To evaluate the cytotoxicity of co‐initiators of polymerization and its influence on cytokine release from human dental pulp cells (hDPCs). Cells were isolated from the dental pulp of sound human third molars. The co‐initiators dimethylaminoethyl amine benzoate—(EDAB), 2‐(dimethylamino)ethyl methacrylate (DMAEMA); 2‐Ethylhexyl 4‐(dimethylamino)benzoate (EHA) and bis(4‐methyl phenyl)iodonium hexafluorophosphate (BPI) were diluted in dimethylsulfoxide (DMSO) at different concentrations. In this way, experimental groups and one control (without treatment) were obtained. hDPCs (10 × 104 cell per well) were seeded on 96 well plates and incubated at 37°C and 5% CO2 for 48 h. After this, the cells were exposed to different concentrations of co‐initiators cited for 24 h. After this time, the culture medium was removed, and the mitochondrial metabolism was evaluated by MTT assay, cell death by flow cytometry, and cytokine released (IL‐1β, IL6, IL‐8, IL‐10, and TNF‐α) was analyzed by MAGPIX assay. The data were analyzed by ANOVA one‐way and Tukey's test. EHA, DMAEMA, and EDAB did not reduce the mitochondrial metabolism. BPI presented high toxicity with remarkable reduction (80%) after exposure to 1 mM. The cell death of all test groups was similar to control. After 24 h treatment, the IL‐8 was up‐regulated by all compounds, while IL‐6 was upregulated after exposure to EHA and downregulated after DMAEMA stimulation. BPI, EHA, EDAB, and DMAEMA can trigger an initial inflammatory response, upregulating the IL‐8 secretion in hDPCs in a compound‐concentration‐dependent manner; however, this was not accompanied by major cytotoxic effects at cell death or mitochondrial‐metabolism levels.