Advanced 3D Platforms for Modeling CNS Neuroinflammation: Cell Integration, Techniques, and Challenges

Neuroinflammation is a multicellular immune response within the central nervous system (CNS) triggered by injury, infection, or disease, and it drives long-term changes in neurons and glia. Understanding this mechanism is important for advancing research into neurodegenerative diseases and trauma-induced CNS damage. Three-dimensional (3D) cell culture models offer enhanced structural and functional capabilities compared to traditional 2D models, yet their implementation remains challenging. Key limitations include cell survival, faithful recapitulation of CNS cellular diversity, simulation of immune responses, and issues of scalability and long-term culture. This review explores state-of-the-art methods for incorporating neurons, astrocytes, microglia, oligodendrocytes, and other CNS-relevant cell types into 3D models to study neuroinflammation. Additionally, it examines emerging trends in bioengineering, co-culture strategies, and dynamic systems that improve model scalability and longevity. By critically evaluating both established and innovative protocols, we aim to provide guidance for researchers in selecting effective methodologies and to accelerate the development of physiologically relevant in vitro models for neuroinflammation research.