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KMID : 1177720150090010023
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Journal of Alternatives to Animal Experiments
2015 Volume.9 No. 1 p.23 ~ p.32
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Development of an Alternative Oral Irritation Test Method Using Three-Dimensional Human Oral Mucosal Models
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Lim Hye-Rim
Yi Jung-Sun
Kim Tae-Sung
Koh Kyoung-Yuk
Ahn Il-Young
Kim Joo-Hwan
Lee Jin-Ha
Yang Song-Yi
Kim Kwang-Man
Shon Soo-Jung
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Abstract
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Ensuring the biological safety of dental materials is important, because people of all ages use them and they stay inside the mouth of a person for years once they are put into it. However, current oral irritation tests bring pain to animals, and there are no internationally accepted alternative test methods. In this study, we have developed a new method to test for oral irritation using three-dimensional (3D) human oral mucosal models in the light of growing needs for alternatives to animal testing in order to ensure the safety of dental materials. The commercially available 3D human oral mucosal models (EpiOral™, HOM™) were cultured, using normal oral epithelial cells, and histologically and phenotypically similar to native buccal tissues. Two controls and three dental materials that are widely used in dentistry were selected for testing. These included a polyethylene film (negative control), 1% Triton X-100 (positive control), resin, denture base resin and impression materials. These three dental materials were prepared according to manufacturers’ instructions, and were turned into 0.7-cm discs in diameter. And then, we directly placed the controls and the three dental materials on the top of each human oral mucosal model for 24 hours and measured tissue viability via the MTT assay and cytokine secretion. When cell viability is less than 50% or cytokine secretion is more than 250 pg/ml, a test material is evaluated as an irritant. The negative control, resin and denture base resin turned out non-irritants while the positive control and the impression materials irritants. Since the results of the oral mucosal irritation test of dental materials using human oral mucosal models matched those of existing cytotoxicity tests, it seems that the oral mucosal test can be a good alternative method. The results of this study suggest that the oral mucosal irritation test employing the 3D human oral mucosal models can be an alternative test method for dental materials. And further validation studies are required in order to take advantage of this method in the future.
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KEYWORD
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Dental materials, Oral mucosal irritation, Oral mucosal model, MTT assay, Cytokine secretion
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