Central nervous system (CNS) injuries in humans are frequently associated with heterotopic ossification (HO) and with enhanced fracture healing. In search for an experimental HO model we tested sera, from an established rat model of closed head injury (CHI), for their osteogenic effects on rat marrow stromal cells. Most normal rat sera increased cell proliferation not discriminating between osteoprogenitors and other stromal cells. Rats followed longitudinally by sequential blood sampling were bled 24 hours before and 24 hours, 48 hours, and 7 days after CHI. Sera obtained 24 and 48 hours after CHI progressively decreased cell proliferation and specific alkaline phosphatase (ALP) activity compared with pre-CHI sera of the same rats. Sera obtained from these rats, 7 days post-CHI, partially recovered proliferation induction, more than recovering induction of specific ALP activity. A positive correlation between day 11 ALP activity and day 21 mineralization was found under stimulation by pre-CHI sera. However, no correlation was found on stimulation with sera obtained 48 hours after CHI. Correlation between ALP and mineralization partially recovered in cultures exposed to sera obtained 7 days after CHI. In cross-sectional experiments where rats were subjected to single blood sampling, sera of 24 hours and 7 days post-CHI induced proliferation, whereas sera of 48 hours and 14 days post-CHI did not. The results indicate that 48 hours post-CHI, the mitogenicity of sera decreased in both cross-sectional and longitudinal blood sampling. In addition, 48 hours post-CHI, the specific ALP activity increases in cultured marrow stromal cells. Thus, changes in bone-seeking factors, causing serum-mediated osteogenic activity in rats, are expected close on 48 hour post-CNS injury.