Although vitamin D deficiency is common at birth, the consequences to growth and bone mass by weaning are unclear. This study was designed to determine whether maternal dietary vitamin D deficiency in pregnancy has a negative impact on the bone mass of full-term neonates and if postnatal supplementation could restore bone mass. Forty guinea pigs were randomized to receive a control (C) or deficient (D) diet (0.03 microg vs. 0.00 microg cholecalciferol/g) during pregnancy. Offspring were randomized at birth to receive 0.25 microg of cholecalciferol supplement (S) or a placebo (P) orally per day for 28 d. Measurements at birth and d 28 included whole body and regional bone mass and serum osteocalcin and deoxypyridinoline, plus biomechanical testing and peripheral quantitative computed tomography of excised tibias and femurs. Main and interactive effects were tested using mixed model ANOVA and post hoc Bonferroni's tests. At birth and d 28, offspring of the D sows had lower serum vitamin D and osteocalcin concentration, lower body weight, length, whole body and total tibia bone mineral content, and lower biomechanical integrity of tibia compared with those of the C sows, regardless of supplementation. Although postnatal vitamin D supplementation improved vitamin D status at d 28 in D offspring, values remained significantly lower than C groups. This study suggests that efforts should be made to optimize maternal vitamin D status in pregnancy, along with maintenance of vitamin D status in infancy, rather than relying on postnatal supplementation to normalize vitamin D status and bone mass.