Effects of substrate stiffness on adipogenic and osteogenic differentiation of human mesenchymal stem cells

Zhao, Wen; Li, Xiaowei; Liu, Xiaoyan; Zhang, Ning; Wen, Xuejun

doi:10.1016/j.msec.2014.03.048

Cited by 107 publications

(67 citation statements)

References 44 publications

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“…Stiffness-mediated down-regulation of the antiangiogenic isoform of VEGF, which would result in the alternative splicing of more proangiogenic isoforms (31), could play an important role in regulating angiogenesis. In addition, adipocyte differentiation has been shown to be independently driven by SRp40-mediated splicing (41) and tissue stiffness (56), hinting at a possible role of splicing in tissue stiffness induced differentiation. Therefore, our findings have important ramifications for the fundamental understanding of how changes in the mechanical microenvironment ultimately regulate cell-ECM interactions in normal physiological responses, such as tissue morphogenesis, and abnormal responses, like tumorigenesis.…”

Section: Discussionmentioning

confidence: 99%

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Bordeleau

Califano

Abril

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Alternative splicing of proteins gives rise to different isoforms that play a crucial role in regulating several cellular processes. Notably, splicing profiles are altered in several cancer types, and these profiles are believed to be involved in driving the oncogenic process. Although the importance of alternative splicing alterations occurring during cancer is increasingly appreciated, the underlying regulatory mechanisms remain poorly understood. In this study, we use both biochemical and physical tools coupled with engineered models, patient samples, and a murine model to investigate the role of the mechanical properties of the tumor microenvironment in regulating the production of the extra domain-B (EDB) splice variant of fibronectin (FN), a hallmark of tumor angiogenesis. Specifically, we show that the amount of EDB-FN produced by endothelial cells increases with matrix stiffness both in vitro and within mouse mammary tumors. Matrix stiffness regulates splicing through the activation of serine/arginine rich (SR) proteins, the splicing factors involved in the production of FN isoforms. Activation of the SR proteins by matrix stiffness and the subsequent production of EDB-FN are dependent on intracellular contractility and PI3K-AKT signaling. Notably, matrix stiffness-mediated splicing is not limited to EDB-FN, but also affects splicing in the production of PKC βII and the VEGF 165b splice variant. Together, these results demonstrate that the mechanical properties of the microenvironment regulate alternative splicing and establish a previously unidentified mechanism by which cells can adapt to their microenvironment.alternative splicing | extracellular matrix | matrix stiffness | angiogenesis | cancer progression D ifferential expression of protein isoforms through alternative splicing is a key element to generating protein diversity and can result in widely different cell phenotypes and behaviors (1-3). Interestingly, tumors exhibit several major differences in protein isoform expression patterns compared with healthy tissue (2, 4), and some of these changes are thought to favor oncogenesis (1). Notably, splicing events are regulated at the pre-mRNA level by splicing regulatory factors that include a family of serine/arginine rich (SR) proteins (5). Elevated SR protein levels have been associated with cancer (5); however, the physiological cues that drive splicing are not well defined.Among the various alternatively spliced proteins present in tumors, the splice variant of fibronectin (FN) that includes the extra domain-B (EDB) type III repeat has been of particular interest in the cancer community because inclusion of the EDB fragment has been proposed as a way to identify and target tumor vasculature (6). EDB-FN is produced by endothelial cells (ECs) in tumors and may favor angiogenesis (7-9). Splicing of FN is regulated by several SR proteins, including SRp40 and SRp20, in several different cell types (10-12). Although the presence of EDB-FN is fairly unique to tumor vasculature, the mechanisms governin...

