The influence of enamel matrix derivative associated with insulin-like growth factor-I on periodontal ligament fibroblasts.
BACKGROUND
Enamel matrix derivative (EMD) has recently been shown to promote periodontal regeneration in vivo. Insulin-like growth factor-I (IGF-I) is a potent modulator of periodontal regeneration stimulating cell proliferation, differentiation, synthesis of type I collagen, and non-collagenous proteins. However, the biochemical effects of these factors on periodontal ligament (PDL) fibroblasts are not completely understood. The objective of the present study was to evaluate the effect of EMD, IGF-I, and the combination of these two factors on the proliferation, adhesion, migration, and expression of type I collagen in PDL fibroblasts.


METHODS
The proliferation rate was measured by automated cell counting and immunohistochemical expression of proliferating cell nuclear antigen (PCNA). The cell adhesion was analyzed by a colorimetric assay and cell migration was measured in Boyden chambers. Type I collagen expression and production was determined by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively.


RESULTS
The results indicated that the proliferation of PDL fibroblasts was significantly stimulated by EMD and EMD plus IGF-I in a dose- and time-dependent manner. EMD, IGF-I, and the combination of both factors had no effects on cellular migration and adhesion or expression and production of type I collagen.


CONCLUSIONS
Our results showed that EMD, IGF-I, and the combination of both factors stimulated PDL fibroblast proliferation, whereas these factors did not affect adhesion, migration, or expression of type I collagen of these cells.
Palioto
Daniela B
DB
Department of Maxillo-Facial Surgery and Periodontics, School of Dentistry of Ribeirão Preto, University of São Paulo, Brazil. dpalioto@forp.usp.br
Coletta
Ricardo D
RD
Graner
Edgard
E
Joly
Julio Cesar
JC
de Lima
Antonio Fernando Martorelli
AF
D000293:Adolescent; D000704:Analysis of Variance; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D004338:Drug Combinations; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric
D024042:Collagen Type I; D003746:Dental Enamel Proteins; D004338:Drug Combinations; D018809:Proliferating Cell Nuclear Antigen; C109069:enamel matrix proteins; D007334:Insulin-Like Growth Factor I