microMESH was fabricated through a top-down soft lithography approach. microMESH consists of a hydrophilic PVA microlayer supporting a hydrophobic PLGA micronetwork, which enabled the precise compartmentalization of multiple therapeutic agents, including small molecules, inhibitors, chemotherapeutic drugs, peptides, proteins, antibodies, and nanomedicines. In this proof of concept, microMESH was loaded with anti-CD47 monoclonal antibodies in the PVA microlayer and docetaxel (DTXL) within the PLGA strands, allowing for controlled and uniform drug distribution with high encapsulation efficiencies. The release profile demonstrated that DTXL was gradually delivered over months, while anti-CD47 was released within the first week, preserving its structural integrity and binding affinity. For in vitro therapeutic efficacy, anti-CD47 blocked the ‘do-not-eat-me’ signal on GBM cells, enabling macrophages to recognize and eliminate them. CT2A glioblastoma cells were treated with free or microMESH-loaded anti-CD47, then co-incubated with macrophages, where bioluminescence assays confirmed that higher anti-CD47 doses enhanced macrophage phagocytosis, reducing tumor cell viability. Confocal imaging supported these findings, showing reduced tumor presence at higher anti-CD47 concentrations. The cytotoxic potential of DTXL was assessed in CT2A cells, revealing dose- and time-dependent tumor cell death, and its combination with anti-CD47 in the presence of macrophages further enhanced tumor clearance. After extensive in vitro characterizations, microMESH was tested in vivo in a rigorous preclinical model of a highly aggressive syngeneic GBM model. Tumor growth was monitored via bioluminescence imaging, and nine experimental groups were evaluated, including control, free drug, and low- or high-dose microMESH treatments with DTXL alone, anti-CD47 alone, and their combination. Kaplan-Meier survival analysis showed that free DTXL and anti-CD47 had limited impact, whereas microMESH achieved the longest median survival, confirming the synergy between anti-CD47 and DTXL.
