Human intraoral harvested mesenchymal stem cells: characterization, multilineage differentiation analysis, and 3-dimensional migration of natural bone mineral and tricalcium phosphate scaffolds

J Oral Maxillofac Surg. 2012 Oct;70(10):2309-15. doi: 10.1016/j.joms.2011.06.216. Epub 2011 Sep 21.

Abstract

Purpose: The 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 methods: Human 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.

Results: MSCs 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.

Conclusions: The 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5'-Nucleotidase / analysis
  • Adipogenesis / physiology
  • Adolescent
  • Animals
  • Antigens, CD / analysis
  • Bone Substitutes / chemistry*
  • Calcium Phosphates / chemistry*
  • Cattle
  • Cell Culture Techniques
  • Cell Differentiation / physiology
  • Cell Lineage
  • Cell Movement / physiology
  • Cell Separation
  • Cell Survival / physiology
  • Chondrogenesis / physiology
  • Endoglin
  • Flow Cytometry
  • GPI-Linked Proteins / analysis
  • Humans
  • Imaging, Three-Dimensional / methods
  • Immunophenotyping
  • Maxilla / cytology*
  • Mesenchymal Stem Cells / physiology*
  • Microscopy, Confocal / methods
  • Minerals / chemistry*
  • Osteogenesis / physiology
  • Receptors, Cell Surface / analysis
  • Thy-1 Antigens / analysis
  • Tissue Scaffolds* / classification
  • Tissue and Organ Harvesting / methods*
  • Young Adult

Substances

  • Antigens, CD
  • Bio-Oss
  • Bone Substitutes
  • Calcium Phosphates
  • ENG protein, human
  • Endoglin
  • GPI-Linked Proteins
  • Minerals
  • Receptors, Cell Surface
  • Thy-1 Antigens
  • beta-tricalcium phosphate
  • 5'-Nucleotidase
  • NT5E protein, human