Enamel matrix-derived protein stimulates attachment of periodontal ligament fibroblasts and enhances alkaline phosphatase activity and transforming growth factor beta1 release of periodontal ligament and gingival fibroblasts.
BACKGROUND
Although it is claimed that enamel matrix-derived proteins (EMP) can be used to promote new attachment formation around periodontally involved teeth, the underlying biological mechanism is not understood. It was the aim of the present study to investigate the effects of EMP on the behavior of human periodontal ligament (HPLF) and gingival fibroblasts (HGF) in vitro, with special focus on their attachment properties, the expression of alkaline phosphatase (ALP) activity, the release of transforming growth factor (TGF)beta1, and their proliferative rate.


METHODS
Fibroblast populations were obtained from 10 individuals with a healthy periodontium and cultured in chemically defined medium on culture plates coated with EMP, purified collagen type I, or their respective vehicles. Experiments were performed in the absence of serum for periods up to 48 hours.


RESULTS
It was shown that HGF barely attached and spread on EMP-coated substrata, whereas HPLF attached and spread within 24 hours. However, when cultured on purified collagen type I, both cell types showed rapid attachment and spreading. Furthermore, the expression of ALP activity was significantly enhanced under the influence of EMP, especially in HPLF. HPLF and HGF both released significantly higher levels of TGFbeta1 in the presence of EMP. EMP did not influence 3H-thymidine incorporation by HPLF and HGF.


CONCLUSIONS
Our results indicate that HPLF and HGF respond differently to EMP. A more rapid attachment of HPLF to this substratum might contribute, during the initial stages of periodontal healing, to selective outgrowth and colonization of exposed root surfaces in vivo.
Van der Pauw
M T
MT
Department of Periodontology, Academic Center for Dentistry Amsterdam, University of Amsterdam, The Netherlands.
Van den Bos
T
T
Everts
V
V
Beertsen
W
W
D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D003094:Collagen; D003470:Culture Media; D016895:Culture Media, Serum-Free; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D005881:Gingiva / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D019275:Radiopharmaceuticals; D013936:Thymidine / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014316:Tritium
D003470:Culture Media; D016895:Culture Media, Serum-Free; D003746:Dental Enamel Proteins; D019275:Radiopharmaceuticals; D016212:Transforming Growth Factor beta; D014316:Tritium; D003094:Collagen; D000469:Alkaline Phosphatase; D013936:Thymidine