Bone regeneration in critical-size calvarial defects using human dental pulp cells in an extracellular matrix-based scaffold

The rat calvarial defect is an established model to evaluate craniofacial bone regeneration using cell-scaffold biocomplexes. Dental pulp harbors stem cells with significant osteogenic properties. Extracellular matrix (ECM)-like scaffolds simulate the environment that cells observe in vivo. In the present study, we evaluated the osteogenic effect of a biocomplex of human dental pulp cells and a hyaluronic-based hydrogel scaffold in calvarial defects of immunocompetent rats. Dental pulp cells at the 2nd passage were characterized by flow cytometry, osteodifferentiated ex vivo for 4 days and the whole population was encapsulated in the synthetic ECM matrix. Cell vitality was verified 24 h upon encapsulation. 5 mm calvarial defects were created in 30 male rats and filled with the biocomplex, the scaffold alone, or left untreated. Histological evaluation at 8 weeks showed incomplete bone regeneration in all groups. The scaffold was not fully degraded and entrapped cells were detected in it. Histomorphometry showed statistically significant superior new bone formation in the biocomplex-treated group, compared to the two other groups. The present study provides evidence that the whole population of human dental pulp cells can advance bone healing when transplanted in immunocompetent animals and highlights the importance of proper scaffold degradation in cell-driven bioengineering treatments.