Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates

J E de Ruijter, P J ter Brugge, S C Dieudonné, S J van Vliet, R Torensma, J A Jansen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Earlier we observed that calcium phosphate (Ca-P)-coated implant substrates stimulated the differentiation of osteoblast-like cells compared to uncoated substrates. This suggests that this difference in osteogenic induction is due to the chemical composition of the substratum. We hypothesized that Ca-P coatings modulate integrin expression patterns, because those receptors are the sensors of the cell. Therefore, in the present study we quantitatively analyzed integrin expression of osteosarcoma cells and their proliferation behavior on various well-defined Ca-P substrates. For this study we used the osteosarcoma cell line U2OS. Five groups of substrates were used: thermanox (Th), uncoated titanium (Ti), dense sintered hydroxyapatite (HA), and two Ca-P-coated titanium discs (TiHA-O% and TiHA-5%). At day 5, cell numbers were significantly lower (p < 0.05) for both types of Ca-P-coated titanium substrates compared to the other substrates. There were no significant differences between HA and uncoated titanium. From day 5 to 8, accumulated cell number was ranking highest to lowest HA > Th = Ti > TiHA-0% > TiHA-5%. Integrin expression at day 5 and day 8 of incubation was analyzed by flow cytometry for integrin subunits beta 1, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v. Fluorescence-activated cell sorting (FACS) analysis showed that the cells express high levels of beta 1, low levels of alpha 4, alpha 5, and alpha 6, and moderate levels of alpha 3 and alpha v integrin subunits on the various biomaterial substrates. Minor differences in integrin expression between the various substrates were seen. Therefore, the observed differences in proliferation between the coatings may reside in modulating the functional properties of integrins.

Original languageEnglish
Pages (from-to)279-89
Number of pages11
JournalTissue engineering part b-reviews
Volume7
Issue number3
DOIs
Publication statusPublished - Jun 2001

Cite this

de Ruijter, J. E., ter Brugge, P. J., Dieudonné, S. C., van Vliet, S. J., Torensma, R., & Jansen, J. A. (2001). Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates. Tissue engineering part b-reviews, 7(3), 279-89. https://doi.org/10.1089/10763270152044143
de Ruijter, J E ; ter Brugge, P J ; Dieudonné, S C ; van Vliet, S J ; Torensma, R ; Jansen, J A. / Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates. In: Tissue engineering part b-reviews. 2001 ; Vol. 7, No. 3. pp. 279-89.
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abstract = "Earlier we observed that calcium phosphate (Ca-P)-coated implant substrates stimulated the differentiation of osteoblast-like cells compared to uncoated substrates. This suggests that this difference in osteogenic induction is due to the chemical composition of the substratum. We hypothesized that Ca-P coatings modulate integrin expression patterns, because those receptors are the sensors of the cell. Therefore, in the present study we quantitatively analyzed integrin expression of osteosarcoma cells and their proliferation behavior on various well-defined Ca-P substrates. For this study we used the osteosarcoma cell line U2OS. Five groups of substrates were used: thermanox (Th), uncoated titanium (Ti), dense sintered hydroxyapatite (HA), and two Ca-P-coated titanium discs (TiHA-O{\%} and TiHA-5{\%}). At day 5, cell numbers were significantly lower (p < 0.05) for both types of Ca-P-coated titanium substrates compared to the other substrates. There were no significant differences between HA and uncoated titanium. From day 5 to 8, accumulated cell number was ranking highest to lowest HA > Th = Ti > TiHA-0{\%} > TiHA-5{\%}. Integrin expression at day 5 and day 8 of incubation was analyzed by flow cytometry for integrin subunits beta 1, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v. Fluorescence-activated cell sorting (FACS) analysis showed that the cells express high levels of beta 1, low levels of alpha 4, alpha 5, and alpha 6, and moderate levels of alpha 3 and alpha v integrin subunits on the various biomaterial substrates. Minor differences in integrin expression between the various substrates were seen. Therefore, the observed differences in proliferation between the coatings may reside in modulating the functional properties of integrins.",
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de Ruijter, JE, ter Brugge, PJ, Dieudonné, SC, van Vliet, SJ, Torensma, R & Jansen, JA 2001, 'Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates' Tissue engineering part b-reviews, vol. 7, no. 3, pp. 279-89. https://doi.org/10.1089/10763270152044143

Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates. / de Ruijter, J E; ter Brugge, P J; Dieudonné, S C; van Vliet, S J; Torensma, R; Jansen, J A.

