Evaluating cytotoxicity of four different dental metal materials toward L-929 cells using three assay systems

doi:10.3969/j.issn.2095-4344.2013.08.011

Abstract

Abstract:

BACKGROUND: Different methods to evaluate cytotoxicity of the same material may produce different results.
OBJECTIVE: To evaluate the cytotoxicity of nickel-chromium alloy, cobalt-chromium alloy, 3Cr13 stainless steel and pure titanium toward L-929 cells by using three assay systems.
METHODS: The L-929 cells were cultivated in vitro in the extracts of four different dental metal materials (nickel-chromium alloy, cobalt-chromium alloy, 3Cr13 stainless steel and pure titanium) at 24 hours and 72 hours, separately. The L-929 cells, cultured in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum, served as the negative control group. And cells, cultured in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum and 0.7% acrylamide, served as the positive control group. The detection methods for the cytotoxicity included MTT, cell counting kit-8 and crystal violet cell proliferation assay system.
RESULTS AND CONCLUSION: (1) Microscopy showed that the L-929 cells, cultivated in the extracts of four different dental materials, were normal and their intracellular structure was clear. They proliferated well with extension of incubation time. There were no significant differences between the test and negative control groups at all times. However, the number of cells in the positive control group significantly reduced as compared with the negative control group. The morphological integrity destroyed and a lot of cell debris formed. (2) After culture for 24 hours, the cell counting kit-8 assay revealed that the relative growth rate of the cobalt-chromium alloy group was significantly lower than that of the negative group (P < 0.05), while the MTT and crystal violet assay revealed that there were no significant differences in cell viability between the cobalt-chromium alloy and negative control group (P > 0.05). After culture for 72 hours, the MTT assay revealed that the relative growth rate of four dental metal materials groups was significantly lower than that of the negative group (P < 0.01), while the cell counting kit-8 and crystal violet assay revealed that there were no significant differences in cell viability between the four dental metal materials and negative control group (P > 0.05). The cytotoxicity of the tested dental metal materials was grade 0-1, which meets the safety standard. These indicate that these four dental metal materials have good biological safety.

Key words: biomaterials, tissue engineering oral materials, nickel-chromium alloy, cobalt-chromium alloy, 3Cr13, pure titanium, dental metal materials, cytotoxicity, L-929 cells, MTT colorimetric assay, cell counting kit-8 assay, crystal violet assay, provincial grants-supported paper, biomaterial photographs-containing paper

CLC Number:

R318

Chen Yao-zhong, Liu Gen-di, Song Ping, Gao Feng. Evaluating cytotoxicity of four different dental metal materials toward L-929 cells using three assay systems[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(8): 1390-1397.

Figures/Tables 5

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