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Section: Discussionmentioning

confidence: 99%

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Bordeleau

Califano

Abril

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…21 The pH was neutralized using 0.1 mol/L NaOH and the scaffolds were lyophilized. The prepared CS/CaPP and CS/ CaPP/Pg scaffolds were subjected to SEM, XRD and EDAX studies, as described previously.…”

Section: Preparation and Characterization Of Cs/capp And Cs/capp/pgmentioning

confidence: 99%

Proliferation and differentiation of mesenchymal stem cells on scaffolds containing chitosan, calcium polyphosphate and pigeonite for bone tissue engineering

et al. 2017

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Objectives: Treatment of critical-sized bone defects with cells and biomaterials offers an efficient alternative to traditional bone grafts. Chitosan (CS) is a natural biopolymer that acts as a scaffold in bone tissue engineering (BTE). Polyphosphate (PolyP), recently identified as an inorganic polymer, acts as a potential bone morphogenetic material, whereas pigeonite (Pg) is a novel iron-containing ceramic. In this study, we prepared and characterized scaffolds containing CS, calcium polyphosphate (CaPP) and Pg particles for bone formation in vitro and in vivo.

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“…7, where the mechanical properties of the nucleus, cytoplasm, and cell are presented. Although the effects of biomaterials' mechanical strength on osteogenic differentiation have been widely analyzed, 32,36,50 the literature contains few reports on the correlation between cellular mechanical properties and differentiation of osteoblastic cells. The results reflected in Fig.…”

Section: Analysis Of Cellular Mechanical Strengthmentioning

confidence: 99%

“…The osteoconductivity of scaffolds is critical in bone regeneration, because scaffolds influence not only cell proliferation but also osteogenic differentiation. According to previous researches, the osteoconductivity of biomaterials is affected by materials' chemical compositions, 26 hydrophilicity, 20 mechanical strength, 32,36,50 roughness, 13 crystallinity, 18 etc. Information related to materials' osteoconductivity is possibly provided by a serious of in vivo or in vitro experiments.…”

Section: Introductionmentioning

confidence: 99%

“…Many studies have indicated that materials' osteoconductivity is related to the mechanical properties of cultured cells. 32,36,50 Previous research has also shown that the cell-material adhesive strength influences cell attachment and proliferation, 16 which may affect subsequent cell differentiation. Nevertheless, AFM techniques have never been applied to measure cell-material adhesive force and the cellular mechanical strength of living osteogenic cells to identify materials' osteoconductivity.…”

Section: Introductionmentioning

confidence: 99%

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Preparation of Chitosan/Hydroxyapatite Substrates with Controllable Osteoconductivity Tracked by AFM

Hsiao

et al. 2014

Ann Biomed Eng

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In this study, cell-material adhesive strength and cellular mechanical properties were measured using atomic force microscopy (AFM) to track cell attachment and osteogenic differentiation. First, chitosan substrates were treated with simulated body fluid (SBF) for various periods, resulting in substrates with different osteoconductivity. The X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS) and in vitro tests revealed that the biomimeticity and osteoconductivity of substrates increased with increasing time of SBF treatment. When the SBF immersion exceeded 14 days, the chitosan substrates exhibited their highest biocompatibility and osteoconductivity. AFM measurements indicated specifically high adhesive forces between SBF-treated chitosan and osteogenic cells, causing better cell attachment. The results demonstrate that cell adhesion was controlled by cell-material adhesive strength, which were in turn controlled via the SBF treatment time. The adhesive strength between cells and material also accounted for the chitosan substrates' specific selectivity toward osteogenic cells. A two-step increase in mechanical strength was observed for the nucleus and cytoplasm of osteogenic cells. The results indicate that through the use of AFM, the real-time cell-material interforce and cellular mechanics can be identified. The adhesive strength was positively correlated to the cell attachment, and the second increase in the Young's modulus of nucleus and cytoplasm was correlated to early osteogenic differentiation.

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Effects of substrate stiffness on adipogenic and osteogenic differentiation of human mesenchymal stem cells

Cited by 107 publications

References 44 publications

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Tissue stiffness regulates serine/arginine-rich protein-mediated splicing of the extra domain B-fibronectin isoform in tumors

Proliferation and differentiation of mesenchymal stem cells on scaffolds containing chitosan, calcium polyphosphate and pigeonite for bone tissue engineering

Preparation of Chitosan/Hydroxyapatite Substrates with Controllable Osteoconductivity Tracked by AFM

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