In vitro mineralization of human mesenchymal stem cells on three-dimensional type I collagen versus PLGA scaffolds: A comparative analysis

Erwin A. Kruger, Daniel D. Im, David S. Bischoff, Clifford Pereira, Weibiao Huang, George H. Rudkin, Dean T. Yamaguchi, Timothy A. Miller

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: Development of a tissue engineered bone graft requires efficient bioactivity screening of biomaterials in clinically relevant three-dimensional systems. The authors analyzed the relative osteogenic potential of two three-dimensional biomaterials-type I collagen and poly(L-lactide-co-glycolide) (PLGA)- to support in vitro mineralization of human mesenchymal stem cells. Methods: Human mesenchymal stem cells were seeded onto three-dimensional PLGA or type I collagen scaffolds; incubated in osteogenic media; and harvested at 1, 4, and 7 days. Messenger RNA expression was analyzed using quantitative real-time reverse-transcriptase polymerase chain reaction for osteogenic (i.e., alkaline phosphatase, osteocalcin, bone sialoprotein, Runx2/core binding factor α-1) and angiogenic (i.e., vascular endothelial growth factor and interleukin-8) markers. Alkaline phosphatase enzyme activity was measured at 4 and 7 days. Mineralization was detected by alizarin red staining and micro-computed tomographic imaging at 8 and 12 weeks. Mineral composition was analyzed by solid-phase nuclear magnetic resonance spectroscopy. Results: Early osteogenic and angiogenic markers, and alkaline phosphatase enzyme activity, were up-regulated on PLGA versus collagen scaffolds. However, long-term mineralization endpoints favored type I collagen. By 8 weeks, human mesenchymal stem cells on collagen exhibited significantly higher mineral density by micro-computed tomographic and alizarin red staining than PLGA scaffolds. Both biomaterials deposited calcium hydroxyapatite as determined by nuclear magnetic resonance spectroscopy. Conclusions: The authors' findings suggest that despite early PLGA induction of osteogenic gene expression, long-term mineralization occurs earlier and to a greater extent on type I collagen, highlighting collagen as a potential bone tissue engineering scaffold in the human mesenchymal stem cell niche. When screening the relative osteoinductive profiles of three-dimensional bone tissue engineering scaffolds in vitro, the authors recommend including long-term endpoints of osteogenesis.

Original languageEnglish (US)
Pages (from-to)2301-2311
Number of pages11
JournalPlastic and Reconstructive Surgery
Volume127
Issue number6
DOIs
StatePublished - Jun 1 2011

Fingerprint

Collagen Type I
Mesenchymal Stromal Cells
Biocompatible Materials
Tissue Scaffolds
Alkaline Phosphatase
Collagen
Tissue Engineering
Bone and Bones
Minerals
Magnetic Resonance Spectroscopy
Core Binding Factors
Integrin-Binding Sialoprotein
Staining and Labeling
Stem Cell Niche
Polyglactin 910
Osteocalcin
Enzymes
Durapatite
Reverse Transcriptase Polymerase Chain Reaction
Interleukin-8

ASJC Scopus subject areas

  • Surgery

Cite this

In vitro mineralization of human mesenchymal stem cells on three-dimensional type I collagen versus PLGA scaffolds : A comparative analysis. / Kruger, Erwin A.; Im, Daniel D.; Bischoff, David S.; Pereira, Clifford; Huang, Weibiao; Rudkin, George H.; Yamaguchi, Dean T.; Miller, Timothy A.

In: Plastic and Reconstructive Surgery, Vol. 127, No. 6, 01.06.2011, p. 2301-2311.

