A bioengineered 3D fibrous plaque microsystem for evaluating multimodal nanotherapy in atherosclerosis

Atherosclerosis progression involves macrophage-mediated inflammation and collagen-associated changes in the fibrous matrix, yet effective therapeutic strategies to modulate plaque-associated macrophage activity remain limited. Here, we established a bioengineered three-dimensional in vitro fibrous plaque (I-FP) microsystem that recapitulates selected inflammatory, foam-cell-associated, and fibrous matrix features of atherosclerotic lesions and enables quantitative evaluation of nanotherapeutic interventions. Using this multicellular platform, we evaluated a multimodal metal-organic framework nanocarrier integrating pH-responsive TCH release, Cu-associated oxidative responses, and nitric oxide-associated signaling. Within the I-FP microsystem, treatment significantly suppressed macrophage-associated inflammatory responses, including a 2.04-fold reduction in IL-6 and a 1.94-fold decrease in matrix metalloproteinase (MMP-1) expression. T-5CuZI also reduced macrophage-associated viability by 1.27-fold while preserving myofibroblast viability, and increased collagen-associated fibrous matrix content, indicating a collagen-retaining fibrous matrix phenotype under atherosclerosis-mimicking conditions. These findings establish the I-FP microsystem as an applied plaque-mimicking in vitro platform for quantitative evaluation of treatment-induced inflammatory, macrophage-associated, and fibrous matrix-related readouts relevant to atherosclerosis.