IADR Abstract Archives

In vitro formation of odontoblast-like cells on dentin

Recent characterization of human dental pulp stem cells has shed new light on the understanding of the odontoblastic lineage and potential for pulp tissue regeneration. Objectives: i) to characterize pulp cells isolated and cultured in vitro on culture dish; and ii) to examine pulp cell differentiation potential grown on dentin. Methods: collected pulp tissues from 3rd molars or pulpectomy were minced and digested in collagenase/dispase and the released cells were seeded in culture dishes containing a-MEM supplemented with 20% fetal bovine serum, 2 mM L-glutamine, 100 μM L-ascorbic acid-2-phospate, and antibiotics. Pulp cells at confluence were exposed to 2 mM KH2PO4, 20 mM HEPES and 100 nM dexamethasone; or the above plus 5 x 10-8 M 1,25 (OH)2 D3 for 6 weeks. Pulp cells were also seeded onto flat human dentin surface treated by mechanical grinding and polishing, chemical etching and sterilization, and at different time points after seeding, processed for morphological examination using scanning electron microscopy.  Results: pulp cells formed colonies of various characteristics in 1-2 wks after enzyme separation. A set of actively growing pulp cells (passage 3) had a similar proliferation rate as the immortal cell line NIH 3T3 (population doubling time-- pulp cells: 22.6 ± 0.5 h; NIH 3T3: 23.1 ± 2.3 h). These pulp cells formed mineral nodules stimulated with dexamethasone or dexamethasone plus 1,25 (OH)2D3 demonstrated by Alizarin red stain. Pulp cells, after seeded onto treated dentin surface, did not proliferate and appeared to establish an odontoblast-like morphology with a cytoplasmic process extending into a dentinal tubule revealed by scanning electron microscopy analysis 16 d after seeding. Conclusions: actively growing pulp cells exhibiting stem cell characteristics (form mineral nodules) may potentially convert to cells of odontoblastic morphologies on existing dentin in vitro. (Supported by an Endodontic Research Grant from American Association of Endodontists Foundation)

 

 


AADR/CADR Annual Meeting
2006 AADR/CADR Annual Meeting (Orlando, Florida)
Orlando, Florida
2006
941
Pulp Biology
  • Shagramanova, Kristina  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Chan, Selina W.  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Aurasteh, Parnaz  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Tam, Irene  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Huang, George T.-j  ( University of Maryland, Baltimore, MD, USA )
    • Oral Session
    Vital Pulp Therapy and Pulp-regenerative Responses
    03/10/2006