Objectives: Enamel matrix derivative (EMD: Emdogain®) is derived from porcine developing enamel matrix and contains mainly amelogenin protein. EMD has been shown to facilitate regeneration of periodontium, although its mechanism of action is unknown. The Protein Kinase-C (PKC) isoforms are involved in a number of processes like growth, differentiation, and cytokine secretion. The purpose of this study was to evaluate EMD effectiveness of the expression of differential markers (i.e. alkaline phosphatase and collagen type I activity) and potential differential markers, PKC isoforms on human periodontal ligament fibroblasts (HPLF).
Methods: HPLF were established from patients with sound extracted third molars. HPLF were seeded into 6-well plates and exposed to three different concentrations (5µg/ml, 25µg/ml, and 50µg/ml) of EMD for 3 days. Alkaline phosphatase activity was assayed in microplate reader and total mRNA was extracted for reverse-transcriptase polymerase chain-reaction (RT-PCR) to assess the gene expression of alkaline phosphatase and collagen type I. Western blotting was done to assess the expression of PKC isoforms. The data were analyzed by mean, standard deviation and ANOVA.
Results: The results showed that EMD had effects on significant increase of alkaline phosphatase activity at 5µg/ml concentration group; 17% higher activity than control group and 15% higher than 50µg/ml concentration group. RT-PCR also showed the increase of alkaline phosphatase and collagen type I gene expression. Western blotting revealed the increased expression of PKC isoforms-α, d, ε, η, θ, ι with the greatest effects at 5μg/ml concentration group.
Conclusion: EMD activated the increase of alkaline phosphatase and collagen type I activity of HPLF and stimulated the expression of PKC isoforms. Therefore, this study indicated that the PKC family can be used as the marker of differential activity of HPLF under the effect of EMD. This study was supported by funds from UB School of Dental Medicine.