Repairing Critical-Sized Rat Calvarial Defects with a Periosteal Cell-Seeded Small Intestinal Submucosal Layer

Background: Small intestinal submucosa was evaluated as a bioscaffold candidate for periosteum-derived osteoblasts, and its suitability as a bone replacement material for cranial defects was investigated. Methods: In the in vitro phase, osteoblasts were expanded in osteogenic medium and were then seeded onto small intestinal submucosa. To confirm osteoblast phenotype, they were tested for alkaline phosphatase, collagen type 1, and calcium expression. In the in vivo phase, calvarial critical-sized defects were created in 35 rats. The defects were either left untreated for surgical control (group 1), treated with small intestinal submucosa alone (group 2), treated with an osteoblast-embedded construct (group 3), or treated with an autogenous bone graft (group 4). The results were evaluated 12 weeks after surgery with radiopacity measurements and with stereologic analysis. Results: Periosteal cells grew successfully in vitro. The percentage radiopaque area at the defect was measured to be 42, 74, 76, and 89 percent for groups 1, 2, 3, and 4, respectively. The pixel intensity of the same site was 36.4, 48.1, 47.5, and 54.5 for the same groups, respectively. Tissue-engineered constructs did not achieve enough bone formation and calcification to be effective as autogenous bone grafts and were not superior to the small intestinal submucosa alone. However, both small intestinal submucosa and cell-seeded small intestinal submucosa showed significantly more bone formation compared with the untreated group. Conclusions: Although it was demonstrated that the small intestinal submucosa itself has osteogenic properties, it was not significantly increased by adding periosteum-derived osteoblasts to it. The osteogenic properties of small intestinal submucosa are promising, and its role as a scaffold should be investigated further.