Chondrocyte response to cyclic hydrostatic pressure in alginate versus pellet culture

Steven H. Elder, Shawn W. Sanders, William R. McCulley, Misti L. Marr, Joon W. Shim, Karen Hasty

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Cells are often cultured at high density (e.g., confluent monolayer and as pellets) to promote chondrogenic differentiation and to maintain the chondrocyte phenotype. They are also frequently suspended in hydrogels such as agarose or alginate for the same purposes. These culture techniques differ markedly with respect to frequency of direct contact between cells and overall intercellular spacing. Because these factors may significantly affect mechanotransduction, the purpose of this study was to determine if the response of articular chondrocytes to cyclic hydrostatic pressure would depend on the culture condition. Primary articular chondrocytes from young and mature pigs were cultured either as pellets or suspended in alginate beads. Both groups were exposed to dynamic hydrostatic pressure (4 MPa, 1 Hz, 5400 cycles per day) for 7 days. Cell proliferation was unaffected by pressure, but pressurized chondrocytes in pellet culture had significantly greater sGAG content and incorporated [3H]proline at a higher rate than nonpressurized controls. Electron microscopy revealed a fibrous extracellular matrix (ECM) surrounding pellets, but not cells in alginate. In addition, expression of Connexin 43 (Cx43) mRNA was slightly lower in alginate than in pellet cultures and was not significantly altered by loading. Thus, metabolic response of chondrocytes to dynamic hydrostatic pressure was affected by culture technique; chondrocytes cultured as pellets exhibited the classical anabolic response to dynamic hydrostatic pressure, but those in alginate did not. Although cell-ECM interaction could be important, the differential response is not likely attributable to differential expression of Cx43 mRNA.

Original languageEnglish (US)
Pages (from-to)740-747
Number of pages8
JournalJournal of Orthopaedic Research
Volume24
Issue number4
DOIs
StatePublished - Apr 1 2006

Fingerprint

Hydrostatic Pressure
Chondrocytes
Culture Techniques
Connexin 43
Extracellular Matrix
Joints
Messenger RNA
Hydrogels
Proline
Sepharose
alginic acid
Cultured Cells
Electron Microscopy
Swine
Cell Proliferation
Phenotype
Pressure

All Science Journal Classification (ASJC) codes

  • Orthopedics and Sports Medicine

Cite this

Chondrocyte response to cyclic hydrostatic pressure in alginate versus pellet culture. / Elder, Steven H.; Sanders, Shawn W.; McCulley, William R.; Marr, Misti L.; Shim, Joon W.; Hasty, Karen.

In: Journal of Orthopaedic Research, Vol. 24, No. 4, 01.04.2006, p. 740-747.

Research output: Contribution to journalArticle

Elder, Steven H. ; Sanders, Shawn W. ; McCulley, William R. ; Marr, Misti L. ; Shim, Joon W. ; Hasty, Karen. / Chondrocyte response to cyclic hydrostatic pressure in alginate versus pellet culture. In: Journal of Orthopaedic Research. 2006 ; Vol. 24, No. 4. pp. 740-747.
@article{c2567db878f346729d15cb693c72b905,
title = "Chondrocyte response to cyclic hydrostatic pressure in alginate versus pellet culture",
abstract = "Cells are often cultured at high density (e.g., confluent monolayer and as pellets) to promote chondrogenic differentiation and to maintain the chondrocyte phenotype. They are also frequently suspended in hydrogels such as agarose or alginate for the same purposes. These culture techniques differ markedly with respect to frequency of direct contact between cells and overall intercellular spacing. Because these factors may significantly affect mechanotransduction, the purpose of this study was to determine if the response of articular chondrocytes to cyclic hydrostatic pressure would depend on the culture condition. Primary articular chondrocytes from young and mature pigs were cultured either as pellets or suspended in alginate beads. Both groups were exposed to dynamic hydrostatic pressure (4 MPa, 1 Hz, 5400 cycles per day) for 7 days. Cell proliferation was unaffected by pressure, but pressurized chondrocytes in pellet culture had significantly greater sGAG content and incorporated [3H]proline at a higher rate than nonpressurized controls. Electron microscopy revealed a fibrous extracellular matrix (ECM) surrounding pellets, but not cells in alginate. In addition, expression of Connexin 43 (Cx43) mRNA was slightly lower in alginate than in pellet cultures and was not significantly altered by loading. Thus, metabolic response of chondrocytes to dynamic hydrostatic pressure was affected by culture technique; chondrocytes cultured as pellets exhibited the classical anabolic response to dynamic hydrostatic pressure, but those in alginate did not. Although cell-ECM interaction could be important, the differential response is not likely attributable to differential expression of Cx43 mRNA.",
author = "Elder, {Steven H.} and Sanders, {Shawn W.} and McCulley, {William R.} and Marr, {Misti L.} and Shim, {Joon W.} and Karen Hasty",
year = "2006",
month = "4",
day = "1",
doi = "10.1002/jor.20086",
language = "English (US)",
volume = "24",
pages = "740--747",
journal = "Journal of Orthopaedic Research",
issn = "0736-0266",
publisher = "John Wiley and Sons Inc.",
number = "4",

