A Tunable, Three-Dimensional in Vitro Culture Model of Growth Plate Cartilage Using Alginate Hydrogel Scaffolds

Alek G. Erickson, Taylor D. Laughlin, Sarah M. Romereim, Catherine N. Sargus-Patino, Angela K Pannier, Andrew T Dudley

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Defining the final size and geometry of engineered tissues through precise control of the scalar and vector components of tissue growth is a necessary benchmark for regenerative medicine, but it has proved to be a significant challenge for tissue engineers. The growth plate cartilage that promotes elongation of the long bones is a good model system for studying morphogenetic mechanisms because cartilage is composed of a single cell type, the chondrocyte; chondrocytes are readily maintained in culture; and growth trajectory is predominately in a single vector. In this cartilage, growth is generated via a differentiation program that is spatially and temporally regulated by an interconnected network composed of long- and short-range signaling mechanisms that together result in the formation of functionally distinct cellular zones. To facilitate investigation of the mechanisms underlying anisotropic growth, we developed an in vitro model of the growth plate cartilage by using neonatal mouse growth plate chondrocytes encapsulated in alginate hydrogel beads. In bead cultures, encapsulated chondrocytes showed high viability, cartilage matrix deposition, low levels of chondrocyte hypertrophy, and a progressive increase in cell proliferation over 7 days in culture. Exogenous factors were used to test functionality of the parathyroid-related protein-Indian hedgehog (PTHrP-IHH) signaling interaction, which is a crucial feedback loop for regulation of growth. Consistent with in vivo observations, exogenous PTHrP stimulated cell proliferation and inhibited hypertrophy, whereas IHH signaling stimulated chondrocyte hypertrophy. Importantly, the treatment of alginate bead cultures with IHH or thyroxine resulted in formation of a discrete domain of hypertrophic cells that mimics tissue architecture of native growth plate cartilage. Together, these studies are the first demonstration of a tunable in vitro system to model the signaling network interactions that are required to induce zonal architecture in growth plate chondrocytes, which could also potentially be used to grow cartilage cultures of specific geometries to meet personalized patient needs.

Original languageEnglish (US)
Pages (from-to)94-105
Number of pages12
JournalTissue Engineering - Part A
Volume24
Issue number1-2
DOIs
StatePublished - Jan 1 2018

Fingerprint

Growth Plate
Hydrogel
Alginate
Cartilage
Chondrocytes
Hydrogels
Scaffolds
Hypertrophy
Growth
Tissue
Cell proliferation
Hedgehog Proteins
Cell Proliferation
Parathyroid Hormone-Related Protein
Benchmarking
Regenerative Medicine
alginic acid
In Vitro Techniques
Thyroxine
Geometry

Keywords

  • 3D culture model
  • Growth plate cartilage
  • IHH/PTHrP signaling
  • alginate hydrogel
  • tissue architecture

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biomaterials
  • Biomedical Engineering

Cite this

A Tunable, Three-Dimensional in Vitro Culture Model of Growth Plate Cartilage Using Alginate Hydrogel Scaffolds. / Erickson, Alek G.; Laughlin, Taylor D.; Romereim, Sarah M.; Sargus-Patino, Catherine N.; Pannier, Angela K; Dudley, Andrew T.

In: Tissue Engineering - Part A, Vol. 24, No. 1-2, 01.01.2018, p. 94-105.

Research output: Contribution to journalArticle

Erickson, Alek G. ; Laughlin, Taylor D. ; Romereim, Sarah M. ; Sargus-Patino, Catherine N. ; Pannier, Angela K ; Dudley, Andrew T. / A Tunable, Three-Dimensional in Vitro Culture Model of Growth Plate Cartilage Using Alginate Hydrogel Scaffolds. In: Tissue Engineering - Part A. 2018 ; Vol. 24, No. 1-2. pp. 94-105.
@article{16f3e927f56e4131a4a168e7edf843a2,
title = "A Tunable, Three-Dimensional in Vitro Culture Model of Growth Plate Cartilage Using Alginate Hydrogel Scaffolds",
abstract = "Defining the final size and geometry of engineered tissues through precise control of the scalar and vector components of tissue growth is a necessary benchmark for regenerative medicine, but it has proved to be a significant challenge for tissue engineers. The growth plate cartilage that promotes elongation of the long bones is a good model system for studying morphogenetic mechanisms because cartilage is composed of a single cell type, the chondrocyte; chondrocytes are readily maintained in culture; and growth trajectory is predominately in a single vector. In this cartilage, growth is generated via a differentiation program that is spatially and temporally regulated by an interconnected network composed of long- and short-range signaling mechanisms that together result in the formation of functionally distinct cellular zones. To facilitate investigation of the mechanisms underlying anisotropic growth, we developed an in vitro model of the growth plate cartilage by using neonatal mouse growth plate chondrocytes encapsulated in alginate hydrogel beads. In bead cultures, encapsulated chondrocytes showed high viability, cartilage matrix deposition, low levels of chondrocyte hypertrophy, and a progressive increase in cell proliferation over 7 days in culture. Exogenous factors were used to test functionality of the parathyroid-related protein-Indian hedgehog (PTHrP-IHH) signaling interaction, which is a crucial feedback loop for regulation of growth. Consistent with in vivo observations, exogenous PTHrP stimulated cell proliferation and inhibited hypertrophy, whereas IHH signaling stimulated chondrocyte hypertrophy. Importantly, the treatment of alginate bead cultures with IHH or thyroxine resulted in formation of a discrete domain of hypertrophic cells that mimics tissue architecture of native growth plate cartilage. Together, these studies are the first demonstration of a tunable in vitro system to model the signaling network interactions that are required to induce zonal architecture in growth plate chondrocytes, which could also potentially be used to grow cartilage cultures of specific geometries to meet personalized patient needs.",
keywords = "3D culture model, Growth plate cartilage, IHH/PTHrP signaling, alginate hydrogel, tissue architecture",
author = "Erickson, {Alek G.} and Laughlin, {Taylor D.} and Romereim, {Sarah M.} and Sargus-Patino, {Catherine N.} and Pannier, {Angela K} and Dudley, {Andrew T}",
year = "2018",
month = "1",
day = "1",
doi = "10.1089/ten.tea.2017.0091",
language = "English (US)",
volume = "24",
pages = "94--105",
journal = "Tissue Engineering - Part A.",
issn = "1937-3341",
publisher = "Mary Ann Liebert Inc.",
number = "1-2",

