical hydrogel microsphere platform, HGM + Multi(+), for spatiotemporally programmed magnetothermal ablation, anti-angiogenic blockade, and targeted chemotherapy. The system integrates T7-modified, pH-responsive temozolomide nanocarriers in GelMA microspheres with bevacizumab and Fe₃O₄ nanoparticles in a HAMA matrix, enabling rapid magnetothermal heating and sequential dual-drug release. Cellular and release studies demonstrated receptor-mediated uptake and tumor-microenvironment-responsive drug release. In orthotopic GBM resection models, a single intracavitary administration under alternating magnetic field markedly suppressed recurrence, reducing tumor volume to approximately 5% of controls and doubling median survival. In the immunocompetent postoperative GL261/C57BL/6J model, longitudinal IVIS, H&E-based tumor area quantification, and Kaplan-Meier analysis further confirmed reduced recurrent tumor burden and prolonged survival. Single-cell transcriptomic profiling of 47,781 cells revealed extensive tumor microenvironment remodeling, including macrophage polarization toward M1-like states, alleviated T cell exhaustion, enhanced cytotoxic programs, and an approximately 37% reduction in cancer stemness. These findings were further supported by ex vivo tumor-sphere assays and reduced Sox9/Nestin expression in recurrent tumors. Intercellular communication analysis indicated strengthened antigen-presentation signaling and intensified interactions between T cells and myeloid cells. Together, the integrated therapeutic, histological, and single-cell transcriptomic data support HGM + Multi(+) as a versatile postoperative strategy for eliminating residual GBM and advancing intelligent biomaterials for oncology.