Substrate stiffness- and topography-dependent differentiation of annulus fibrosus-derived stem cells is regulated by Yes-associated protein

Chu, Genglei; Yuan, Zhangqin; Zhu, Caihong; Zhou, Pinghui; Wang, Huan; Zhang, Weidong; Cai, Yan; Zhang, Yijian; Li, Bin

doi:10.1016/j.actbio.2019.05.013

Cited by 76 publications

(46 citation statements)

References 64 publications

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“…The mechanotransduction effects can further affect intracellular and intercellular signalling pathways and ultimately result in stiffness-regulated stem cell behaviour [ 36 , 37 ]. YAP is a key protein in the Hippo pathway of mechano-transduction [ 45 ], and many studies have shown that the mechano-transduction triggered by matrix stiffness in stem cells is mediated by YAP [ 26 , 27 , 46 , 47 ]. Therefore, we further selected two YAP inhibitors (VP and GW5074) to verify whether the effect of matrix stiffness on the osteogenic and adipogenic differentiation of MSCs was transduced by YAP.…”

Section: Discussionmentioning

confidence: 99%

Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels

Liu

Zhao

et al. 2020

Burns &Amp; Trauma

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Background Hydrogels with tuneable mechanical properties are an attractive material platform for 3D bioprinting. Thus far, numerous studies have confirmed that the biophysical cues of hydrogels, such as stiffness, are known to have a profound impact on mesenchymal stem cell (MSC) differentiation; however, their differentiation potential within 3D-bioprinted hydrogels is not completely understood. Here, we propose a protocol for the exploration of how the stiffness of alginate–gelatin (Alg-Gel) composite hydrogels (the widely used bioink) affects the differentiation of MSCs in the presence or absence of differentiation inducing factors. Methods Two types of Alg-Gel composite hydrogels (Young’s modulus: 50 kPa vs. 225 kPa) were bioprinted independently of porosity. Then, stiffness-induced biases towards adipogenic and osteogenic differentiation of the embedded MSCs were analysed by co-staining with alkaline phosphatase (ALP) and oil red O. The expression of specific markers at the gene level was detected after a 3-day culture. Results Confocal microscopy indicated that all tested hydrogels supported MSC growth and viability during the culture period. Higher expression of adipogenic and osteogenic markers (ALP and lipoprotein lipase (LPL)) in stiffer 3D-bioprinted matrices demonstrated a more significant response of MSCs to stiffer hydrogels with respect to differentiation, which was more robust in differentiation-inducing medium. However, the LPL expression in stiffer 3D-bioprinted constructs was reduced at day 3 regardless of the presence of differentiation-inducing factors. Although MSCs embedded in softer hydrogels to some extent proceeded toward adipogenic and osteogenic lineages within a few days, their differentiation seemed to be slower and more limited. Interestingly, the hydrogel itself (without differentiation-inducing factors) exhibited a slight effect on whether MSCs differentiated towards an adipogenic or an osteogenic fate. Considering that the mechano-regulated protein Yes-associated protein (YAP) is involved in MSC fate decisions, we further found that inhibition of YAP significantly downregulated the expression of ALP and LPL in MSCs in stiffer constructs regardless of the induced growth factors present. Conclusions These results demonstrate that the differentiation of MSCs in 3D-bioprinted matrices is dependent on hydrogel stiffness, which emphasizes the importance of biophysical cues as a determinant of cellular behaviour.

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Section: Discussionmentioning

confidence: 99%

Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels

Liu

Zhao

et al. 2020

Burns &Amp; Trauma

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“…In our study, we chose to focus on the prolonged inhibition of YAP activity to observe the involvement of YAP in a timely fashion on the OL differentiation and myelination as assessed on the various axon stiffnesses. While verteporfin has been widely used to elucidate the involvement of YAP in cell mechanotransduction, [ 25,28,29 ] its mode of inhibition and mechanism of interaction with YAP remain largely to be elucidated. [ 30 ] One known mechanism describes that verteporfin blocks the YAP‐TEAD regulation complex formation, which locates within the cell nucleus.…”

