Analysis of proteins showing differential changes during ATP oscillations in chondrogenesis

Kwon, Hyuck Joon; Kurono, Sadamu; Kaneko, Yuka; Ohmiya, Yoshihiro; Yasuda, Kazunori

doi:10.1002/cbf.3033

Cited by 2 publications

(3 citation statements)

References 32 publications

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“…Our previous results demonstrated that ATP oscillations driven by chondrogenic growth factors such as TGF beta and insulin play essential roles for prechondrogenic condensation that is the initial step of chondrogenesis by inducing oscillatory expression of proteins involved in actin dynamics, cell migration, and adhesion which leads to collective migration and adhesion3856. The present results demonstrate that ES generates Ca 2+ /ATP oscillations in MSCs even in the absence of exogenous growth factors (Fig.…”

Section: Discussionsupporting

confidence: 68%

Electrical stimulation drives chondrogenesis of mesenchymal stem cells in the absence of exogenous growth factors

Kwon

Lee

Chun

2016

Sci Rep

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Electrical stimulation (ES) is known to guide the development and regeneration of many tissues. However, although preclinical and clinical studies have demonstrated superior effects of ES on cartilage repair, the effects of ES on chondrogenesis remain elusive. Since mesenchyme stem cells (MSCs) have high therapeutic potential for cartilage regeneration, we investigated the actions of ES during chondrogenesis of MSCs. Herein, we demonstrate for the first time that ES enhances expression levels of chondrogenic markers, such as type II collagen, aggrecan, and Sox9, and decreases type I collagen levels, thereby inducing differentiation of MSCs into hyaline chondrogenic cells without the addition of exogenous growth factors. ES also induced MSC condensation and subsequent chondrogenesis by driving Ca2+/ATP oscillations, which are known to be essential for prechondrogenic condensation. In subsequent experiments, the effects of ES on ATP oscillations and chondrogenesis were dependent on extracellular ATP signaling via P2X4 receptors, and ES induced significant increases in TGF-β1 and BMP2 expression. However, the inhibition of TGF-β signaling blocked ES-driven condensation, whereas the inhibition of BMP signaling did not, indicating that TGF-β signaling but not BMP signaling mediates ES-driven condensation. These findings may contribute to the development of electrotherapeutic strategies for cartilage repair using MSCs.

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Section: Discussionsupporting

confidence: 68%

Electrical stimulation drives chondrogenesis of mesenchymal stem cells in the absence of exogenous growth factors

Kwon

Lee

Chun

2016

Sci Rep

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“…The supernatants were concentrated and desalted using the Amicon Ultra Centrifugal Filters (3,000 MWCO, EMD Millipore, Darmstadt, Germany). Nano liquid chromatography–mass spectrometry (LC–MS) was performed as described previously (Kwon, Kurono, Kaneko, Ohmiya, & Yasuda, 2014).…”

Section: Methodsmentioning

confidence: 99%

“…(3,000 MWCO, EMD Millipore, Darmstadt, Germany). Nano liquid chromatography-mass spectrometry (LC-MS) was performed as described previously (Kwon, Kurono, Kaneko, Ohmiya, & Yasuda, 2014).…”

Section: Real-time Pcr Analysismentioning

confidence: 99%

Synthetic poly(2‐acrylamido‐2‐methylpropanesulfonic acid) gel induces chondrogenic differentiation of ATDC5 cells via a novel protein reservoir function

Semba

Kitamura

Tsuda

et al. 2020

J Biomedical Materials Res

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We previously demonstrated that a synthetic negatively charged poly(2‐acrylamido‐2‐methylpropanesulfonic acid) (PAMPS) gel induced chondrogenic differentiation of ATDC5 cells. In this study, we clarified the underlying molecular mechanism, in particular, focusing on the events that occurred at the interface between the gel and the cells. Gene expression profiling revealed that the expression of extracellular components was enhanced in the ATDC5 cells that were cultured on the PAMPS gel, suggesting that extracellular proteins secreted from the ATDC5 cells might be adsorbed in the PAMPS gel, thereby contributing to the induction of chondrogenic differentiation. Therefore, we created “Treated‐PAMPS gel,” which adsorbed various proteins secreted from the cultured ATDC5 cells during 7 days. Proteomic analysis identified 27 proteins, including extracellular matrix proteins such as Types I, III, and V collagens and thrombospondin (THBS) in the Treated‐PAMPS gel. The Treated‐PAMPS gel preferentially induced expression of chondrogenic markers, namely, aggrecan and Type II collagen, in the ATDC5 cells compared with the untreated PAMPS gel. Addition of recombinant THBS1 to the ATDC5 cells significantly enhanced the PAMPS‐induced chondrogenic differentiation, whereas knockdown of THBS1 completely abolished this response. In conclusion, we demonstrated that the PAMPS gel has the potential to induce chondrogenic differentiation through novel reservoir functions, and the adsorbed THBS plays a significant role in the induction.

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Analysis of proteins showing differential changes during ATP oscillations in chondrogenesis

Cited by 2 publications

References 32 publications

Electrical stimulation drives chondrogenesis of mesenchymal stem cells in the absence of exogenous growth factors

Electrical stimulation drives chondrogenesis of mesenchymal stem cells in the absence of exogenous growth factors

Synthetic poly(2‐acrylamido‐2‐methylpropanesulfonic acid) gel induces chondrogenic differentiation of ATDC5 cells via a novel protein reservoir function

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