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

doi:10.1186/s13018-020-02153-z

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 journal › Article › Academic › peer-review

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.

Original language	English
Article number	13
Number of pages	7
Journal	Journal of Orthopaedic Surgery and Research
Volume	16
Issue number	1
DOIs	https://doi.org/10.1186/s13018-020-02153-z
Publication status	Published - 6 Jan 2021

Keywords

2D
3D cultivation
Donor age
Osteoblasts
RNA
CELL CULTURES
GROWTH
LINEAGE CELLS
DIFFERENTIATION
PROTEINS

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@article{27ebcb280f834559b2d3d704c7b7f89d,

title = "Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D",

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, RNA, CELL CULTURES, GROWTH, LINEAGE CELLS, DIFFERENTIATION, PROTEINS",

author = "Stephan Payr and Elizabeth Rosado-Balmayor and Thomas Tiefenboeck and Tim Schuseil and Marina Unger and Claudine Seeliger and {van Griensven}, Martijn",

note = "Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

month = jan,

day = "6",

doi = "10.1186/s13018-020-02153-z",

language = "English",

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TY - JOUR

T1 - Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D

AU - Payr, Stephan

AU - Rosado-Balmayor, Elizabeth

AU - Tiefenboeck, Thomas

AU - Schuseil, Tim

AU - Unger, Marina

AU - Seeliger, Claudine

AU - van Griensven, Martijn

PY - 2021/1/6

Y1 - 2021/1/6

N2 - 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.

AB - 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.

KW - 2D

KW - 3D cultivation

KW - Donor age

KW - Osteoblasts

KW - RNA

KW - CELL CULTURES

KW - GROWTH

KW - LINEAGE CELLS

KW - DIFFERENTIATION

KW - PROTEINS

U2 - 10.1186/s13018-020-02153-z

DO - 10.1186/s13018-020-02153-z

M3 - Article

C2 - 33407623

SN - 1749-799X

VL - 16

JO - Journal of Orthopaedic Surgery and Research

JF - Journal of Orthopaedic Surgery and Research

IS - 1

M1 - 13

ER -