Mesenchymal stem cells with osteogenic potential in human maxillary sinus membrane: an in vitro study

Clin Oral Investig. 2017 Jun;21(5):1599-1609. doi: 10.1007/s00784-016-1945-6. Epub 2016 Sep 1.

Abstract

Objectives: The aim of our study is to prove and validate the existence of an osteogenic progenitor cell population within the human maxillary Schneiderian sinus membrane (hMSSM) and to demonstrate their potential for bone formation.

Materials and methods: Ten hMSSM samples of approximately 2 × 2 cm were obtained during a surgical nasal approach for treatment of chronic rhinosinusitis and were retained for this study. The derived cells were isolated, cultured, and assayed at passage 3 for their osteogenic potential using the expression of Alkaline phosphatase, alizarin red and Von Kossa staining, flow cytometry, and quantitative real-time polymerase chain reaction.

Results: hMSSM-derived cells were isolated, showed homogenous spindle-shaped fibroblast-like morphology, characteristic of mesenchymal progenitor cells (MPCs), and demonstrated very high expression of MPC markers such as STRO-1, CD44, CD90, CD105, and CD73 in all tested passages. In addition, von Kossa and Alizarin red staining showed significant mineralization, a typical feature of osteoblasts. Moreover, alkaline phosphatase (ALP) activity was significantly increased at days 7, 14, 21, and 28 of culture in hMSSM-derived cells grown in osteogenic medium, in comparison to controls. Furthermore, osteogenic differentiation significantly upregulated the transcriptional expression of osteogenic markers such as ALP, Runt-related transcription factor 2 (Runx-2), bone morphogenetic protein (BMP)-2, osteocalcin (OCN), osteonectin (ON), and osteopontin (OPN), confirming that hMSSM-derived cells are of osteoprogenitor origin. Finally, hMSSM-derived cells were also capable of producing OPN proteins upon culturing in an osteogenic medium.

Conclusion: Our data showed that hMSSM holds mesenchymal osteoprogenitor cells capable of differentiating to the osteogenic lineage.

Clinical relevance: hMSSM contains potentially multipotent postnatal stem cells providing a promising clinical application in preimplant and implant therapy.

Keywords: Maxillary sinus; Mesenchymal stem cells; Osteogenic potential; Progenitor cells; Schneiderian membrane.

MeSH terms

  • Alkaline Phosphatase
  • Anthraquinones
  • Biomarkers / metabolism
  • Cell Differentiation
  • Cells, Cultured
  • Flow Cytometry
  • Humans
  • In Vitro Techniques
  • Maxillary Sinus / cytology*
  • Mesenchymal Stem Cells / cytology*
  • Osteoblasts / cytology
  • Osteogenesis / physiology*
  • Real-Time Polymerase Chain Reaction

Substances

  • Anthraquinones
  • Biomarkers
  • alizarin
  • Alkaline Phosphatase