Osteogenic potential of platelet-rich plasma in dental stem-cell cultures

Abstract

The purpose of this study was to analyse the potential of platelet-rich plasma (PRP) culture media to induce osteogenic differentiation of periodontal ligament stem cells and dental pulp stem cells compared with four other methods of culture. Both types of cell were collected from 35 healthy patients and cultured in five different media (Dulbecco's modified eagle's medium (DMEM); DMEM and melatonin; DMEM and PRP; DMEM and ascorbic acid 200μmol; DMEM and l-ascorbate 2-phosphate 50μmol). Cells were characterised by flow cytometry. Alizarin Red stain, alkaline phosphatase stain, and the expression of collagen type 1 (Col-1), runt-related transcription factor (RUNX2), osteoprotegerin, and osteopontin (quantified by qRT-PCR) were used to detect the osteogenic profile in each culture. Flow cytometry showed that both types of stem cell were a homogeneous mixture of CD90(+), CD105(+), STRO-1(+), CD34 (-), and CD45 (-) cells. Dental pulp stem cells that were cultured with PRP showed the best osteogenic profile (RUNX2 p=0.0002; osteoprotegerin p=0.001). The group of these stem cells that showed the best osteogenic profile was also cultured with PRP (osteoprotegerin p=0.001). Medium five (with l-ascorbate 2-phosphate 50μmol added) showed an increase in all osteogenic markers for periodontal ligament stem cells after PRP, while the best culture conditions for osteogenic expression of dental pulp stem cells after PRP was in medium four (ascorbic acid 200μmol added). These results suggested that culture in PRP induces osteogenic differentiation of both types of stem cell, modulating molecular pathways to promote bony formation.

Keywords: Bone regeneration; Platelet-rich plasma; cell culture; dental pulp stem cells; periodontal dental ligament stem cells.