A comparative investigation of fetal skeletal anomalies in rats induced by acetylsalicylic acid with single- and double-staining techniques.
Acetylsalicylic acid (ASA) at single doses of 125, 250, and 500mg/kg was administered to pregnant rats on Gestation Day (GD) 10, and skeletal changes in fetuses harvested on GD 20 and pups on post-natal (PN) Day 21 were evaluated. Changes in cartilage and ossified bones identified by Alizarin Red S single-staining were compared with Alizarin Red S and Alcian Blue double-staining. By the single-staining technique, skeletal abnormalities including fused rib, incomplete ossification of the cervical arch, absent/hemicentric body of thoracic or lumbar vertebra, deformation of lumbar arch, and absent sacral arch were demonstrated in at 250 and 500mg/kg ASA on GD 20. The double-staining technique facilitated identification of additional cartilaginous changes in the vertebrae, paws, and ribs: including discontinuous rib cartilage, fused carpus, and split cartilage of thoracic centrum at same doses. Discontinuous rib cartilage and fused carpus persisted in pups until PN Day 21 demonstrating that these changes were irreversible. With use of the double-staining technique, the incidence of abnormalities at 250mg/kg were dramatically increased, thus this technique was more sensitive for identifying fetal cartilaginous and ossified skeletal changes.
Dodo
Tetsushi
T
Drug Safety Research Laboratories, Eisai Co., Ltd., 1 Kawashimatakehaya-machi Kakamigahara-shi, Gifu 501-6195, Japan. t-dodo@hhc.eisai.co.jp
Fukuta
Tetsuya
T
Uchida
Keiji
K
Mineshima
Hiroshi
H
Okuda
Yasuyuki
Y
Okada
Fumihiro
F
Katsutani
Naruo
N
Hirano
Kazuyuki
K
Aoki
Toyohiko
T
D000014:Abnormalities, Drug-Induced / Q000473:pathology*; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000633:toxicity*; D001241:Aspirin / Q000633:toxicity*; D001842:Bone and Bones / Q000002:abnormalities*; D004305:Dose-Response Relationship, Drug; D005260:Female; D008297:Male; D011247:Pregnancy; D011297:Prenatal Exposure Delayed Effects / Q000473:pathology*; D051381:Rats; D013194:Staining and Labeling
D000894:Anti-Inflammatory Agents, Non-Steroidal; D001241:Aspirin