Research output: Contribution to journalArticle

Kruger, Erwin A. ; Im, Daniel D. ; Bischoff, David S. ; Pereira, Clifford ; Huang, Weibiao ; Rudkin, George H. ; Yamaguchi, Dean T. ; Miller, Timothy A. / In vitro mineralization of human mesenchymal stem cells on three-dimensional type I collagen versus PLGA scaffolds : A comparative analysis. In: Plastic and Reconstructive Surgery. 2011 ; Vol. 127, No. 6. pp. 2301-2311.
@article{7cbe11291c01476dbf017876ca2cd206,
title = "In vitro mineralization of human mesenchymal stem cells on three-dimensional type I collagen versus PLGA scaffolds: A comparative analysis",
abstract = "Background: Development of a tissue engineered bone graft requires efficient bioactivity screening of biomaterials in clinically relevant three-dimensional systems. The authors analyzed the relative osteogenic potential of two three-dimensional biomaterials-type I collagen and poly(L-lactide-co-glycolide) (PLGA)- to support in vitro mineralization of human mesenchymal stem cells. Methods: Human mesenchymal stem cells were seeded onto three-dimensional PLGA or type I collagen scaffolds; incubated in osteogenic media; and harvested at 1, 4, and 7 days. Messenger RNA expression was analyzed using quantitative real-time reverse-transcriptase polymerase chain reaction for osteogenic (i.e., alkaline phosphatase, osteocalcin, bone sialoprotein, Runx2/core binding factor α-1) and angiogenic (i.e., vascular endothelial growth factor and interleukin-8) markers. Alkaline phosphatase enzyme activity was measured at 4 and 7 days. Mineralization was detected by alizarin red staining and micro-computed tomographic imaging at 8 and 12 weeks. Mineral composition was analyzed by solid-phase nuclear magnetic resonance spectroscopy. Results: Early osteogenic and angiogenic markers, and alkaline phosphatase enzyme activity, were up-regulated on PLGA versus collagen scaffolds. However, long-term mineralization endpoints favored type I collagen. By 8 weeks, human mesenchymal stem cells on collagen exhibited significantly higher mineral density by micro-computed tomographic and alizarin red staining than PLGA scaffolds. Both biomaterials deposited calcium hydroxyapatite as determined by nuclear magnetic resonance spectroscopy. Conclusions: The authors' findings suggest that despite early PLGA induction of osteogenic gene expression, long-term mineralization occurs earlier and to a greater extent on type I collagen, highlighting collagen as a potential bone tissue engineering scaffold in the human mesenchymal stem cell niche. When screening the relative osteoinductive profiles of three-dimensional bone tissue engineering scaffolds in vitro, the authors recommend including long-term endpoints of osteogenesis.",
author = "Kruger, {Erwin A.} and Im, {Daniel D.} and Bischoff, {David S.} and Clifford Pereira and Weibiao Huang and Rudkin, {George H.} and Yamaguchi, {Dean T.} and Miller, {Timothy A.}",
year = "2011",
month = "6",
day = "1",
doi = "10.1097/PRS.0b013e318213a004",
language = "English (US)",
volume = "127",
pages = "2301--2311",
journal = "Plastic and Reconstructive Surgery",
issn = "0032-1052",
publisher = "Lippincott Williams and Wilkins",
number = "6",

}

TY - JOUR

T1 - In vitro mineralization of human mesenchymal stem cells on three-dimensional type I collagen versus PLGA scaffolds

T2 - A comparative analysis

AU - Kruger, Erwin A.

AU - Im, Daniel D.

AU - Bischoff, David S.

AU - Pereira, Clifford

AU - Huang, Weibiao

AU - Rudkin, George H.

AU - Yamaguchi, Dean T.

AU - Miller, Timothy A.

