The ectopic bone formation of recombinant human bone morphogenetic protein-2(rhBMP-2) was evaluated using absorbable collagen sponges (ACS) and beta tricalcium phosphate (beta-TCP) as carriers in a rat subcutaneous assay model. Subcutaneous pockets were created on the back of rats. The pockets were implanted with rhBMP-2/ACS, rhBMP-2/beta-TCP, ACS alone, and beta-TCP alone. The rats were sacrificed at 2 or 8 weeks for histological and immunohistochemical evaluation. At 2 weeks, bone formation was evident in both the rhBMP-2/ACS and rhBMP-2/beta-TCP sites. At 8 weeks, the quantity of the new bone with a more advanced stage of remodeling had increased further in the rhBMP-2/beta-TCP sites. However, the newly formed bone observed at 2 weeks was not found in the rhBMP-2/ACS sites. On immunohistochemical observation, osteopontin staining was observed on both the rhBMP-2/ACS (2 weeks) and rhBMP-2/beta-TCP (2 and 8 weeks) sites. Osteocalcin was not detected in any of the samples. The lack of space-providing capacity of ACS may be one of the major factors responsible for its failure to maintain the newly induced bone. Therefore, a carrier for BMPs should provide space for bone formation and maturation during the more advanced healing stages.