Evaluación in vitro del efecto de epigalocatequina-3-galato (EGCG) en células con fenotipo de osteoblasto y células madre de pulpa dental tratada con biomateriales de regeneración ósea

Resumo

Objective: A study was made of the in vitro effect of ECGC on SAOS-2 osteoblast-like cells and upon cultured dental pulp stem cells (DPSCs) treated with a bovine (Geistlich BioOss®), porcine (OsteoBiol Gen-Os®) and beta-tricalcium phosphate (Cerasorb M®) bone substitutes. Material and methods: The DPSCs were isolated from third molars extracted from healthy individuals. Cytotoxicity assays were performed using concentrations of 1, 2.5, 5, and 10 μM of EGCG on Saos-2 cell lines and DPSCs. Subsequently, the DPSCs treated with 5 mg/ml of Bio-Oss® (BO), Gen-Os® (GO) and Cerasorb® (CE) in combination with EGCG 1 μM. The effects were evaluated based on cell viability / cytotoxicity assay (MTT, cell viability staining test), cell migration, scanning electron microscopy (SEM), and alkaline phosphatase (ALP) activity. Results: EGCG exerted no cytotoxic effects at any of the doses evaluated on Saos-2 and DPSCs, with respect to control group. Furthermore, an increase in viability was observed with 5μM in Saos-2 and with 1μM in DPSCs. The results of the MTT assay of the DPSCs treated with the biomaterials alone and in combination with EGCG 1 μM showed BO and CE to produced negative effects upon cell viability and migration, and GO and CE resulted in deficient cell adhesion. On the other hand, the biomaterials exerted no adverse effects upon ALP activity. The addition of EGCG reverted the cytotoxic effect and the loss of migration capacity in the BO and CE groups, and improved cell adhesion in the GO and CE groups. Furthermore, EGCG promoted increased ALP activity. Conclusions: EGCG is biocompatible with SAOS-2 osteoblast-like cells and dental pulp mesenchymal cells (DPSCs). In addition, its viability increases in a dose-dependent manner. The addition of EGCG to the biomaterials BO, GO and CE reverts their negative effects and improves their biocompatibility with cultured DPSCs. The use of EGCG thus appears to be a promising strategy for restoring and enhancing the osteoconductive properties of these biomaterials in bone defect regeneration treatments.