Bruzzesi, Giacomo

De Pol, Anto

Ferrari, Adriano

Maraldi, Tullia

Migliaresi, Claudio

Motta, Antonella

Resca, Elisa

Riccio, Massimo

Firenze University Press: E-Journals

IJAE Vol. 115, n. 1/2 (Supplement), 2010
147
Osteogenic differentiation of human dental pulp stem 
cells in 3D scaffolds
Massimo Riccio1, Elisa Resca1, Tullia Maraldi1, Claudio Migliaresi2, Antonella Motta2, Adriano Ferrari3, 
Giacomo Bruzzesi4, Anto de Pol1
1 Department of Anatomy and Histology, University of Modena and Reggio Emilia, Italy
2 Department of Materials Engineering and Industrial Technologies, University of Trento, Italy
3 Department of neuroscience, Ospedale Santa Maria Nuova, Reggio Emilia, Italy
4 Oro-maxillo-facial Department, AUSL Baggiovara, Modena, Italy
The aim of this study was to characterize the in vitro osteogenic differentiation of 
dental pulp stem cells (DPSCs) in 2D and 3D cultures. DPSCs were isolated by mag-
netic cell sorting using antibodies against c-Kit, CD34 and STRO-1 surface antigens and 
then differentiated toward osteogenic lineage on 2D surface of culture flask by using an 
osteogenic medium. Differentiated cells express specific bone proteins such as Runx-2, 
Osx, OPN and OCN with a sequential expression analogous to those occurring dur-
ing osteoblast differentiation and produce extracellular calcium deposits. In a second 
phase, DPSCs were cultured in MatrigelTM, Collagen and Fibroin 3D scaffolds in order 
to differentiate cells in a 3D space that mimes the physiological environment. In DP-
SC-MatrigelTM complexes, cells differentiate in osteoblast phenotype and form calcified 
nodules. DPSCs differentiated in collagen sponge actively secrete human type I collagen 
and after implant in immuno-suppressed rats were enveloped in a highly vascularised 
connectival capsule. DPSC-Collagen complex presents small areas of mineralization but 
appears infiltrated by inflammatory cells indicating that an immuno-response occurred. 
Fibroin scaffolds colonized with DPSCs after implant appear enveloped in a connecti-
val capsule highly vascularised similarly to collagen scaffolds. In contrast the immune 
system invasion did not occur. DPSC are present in the whole scaffold thickness and 
in intimate contact with the scaffold fibers. After 20 days of implant extended areas of 
mineralization were stained by alizarin red. Fibroin scaffold appears more suitable for 
future application in regenerative medicine because it is highly histo-compatible, do not 
produces immune-response and offers the optimal environment for DPSCs growth and 
differentiation. These neo-formed DPSCs-scaffold devices may be used in regenerative 
surgical applications to resolve pathologies and traumas characterized by critical size 
bone defects.
Key words
DPSC, Stem Cells, Osteogenic Differentiation, 3D Scaffold, Fibroin


Osteogenic differentiation of human dental pulp stem cells in 3D scaffolds

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Osteogenic differentiation of human dental pulp stem cells in 3D scaffolds

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