Silk scaffolds for musculoskeletal tissue engineering

Yao, Danyu; Liu, Haifeng; Fan, Yubo

doi:10.1177/1535370215606994

Cited by 51 publications

(36 citation statements)

References 85 publications

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“…Furthermore renal mesangial cells show phenotype loss during the first five passages 24 . As known from our own prior experiments 25 and investigated by Yao et al in detail 4 , dedifferentiation of human tenocytes occurs after early passaging. Three-dimensional culture models have been shown to counteract in vitro dedifferentiation to a certain extent.…”

Section: Discussionmentioning

confidence: 89%

“…However, tenocytes are difficult to obtain in sufficient number, and when expanded in vitro, they quickly lose their original phenotype. Yao et al 4 observed changes in morphology and rapid decrease in cell proliferation, collagen 1 production and decorin expression during 2D in vitro culture after as few as four passages. Attempts with 3D culture models have been undertaken to overcome this problem of in vitro senescence.…”

Section: Introductionmentioning

confidence: 99%

“…These contructs consist of an acellular scaffold providing mechanical stability, and a seeded cell line providing regeneration and tissue function. Several scaffold materials have been used with some success so far, such as chitosan, silk fibroin, PLGA or decellularized cadaver tendons [1][2][3][4] . Compared to other cell lines such as skin fibroblast and adipose stem cells 5,6 , tenocytes seem highly suitable or scaffold seeding as the most original cell line 7 .…”

Section: Introductionmentioning

confidence: 99%

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Spheroid formation and modulation of tenocyte-specific gene expression under simulated microgravity

Kraus

Luetzenberg

Abuagela

et al. 2017

MLTJ

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SummaryBackground: For tendon tissue engineering, tenocyte-seeded scaffolds are a promising approach. Under conventional 2D culture however, tenocytes show rapid senescene and phenotype loss. We hypothesized that phenotype loss could be counteracted by simulated microgravity conditions. Methods: Human tenocytes were exposed to microgravity for 9 days on a Random Positioning Machine (RPM). Formation of 3D-structures (spheroids) was observed under light microscopy, gene expression was measured by realtime PCR. Cells under conventional 2D-culture served as control group. Results: Simulated microgravity reached a value of as low as 0.003g. Spheroid formation was observed after 4 days, and spheroids showed stable existance to the end of the observation period. After 9 days, spheroids showed a significantly higher gene expression of collagen 1 (Col1A1) compared to adherent cells under microgravity (4.4x, p=0.04) and compared to the control group (5.6x, p=0.02). Gene expression of collagen 3 (COL3A1) was significantly increased in spheroids compared to the control group (2.3x, p=0.03). Gene expressions of the extracellular matrix genes Tenascin C und Fibronectin (TNC and FN) were increased in adherent cells under microgravity compared to the 1g-control group, not reaching statistical significance (p=0.1 and p=0.3). For the gene expression of vimentin, no significant alteration was observed both in the adherent cells and in the spheroids compared to the 1g control group. Gene expression of the tenocyte-specific transcription factor scleraxis (SCX) was significantly increased in spheroids compared to the control group (3.7x, p=0.03). Conclusion: Simulated microgravity could counteract tenocyte senescence in vitro and serve as a promising model for scaffold-free 3D cell culturing and tissue engineering. Level of evidence: V (laboratory study).

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Section: Discussionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Spheroid formation and modulation of tenocyte-specific gene expression under simulated microgravity

Kraus

Luetzenberg

Abuagela

et al. 2017

MLTJ

View full text Add to dashboard Cite

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“…Natural-based polymers originate from natural sources and are being evaluated in tendon regeneration due to their high availability and low cost. These include silk fibroin (Yao et al 2016 ) collagen (Purcel, 2016 ), gelatin (Selle et al 2015 ), and hyaluronan (Liang et al 2014 ). They have shown promising results in vitro and in vivo.…”

Section: Treatment Optionsmentioning

confidence: 99%

Current trends in tendinopathy: consensus of the ESSKA basic science committee. Part II: treatment options

Abat¹,

Alfredson

Cucchiarini

et al. 2018

J EXP ORTOP

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The treatment of painful chronic tendinopathy is challenging. Multiple non-invasive and tendon-invasive methods are used. When traditional non-invasive treatments fail, the injections of platelet-rich plasma autologous blood or cortisone have become increasingly favored. However, there is little scientific evidence from human studies supporting injection treatment. As the last resort, intra- or peritendinous open or endoscopic surgery are employed even though these also show varying results. This ESSKA basic science committee current concepts review follows the first part on the biology, biomechanics and anatomy of tendinopathies, to provide a comprehensive overview of the latest treatment options for tendinopathy as reported in the literature.

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“…These findings suggest that the addition of bioactive molecules to alginate will improve their ability to act as cell carriers [ 9 ]. In this regard, silk-fibroin, a fibrous polymer derived from different silkworm species, has been widely used as suitable matrix/substrate due to its high biocompatibility, excellent mechanical properties and abundance of cell binding motifs (arginine-glycine-aspartic acid, RGD) in its structure, which enhance cell attachment and proliferation [ 10 ][ 11 ][ 12 ]. However, only a few papers demonstrated the improved adhesion of the cells onto silk-fibroin coated alginate microcarriers.…”

Section: Introductionmentioning

confidence: 99%

A new holistic 3D non-invasive analysis of cellular distribution and motility on fibroin-alginate microcarriers using light sheet fluorescent microscopy

et al. 2017

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Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading. In this work, we propose a non-invasive and non-destructive imaging analysis for investigating motility, viability and distribution of Mesenchymal Stem Cells (MSCs) on silk fibroin-based alginate microcarriers, to test the adhesion capacity of the fibroin coating onto alginate which is known to be unsuitable for cell adhesion. However, in depth characterization of the biomaterial is beyond the scope of this paper. Scaffold-loaded MSCs were stained with Calcein-AM and Ethidium homodimer-1 to detect live and dead cells, respectively, and counterstained with Hoechst to label cell nuclei. Time-lapse Light Sheet Fluorescent Microscopy (LSFM) was then used to produce three-dimensional images of the entire cells-loaded fibroin/alginate microcarriers. In order to quantitatively track the cell motility over time, we also developed an open source user friendly software tool called Fluorescent Cell Tracker in Three-Dimensions (F-Tracker3D). Combining LSFM with F-Tracker3D we were able for the first time to assess the distribution and motility of stem cells in a non-invasive, non-destructive, quantitative, and three-dimensional analysis of the entire surface of the cell-loaded scaffold. We therefore propose this imaging technique as an innovative holistic tool for monitoring cell-biomaterial interactions, and as a tool for the design, fabrication and functionalization of a scaffold as a medical device.

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Silk scaffolds for musculoskeletal tissue engineering

Cited by 51 publications

References 85 publications

Spheroid formation and modulation of tenocyte-specific gene expression under simulated microgravity

Spheroid formation and modulation of tenocyte-specific gene expression under simulated microgravity

Current trends in tendinopathy: consensus of the ESSKA basic science committee. Part II: treatment options

A new holistic 3D non-invasive analysis of cellular distribution and motility on fibroin-alginate microcarriers using light sheet fluorescent microscopy

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