In: Tissue engineering part b-reviews, Vol. 7, No. 3, 06.2001, p. 279-89.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates

AU - de Ruijter, J E

AU - ter Brugge, P J

AU - Dieudonné, S C

AU - van Vliet, S J

AU - Torensma, R

AU - Jansen, J A

PY - 2001/6

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N2 - Earlier we observed that calcium phosphate (Ca-P)-coated implant substrates stimulated the differentiation of osteoblast-like cells compared to uncoated substrates. This suggests that this difference in osteogenic induction is due to the chemical composition of the substratum. We hypothesized that Ca-P coatings modulate integrin expression patterns, because those receptors are the sensors of the cell. Therefore, in the present study we quantitatively analyzed integrin expression of osteosarcoma cells and their proliferation behavior on various well-defined Ca-P substrates. For this study we used the osteosarcoma cell line U2OS. Five groups of substrates were used: thermanox (Th), uncoated titanium (Ti), dense sintered hydroxyapatite (HA), and two Ca-P-coated titanium discs (TiHA-O% and TiHA-5%). At day 5, cell numbers were significantly lower (p < 0.05) for both types of Ca-P-coated titanium substrates compared to the other substrates. There were no significant differences between HA and uncoated titanium. From day 5 to 8, accumulated cell number was ranking highest to lowest HA > Th = Ti > TiHA-0% > TiHA-5%. Integrin expression at day 5 and day 8 of incubation was analyzed by flow cytometry for integrin subunits beta 1, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v. Fluorescence-activated cell sorting (FACS) analysis showed that the cells express high levels of beta 1, low levels of alpha 4, alpha 5, and alpha 6, and moderate levels of alpha 3 and alpha v integrin subunits on the various biomaterial substrates. Minor differences in integrin expression between the various substrates were seen. Therefore, the observed differences in proliferation between the coatings may reside in modulating the functional properties of integrins.

AB - Earlier we observed that calcium phosphate (Ca-P)-coated implant substrates stimulated the differentiation of osteoblast-like cells compared to uncoated substrates. This suggests that this difference in osteogenic induction is due to the chemical composition of the substratum. We hypothesized that Ca-P coatings modulate integrin expression patterns, because those receptors are the sensors of the cell. Therefore, in the present study we quantitatively analyzed integrin expression of osteosarcoma cells and their proliferation behavior on various well-defined Ca-P substrates. For this study we used the osteosarcoma cell line U2OS. Five groups of substrates were used: thermanox (Th), uncoated titanium (Ti), dense sintered hydroxyapatite (HA), and two Ca-P-coated titanium discs (TiHA-O% and TiHA-5%). At day 5, cell numbers were significantly lower (p < 0.05) for both types of Ca-P-coated titanium substrates compared to the other substrates. There were no significant differences between HA and uncoated titanium. From day 5 to 8, accumulated cell number was ranking highest to lowest HA > Th = Ti > TiHA-0% > TiHA-5%. Integrin expression at day 5 and day 8 of incubation was analyzed by flow cytometry for integrin subunits beta 1, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v. Fluorescence-activated cell sorting (FACS) analysis showed that the cells express high levels of beta 1, low levels of alpha 4, alpha 5, and alpha 6, and moderate levels of alpha 3 and alpha v integrin subunits on the various biomaterial substrates. Minor differences in integrin expression between the various substrates were seen. Therefore, the observed differences in proliferation between the coatings may reside in modulating the functional properties of integrins.

KW - Alkaline Phosphatase/analysis

KW - Biomedical Engineering/methods

KW - Calcium Phosphates/chemistry

KW - Cell Adhesion/physiology

KW - Cell Division/physiology

KW - Ceramics

KW - Coated Materials, Biocompatible/chemistry

KW - Extracellular Matrix/metabolism

KW - Flow Cytometry

KW - Humans

KW - Hydroxyapatites/analysis

KW - Integrins/analysis

KW - Microscopy, Electron, Scanning

KW - Osteoblasts/metabolism

KW - Surface Properties

KW - Time Factors

KW - Titanium/analysis

KW - Tumor Cells, Cultured

U2 - 10.1089/10763270152044143

DO - 10.1089/10763270152044143

M3 - Article

VL - 7

SP - 279

EP - 289

JO - Tissue engineering part b-reviews

JF - Tissue engineering part b-reviews

SN - 1937-3368

IS - 3

ER -