Stimulation of plasma membrane and matrix vesicle enzyme activity by transforming growth factor-β in osteosarcoma cell cultures

Lynda Bonewald, Z. Schwartz, L. D. Swain, V. Ramirez, J. Poser, B. D. Boyan

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Transforming growth factor-β (TGFβ) serves an important role in extracellular matrix formation by stimulating the production of numerous extracellular matrix proteins by connective tissue cells and by osteoblasts or bone-forming cells. TGFβ has been shown to stimulate alkaline phosphatase (ALPase) activity in the rat osteoblast-like osteosarcoma cell line ROS 17/2.8. Previous studies have shown that this enzyme is elevated during calcification of bone and that it is enriched in matrix vesicles, an extracellular organelle associated with initial hydroxyapatite formation. To test the hypothesis that TGFβ plays a role in regulating mineral deposition in the matrix, the effects of TGFβ on ALPase and phospholipase A2, two enzymes associated with mineralization, were examined. ROS 17/2.8 cells were cultured at high and low density with recombinant human TGFβ (0.1-10 ng/ml) to examine the influence of cell maturation on response to TGFβ. Maximal stimulation of ALPase activity in the low density cultures was seen at 5 ng/ml; in high-density cultures, there was further stimulation at 10 ng/ml. There was a dose-dependent increase in ALPase activity seen in the matrix vesicles and plasma membranes in both types of cultures. Matrix vesicle ALPase exhibited a greater response to factor than did the plasma membrane enzyme. However, in low-density cultures, the two membrane fractions exhibited a parallel response with greatest activity consistently in the matrix vesicles. There was a dose-dependent increase in phospholipase A2-specific activity in the plasma membranes and matrix vesicles of both high- and low-density cultures. In agreement with previous studies, TGFβ inhibited cellular proliferation 50%. The results show that addition of TGFβ stimulates the activity of enzymes associated with calcification. The effect of TGFβ is dependent on the stage of maturation of the cell. This study indicates that TGFβ may play an important role in induced bone formation, calcification, and fracture repair in addition to its role in promoting chondrogenesis.

Original languageEnglish (US)
Pages (from-to)200-206
Number of pages7
JournalJournal of Cellular Physiology
Volume145
Issue number2
StatePublished - Nov 1990
Externally publishedYes

Fingerprint

Enzyme activity
Transforming Growth Factors
Osteosarcoma
Cell membranes
Cell culture
Cell Culture Techniques
Cell Membrane
Enzymes
Alkaline Phosphatase
Bone
Phospholipases A2
Osteoblasts
Extracellular Matrix
Connective Tissue Cells
Chondrogenesis
Bone and Bones
Extracellular Matrix Proteins
Bone Fractures
Durapatite
Osteogenesis

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Stimulation of plasma membrane and matrix vesicle enzyme activity by transforming growth factor-β in osteosarcoma cell cultures. / Bonewald, Lynda; Schwartz, Z.; Swain, L. D.; Ramirez, V.; Poser, J.; Boyan, B. D.

In: Journal of Cellular Physiology, Vol. 145, No. 2, 11.1990, p. 200-206.

Research output: Contribution to journalArticle

Bonewald, Lynda ; Schwartz, Z. ; Swain, L. D. ; Ramirez, V. ; Poser, J. ; Boyan, B. D. / Stimulation of plasma membrane and matrix vesicle enzyme activity by transforming growth factor-β in osteosarcoma cell cultures. In: Journal of Cellular Physiology. 1990 ; Vol. 145, No. 2. pp. 200-206.
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