Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides

Heungsoo Shin, Kyriacos Zygourakis, Mary C. Farach-Carson, Michael J Yaszemski, Antonios G. Mikos

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 ± 4.2 pmol/cell/h at day 10 for the concentration of 2 μmol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 ± 2.3 ng/cell for the peptide concentration of 2 μmol/g and 1.0 ± 0.3 ng/cell for 1.0 μmol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering.

Original languageEnglish (US)
Pages (from-to)535-543
Number of pages9
JournalJournal of Biomedical Materials Research - Part A
Volume69
Issue number3
StatePublished - Jun 1 2004

Fingerprint

Hydrogels
Biomimetics
Peptides
Modulation
Osteopontin
Hydrogel
Osteoblasts
Phosphatases
Alkaline Phosphatase
Calcium
glycyl-arginyl-glycyl-aspartyl-serine
Tissue culture
Polystyrenes
Differentiation Antigens
Scaffolds (biology)
Tissue engineering
Cell culture
Culture Media
Assays
Bone

Keywords

  • Biomimetic hydrogel
  • Differentiation
  • Marrow stromal cells
  • Osteoblast
  • Osteopontin

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials

Cite this

Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides. / Shin, Heungsoo; Zygourakis, Kyriacos; Farach-Carson, Mary C.; Yaszemski, Michael J; Mikos, Antonios G.

In: Journal of Biomedical Materials Research - Part A, Vol. 69, No. 3, 01.06.2004, p. 535-543.

Research output: Contribution to journalArticle

@article{5313a9538cca496ebee5e08abcebf5e6,
title = "Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides",
abstract = "We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 ± 4.2 pmol/cell/h at day 10 for the concentration of 2 μmol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 ± 2.3 ng/cell for the peptide concentration of 2 μmol/g and 1.0 ± 0.3 ng/cell for 1.0 μmol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering.",
keywords = "Biomimetic hydrogel, Differentiation, Marrow stromal cells, Osteoblast, Osteopontin",
author = "Heungsoo Shin and Kyriacos Zygourakis and Farach-Carson, {Mary C.} and Yaszemski, {Michael J} and Mikos, {Antonios G.}",
year = "2004",
month = "6",
day = "1",
language = "English (US)",
volume = "69",
pages = "535--543",
journal = "Journal of Biomedical Materials Research - Part A",
issn = "1549-3296",
publisher = "John Wiley and Sons Inc.",
number = "3",

}

TY - JOUR

T1 - Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides

AU - Shin, Heungsoo

AU - Zygourakis, Kyriacos

AU - Farach-Carson, Mary C.

AU - Yaszemski, Michael J

AU - Mikos, Antonios G.

PY - 2004/6/1

Y1 - 2004/6/1

N2 - We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 ± 4.2 pmol/cell/h at day 10 for the concentration of 2 μmol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 ± 2.3 ng/cell for the peptide concentration of 2 μmol/g and 1.0 ± 0.3 ng/cell for 1.0 μmol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering.

AB - We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 ± 4.2 pmol/cell/h at day 10 for the concentration of 2 μmol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 ± 2.3 ng/cell for the peptide concentration of 2 μmol/g and 1.0 ± 0.3 ng/cell for 1.0 μmol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering.

KW - Biomimetic hydrogel

KW - Differentiation

KW - Marrow stromal cells

KW - Osteoblast

KW - Osteopontin

UR - http://www.scopus.com/inward/record.url?scp=2442712725&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2442712725&partnerID=8YFLogxK

M3 - Article

C2 - 15127400

AN - SCOPUS:2442712725

VL - 69

SP - 535

EP - 543

JO - Journal of Biomedical Materials Research - Part A

JF - Journal of Biomedical Materials Research - Part A

SN - 1549-3296

IS - 3

ER -