Expression of the pleiotrophin–midkine axis in a sheep tooth socket model of bone healing

OBJECTIVE AND BACKGROUND: Resorption of alveolar bone after tooth extraction is a common problem often requiring bone grafting. The success of the grafting procedures is dependent on multiple factors including the presence of growth factors. This is the first in vivo study to investigate the role of the pleiotrophin family of cytokines in alveolar bone regeneration. This research investigated the role of the pleiotrophin–midkine (PTN–MDK) axis during osteogenesis, with and without a grafting material, after tooth extraction in a sheep model. METHODS: Thirty Romney‐cross ewes were anesthetized, and all premolar teeth on the right side were extracted. The sockets were randomized to controls sites with no treatment and test sites with Bio‐Oss® graft material and Bio‐Gide® membrane. Samples were harvested after sacrificing animals 4, 8, and 16 weeks post‐grafting (n = 10 per time‐point). Tissue for qRT(2)‐PCR gene analysis was recovered from the socket next to the first molar using a trephine (Ø = 2 mm). Each socket was fixed, decalcified, paraffin‐embedded, and sectioned. Immunohistochemistry was conducted to localize both PTN and MDK along with their receptors, protein tyrosine phosphatase receptor type Z1 (PTPRZ1), ALK receptor tyrosine kinase (ALK), and notch receptor 2 (NOTCH2). RESULTS: Within the healing sockets, high expression of genes for PTN, MDK, NOTCH2, and ALK was found at all time‐points and in both grafted and non‐grafted sites, while PTPRZ1 was only expressed at low levels. The relative gene expression of the PTN family of cytokines was not statistically different at the three time‐points between test and control groups (p > .05). Immunohistochemistry found PTN and MDK in association with new bone, NOTCH2 in the connective tissue, and PTPRZ1 and ALK in association with cuboidal osteoblasts involved in bone formation. CONCLUSIONS: The PTN–MDK axis was highly expressed in both non‐grafted and grafted sockets during osteogenesis in a sheep model of alveolar bone regeneration with no evidence that grafting significantly affected expression. The activation of NOTCH2 and PTPRZ1 receptors may be important during bone regeneration in vivo. The discovery of the PTN–MDK axis as important during alveolar bone regeneration is novel and opens up new avenues of research into these stably expressed highly active cytokines. Growth factor supplementation with PTN and/or MDK during healing may be an approach for enhanced regeneration or to initiate healing where delayed.