Human Intraoral Harvested Mesenchymal Stem Cells: Characterization, Multilineage Differentiation Analysis, and 3-Dimensional Migration of Natural Bone Mineral and Tricalcium Phosphate Scaffolds

PurposeThe aim of this study was the establishment of a minimally invasive technique of mesenchymal stem cell (MSC) harvesting and a predictable isolation and cultivation method on 2 different bone substitutes used as potential scaffolds.Materials and MethodsHuman MSCs isolated from the posterior maxilla were characterized by flow cytometric analysis. After in vitro expansion, cells were cultured and differentiated toward osteogenic, adipogenic, and chondrogenic lineages in 2-dimensional cultures and on natural bone mineral of bovine origin and β-tricalcium phosphate scaffolds. Three-dimensional growth was analyzed using live cell staining and confocal laser scanning microscopy.ResultsMSCs from all patients demonstrated the same immunophenotype, with expression of CD73, CD90, and CD105 but no expression of CD45, CD34, CD14, CD11, and HLA-DR. The potential of MSCs for multilineage differentiation along osteogenic, adipogenic, and chondrogenic lines was shown. Based on knowledge of the characteristics of the cells, a method was established to increase MSC expansion efficiency and seeding conditions on each scaffold. Results of the in vitro characterization and laser scanning microscopy visualized the 3-dimensional growth of MSCs on the 2 scaffold types.ConclusionsThe present data showed that intraoral MSCs can be cultured predictably under 2- and 3-dimensional conditions, have proved multiple potencies, and thus seem to be potential candidates for tissue engineering approaches in maxillofacial reconstructions.