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Growth differentiation factor 5 modulation of chondrogenesis of self‐assembled constructs involves gap junction‐mediated intercellular communication
Author(s) -
Sun Zhibo,
Zhang Yukun,
Yang Shuhua,
Jia Jie,
Ye Shunan,
Chen Dong,
Mo Fengbo
Publication year - 2012
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/dgd.12009
Subject(s) - chondrogenesis , microbiology and biotechnology , chemistry , chondrocyte , mesenchymal stem cell , gap junction , transcription factor , intracellular , cartilage , cellular differentiation , cell signaling , biophysics , signal transduction , anatomy , biology , in vitro , biochemistry , gene
A novel scaffold‐free self‐assembled cartilage construct has been generated and used to repair particular chondral defects effectively. However, the mechanisms related to the construction of these self‐assembled cartilages have not yet been fully elucidated. We hypothesize that gap junction intercellular communication ( GJIC ) plays a critical role in the development of self‐assembled constructs upon GDF ‐5 induction. In this study, we investigated the effect of connexin 43 (C×43) mediated GJIC on GDF ‐5 modulation of chondrogenesis from two aspects, cell monolayer culture and 3‐D self‐assembly culture. We induced cells or self‐assembled constructs with chondrogenic media ( CM ), growth differentiation factor 5 ( GDF ‐5) or 1‐heptanol for 3 weeks. At the end of that time, the results of quantitative fluorescence redistribution after photobleaching ( FRAP ) assay and immunofluorescence demonstrated that GDF ‐5 improved both GJIC and chondrogenic differentiation to a significant degree while 1‐heptanol nearly offset the expected improvements in chondrogenesis. Biochemical assay and histology showed that GDF ‐5 can obviously enhance GAG , C×43 and type II collagen expressions. Conversely, we also showed that while 1‐heptanol weakened GAG and type II collagen expression in self‐assembled constructs, it had no effect on C×43 expression. Furthermore, real‐time polymerase chain reaction showed that GDF ‐5 enhanced GAG and type II collagen transcription while 1‐heptanol reduced them, but was affectless on C×43 transcription. This suggests that the generation of scaffold‐free self‐assembled cartilage from human mesenchymal stem cells upon GDF ‐5 induction may be mediated, at least in part, via the modulation of GJIC .