Dentinogenic potential of human adult dental pulp cells during the extended primary culture

Abstract

Despite the frequent use of primary dental pulp cells in dental regenerative research, few systematic studies of stemness for osteogenic and dentinogenic differentiation of human adult pulp cells have been reported. To investigate the stemness of human adult dental pulp cells, pulp tissues were obtained from extracted third molars and used as a source of pulp cells. In FACS analysis and immunophenotyping, the general mesenchymal stem cell markers CD44, CD90, and CD146 were highly expressed in early passages of the pulp cell culture. The stem cell population was dramatically decreased in an expansion culture of human dental pulp cells. When pulp cells were treated with additives such as β-glycerophosphate, ascorbic acid, and dexamethasone, nodule formation was facilitated and mineralization occurred within 2 weeks. Expression of osteogenic markers such as alkaline phosphatase, osteocalcin, and osteonectin was relatively low in undifferentiated cells, but increased significantly under differentiation conditions in whole passages. Dentinogenic markers such as dentin sialophosphoprotein and dentin matrix protein-1 appeared to decrease in their expression with increasing passage number; however, peak levels of expression occurred at around passage 5. These data suggested that stem cells with differentiation potential might exist in the dental pulp primary culture, and that their phenotypes were changed during expansion culture over 8-9 passages. Under these conditions, a dentinogenic population of pulp cells occurred in limited early passages, whereas osteogenic cells occurred throughout the whole passage range.

Dose-responses of Stem Cells from Human Exfoliated Teeth to Infrared LED Irradiation

“…The evaluation of mineral nodules was performed using the alizarin red staining (19). Cells were seeded (10 4 cells/cm 2 ) and incubated for 48 h in basal medium, then kept in contact with an osteogenic medium containing 5 mM α-glycerophosphate and 100 mM ascorbic acid (Sigma-Aldrich) (19).…”

“…Cells were seeded (10 4 cells/cm 2 ) and incubated for 48 h in basal medium, then kept in contact with an osteogenic medium containing 5 mM α-glycerophosphate and 100 mM ascorbic acid (Sigma-Aldrich) (19). Irradiation was performed at 12 or 72 h before the evaluation time.…”

Optimization of treatment with recombinant FGF-2 for proliferation and differentiation of human dental stem cells, mesenchymal stem cells, and osteoblasts

“…Generally, stem cell populations of various oral cells decreased in a narrow range during the culture period (Min et al 2011). For evaluation of the expression amount of stem cell surface markers, FITC-labeled anti-CD44, -CD73, -CD90, -CD146, and -CD166 antibodies were used, and their binding intensities were analyzed by FACS and quantified using mean fluorescence values.…”

Characteristic expression of MSX1, MSX2, TBX2 and ENTPD1 in dental pulp cells

“…Dental pulp cells (DPCs) show odontoblast-like differentiation, with increased expression of alkaline phosphatase and osteocalcin and matrix calcification in vitro following exposure to osteogenesis induction medium (1)(2)(3). In addition, transplantation of DPCs enhances the regeneration of various tissues, including dental, skeletal and nerve tissues (4,5).…”