In Vitro Embryotoxicity Testing of Polymeric Substances for Dental Use by Differentiation of Embryonic Stem Cells
Koichi Imai1, Horst Spielmann2, Gabi Scholz2, Ingeborg pohl2 and Masaaki Nakamura1
1 Osaka Dental University, 2 ZEBET (National German Center for the Documentation and Evaluation of Alternatives to Testing in Animals) at BgVV (National Center for Health Protection of Consumers and Veterinary Medicine )
Correspondence: Koichi Imai, Ph.D.
Department of Biomaterials, Osaka Dental University
8-1, Kuzuhahanazono-cho, Hirakara-city, Osaka573-1121, Japan
Tel: 072-864-3056, Fax: 072-864-3156,

Original paper :AATEX 8(1):31-39
2,2-bis [4-(2-hydroxy-3-methacryloy-loxypropoxy) phenyl] propane (Bis-GMA), di(methacryloyloxyethyl) trimethyl hexamethylene-diurethane (UDMA), 2,2-bis (4-methacryloyloxypoly ethoxy phenyl) propane (Bis-MPEPP) and triethylen glycoldimethacrylate (TEGDMA) were used as main component monomers for the tooth restorative composites resin or preventive filling sealant. These monomers absorbed could possibly be metabolized in the living body. The effects of four dental monomers on the capability of embryonic stem (ES) cells of the mouse cell-line D3 to differentiate into contracting myocard was examined using an embryonic stem cell test (EST), together with the cytotoxic effect on differentiated mouse 3T3 fibroblasts and on differentiated ES cells. The half-inhibition concentration for differentiation (ID50) on ES cells was 10.5ug/mL for Bis-GMA, 9.8ug/mL for UDMA, 7.9ug/mL for Bis-MPEPP was and 15.4ug/mL for TEGDMA. The IC50 ES for Bis-GMA was 23.6ug/mL and declined in the order of Bis-MPEPP, UDMA and TEGDMA. On the contrary, the IC503T3 for Bis-GMA was 19.8ug/mL and declined in the order of UDMA, TEGDMA and Bis-MPEPP in 3T3 cells. The embryotoxic potential was obtained from a biostatistically based prediction model of EST, which was based upon the three endpoints, ID50, IC50ES and IC503T3. It is considered that the four dental monomers are classified as "weak embryotoxic". The in vitro test using ES cells holds promise to be used to examine in vitro embryotoxicity of dental biomaterials.

Keywords: embryonic stem cell, EST, in vitro embryotoxicity test, dental, monomer