The cytotoxicity assessment of novel formulation developed to reduce dentin hypersensitivity utilizing lactate dehydrogenase assay

Abstract

The problem of real treatment of teeth hypersensitivity is still important and unsolved. The main goal of the experiment was to calculate the possible toxic effects on the fibroblasts cells CCL-1™ (NCTC clone 929) caused by original preparation to reduce tooth surfaces’ hypersensitivity, compared to the marketable preparation Seal & Protect (Dentsply). The assessment was made through measuring lactate dehydrogenase (LDH assay). Lactate dehydrogenase releases from the cell’s cytoplasm to the culture medium as a result of cell membrane damage and lysis of the cells. The measurement is based on an assessment of the ability of LDH to oxidize lactic acid to pyruvic acid, which is dependent on the increase of the release level. The increase of LDH activity in the supernatants of cell cultures shows a relationship with the percentage of dead cells (increased cytotoxicity correlates with the increasing content of dead cells). In the LDH assay, both formulations evaluated after 24 h obtained results which were located below the control values. After seven days, the mean values obtained in cytotoxicity assay LDH are measurable and lower for the original formulation in comparison to the commercial one at the dilution of 1:5. At the dilution with 1:10 ratio, they are comparable and within the range of accepted values. At the maximal dilution of 1:15, the results are higher for the experimental formulation in comparison to the marketable formulation. The polymerization process is beneficial for the cytotoxicity test results in case of both tested preparations. Average values of cytotoxicity of both preparations attain an acceptable level of less than 22.6% ± 8.1, reliant on the degree of dilution and the remark time. Original formulation is characterized by a greater homogeneity of results. The marketable preparation has a larger diversity of effects, dependent on the time of observation and attenuation; however, the cytotoxicity values are lower when paralleled to the experimental formulation in the test conducted after seven days. This should not have a disastrous effect on the pulp, as the values of both as the values of both preparations are within expected ranges. The obtained results allows to assume that will be possible to introduce the original formulation to the stage of clinical trials in the future.