The efficiency of the in vitro osteo/dentinogenic differentiation of human dental pulp cells, periodontal ligament cells and gingival fibroblasts

Abstract

Although the primary cell cultures from dental pulp and other oral tissue are frequently used to study osteogenic potential and stem cell responses, few systematic and comparative studies on stemness for the dentinogenic differentiation of these cells have been conducted. In the present study, to investigate the stemness of oral primary cells during extended culture, human adult dental pulp cells (hDPCs), periodontal ligament stem cells (hPDLSCs) and gingival fibroblasts (hGFs) were obtained and cultured from pulp tissue, periodontal ligaments, and marginal and attached gingival tissue of extracted third molars, respectively. As shown by fluorescence-activated cell sorting analysis and immunophenotyping, the mesenchymal stem cell markers, CD44, CD73, CD90, CD146 and CD166, were highly expressed in early passage hDPCs, hPDLSCs and hGFs. However, when the cells were treated with osteogenic additives, mineralization markedly increased in the hDPCs and hPDLSCs, but not in the hGFs. Moreover, the expression of dentinogenic markers, such as dentin sialophosphoprotein and dentin matrix protein-1, appeared to decrease during extended culture past passage number 8 of the hDPCs and hPDLSCs. These data suggest that hDPCs and hPDLSCs may have differentiation potential during the early passages, and that their progenitor potential is diminished during extended culture. The hGFs did not show differentiation capability during culture, even though they contained general mesenchymal stem cell surface proteins. The transcriptional expression of dentinogenic markers in hDPCs was not affected by co-culture with hPDLSCs and/or hGFs.

Low‑intensity pulsed ultrasound promotes periodontal ligament stem cell migration through TWIST1‑mediated SDF‑1 expression

“…Seo et al ( 43 ) isolated MSCs and used them to generate cementum and periodontal ligament in vivo . The present findings suggest that the PDLSCs that were isolated possess MSC properties, such as multipotency, and express MSC markers, which is consistent with previous studies ( 44 ). MSC mobilization has been reported to participate in periodontal tissue homeostasis ( 24 , 45 ).…”

Indirect co-culture of stem cells from human exfoliated deciduous teeth and oral cells in a microfluidic platform

“…Cell suspension was obtained by passing through a 70-µM strainer, and incubated in α-MEM containing 20% fetal bovine serum (Hyclone, GE Healthcare, Buckinghamshire, UK) and antibiotics at 37°C in 5% CO2. For differentiation and mineralization, cells were incubated in α-MEM containing 10% fetal bovine serum, 5 mM β-glycerophosphate, 500 nM dexamethasone, and 100 µM ascorbic acid for 15 days [33,34].…”

Effect of FGF-2, TGF-β-1, and BMPs on Teno/Ligamentogenesis and Osteo/Cementogenesis of Human Periodontal Ligament Stem Cells

“…These cells can differentiate into osteoblasts, cementoblasts, and PDL fibroblasts, and are a promising source for regeneration of periodontium ( Maeda et al, 2011 ). The MSC markers such as CD44, CD73, CD90, CD146, CD166, and STRO-1, are highly expressed in these cells, whereas the population of hematopoietic marker (CD34)-positive cells is extremely small ( Choi et al, 2015 ; Seo et al, 2004 ). Although PDL stem cells (PDLSCs) are thought to play key roles for not only bone remodeling but also wound healing and regeneration of the tissues ( Lekic and McCulloch, 1996 ), the regulation of differentiation mechanism are not fully understood yet and remain unclear.…”