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Resin-modified glass ionomer cement (RMGIC) has important properties. However, like other restorative materials, it has limitations such as decreased biocompatibility. The incorporation of nanoparticles (NP) in the RMGIC resulted in improvements in some of its properties. The aim of this study was to evaluate the physical-biological properties of RMGIC with the addition of nanohydroxyapatite (HANP). Material and Methods: Vitremer RMGIC was used, incorporating HANP by amalgamator, vortex and manual techniques, totaling ten experimental groups. The distribution and dispersion of the HANP were evaluated qualitatively by field emission scanning electron microscope (SEM-FEG). The evaluation of image porosity (SEM-FEG) with the help of imageJ. Cell viability 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazoline bromide (MTT) and cell morphology analyses were performed on MDPC-23 odontoblastoid cells at 24 and 72 h. Results: It was possible to observe good dispersion and distribution of HANP in the samples in all experimental groups. The incorporation of 5% HANP into the vortex stirred RMGIC resulted in fewer pores. The increase in the concentration of HANP was directly proportional to the decrease in cytotoxicity. Conclusions: It is concluded that the use of a vortex with the incorporation of 5% HANP is the most appropriate mixing technique when considering the smallest number of pores inside the material. A higher concentration of HANP resulted in better cell viability, suggesting that this association is promising for future studies of new restorative materials.