Cytotoxicity of Chelating Agents Used In Endodontics and Their Influence on MMPs of Cell Membranes

Abstract

This study evaluated the cytotoxic effect and the ability to inhibit matrix metalloproteinases (MMP-2 and MMP-9) of 0.2% chitosan (CH) and 1% acetic acid (AA) compared with 17% ethylenediaminetetraacetic acid (EDTA). Cell viability assay was performed according to ISO 10993-5 with mouse fibroblasts (L929). The culture was exposed to 0.2% CH, 1% AA, and 17% EDTA. The chelating agents were evaluated immediately after contact with the cells and after 6 h, 12 h, and 24 h of incubation. Cell viability was analyzed using the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Inhibition of the gelatinolytic activity of MMP-2 and MMP-9 was evaluated by gelatin zymography. Different concentrations of CH were evaluated: 50 mM, 5 mM, 0.5 mM, and 0.05 mM. EDTA (0.5 mM) was used as a positive control. The results demonstrated that CH and AA had an initial cytotoxic effect, which decreased after 6 h, 12 h, and 24 h, being statistically similar to EDTA (P > 0.05). Additionally, CH at concentrations of 50 mM, 5 mM, and 0.5 mM had an inhibitory effect on MMP-2 and MMP-9, similar to that of the control with EDTA. The chelating agents had no cytotoxic effects after 24 h. MMP-2 and MMP-9 were inhibited by the experimental solutions.

Cytotoxic Effect of Different Concentrations of Chitosan and Propolis Nanoparticles on Periodontal Ligament Stem Cells

“…(29) The MTT assay was chosen for this study because it is a standardised method for determining the effect on cell viability, safe, simple, easily repeatable, rapid, minimally toxic, and superior for determining the activity of mitochondrial dehydrogenase in living cells. It measures mitochondrial dehydrogenase activity, depending on the water soluble methylthiazol tetrazolium being converted to an insoluble purple formazan where the amount of formazan crystals formed is proportional to the activity of the living cells as these crystals are impermeable to cell membranes (30) . Tetrazolium salt reduction is now widely recognised as a reliable method for determining cell proliferation.…”

Efficacy of Chitosan as Final Irrigating Solution on TGF-β1 Release from Root Canal Dentin and its Effect on Dental Pulp Stem Cells Response

“…Another recent study revealed conflicting results, where 0.2% Chitosan and 17% EDTA showed similar increase in the number of viable cells. This might be explained by the use of lower concentration of chitosan, besides using cytotoxic test model other than DPSCs that might have different sensitivity towards the tested solutions (33) . Within this study, the released TGF-β1 in the concentration range of 82-517 pg.…”

Nanoparticles in Dentistry: A Comprehensive Review

“…Chitosan has many applications in endodontics. It can be successfully used as a chelating substance [ 51 , 52 , 53 , 54 , 55 ], as a scaffold for the delivery of growth factors [ 56 ], or medications [ 57 ]. Moreover, it can be used as an irrigant solution [ 47 , 58 , 59 , 60 ] with less toxicity than sodium hypochlorite or chlorhexidine [ 58 , 59 ], as an intracanal medication by itself [ 61 ], or as an adjunct to calcium hydroxide [ 62 , 63 ].…”