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Effects of caffeine and related methylxanthines on fetal mouse palates cultured in vitro
1Institute of Applied Medicine, Hokkaido Radio-isotope Center, Ltd., Japan.
Abstract:
The maxillary regions of day-12.5 ICR mouse fetuses were cultured in a chemically-defined serumless medium and the effects of methylxanthine derivatives on cultured palates were studied. Explanted palates were treated for 72 hr in vitro with 0.5-2 mM caffeine (CA), 1-2 mM theophylline (TP), or 1-2 mM theobromine (TB). Although the three compounds tested did not prevent the contact of opposing palatal shelves, palatal fusion was inhibited by CA and TP at a concentration of 2 mM, and the inhibitory effect of CA was more evident than that of TP. On the other hand, TB did not exert an inhibitory effect on palatogenesis at 2 mM. Since the in vitro toxicity of the methylxanthine compounds appeared to correlate well with their in vivo teratogenic potential, the organ culture method of fetal rodent palates should be a useful tool for screening teratogenic agents, especially those causing cleft palate.
Insights
Caffeine and theophylline inhibited mouse palate fusion in vitro, while theobromine did not. This suggests organ culture can screen for cleft palate-causing agents.
Area of Science:
- Developmental biology
- Pharmacology
- Toxicology
Background:
- Palatogenesis is a critical developmental process.
- Cleft palate is a common birth defect.
- Methylxanthine derivatives like caffeine are widely consumed.
Purpose of the Study:
- To investigate the effects of methylxanthine derivatives on palate fusion using an in vitro organ culture system.
- To assess the potential of these compounds to cause cleft palate.
- To evaluate the utility of rodent palate organ culture for teratogenicity screening.
Main Methods:
- Maxillary regions from day-12.5 ICR mouse fetuses were cultured in a chemically-defined, serumless medium.
- Explanted palates were treated with varying concentrations of caffeine, theophylline, and theobromine for 72 hours.
- Inhibition of palatal fusion and compound toxicity were assessed.
Main Results:
- Caffeine (2 mM) and theophylline (2 mM) inhibited palatal fusion, with caffeine showing a stronger effect.
- Theobromine (2 mM) did not inhibit palatogenesis.
- No tested compound prevented initial contact between palatal shelves.
- In vitro toxicity correlated with known in vivo teratogenic potential.
Conclusions:
- Methylxanthine derivatives can interfere with palatal fusion in a concentration-dependent manner.
- In vitro organ culture of fetal rodent palates is a promising method for screening teratogenic agents causing cleft palate.
- Caffeine and theophylline exhibit teratogenic potential relevant to cleft palate formation.