This study assessed the pH, radiopacity, antimicrobial effect, cytotoxicity and biocompatibility of endodontic filling materials for primary teeth. Zinc oxide eugenol (ZOE), Vitapex and Calen paste thickened with zinc oxide (ZO) were evaluated in comparison to an experimental MTA-based material. Radiopacity was tested using a graduated aluminum stepwedge with a digital sensor (n=5). The materials pH was recorded at 1, 4, 12 h; 1, 3, 7, 15 and 30 days (n=5). Direct contact test was used to assess the antimicrobial efficacy against Enterococcus faecalis after 1, 4, 12, 24 h (n=5). Cytotoxicity assay used MTT test for cell viability after incubation for 1, 3 and 7 days (n=5). For biocompatibility test, Wistar rats had received implants containing each material (n=5). The biopsied tissues were histologically analyzed after 15, 30 and 60 days. The results of radiopacity, pH, antimicrobial capacity and cytotoxicity were analyzed using ANOVA and Tukey tests. The histological data were submitted to Kruskal-Wallis test. The experimental material presented the lowest radiopacity (3.28 mm Al) and had a pH>12.0 throughout the test period. The experimental material showed the highest antibacterial effect, killing over 99.97% bacteria in 4 h. Vitapex presented the highest cell viability. Initially, biocompatibility test showed moderate to severe inflammation in all groups. After 60 days, Calen+ZO group showed moderate inflammation, while the others showed predominantly mild inflammatory reaction. The present results demonstrated that the experimental MTA-based material exhibited satisfactory behavior regarding the studied properties. Additional in vivo studies are necessary for a better evaluation of the material.