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J Craniofac Surg. 2012 Nov;23(6):e571-5. doi: 10.1097/SCS.0b013e31825e4e16.

Tissue engineering with dental pulp stem cells: isolation, characterization, and osteogenic differentiation.

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  • 1Cell Therapy Unit, Hospital Universitario Virgen de la Arrixaca, Faculty of Medicine, University of Murcia, Spain.



Dental pulp stem cells (DPSCs) are adult mesenchymal stem cells that have the ability to differentiate into osteoblasts, a fact that is very interesting in the context of tissue engineering. Our purpose was to isolate and characterize DPSCs and to compare the differentiation potential of 3 different osteogenic media.


Human dental pulp extracted from healthy young adults was placed in flasks with a mesenchymal expansion medium. At passage 4 DPSCs were analyzed for cell-cycle stage, proliferation, viability, and immunophenotype. DPSCs were grown in 3 different osteogenic media for 40 days. Flasks were incubated at 37 °C in 5% CO2, and the medium was changed twice a week. At day 40, the mineralization of the matrix was determined with Alizarin Red S dye.


After osteogenic induction, DPSCs developed mineralization nodules (clusters), as revealed by Alizarin Red staining. This staining was stronger in the Osteodiff (Miltenyi) medium when compared to the other osteogenic media.


This study demonstrates the ability of DPSC to differentiate into osteoblasts, especially in the presence of Osteodiff (Miltenyi). DPSCs are therefore a good candidate model for the study of hard-tissue mineralization.

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