Integrin expression and osteopontin regulation in human fetal osteoblastic cells mediated by substratum surface characteristics

Jung Yul Lim, Amanda F. Taylor, Zhongyong Li, Erwin A. Vogler, Henry J. Donahue

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Integrin-mediated adhesion of anchorage-dependent cells to scaffolds is a critical component of tissue engineering. We investigated integrin expression by the human fetal osteoblastic cell line, hFOB 1.19 (hFOB), as a function of substratum surface wettability. The influence of surface wettability on bone cell phenotype was also examined. Plasma-treated quartz (PTQ) and glass (PTG) (hydrophilic, contact angles of 0°), octadecyltrichlorosilane-treated quartz (STQ) and glass (STG) (hydrophobic, contact angles above about 100°), and tissue culture polystyrene were used for cell culture. hFOB cells cultured on hydrophilic substrata displayed well-developed actin stress fibers relative to cells on hydrophobic substrata. Western blot analysis revealed that hFOB cells cultured on hydrophobic substrata (STQ or STG) express lower levels of αv and β3 integrin subunits than do cells on hydrophilic substrata (PTQ or PTG). This effect was more pronounced in cells on STQ than on STG. These variations in integrin expression were lessened by extended culture time. Double-labeled integrin/actin immunofluorescence confirmed Western blot results, that is, cells cultured on PTQ displayed distinct, large plaques of αv and β3 subunits and integrin αvβ3, as well as their colocalization with actin stress fiber ends, whereas cells on STQ did not display integrin plaques after 24 h and displayed only minimal plaque formation after 3 days. Vinculin, a focal adhesion protein that mediates binding between the integrin and actin cytoskeleton, appeared in Western blots to mimic the variations of αv and β3 expression with respect to surface wettability. Interestingly, real-time RT-PCR analysis showed that hFOB cultured on hydrophobic substrata, which have down-regulated αv and β3 integrin subunits, displayed greater steady state mRNA levels of osteopontin, an extracellular matrix (ECM) protein containing the Arg-Gly-Asp (RGD) integrin recognition sequence, than did cells cultured on hydrophilic substrata. Our results imply that substratum surface wettability regulates integrin-mediated bone cell adhesion and further influences the expression of bone cell-ECM complexes.

Original languageEnglish (US)
Pages (from-to)19-29
Number of pages11
JournalTissue Engineering
Volume11
Issue number1-2
DOIs
StatePublished - Jan 1 2005

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Wetting
Quartz
Glass
Bone
Plasmas
Contact angle
Adhesion
Cells
Proteins
Tissue culture
Fibers
Cell adhesion
Scaffolds (biology)
Tissue engineering
Cell culture
Polystyrenes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biophysics
  • Cell Biology

Cite this

Lim, Jung Yul ; Taylor, Amanda F. ; Li, Zhongyong ; Vogler, Erwin A. ; Donahue, Henry J. / Integrin expression and osteopontin regulation in human fetal osteoblastic cells mediated by substratum surface characteristics. In: Tissue Engineering. 2005 ; Vol. 11, No. 1-2. pp. 19-29.
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abstract = "Integrin-mediated adhesion of anchorage-dependent cells to scaffolds is a critical component of tissue engineering. We investigated integrin expression by the human fetal osteoblastic cell line, hFOB 1.19 (hFOB), as a function of substratum surface wettability. The influence of surface wettability on bone cell phenotype was also examined. Plasma-treated quartz (PTQ) and glass (PTG) (hydrophilic, contact angles of 0°), octadecyltrichlorosilane-treated quartz (STQ) and glass (STG) (hydrophobic, contact angles above about 100°), and tissue culture polystyrene were used for cell culture. hFOB cells cultured on hydrophilic substrata displayed well-developed actin stress fibers relative to cells on hydrophobic substrata. Western blot analysis revealed that hFOB cells cultured on hydrophobic substrata (STQ or STG) express lower levels of αv and β3 integrin subunits than do cells on hydrophilic substrata (PTQ or PTG). This effect was more pronounced in cells on STQ than on STG. These variations in integrin expression were lessened by extended culture time. Double-labeled integrin/actin immunofluorescence confirmed Western blot results, that is, cells cultured on PTQ displayed distinct, large plaques of αv and β3 subunits and integrin αvβ3, as well as their colocalization with actin stress fiber ends, whereas cells on STQ did not display integrin plaques after 24 h and displayed only minimal plaque formation after 3 days. Vinculin, a focal adhesion protein that mediates binding between the integrin and actin cytoskeleton, appeared in Western blots to mimic the variations of αv and β3 expression with respect to surface wettability. Interestingly, real-time RT-PCR analysis showed that hFOB cultured on hydrophobic substrata, which have down-regulated αv and β3 integrin subunits, displayed greater steady state mRNA levels of osteopontin, an extracellular matrix (ECM) protein containing the Arg-Gly-Asp (RGD) integrin recognition sequence, than did cells cultured on hydrophilic substrata. Our results imply that substratum surface wettability regulates integrin-mediated bone cell adhesion and further influences the expression of bone cell-ECM complexes.",
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Integrin expression and osteopontin regulation in human fetal osteoblastic cells mediated by substratum surface characteristics. / Lim, Jung Yul; Taylor, Amanda F.; Li, Zhongyong; Vogler, Erwin A.; Donahue, Henry J.

In: Tissue Engineering, Vol. 11, No. 1-2, 01.01.2005, p. 19-29.

Research output: Contribution to journalArticle

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T1 - Integrin expression and osteopontin regulation in human fetal osteoblastic cells mediated by substratum surface characteristics

AU - Lim, Jung Yul

AU - Taylor, Amanda F.

AU - Li, Zhongyong

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AU - Donahue, Henry J.

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