Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D

  • J Orthop Surg Res. 2021 Jan 6;16(1):13. doi: 10.1186/s13018-020-02153-z.
Stephan Payr  1  2 ,  Elizabeth Rosado-Balmayor  3  4 ,  Thomas Tiefenboeck  5 ,  Tim Schuseil  3 ,  Marina Unger  3 ,  Claudine Seeliger  3 ,  Martijn van Griensven  3  6
Affiliations
  • 1. Department of Experimental Trauma Surgery, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany. [email protected].
  • 2. Department of Orthopedics and Trauma Surgery, Division of Trauma Surgery, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria. [email protected].
  • 3. Department of Experimental Trauma Surgery, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany.
  • 4. Department IBE, MERLN Institute, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, The Netherlands.
  • 5. Department of Orthopedics and Trauma Surgery, Division of Trauma Surgery, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
  • 6. Department cBITE, MERLN Institute, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, The Netherlands.
PMID: 33407623 DOI: 10.1186/s13018-020-02153-z
Abstract

Background: The aim of this study was the investigation of the osteogenic potential of human osteoblasts of advanced donor age in 2D and 3D culture.

Methods: Osteoblasts were induced to osteogenic differentiation and cultivated, using the same polystyrene material in 2D and 3D culture for 2 weeks. Samples were taken to evaluate Alkaline Phosphatase (ALP) activity, mineralization and gene expression.

Results: Osteoprotegerin (OPG) levels were significantly increased (8.2-fold) on day 7 in 3D compared to day 0 (p < 0.0001) and 11.6-fold higher in 3D than in 2D (p < 0.0001). Both culture systems showed reduced osteocalcin (OC) levels (2D 85% and 3D 50% of basic value). Collagen type 1 (Col1) expression was elevated in 3D on day 7 (1.4-fold; p = 0.009). Osteopontin (OP) expression showed 6.5-fold higher levels on day 7 (p = 0.002) in 3D than in 2D. Mineralization was significantly higher in 3D on day 14 (p = 0.0002).

Conclusion: Advanced donor age human primary osteoblasts reveal significantly higher gene expression levels of OPG, Col1 and OP in 3D than in monolayer. Therefore, it seems that a relatively high potential of bone formation in a natural 3D arrangement is presumably still present in osteoblasts of elderly people.

Trial registration: 5217/11 on the 22nd of Dec. 2011.

Keywords
2D/3D cultivation; Donor age; Osteoblasts.
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