Appendix A
CYTOTOXICITY
The biocompatibility of the obtained nanoparticle was carried out using an MTT assay.Isolation of primary cells, Cell Culture, and Caffeic acid phenethyl ester (CAPE) TreatmentPrimary cells like human gingival fibroblasts (HGF), dental pulp stem cells (DPSCc), and periodontal ligaments (PDL) cells were isolated through mechanical digestion and cultured in Dulbecco’s Modified Eagle’s Medium supplemented with a nutrient mixture, which comprised F-12 Ham’s medium (Life Technologies, Grand Island, NY, USA), 10% fetal bovine serum (Hyclone Laboratories, Logan, UT, USA), 2 mM glutamine,100 U/mL penicillin, and 100 μg/mL streptomycin. All cell cultures were maintained at 37 °C in a humidified atmosphere of 5% CODetermination of Cell Viability (MTT Assay)For the cell viability experiment, a microculture tetrazolium (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colorimetric assay was performed to determine the cytotoxicity of CAPE. Primary cells were seeded in 24-well plates at a density of 5 × 10The biocompatibility of the propolis nanoparticle was found to be high with more than 80% cell viability with HGF, DPSCc, and PDL cells at all given drug concentrations of the prepared solution compared with saline as a control (The cell viability of control and experimental groups was tested by MTT assay for 24 h, respectively, and absorbance was taken at 570 nm. Data are shown as means ± SD (