Section: Discussionmentioning

confidence: 99%

Biomimicking Fiber Platform with Tunable Stiffness to Study Mechanotransduction Reveals Stiffness Enhances Oligodendrocyte Differentiation but Impedes Myelination through YAP‐Dependent Regulation

Ong

Marinval

Lin

et al. 2020

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A key hallmark of many diseases, especially those in the central nervous system (CNS), is the change in tissue stiffness due to inflammation and scarring. However, how such changes in microenvironment affect the regenerative process remains poorly understood. Here, a biomimicking fiber platform that provides independent variation of fiber structural and intrinsic stiffness is reported. To demonstrate the functionality of these constructs as a mechanotransduction study platform, these substrates are utilized as artificial axons and the effects of axon structural versus intrinsic stiffness on CNS myelination are independently analyzed. While studies have shown that substrate stiffness affects oligodendrocyte differentiation, the effects of mechanical stiffness on the final functional state of oligodendrocyte (i.e., myelination) has not been shown prior to this. Here, it is demonstrated that a stiff mechanical microenvironment impedes oligodendrocyte myelination, independently and distinctively from oligodendrocyte differentiation. Yes‐associated protein is identified to be involved in influencing oligodendrocyte myelination through mechanotransduction. The opposing effects on oligodendrocyte differentiation and myelination provide important implications for current work screening for promyelinating drugs, since these efforts have focused mainly on promoting oligodendrocyte differentiation. Thus, the platform may have considerable utility as part of a drug discovery program in identifying molecules that promote both differentiation and myelination.

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“…Though many researches have investigated biomaterial‐induced AF cell‐like differentiation (Chu et al, ; Ma et al, ), no final standards have been clarified, considering the specific AF cells phenotype markers. Col I is the primary component of AF tissues, and COL1A1 is also the most broadly used AF cells gene markers (Li et al, ).…”

Section: Discussionmentioning

confidence: 99%

Genipin‐crosslinked decellularized annulus fibrosus hydrogels induces tissue‐specific differentiation of bone mesenchymal stem cells and intervertebral disc regeneration

Peng

Huang

et al. 2020

J Tissue Eng Regen Med

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Biomaterial‐based therapy that can restore annulus fibrosus (AF) function in early stage and promote endogenous repair of AF tissues is a promising approach for AF tissue repair. In this study, we established a genipin‐crosslinked decellularized AF hydrogels (g‐DAF‐G) that are injectable and could manifest better in situ formability than noncrosslinked decellularized AF hydrogel, while preserving the capacity of directing differentiation of human bone mesenchymal stem cells (hBMSCs) towards AF cells. Hematoxylin and eosin staining, 4',6‐diamidino‐2‐phenylindole staining, and so forth showed that the majority of cellular components were removed, whereas extracellular matrix and microstructure were largely preserved. The storage modulus increased from 465.5 ± 9.4 Pa to 3.29 ± 0.24 MPa after 0.02% genipin crosslinking of decellularized AF hydrogels (DAF‐G) to form g‐DAF‐G. AF‐specific genes (COL1A1, COL5A1, TNMD, IBSP, FBLN1) were significantly higher in DAF‐G and g‐DAF‐G groups than that in control group after 21 days of culturing. g‐DAF‐G significantly restored nucleus pulposus water content and preserved intervertebral structure in vivo. Summarily, we produced a novel AF regeneration biomaterial, g‐DAF‐G, which exhibited well biocompatibility, great bioactivity, and much higher mechanical strength than DAF‐G. This study will provide an easy and fast therapeutic alternative to repair AF injuries or tears.

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Substrate stiffness- and topography-dependent differentiation of annulus fibrosus-derived stem cells is regulated by Yes-associated protein

Cited by 76 publications

References 64 publications

Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels

Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels

Biomimicking Fiber Platform with Tunable Stiffness to Study Mechanotransduction Reveals Stiffness Enhances Oligodendrocyte Differentiation but Impedes Myelination through YAP‐Dependent Regulation

Genipin‐crosslinked decellularized annulus fibrosus hydrogels induces tissue‐specific differentiation of bone mesenchymal stem cells and intervertebral disc regeneration

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