In vitro effects of Pepgen biomaterial on periodontal ligament fibroblasts

Objectives: The aim of this study is to evaluate the effects of PepGen (P-15) a synthetic bone grafting biomaterial that mimic the organic and inorganic components of the autogenous bone, on proliferation, attachment and mineralization of periodontal ligament (PDL) fibroblasts. Methods: To analyze the effects of P-15 on cell proliferation, 5x10⁴ PDL fibroblasts were plated on 50mg P-15 biomaterial and counted on days 1, 3, 6, 8, 10 by hemacytometer, in which the cells plated on the surface of the cell plate served as control group. In the attachment assay, %3 agarose was used to stabilize P-15 in the cell plate surface. The morphology of PDL fibroblasts and interactions between the PDL fibroblasts and P-15 was visualized by the scanning electron microscopy. For mineralization assay, PDL were incubated with mineralization media (10mM β-glycerophosphate + 50mg/ml Ascorbic acid) for 4 weeks. Von Kossa method was used to observe mineralized-like nodule formation on weeks 1, 2, 3 and 4. Results: Both P-15 and control groups showed an increase through the proliferation assay. While the increase in control group did not reach to any significance, the increase in P-15 group between day 3 and 6 was statistically significant (p<0.01). The comparision of P-15 and control group indicated that there was a significant difference between groups at days 6, 8, 10 (p<0.05) Neither P-15 nor control groups showed mineralized-like nodule formation during the period examined. The morphology and also attachment of PDL cells on P-15 material in tissue culture model was visualised by scanning electron microscopy. Conclusion: The findings demonstrate that P-15 enhance proliferation of PDL fibroblasts, but did not effect the mineralized-like nodule formation in vitro. Thus, P-15 may enhance periodontal regeneration in the early stage of wound healing by enhancing cell proliferation. (This study is supported by the Hacettepe University Research Foundation).