}

TY - JOUR

T1 - Chondrocyte response to cyclic hydrostatic pressure in alginate versus pellet culture

AU - Elder, Steven H.

AU - Sanders, Shawn W.

AU - McCulley, William R.

AU - Marr, Misti L.

AU - Shim, Joon W.

AU - Hasty, Karen

PY - 2006/4/1

Y1 - 2006/4/1

N2 - Cells are often cultured at high density (e.g., confluent monolayer and as pellets) to promote chondrogenic differentiation and to maintain the chondrocyte phenotype. They are also frequently suspended in hydrogels such as agarose or alginate for the same purposes. These culture techniques differ markedly with respect to frequency of direct contact between cells and overall intercellular spacing. Because these factors may significantly affect mechanotransduction, the purpose of this study was to determine if the response of articular chondrocytes to cyclic hydrostatic pressure would depend on the culture condition. Primary articular chondrocytes from young and mature pigs were cultured either as pellets or suspended in alginate beads. Both groups were exposed to dynamic hydrostatic pressure (4 MPa, 1 Hz, 5400 cycles per day) for 7 days. Cell proliferation was unaffected by pressure, but pressurized chondrocytes in pellet culture had significantly greater sGAG content and incorporated [3H]proline at a higher rate than nonpressurized controls. Electron microscopy revealed a fibrous extracellular matrix (ECM) surrounding pellets, but not cells in alginate. In addition, expression of Connexin 43 (Cx43) mRNA was slightly lower in alginate than in pellet cultures and was not significantly altered by loading. Thus, metabolic response of chondrocytes to dynamic hydrostatic pressure was affected by culture technique; chondrocytes cultured as pellets exhibited the classical anabolic response to dynamic hydrostatic pressure, but those in alginate did not. Although cell-ECM interaction could be important, the differential response is not likely attributable to differential expression of Cx43 mRNA.

AB - Cells are often cultured at high density (e.g., confluent monolayer and as pellets) to promote chondrogenic differentiation and to maintain the chondrocyte phenotype. They are also frequently suspended in hydrogels such as agarose or alginate for the same purposes. These culture techniques differ markedly with respect to frequency of direct contact between cells and overall intercellular spacing. Because these factors may significantly affect mechanotransduction, the purpose of this study was to determine if the response of articular chondrocytes to cyclic hydrostatic pressure would depend on the culture condition. Primary articular chondrocytes from young and mature pigs were cultured either as pellets or suspended in alginate beads. Both groups were exposed to dynamic hydrostatic pressure (4 MPa, 1 Hz, 5400 cycles per day) for 7 days. Cell proliferation was unaffected by pressure, but pressurized chondrocytes in pellet culture had significantly greater sGAG content and incorporated [3H]proline at a higher rate than nonpressurized controls. Electron microscopy revealed a fibrous extracellular matrix (ECM) surrounding pellets, but not cells in alginate. In addition, expression of Connexin 43 (Cx43) mRNA was slightly lower in alginate than in pellet cultures and was not significantly altered by loading. Thus, metabolic response of chondrocytes to dynamic hydrostatic pressure was affected by culture technique; chondrocytes cultured as pellets exhibited the classical anabolic response to dynamic hydrostatic pressure, but those in alginate did not. Although cell-ECM interaction could be important, the differential response is not likely attributable to differential expression of Cx43 mRNA.

UR - http://www.scopus.com/inward/record.url?scp=33646170273&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33646170273&partnerID=8YFLogxK

U2 - 10.1002/jor.20086

DO - 10.1002/jor.20086

M3 - Article

VL - 24

SP - 740

EP - 747

JO - Journal of Orthopaedic Research

JF - Journal of Orthopaedic Research

SN - 0736-0266

IS - 4

ER -