In this study, a customized biomimetic brain-mimetic hydrogel model was developed using Digital Light Processing (DLP) 3D printing technology. This model aims to optimize the cellular microenvironment for the growth of neural stem cells and to evaluate the neurotoxicity of various drugs effectively. By precisely controlling the material composition, printing process, and structural design, the pore structure, water absorption, mechanical properties, and formability of the hydrogel were optimized, creating a more biomimetic microenvironment for the survival and proliferation of neural stem cells. Our developed composite hydrogel (GelMA-HAMA-SilMA-Gelatin) achieved a relatively low elastic modulus, excellent water absorption, good biocompatibility, and superior printability by adjusting printing parameters. This makes it better suited to approximate the optimal microenvironment for neural stem cell growth, enhancing cellular proliferation and vitality. Furthermore, our research demonstrated that the 3D hollow brain-mimetic hydrogel model, which features macronutrient channels and microporous networks, significantly improved the survival and proliferation of neural stem cells. Using this model, the neurotoxicity of acrylamide and oxaliplatin was evaluated, confirming the model's effectiveness in evaluating drug neurotoxicity and its ability to demonstrate cellular sensitivity to drug dosages. Compared to conventional two-dimensional adherent cell culture models, our three-dimensional brain-mimetic model can more accurately simulate the complex in vivo environment, offering new perspectives for drug screening and neurotoxicity evaluation. This study not only demonstrates the application potential of 3D printing technology in optimizing cellular microenvironments and drug screening but also emphasizes the importance of considering the cellular microenvironment during drug development. Ultimately, it provides new strategies and tools for evaluating drug safety.