Human cementoblasts express enamel-associated molecules in vitro and in vivo.
OBJECTIVE
Cementum is a mineralized tissue that facilitates the attachment of periodontal ligament to the root and surrounding alveolar bone and plays a key role in the regeneration of periodontal tissues. The molecular mechanisms that regulate the proliferation and differentiation of cementoblasts, however, have not been elucidated to date. Enamel molecules are believed to regulate cementoblast differentiation and to initiate the formation of acellular extrinsic fiber cementum. The purpose of this study was therefore to isolate and culture human root-derived cells (HRDC) in order to determine whether they are able to express both cementum and specific enamel proteins and subsequently to confirm these findings in vivo.


METHODS
Human root-derived cells were isolated and expanded in vitro. Cells were characterized using RT-PCR, immunostaining, western blotting and by examination of total mRNA to determine the expression of cementum and enamel markers. Human periodontal tissues were also examined for the expression of enamel-related proteins by immunostaining.


RESULTS
We showed that HRDC express mRNA corresponding to ameloblastin (AMBN), amelogenin (AMEL), enamelin (ENAM), tuftelin (TUFT) and cementum-associated molecules such as cementum protein 1 (CEMP1) and cementum attachment protein (CAP). Western blotting revealed that HRDC express both AMEL and AMBN gene products, as well as the cementum markers CEMP1 and CAP. In vivo, we have showed that AMBN and AMEL are expressed by cementoblasts lining cementum, paravascular cells and periodontal ligament cells.


CONCLUSIONS
These results suggest that enamel-associated and cementum-associated proteins could act synergistically in regulating cementoblast differentiation and cementum deposition and offer new approaches on how the cementogenesis process is regulated.
Nuñez
J
J
Facultad de Odontologa, Universidad Complutense de Madrid, España.
Sanz
M
M
Hoz-Rodríguez
L
L
Zeichner-David
M
M
Arzate
H
H
D053523:Amelogenin / Q000096:biosynthesis; D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D025021:Cementogenesis / Q000502:physiology*; D003739:Dental Cementum / Q000166:cytology* / Q000378:metabolism*; D003746:Dental Enamel Proteins / Q000096:biosynthesis*; D006801:Humans; D017027:Protein Tyrosine Phosphatases / Q000096:biosynthesis; D011506:Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014092:Tooth Root / Q000166:cytology
C490889:AMBN protein, human; D053523:Amelogenin; C510886:CEMP1 protein, human; D003746:Dental Enamel Proteins; D011506:Proteins; C038456:tuftelin; C404489:HACD1 protein, human; D017027:Protein Tyrosine Phosphatases