Organic-solvent extraction of model biomaterials for use in the in vitro chromosome aberration test.
We prepared polyurethane (PU) containing 0.4% or 4% 4,4'-methylenedianiline (MDA) as model materials to investigate the effectiveness of sample preparation by organic-solvent extraction for the in vitro chromosome aberration (CA) test. MDA itself (0.4 mg/mL) was positive only in the presence of an exogenous metabolizing system (S9 mix). The culture medium extract of PU containing 4% MDA (PU/4% MDA) was negative with and without S9 mix. Methanol and acetone extracts, on the other hand, induced structural CAs without S9 mix, which we did not expect because MDA requires S9 mix for activity. On chemical analysis, however, we found that the ratio of MDA extracted by the organic solvents to that extracted by the culture medium of PU/4% MDA was about 15:1. Interestingly, oligomers consisting of poly(tetramethyleneglycol) derivatives (OTMG) were also extracted by the organic solvents. The data suggest that the induction of structural CAs in the absence of S9 mix may have been partly due to synergism of MDA and OTMG. CA tests of MDA and PTMG-1000 in combination confirmed that to be the case. Thus, organic-solvent extraction may be more effective than medium extraction in evaluating the biological safety of biomaterials. Detailed chemical analysis of extracts was performed.
Matsuoka
Atsuko
A
Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. matsuoka@nihs.go.jp
Haishima
Yuji
Y
Hasegawa
Chie
C
Matsuda
Yoshie
Y
Tsuchiya
Toshie
T
D000096:Acetone / Q000737:chemistry; D000814:Aniline Compounds / Q000737:chemistry; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods; D002869:Chromosome Aberrations; D006224:Cricetinae; D003470:Culture Media / Q000737:chemistry; D006018:Glycols / Q000737:chemistry; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods; D000432:Methanol / Q000737:chemistry; D011140:Polyurethanes / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D001559:Benzhydryl Compounds
D000096:Acetone; D000814:Aniline Compounds; D001672:Biocompatible Materials; D003470:Culture Media; D006018:Glycols; D000432:Methanol; D011140:Polyurethanes; D012997:Solvents; C047554:polytetramethylene glycol; C009505:4,4'-diaminodiphenylmethane; D001559:Benzhydryl Compounds