Extracellular matrix protein production in human adipose-derived mesenchymal stem cells on three-dimensional polycaprolactone (PCL) scaffolds responds to GDF5 or FGF2

Yan Su, Janet M. Denbeigh, Emily T. Camilleri, Scott M. Riester, Joshua A. Parry, Eric R. Wagner, Michael J Yaszemski, Allan B Dietz, Simon M. Cool, Andre J van Wijnen, Sanjeev Kakar

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The poor healing potential of intra-articular ligament injuries drives a need for the development of novel, viable ‘neo-ligament’ alternatives. Ex vivo approaches combining stem cell engineering, 3-dimensional biocompatible scaffold design and enhancement of biological and biomechanical functionality via the introduction of key growth factors and morphogens, represent a promising solution to ligament regeneration. We investigated growth, differentiation and extracellular matrix (ECM) protein production of human adipose-derived mesenchymal stem/stromal cells (MSCs), cultured in 5% human platelet lysate (PL) and seeded on three-dimensional polycaprolactone (PCL) scaffolds, in response to the connective-tissue related ligands fibroblast growth factor 2 (basic) (FGF2) and growth and differentiation factor-5 (GDF5). Phenotypic alterations of MSCs under different biological conditions were examined using cell viability assays, real time qPCR analysis of total RNA, as well as immunofluorescence microscopy. Phenotypic conversion of MSCs into ECM producing fibroblastic cells proceeds spontaneously in the presence of human platelet lysate. Administration of FGF2 and/or GDF5 enhances production of mRNAs for several ECM proteins including Collagen types I and III, as well as Tenomodulin (e.g., COL1A1, TNMD), but not Tenascin-C (TNC). Differences in the in situ deposition of ECM proteins Collagen type III and Tenascin-C were validated by immunofluorescence microscopy. Treatment of MSCs with FGF2 and GDF5 was not synergistic and occasionally antagonistic for ECM production. Our results suggest that GDF5 alone enhances the conversion of MSCs to fibroblastic cells possessing a phenotype consistent with that of connective-tissue fibroblasts.

Original languageEnglish (US)
Pages (from-to)149-156
Number of pages8
JournalGene Reports
Volume10
DOIs
StatePublished - Mar 1 2018

Fingerprint

Growth Differentiation Factor 5
Extracellular Matrix Proteins
Fibroblast Growth Factor 2
Mesenchymal Stromal Cells
Tenascin
Collagen Type III
Fluorescence Microscopy
Ligaments
Connective Tissue
Extracellular Matrix
Articular Ligaments
Blood Platelets
polycaprolactone
Cell Engineering
Collagen Type I
Regeneration
Cell Survival
Intercellular Signaling Peptides and Proteins

Keywords

  • FGF2
  • GDF5
  • Growth factors
  • MSC
  • Neo-ligament
  • Polymer scaffolds

ASJC Scopus subject areas

  • Genetics

Cite this

Extracellular matrix protein production in human adipose-derived mesenchymal stem cells on three-dimensional polycaprolactone (PCL) scaffolds responds to GDF5 or FGF2. / Su, Yan; Denbeigh, Janet M.; Camilleri, Emily T.; Riester, Scott M.; Parry, Joshua A.; Wagner, Eric R.; Yaszemski, Michael J; Dietz, Allan B; Cool, Simon M.; van Wijnen, Andre J; Kakar, Sanjeev.

In: Gene Reports, Vol. 10, 01.03.2018, p. 149-156.

Research output: Contribution to journalArticle

Su, Yan ; Denbeigh, Janet M. ; Camilleri, Emily T. ; Riester, Scott M. ; Parry, Joshua A. ; Wagner, Eric R. ; Yaszemski, Michael J ; Dietz, Allan B ; Cool, Simon M. ; van Wijnen, Andre J ; Kakar, Sanjeev. / Extracellular matrix protein production in human adipose-derived mesenchymal stem cells on three-dimensional polycaprolactone (PCL) scaffolds responds to GDF5 or FGF2. In: Gene Reports. 2018 ; Vol. 10. pp. 149-156.
@article{56612f2785b54ae2b8b284ea767ed897,
title = "Extracellular matrix protein production in human adipose-derived mesenchymal stem cells on three-dimensional polycaprolactone (PCL) scaffolds responds to GDF5 or FGF2",
abstract = "The poor healing potential of intra-articular ligament injuries drives a need for the development of novel, viable ‘neo-ligament’ alternatives. Ex vivo approaches combining stem cell engineering, 3-dimensional biocompatible scaffold design and enhancement of biological and biomechanical functionality via the introduction of key growth factors and morphogens, represent a promising solution to ligament regeneration. We investigated growth, differentiation and extracellular matrix (ECM) protein production of human adipose-derived mesenchymal stem/stromal cells (MSCs), cultured in 5{\%} human platelet lysate (PL) and seeded on three-dimensional polycaprolactone (PCL) scaffolds, in response to the connective-tissue related ligands fibroblast growth factor 2 (basic) (FGF2) and growth and differentiation factor-5 (GDF5). Phenotypic alterations of MSCs under different biological conditions were examined using cell viability assays, real time qPCR analysis of total RNA, as well as immunofluorescence microscopy. Phenotypic conversion of MSCs into ECM producing fibroblastic cells proceeds spontaneously in the presence of human platelet lysate. Administration of FGF2 and/or GDF5 enhances production of mRNAs for several ECM proteins including Collagen types I and III, as well as Tenomodulin (e.g., COL1A1, TNMD), but not Tenascin-C (TNC). Differences in the in situ deposition of ECM proteins Collagen type III and Tenascin-C were validated by immunofluorescence microscopy. Treatment of MSCs with FGF2 and GDF5 was not synergistic and occasionally antagonistic for ECM production. Our results suggest that GDF5 alone enhances the conversion of MSCs to fibroblastic cells possessing a phenotype consistent with that of connective-tissue fibroblasts.",
keywords = "FGF2, GDF5, Growth factors, MSC, Neo-ligament, Polymer scaffolds",
author = "Yan Su and Denbeigh, {Janet M.} and Camilleri, {Emily T.} and Riester, {Scott M.} and Parry, {Joshua A.} and Wagner, {Eric R.} and Yaszemski, {Michael J} and Dietz, {Allan B} and Cool, {Simon M.} and {van Wijnen}, {Andre J} and Sanjeev Kakar",
year = "2018",
month = "3",
day = "1",
doi = "10.1016/j.genrep.2017.12.004",
language = "English (US)",
volume = "10",
pages = "149--156",
journal = "Gene Reports",
issn = "2452-0144",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Extracellular matrix protein production in human adipose-derived mesenchymal stem cells on three-dimensional polycaprolactone (PCL) scaffolds responds to GDF5 or FGF2

