In vitro clonogenic capacity of PDL fibroblasts cultured with Emdogain

Objective: The aim of the present study was to evaluate the efficiency of Emdogain (EMD) in preserving the clonogenic capacity of periodontal ligament fibroblasts (PDLF). Methods:PDLF were obtained from explants of young permanent healthy tooth. After initial outgrow, the culture medium of experimental flasks was replaced with medium supplemented with 100 ug/ml EMD, whereas the other served as controls . Following five weeks, the cells were washed (3X), harvested (trypsin + EDTA), and evaluated for their viability. Viable cells from were inoculated into six 96-wells-plate at a concentration of 1 viable cell/ 2 wells and were allowed to grow for 5 weeks. The percentage of cells with clonogenic capacity was determined as the number of colonies formed/number of cells seeded x100 in the experimental and control groups. Three degrees of dish area coverage were utilized: up to 25%; between 25– 75% and higher then 75%. This experiment was repeated four times from four different donors. Results:A total of 2328 cells were evaluated, half of which, were cultured with Emdogain. The mean percentage of cells that exhibited any clonogenic capacity in the presence of EMD was comparable to that of cells cultured in the absence of EMD:26.6%+14.3 as compared with 34.6%+20.6, respectively (p=0.186). In contrast, the percentage of clones that covered more then 75% of the well area was significantly lower in the EMD-treated clones as compared to the untreated ones 10.9+11.1 vs. 23.8+14.7, respectively (p= 0.022). In conclusion: EMD decreased the percentage of PDLF with capabilities of arising colonies with 75%-100% confluency probably by increasing their differentiation.