Bone Morphogenetic Proteins and Smad Expression in Ovine Tendon-Bone Healing

Purpose: Bone morphogenetic proteins (BMPs) are being developed to improve tendon-bone healing. To do this, it is essential to understand the endogenous expression of BMPs and their downstream signal transduction factors, Smads, during tendon-bone healing. Methods: An extra-articular patellar tendon-bone healing ovine model was set up, and histologic evaluation of the healing progress at the tendon-bone interface at 1, 2, 3, and 6 weeks was performed. Immunohistochemical staining of BMP-2, BMP-7, Smad1, Smad4, and Smad5 was carried out in all sections. Results: The model revealed formation of a loose granuloma tissue layer between the tendon and bone at 1 week, remodeling starting at 2 weeks, and Sharpey-like collagen fiber formation at 3 and 6 weeks. All detected factors were elevated at the tendon-bone interface during healing, and the expression peaked at 2 to 3 weeks. The cells involved were osteoblastic-like cells, osteoclastic-like cells, mesenchymal cells, and fibroblasts. BMP-7 staining was mainly at the interface close to the bony side, whereas BMP-2 expression shifted to the tendon side at 6 weeks. The expression pattern of Smad1 and Smad5 was similar to that of BMP-7. Smad1 was also found to be expressed in osteoclastic-like cells at 1 and 2 weeks. Smad4 expression was the highest among all of the factors at all time points. Conclusions: The data suggest that endogenous BMP-2 and BMP-7 participate in tendon-bone healing and their functions involve their downstream signal transduction mediators, Smad1, Smad4, and Smad5. Clinical Relevance: The temporal expression of BMPs should be considered when setting up therapeutic strategies using BMPs.