Title: 1723 - Smad4 Knockout of iMDP-3 Restrains Odontogenic Potential


Christopher Adams (Presenter)
UT Health San Antonio


Objectives: Dental pulp stem cells have the capacity to react to a variety of stimuli by differentiating into odontoblast-like cells capable of producing large amounts of dentin sialoprotein (DSP), dentin sialophosphoprotein (DSPP) and other proteins necessary for dentinogenesis. The objective of this study was to investigate the role of Smad4 in the regulation of proliferation and odontogenic differentiation of mouse dental papilla mesenchymal cells from the iMDP-3 cell line.

Methods: The Smad4 gene of iMDP-3 cells was knocked out using CRISPR/cas 9 and the biological effects were investigated. Proliferation of the iMDP-3 cells was measured by direct cell counting and 5-ethynyl-20-deoxyuridine incorporation assay. Odontogenic differentiation of iMDP-3 cells was evaluated by von Kossa staining and alkaline phosphatase (ALP) activity assay. Expression of important mineralization-related genes such as DSPP, dentin matrix acidic phosphoprotein 1 (DMP1) and ALP were determined by real-time polymerase chain reaction. Western blot analysis was performed to determine the expressions of DSPP, DMP1, Osterix (OSX), Distal-less 3 (Dlx3), Distal-less 5 (Dlx5), Runt-related transcription factor 2 (Runx2), and osteocalcin (OCN).

Results: Smad4 knockouts showed decreased proliferation and decreased odontogenic differentiation, but higher mineralization gene expression than control iMDP-3 cells.

Conclusions: By showing the effect of Smad4 knockout on the protein expression of iMDP-3 cells we conclude that Smad4 has an important role as a transcription factor in the dentinogenesis signaling pathway.

Student Presenter

This abstract is based on research that was funded entirely or partially by an outside source:
NIH grants DE019892 and HL075360.

Disclosure Statement:
The submitter must disclose the names of the organizations with which any author have a relationship, the nature of the relationship, and the clinical or research area involved. The following is submitted: NONE

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