Introduction: The use of animal models is widely spread in the scientific field, because the use of animals is essential for the transition from basic research to clinical research. Furthermore, there is no denying that the scientific advances achieved to date are related to the use of animal models. However, diminishing the usage of animals is desirable. One alternative to the use of animals are three-dimensional in vitro models that better mimic the tissue environment, allowing for more reliable results. The aim of this study was to produce and validate a 3D neuroinflammation model (3DNM) as an alternative for drug testing and alternative treatment approaches. Methods: The 3DNM was produced combining 5% alginate, 4% gelatin, 2x10⁵ PC12 cells, 4x10⁵ BV2 cells and 8x10⁵ C6 cells. Cell viability was evaluated with MTT assay. To test the anti-inflammatory potential of a bioink containing 1.5% decellularized spinal cord tissue, 3% gelatin, 4% sodium alginate and 7.5x10⁵ mesenchymal cells, the 3DNM was stressed with lipopolysaccharide (LPS) and treated with the bioink for three days. Reactive oxygen species (ROS) production and lipid peroxidation were measured. IL-6, IL-10 and IL-1B were quantified by ELISA. Non-enzymatic antioxidant defenses were evaluated by the quantification of Thiols. Results: MTT results demonstrated that the model was not cytotoxic to the cells because the viability increased when compared to the model stressed with the LPS. The bioink reduced the levels of ROS and pro-inflammatory cytokines on the 3DNM. TBARS assay showed that 3DNM lipid degradation was reduced when it was in contact with the bioink and was very close to that of the control. Thiols indicated that the bioink increased the non-enzymatic antioxidant defense in the 3DNM. Conclusion: The 3DNM showed promising results as an alternative model to study neural inflammation. This biomaterial may, therefore, be a solution for decreasing the laboratory use of animals.