AU - Su, Yan

AU - Denbeigh, Janet M.

AU - Camilleri, Emily T.

AU - Riester, Scott M.

AU - Parry, Joshua A.

AU - Wagner, Eric R.

AU - Yaszemski, Michael J

AU - Dietz, Allan B

AU - Cool, Simon M.

AU - van Wijnen, Andre J

AU - Kakar, Sanjeev

PY - 2018/3/1

Y1 - 2018/3/1

N2 - The poor healing potential of intra-articular ligament injuries drives a need for the development of novel, viable ‘neo-ligament’ alternatives. Ex vivo approaches combining stem cell engineering, 3-dimensional biocompatible scaffold design and enhancement of biological and biomechanical functionality via the introduction of key growth factors and morphogens, represent a promising solution to ligament regeneration. We investigated growth, differentiation and extracellular matrix (ECM) protein production of human adipose-derived mesenchymal stem/stromal cells (MSCs), cultured in 5% human platelet lysate (PL) and seeded on three-dimensional polycaprolactone (PCL) scaffolds, in response to the connective-tissue related ligands fibroblast growth factor 2 (basic) (FGF2) and growth and differentiation factor-5 (GDF5). Phenotypic alterations of MSCs under different biological conditions were examined using cell viability assays, real time qPCR analysis of total RNA, as well as immunofluorescence microscopy. Phenotypic conversion of MSCs into ECM producing fibroblastic cells proceeds spontaneously in the presence of human platelet lysate. Administration of FGF2 and/or GDF5 enhances production of mRNAs for several ECM proteins including Collagen types I and III, as well as Tenomodulin (e.g., COL1A1, TNMD), but not Tenascin-C (TNC). Differences in the in situ deposition of ECM proteins Collagen type III and Tenascin-C were validated by immunofluorescence microscopy. Treatment of MSCs with FGF2 and GDF5 was not synergistic and occasionally antagonistic for ECM production. Our results suggest that GDF5 alone enhances the conversion of MSCs to fibroblastic cells possessing a phenotype consistent with that of connective-tissue fibroblasts.

AB - The poor healing potential of intra-articular ligament injuries drives a need for the development of novel, viable ‘neo-ligament’ alternatives. Ex vivo approaches combining stem cell engineering, 3-dimensional biocompatible scaffold design and enhancement of biological and biomechanical functionality via the introduction of key growth factors and morphogens, represent a promising solution to ligament regeneration. We investigated growth, differentiation and extracellular matrix (ECM) protein production of human adipose-derived mesenchymal stem/stromal cells (MSCs), cultured in 5% human platelet lysate (PL) and seeded on three-dimensional polycaprolactone (PCL) scaffolds, in response to the connective-tissue related ligands fibroblast growth factor 2 (basic) (FGF2) and growth and differentiation factor-5 (GDF5). Phenotypic alterations of MSCs under different biological conditions were examined using cell viability assays, real time qPCR analysis of total RNA, as well as immunofluorescence microscopy. Phenotypic conversion of MSCs into ECM producing fibroblastic cells proceeds spontaneously in the presence of human platelet lysate. Administration of FGF2 and/or GDF5 enhances production of mRNAs for several ECM proteins including Collagen types I and III, as well as Tenomodulin (e.g., COL1A1, TNMD), but not Tenascin-C (TNC). Differences in the in situ deposition of ECM proteins Collagen type III and Tenascin-C were validated by immunofluorescence microscopy. Treatment of MSCs with FGF2 and GDF5 was not synergistic and occasionally antagonistic for ECM production. Our results suggest that GDF5 alone enhances the conversion of MSCs to fibroblastic cells possessing a phenotype consistent with that of connective-tissue fibroblasts.

KW - FGF2

KW - GDF5

KW - Growth factors

KW - MSC

KW - Neo-ligament

KW - Polymer scaffolds

UR - http://www.scopus.com/inward/record.url?scp=85039844211&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85039844211&partnerID=8YFLogxK

U2 - 10.1016/j.genrep.2017.12.004

DO - 10.1016/j.genrep.2017.12.004

M3 - Article

VL - 10

SP - 149

EP - 156

JO - Gene Reports

JF - Gene Reports

SN - 2452-0144

ER -