Quantitative analysis of temporal and spatial variations of chondrocyte behavior in engineered cartilage during long-term culture

Kwideok Park, Byoung Hyun Min, Dong Keun Han, Karen Hasty

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

In this work, we present the fact that chondrocyte activity differs in relation to their position in an engineered cartilage construct. Chondrocytes from porcine articular cartilage were cultured in a monolayer. Then the cell/extracellular matrix (ECM) membrane was peeled off and centrifuged into a three-dimensional (3D) pellet-type construct. Cultivated in a static condition, the constructs were harvested at specific time intervals (1, 2, 3, and 5 weeks) and manually cored using a biopsy punch to separate the core from the remaining construct. The resultant parts, core and peripheral remnant were thus obtained and subjected to analysis individually. Cell density (106cells/ cm3) of the core was significantly higher at 1 week than that of the periphery but this trend was reversed at later time points. Cell viability was remarkably better in the peripheral tissue. Alcian blue staining of glycosaminoglycan (GAG) revealed an intense blue staining from the periphery, exhibiting a steep gradient in distribution of GAG concentration. The gene expression ratio of collagen type II to I appeared to be more altered in the periphery, possibly suggesting cell dedifferentiation, especially at later time points (>2 weeks). The mRNA levels of matrix metalloproteinase-1 (MMP-1) and MMP-13 remained in the normal range, whereas collagen type X expression was more significantly upregulated at the periphery. This study showed that chondrocyte behavior could be highly variable in the extent of their proliferation, differentiation and dedifferentiation, depending on their physical location within 3D engineered cartilage construct.

Original languageEnglish (US)
Pages (from-to)419-428
Number of pages10
JournalAnnals of Biomedical Engineering
Volume35
Issue number3
DOIs
StatePublished - Mar 1 2007

Fingerprint

Cartilage
Collagen
Chemical analysis
Biopsy
Gene expression
Monolayers
Cells
Tissue
Membranes
Glycosaminoglycans

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering

Cite this

Quantitative analysis of temporal and spatial variations of chondrocyte behavior in engineered cartilage during long-term culture. / Park, Kwideok; Min, Byoung Hyun; Han, Dong Keun; Hasty, Karen.

In: Annals of Biomedical Engineering, Vol. 35, No. 3, 01.03.2007, p. 419-428.

Research output: Contribution to journalArticle

@article{12f9bbc5d2e54ee499140b58f98fa98f,
title = "Quantitative analysis of temporal and spatial variations of chondrocyte behavior in engineered cartilage during long-term culture",
abstract = "In this work, we present the fact that chondrocyte activity differs in relation to their position in an engineered cartilage construct. Chondrocytes from porcine articular cartilage were cultured in a monolayer. Then the cell/extracellular matrix (ECM) membrane was peeled off and centrifuged into a three-dimensional (3D) pellet-type construct. Cultivated in a static condition, the constructs were harvested at specific time intervals (1, 2, 3, and 5 weeks) and manually cored using a biopsy punch to separate the core from the remaining construct. The resultant parts, core and peripheral remnant were thus obtained and subjected to analysis individually. Cell density (106cells/ cm3) of the core was significantly higher at 1 week than that of the periphery but this trend was reversed at later time points. Cell viability was remarkably better in the peripheral tissue. Alcian blue staining of glycosaminoglycan (GAG) revealed an intense blue staining from the periphery, exhibiting a steep gradient in distribution of GAG concentration. The gene expression ratio of collagen type II to I appeared to be more altered in the periphery, possibly suggesting cell dedifferentiation, especially at later time points (>2 weeks). The mRNA levels of matrix metalloproteinase-1 (MMP-1) and MMP-13 remained in the normal range, whereas collagen type X expression was more significantly upregulated at the periphery. This study showed that chondrocyte behavior could be highly variable in the extent of their proliferation, differentiation and dedifferentiation, depending on their physical location within 3D engineered cartilage construct.",
author = "Kwideok Park and Min, {Byoung Hyun} and Han, {Dong Keun} and Karen Hasty",
year = "2007",
month = "3",
day = "1",
doi = "10.1007/s10439-006-9219-0",
language = "English (US)",
volume = "35",
pages = "419--428",
journal = "Annals of Biomedical Engineering",
issn = "0090-6964",
publisher = "Springer Netherlands",
number = "3",

