Probing the microenvironmental conditions for induction of superficial zone protein expression

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Author(s) Mhanna, R., Öztürk, Ece, Schlink, P., Zenobi-Wong, Marcy
Publication Type Journal Items, Publication Status: Published
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Title Probing the microenvironmental conditions for induction of superficial zone protein expression
Author(s) Mhanna, R.
Öztürk, Ece
Schlink, P.
Zenobi-Wong, Marcy
Journal or Series Title Osteoarthritis and cartilage
Volume Number 21
Issue Number 12
Start Page 1924
End Page 1932
ISSN 1063-4584
Publisher Elsevier
Publication Place Amsterdam
Publication Date 2013-12
Keyword(s) Superficial zone protein
Chondrocytes dedifferentiation
Mechanical strain
Oxygen tension
Abstract Objective: To determine the in vitro conditions which promote expression of superficial zone protein (SZP).
Methods: Chondrocytes from 6-month-old calves were expanded in monolayer culture and the expression of SZP in alginate bead and monolayer culture was quantified with quantitative real time-polymerase chain reaction (qRT-PCR) and immunostaining. The effect of oxygen tension on SZP expression was determined by qRT-PRC analysis of cells cultured in two dimension (2D) and three dimension (3D) under hypoxic (1% pO2) or normoxic (21% pO2) conditions. Finally, to examine the effect of cyclic tensile strain on expression of SZP in 2D and 3D cultures, chondrocytes encapsulated in alginate beams or seeded on type I collagen coated polydimethylsiloxane (PDMS) chambers were subjected to 5% strain at 1 Hz, 2 h/day for 4 days or 2 h at the fourth day of culture and mRNA levels were quantified.
Results: Bovine chondrocytes in monolayer showed a drastic decrease in SZP expression, similar in trend to the commonly reported downregulation of type II collagen (Col2). Chondrocytes embedded in alginate beads for 4 days re-expressed SZP but not Col2. SZP expression was higher under normoxic conditions whereas Col2 was upregulated only in alginate beads under hypoxic conditions. Cyclic mechanical strain showed a tendency to upregulate mRNA levels of SZP.
Conclusions: A microenvironment encompassing a soft encapsulation material and 21% oxygen is sufficient for fibroblastic chondrocytes to re-express SZP. These results serve as a guideline for the design of stratified engineered articular cartilage and suggest that microenvironmental cues (oxygen tension level) strongly influence the pattern of SZP expression in vivo.
DOI 10.1016/j.joca.2013.08.017
Additional Notes Received 24 January 2013, Accepted 14 August 2013
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03949
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NEBIS System Number 004530868
Source Database ID SCOPUS-84888870125
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  author = "Mhanna, R. and {\"{O}}zt{\"{u}}rk, Ece and Schlink, P. and Zenobi-Wong, Marcy",
  title = "{P}robing the microenvironmental conditions for induction of superficial zone protein expression",
  journal = "Osteoarthritis and cartilage",
  year = 2013,
  volume = "21",
  number = "12",
  pages = "1924--1932",
  month = dec,

E-Citations record created: Tue, 01 Oct 2013, 08:43:12 CET