IADR Abstract Archives
Human-Dental-Pulp-Stem-Cells Osteogenic Differentiation Seeded on Equine-Bone-Block with Graphene and Melatonin
Objectives: Equine Bone Blocks showed biocompatibility and osteconductivity. Therefore, loading of scaffolds with osteogenic cells, like human dental pulp stem cells (hDPSCs), has been proposed to develop a biocomplex for bone defect repair, overcoming the limits of the graft materials.
Graphene-based materials have raised a growing interest in many research areas, according to their effects on cell adhesion, proliferation and differentiation. Melatonin is principally synthesized in the pineal gland and it is an important mediator in bone formation.
The aim of this in vitro investigation was to evaluate hDPSCs cultured in the presence of Equine Bone Blocks coated with Graphene Oxide (GO/NH3) and of Melatonin.
Methods: hDPSCs were cultured in growth medium (GM) and in differentiation medium (DM) with Blocks coated or not with GO/NH3 at different concentration (2 and 10 mg/ml) and with or w/o 5% Melatonin. After 7, 14 and 21 days of culture, miRNAs, gene expression (Runx-2, Smad-5), and Osteocalcin levels were evaluated. Histological and vitality tests were also performed.
Results: miR-133a and miR-135a were downregulated already at 7 days in GM in the presence of GO/NH3 at different concentration and of 5% Melatonin and this decrease seems related with the higher expression of Smad5 and RUNX2. At 14 days, the increase of Osteocalcin levels confirmed the onset of differentiation also in GM. Histological evaluation showed increased cells adhesion in G2 and G10 cultures in the presence of Melatonin. This trend is similar to that found in DM (data not shown), confirming an activation of differentiation process also in GM condition.
Conclusions: hDPSCs/Equine Bone Block, Graphene and Melatonin seemed to represent a promising useful tool in bone regeneration. The present data seemed to suggest that the Blocks, synergized by the simultaneous presence of Graphene and Melatonin, stimulated the early stages of cell differentiation.
Graphene-based materials have raised a growing interest in many research areas, according to their effects on cell adhesion, proliferation and differentiation. Melatonin is principally synthesized in the pineal gland and it is an important mediator in bone formation.
The aim of this in vitro investigation was to evaluate hDPSCs cultured in the presence of Equine Bone Blocks coated with Graphene Oxide (GO/NH3) and of Melatonin.
Methods: hDPSCs were cultured in growth medium (GM) and in differentiation medium (DM) with Blocks coated or not with GO/NH3 at different concentration (2 and 10 mg/ml) and with or w/o 5% Melatonin. After 7, 14 and 21 days of culture, miRNAs, gene expression (Runx-2, Smad-5), and Osteocalcin levels were evaluated. Histological and vitality tests were also performed.
Results: miR-133a and miR-135a were downregulated already at 7 days in GM in the presence of GO/NH3 at different concentration and of 5% Melatonin and this decrease seems related with the higher expression of Smad5 and RUNX2. At 14 days, the increase of Osteocalcin levels confirmed the onset of differentiation also in GM. Histological evaluation showed increased cells adhesion in G2 and G10 cultures in the presence of Melatonin. This trend is similar to that found in DM (data not shown), confirming an activation of differentiation process also in GM condition.
Conclusions: hDPSCs/Equine Bone Block, Graphene and Melatonin seemed to represent a promising useful tool in bone regeneration. The present data seemed to suggest that the Blocks, synergized by the simultaneous presence of Graphene and Melatonin, stimulated the early stages of cell differentiation.
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