Stage-, dose-, and course-dependent inhibition of prenatal amoxicillin exposure on fetal articular cartilage development in fetal mice

Amoxicillin is widely used in the treatment of infectious diseases during pregnancy; however, the effects of prenatal amoxicillin exposure (PAE) on fetal development remain largely unknown. Therefore, this study aimed to investigate the toxic effects of PAE on fetal cartilage at different stage-, dose-, and course. Pregnant Kunming mice were orally administered 300 mg/kg·d (converted from clinical dose) amoxicillin on gestational days (GD) 10-12 or 16-18 (mid or late pregnancy stage), 150 or 300 mg/kg.d amoxicillin on GD16-18 (different doses), 300 mg/kg·d amoxicillin on GD16 (single course) or 16-18 (multiple courses), respectively. The fetal articular cartilage of the knee was collected on GD18. The number of chondrocytes and the expression of matrix synthesis/degradation, proliferation/apoptosis-related markers, and the TGF-β signaling pathway were detected. The results showed that the number of chondrocytes and the expression of matrix synthesis markers were reduced in male fetal mice treated with PAE (GD16-18, 300 mg/kg.d, single course and multiple courses), whereas the above indices in female mice showed no changes. The inhibited expression of PCNA, increased expression of Caspase-3, and down-regulated expression of the TGF-β signaling pathway were found in male PAE fetal mice. Accordingly, PAE exerted its "toxic effect window" on the knee cartilage development in male fetal mice, which manifested as reduced chondrocyte number and inhibited expression of matrix synthesis at a clinical dose of multiple courses in the late pregnancy stage. This study provides a theoretical and experimental basis for elucidating the risk of chondrodevelopmental toxicity associated with amoxicillin during pregnancy.