Reakcja ludzkich komórek struktur jamy ustnej na różne materiały endodontyczne – badanie in vitro
The human periodontal ligaments cells were harvested from the periodontal membrane of impacted, surgically removed wisdom teeth, which therefore did not have any contact with the oral cavity at any time. The PDL cells were cultured in Dulbeco’s Modified Eagle Medium 1X (Lot 1012067, 4,5 g/l Glucose, L-Glutamine, Pyruvate; gibco by life technologies, Darmstadt, Germany) with fetal bovine serum, Penicillin (10.000 U/ml), Streptomycin (10.000 μg/ml) and Amphotericin B 250 μg/ml (Biochrom, Berlin, Germany).
The material samples were placed in 6-well-dishes (TPP, Trasadingen, Switzerland) and brought in direct contact to the harvested cell. The cells were plated at a density of 5,000 cells/cm2 and cultured in their respective cell culture medium (PDL cells in Dulbeco’s Modified Eagle Medium 1X and osteoblasts in MM0, High Growth Enhancement Medium with dexamethasone) and in the presence of the samples. Cells at passage P2 were used for the study. All experiments were performed in triplets. The in-vitro investigations were performed after defined intervals of one, eight, thirteen and twenty days. To control the growth and proliferation of the cells also cultures without contact to any test material were assessed (control group).
Morphological analysis
For histological evaluation the cell cultures were fixed in methyl alcohol (Merck, Darmstadt, Germany) at 21°C room temperature, air dried and a Richardson staining was performed. The Richardson solution ready for use consisted of two filtered stock solutions. Stock [...]
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