Effects of morphogen and scaffold porogen on the differentiation of dental pulp stem cells.
BACKGROUND
Dental pulp tissue engineering is an emerging field that can potentially have a major impact on oral health. However, the source of morphogens required for stem cell differentiation into odontoblasts and the scaffold characteristics that are more conducive to odontoblastic differentiation are still unclear. This study investigated the effect of dentin and scaffold porogen on the differentiation of human dental pulp stem cells (DPSCs) into odontoblasts.


METHODS
Poly-L-lactic acid (PLLA) scaffolds were prepared in pulp chambers of extracted human third molars using salt crystals or gelatin spheres as porogen. DPSCs seeded in tooth slice/scaffolds or control scaffolds (without tooth slice) were either cultured in vitro or implanted subcutaneously in immunodefficient mice.


RESULTS
DPSCs seeded in tooth slice/scaffolds but not in control scaffolds expressed putative odontoblastic markers (DMP-1, DSPP, and MEPE) in vitro and in vivo. DPSCs seeded in tooth/slice scaffolds presented lower proliferation rates than in control scaffolds between 7 and 21 days (p < 0.05). DPSCs seeded in tooth slice/scaffolds and transplanted into mice generated a tissue with morphological characteristics similar to those of human dental pulps. Scaffolds generated with gelatin or salt porogen resulted in similar DPSC proliferation. The porogen type had a relatively modest impact on the expression of the markers of odontoblastic differentiation.


CONCLUSIONS
Collectively, this work shows that dentin-related morphogens are important for the differentiation of DPSC into odontoblasts and for the engineering of dental pulp-like tissues and suggest that environmental cues influence DPSC behavior and differentiation potential.
Department of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, USA.
Flavio F
FF
Demarco
Luciano
L
Casagrande
Zhaocheng
Z
Zhang
Zhihong
Z
Dong
Sandra B
SB
Tarquinio
Benjamin D
BD
Zeitlin
Songtao
S
Shi
Anthony J
AJ
Smith
Jacques E
JE
Nör
D000293:Adolescent; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation; D003782:Dental Pulp / Q000166:cytology*; D003786:Dental Pulp Cavity / Q000033:anatomy & histology; D003804:Dentin / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005780:Gelatin / Q000737:chemistry; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry; D008297:Male; D051379:Mice; D016513:Mice, SCID; D009804:Odontoblasts / Q000166:cytology; D010750:Phosphoproteins / Q000032:analysis; D011091:Polyesters; D011108:Polymers / Q000737:chemistry; D016062:Porosity; D012795:Sialoglycoproteins / Q000032:analysis; D012965:Sodium Chloride / Q000737:chemistry; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*; D040521:Subcutaneous Tissue / Q000601:surgery; D013499:Surface Properties; D046509:Tissue Culture Techniques; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein
D001672:Biocompatible Materials; D016326:Extracellular Matrix Proteins; D005780:Gelatin; D006023:Glycoproteins; D019344:Lactic Acid; D010750:Phosphoproteins; D011091:Polyesters; D011108:Polymers; D012795:Sialoglycoproteins; D012965:Sodium Chloride; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C486579:MEPE protein, human; C033616:poly(lactide)