}

TY - JOUR

T1 - Quantitative analysis of temporal and spatial variations of chondrocyte behavior in engineered cartilage during long-term culture

AU - Park, Kwideok

AU - Min, Byoung Hyun

AU - Han, Dong Keun

AU - Hasty, Karen

PY - 2007/3/1

Y1 - 2007/3/1

N2 - In this work, we present the fact that chondrocyte activity differs in relation to their position in an engineered cartilage construct. Chondrocytes from porcine articular cartilage were cultured in a monolayer. Then the cell/extracellular matrix (ECM) membrane was peeled off and centrifuged into a three-dimensional (3D) pellet-type construct. Cultivated in a static condition, the constructs were harvested at specific time intervals (1, 2, 3, and 5 weeks) and manually cored using a biopsy punch to separate the core from the remaining construct. The resultant parts, core and peripheral remnant were thus obtained and subjected to analysis individually. Cell density (106cells/ cm3) of the core was significantly higher at 1 week than that of the periphery but this trend was reversed at later time points. Cell viability was remarkably better in the peripheral tissue. Alcian blue staining of glycosaminoglycan (GAG) revealed an intense blue staining from the periphery, exhibiting a steep gradient in distribution of GAG concentration. The gene expression ratio of collagen type II to I appeared to be more altered in the periphery, possibly suggesting cell dedifferentiation, especially at later time points (>2 weeks). The mRNA levels of matrix metalloproteinase-1 (MMP-1) and MMP-13 remained in the normal range, whereas collagen type X expression was more significantly upregulated at the periphery. This study showed that chondrocyte behavior could be highly variable in the extent of their proliferation, differentiation and dedifferentiation, depending on their physical location within 3D engineered cartilage construct.

AB - In this work, we present the fact that chondrocyte activity differs in relation to their position in an engineered cartilage construct. Chondrocytes from porcine articular cartilage were cultured in a monolayer. Then the cell/extracellular matrix (ECM) membrane was peeled off and centrifuged into a three-dimensional (3D) pellet-type construct. Cultivated in a static condition, the constructs were harvested at specific time intervals (1, 2, 3, and 5 weeks) and manually cored using a biopsy punch to separate the core from the remaining construct. The resultant parts, core and peripheral remnant were thus obtained and subjected to analysis individually. Cell density (106cells/ cm3) of the core was significantly higher at 1 week than that of the periphery but this trend was reversed at later time points. Cell viability was remarkably better in the peripheral tissue. Alcian blue staining of glycosaminoglycan (GAG) revealed an intense blue staining from the periphery, exhibiting a steep gradient in distribution of GAG concentration. The gene expression ratio of collagen type II to I appeared to be more altered in the periphery, possibly suggesting cell dedifferentiation, especially at later time points (>2 weeks). The mRNA levels of matrix metalloproteinase-1 (MMP-1) and MMP-13 remained in the normal range, whereas collagen type X expression was more significantly upregulated at the periphery. This study showed that chondrocyte behavior could be highly variable in the extent of their proliferation, differentiation and dedifferentiation, depending on their physical location within 3D engineered cartilage construct.

UR - http://www.scopus.com/inward/record.url?scp=33847284633&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33847284633&partnerID=8YFLogxK

U2 - 10.1007/s10439-006-9219-0

DO - 10.1007/s10439-006-9219-0

M3 - Article

VL - 35

SP - 419

EP - 428

JO - Annals of Biomedical Engineering

JF - Annals of Biomedical Engineering

SN - 0090-6964

IS - 3

ER -