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

Stephan Payr*, Elizabeth Rosado-Balmayor, Thomas Tiefenboeck, Tim Schuseil, Marina Unger, Claudine Seeliger, Martijn van Griensven

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


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.

Original languageEnglish
Article number13
Number of pages7
JournalJournal of Orthopaedic Surgery and Research
Issue number1
Publication statusPublished - 6 Jan 2021


  • 2D
  • 3D cultivation
  • Donor age
  • Osteoblasts
  • RNA

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