PY - 2011/6/1

Y1 - 2011/6/1

N2 - Background: Development of a tissue engineered bone graft requires efficient bioactivity screening of biomaterials in clinically relevant three-dimensional systems. The authors analyzed the relative osteogenic potential of two three-dimensional biomaterials-type I collagen and poly(L-lactide-co-glycolide) (PLGA)- to support in vitro mineralization of human mesenchymal stem cells. Methods: Human mesenchymal stem cells were seeded onto three-dimensional PLGA or type I collagen scaffolds; incubated in osteogenic media; and harvested at 1, 4, and 7 days. Messenger RNA expression was analyzed using quantitative real-time reverse-transcriptase polymerase chain reaction for osteogenic (i.e., alkaline phosphatase, osteocalcin, bone sialoprotein, Runx2/core binding factor α-1) and angiogenic (i.e., vascular endothelial growth factor and interleukin-8) markers. Alkaline phosphatase enzyme activity was measured at 4 and 7 days. Mineralization was detected by alizarin red staining and micro-computed tomographic imaging at 8 and 12 weeks. Mineral composition was analyzed by solid-phase nuclear magnetic resonance spectroscopy. Results: Early osteogenic and angiogenic markers, and alkaline phosphatase enzyme activity, were up-regulated on PLGA versus collagen scaffolds. However, long-term mineralization endpoints favored type I collagen. By 8 weeks, human mesenchymal stem cells on collagen exhibited significantly higher mineral density by micro-computed tomographic and alizarin red staining than PLGA scaffolds. Both biomaterials deposited calcium hydroxyapatite as determined by nuclear magnetic resonance spectroscopy. Conclusions: The authors' findings suggest that despite early PLGA induction of osteogenic gene expression, long-term mineralization occurs earlier and to a greater extent on type I collagen, highlighting collagen as a potential bone tissue engineering scaffold in the human mesenchymal stem cell niche. When screening the relative osteoinductive profiles of three-dimensional bone tissue engineering scaffolds in vitro, the authors recommend including long-term endpoints of osteogenesis.

AB - Background: Development of a tissue engineered bone graft requires efficient bioactivity screening of biomaterials in clinically relevant three-dimensional systems. The authors analyzed the relative osteogenic potential of two three-dimensional biomaterials-type I collagen and poly(L-lactide-co-glycolide) (PLGA)- to support in vitro mineralization of human mesenchymal stem cells. Methods: Human mesenchymal stem cells were seeded onto three-dimensional PLGA or type I collagen scaffolds; incubated in osteogenic media; and harvested at 1, 4, and 7 days. Messenger RNA expression was analyzed using quantitative real-time reverse-transcriptase polymerase chain reaction for osteogenic (i.e., alkaline phosphatase, osteocalcin, bone sialoprotein, Runx2/core binding factor α-1) and angiogenic (i.e., vascular endothelial growth factor and interleukin-8) markers. Alkaline phosphatase enzyme activity was measured at 4 and 7 days. Mineralization was detected by alizarin red staining and micro-computed tomographic imaging at 8 and 12 weeks. Mineral composition was analyzed by solid-phase nuclear magnetic resonance spectroscopy. Results: Early osteogenic and angiogenic markers, and alkaline phosphatase enzyme activity, were up-regulated on PLGA versus collagen scaffolds. However, long-term mineralization endpoints favored type I collagen. By 8 weeks, human mesenchymal stem cells on collagen exhibited significantly higher mineral density by micro-computed tomographic and alizarin red staining than PLGA scaffolds. Both biomaterials deposited calcium hydroxyapatite as determined by nuclear magnetic resonance spectroscopy. Conclusions: The authors' findings suggest that despite early PLGA induction of osteogenic gene expression, long-term mineralization occurs earlier and to a greater extent on type I collagen, highlighting collagen as a potential bone tissue engineering scaffold in the human mesenchymal stem cell niche. When screening the relative osteoinductive profiles of three-dimensional bone tissue engineering scaffolds in vitro, the authors recommend including long-term endpoints of osteogenesis.

UR - http://www.scopus.com/inward/record.url?scp=79958700738&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79958700738&partnerID=8YFLogxK

U2 - 10.1097/PRS.0b013e318213a004

DO - 10.1097/PRS.0b013e318213a004

M3 - Article

C2 - 21617464

AN - SCOPUS:79958700738

VL - 127

SP - 2301

EP - 2311

JO - Plastic and Reconstructive Surgery

JF - Plastic and Reconstructive Surgery

SN - 0032-1052

IS - 6

ER -