}

TY - JOUR

T1 - A Tunable, Three-Dimensional in Vitro Culture Model of Growth Plate Cartilage Using Alginate Hydrogel Scaffolds

AU - Erickson, Alek G.

AU - Laughlin, Taylor D.

AU - Romereim, Sarah M.

AU - Sargus-Patino, Catherine N.

AU - Pannier, Angela K

AU - Dudley, Andrew T

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Defining the final size and geometry of engineered tissues through precise control of the scalar and vector components of tissue growth is a necessary benchmark for regenerative medicine, but it has proved to be a significant challenge for tissue engineers. The growth plate cartilage that promotes elongation of the long bones is a good model system for studying morphogenetic mechanisms because cartilage is composed of a single cell type, the chondrocyte; chondrocytes are readily maintained in culture; and growth trajectory is predominately in a single vector. In this cartilage, growth is generated via a differentiation program that is spatially and temporally regulated by an interconnected network composed of long- and short-range signaling mechanisms that together result in the formation of functionally distinct cellular zones. To facilitate investigation of the mechanisms underlying anisotropic growth, we developed an in vitro model of the growth plate cartilage by using neonatal mouse growth plate chondrocytes encapsulated in alginate hydrogel beads. In bead cultures, encapsulated chondrocytes showed high viability, cartilage matrix deposition, low levels of chondrocyte hypertrophy, and a progressive increase in cell proliferation over 7 days in culture. Exogenous factors were used to test functionality of the parathyroid-related protein-Indian hedgehog (PTHrP-IHH) signaling interaction, which is a crucial feedback loop for regulation of growth. Consistent with in vivo observations, exogenous PTHrP stimulated cell proliferation and inhibited hypertrophy, whereas IHH signaling stimulated chondrocyte hypertrophy. Importantly, the treatment of alginate bead cultures with IHH or thyroxine resulted in formation of a discrete domain of hypertrophic cells that mimics tissue architecture of native growth plate cartilage. Together, these studies are the first demonstration of a tunable in vitro system to model the signaling network interactions that are required to induce zonal architecture in growth plate chondrocytes, which could also potentially be used to grow cartilage cultures of specific geometries to meet personalized patient needs.

AB - Defining the final size and geometry of engineered tissues through precise control of the scalar and vector components of tissue growth is a necessary benchmark for regenerative medicine, but it has proved to be a significant challenge for tissue engineers. The growth plate cartilage that promotes elongation of the long bones is a good model system for studying morphogenetic mechanisms because cartilage is composed of a single cell type, the chondrocyte; chondrocytes are readily maintained in culture; and growth trajectory is predominately in a single vector. In this cartilage, growth is generated via a differentiation program that is spatially and temporally regulated by an interconnected network composed of long- and short-range signaling mechanisms that together result in the formation of functionally distinct cellular zones. To facilitate investigation of the mechanisms underlying anisotropic growth, we developed an in vitro model of the growth plate cartilage by using neonatal mouse growth plate chondrocytes encapsulated in alginate hydrogel beads. In bead cultures, encapsulated chondrocytes showed high viability, cartilage matrix deposition, low levels of chondrocyte hypertrophy, and a progressive increase in cell proliferation over 7 days in culture. Exogenous factors were used to test functionality of the parathyroid-related protein-Indian hedgehog (PTHrP-IHH) signaling interaction, which is a crucial feedback loop for regulation of growth. Consistent with in vivo observations, exogenous PTHrP stimulated cell proliferation and inhibited hypertrophy, whereas IHH signaling stimulated chondrocyte hypertrophy. Importantly, the treatment of alginate bead cultures with IHH or thyroxine resulted in formation of a discrete domain of hypertrophic cells that mimics tissue architecture of native growth plate cartilage. Together, these studies are the first demonstration of a tunable in vitro system to model the signaling network interactions that are required to induce zonal architecture in growth plate chondrocytes, which could also potentially be used to grow cartilage cultures of specific geometries to meet personalized patient needs.

KW - 3D culture model

KW - Growth plate cartilage

KW - IHH/PTHrP signaling

KW - alginate hydrogel

KW - tissue architecture

UR - http://www.scopus.com/inward/record.url?scp=85040614534&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85040614534&partnerID=8YFLogxK

U2 - 10.1089/ten.tea.2017.0091

DO - 10.1089/ten.tea.2017.0091

M3 - Article

C2 - 28525313

AN - SCOPUS:85040614534

VL - 24

SP - 94

EP - 105

JO - Tissue Engineering - Part A.

JF - Tissue Engineering - Part A.

SN - 1937-3341

IS - 1-2

ER -