BMP activation and Wnt-signalling affect biochemistry and functional biomechanical properties of cartilage tissue engineering constructs

Krase, A.; Abedian, Reza; Steck, Eric; Hurschler, Christof; Richter, Wiltrud

doi:10.1016/j.joca.2013.11.011

Cited by 31 publications

(33 citation statements)

References 47 publications

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“…As such, the sGAG production and proliferation on the construct surface may be caused by the greater exposure of nutrients from the culture medium. This result has been seen in a previous porcine chondrocyte seeded collagen scaffold grown with growth factors . In this porcine study histology images revealed early sGAG production was concentrated on the scaffold surface.…”

Section: Discussionsupporting

confidence: 85%

“…Tissue engineered studies using human articular chondrocytes grown in vitro produced compressive values about 50% of native tissue . In contrast, previous studies using animal derived chondrocytes (porcine, bovine, and canine) have shown compressive properties that range from 14 to 90% of native cartilage . The incorporation of growth factors and mechanical stimulation is known to enhance cartilage production and contributes to the improved compressive properties seen in some of these studies .…”

Section: Discussionmentioning

confidence: 97%

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Mechanical properties and structure‐function relationships of human chondrocyte‐seeded cartilage constructs after in vitro culture

Middendorf

Griffin

Shortkroff

et al. 2017

Journal Orthopaedic Research

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Autologous Chondrocyte Implantation (ACI) is a widely recognized method for the repair of focal cartilage defects. Despite the accepted use, problems with this technique still exist, including graft hypertrophy, damage to surrounding tissue by sutures, uneven cell distribution, and delamination. Modified ACI techniques overcome these challenges by seeding autologous chondrocytes onto a 3D scaffold and securing the graft into the defect. Many studies on these tissue engineered grafts have identified the compressive properties, but few have examined frictional and shear properties as suggested by FDA guidance. This study is the first to perform three mechanical tests (compressive, frictional, and shear) on human tissue engineered cartilage. The objective was to understand the complex mechanical behavior, function, and changes that occur with time in these constructs grown in vitro using compression, friction, and shear tests. Safranin-O histology and a DMMB assay both revealed increased sulfated glycosaminoglycan (sGAG) content in the scaffolds with increased maturity. Similarly, immunohistochemistry revealed increased lubricin localization on the construct surface. Confined compression and friction tests both revealed improved properties with increased construct maturity. Compressive properties correlated with the sGAG content, while improved friction coefficients were attributed to increased lubricin localization on the construct surfaces. In contrast, shear properties did not improve with increased culture time. This study suggests the various mechanical and biological properties of tissue engineered cartilage improve at different rates, indicating thorough mechanical evaluation of tissue engineered cartilage is critical to understanding the performance of repaired cartilage. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:2298-2306, 2017.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 97%

Mechanical properties and structure‐function relationships of human chondrocyte‐seeded cartilage constructs after in vitro culture

Middendorf

Griffin

Shortkroff

et al. 2017

Journal Orthopaedic Research

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“…Although BMP-4/7 heterodimer strongly stimulated cartilage matrix deposition in chondrocytes in a previous study21, BMP-4/7 heterodimer neither enhanced proteoglycan deposition in MSC-derived pellets relative to TGF-β alone (Fig. 5a,b), nor did it stimulate collagen type II and/or collagen type X deposition in day 42 pellets according to Western blotting (Fig.…”

Section: Resultsmentioning

confidence: 55%

Differential expression of TGF-β superfamily members and role of Smad1/5/9-signalling in chondral versus endochondral chondrocyte differentiation

Dexheimer

Gabler

Bomans

et al. 2016

Sci Rep

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Proteins of the transforming-growth-factor-β (TGF-β)-superfamily have a remarkable ability to induce cartilage and bone and the crosstalk of TGF-β - and BMP-signalling pathways appears crucial during chondrocyte development. Aim was to assess the regulation of TGF-β-superfamily members and of Smad2/3- and Smad1/5/9-signalling during endochondral in vitro chondrogenesis of mesenchymal stromal cells (MSC) relative to chondral redifferentiation of articular chondrocytes (AC) to adjust chondrocyte development of MSC towards a less hypertrophic phenotype. While MSC increased BMP4 and BMP7 and reduced TGFBR2 and TGFBR3-expression during chondrogenesis, an opposite regulation was observed during AC-redifferentiation. Antagonists CHRD and CHL2 rose significantly only in AC-cultures. AC showed higher initial BMP4, pSmad1/5/9 and SOX9 protein levels, a faster (re-)differentiation but a similar decline of pSmad2/3- and pSmad1/5/9-signalling versus MSC-cultures. BMP-4/7-stimulation of MSC-pellets enhanced SOX9 and accelerated ALP-induction but did not shift differentiation towards osteogenesis. Inhibition of BMP-signalling by dorsomorphin significantly reduced SOX9, raised RUNX2, maintained collagen-type-II and collagen-type-X lower and kept ALP-activity at levels reached at initiation of treatment. Conclusively, ALK1,2,3,6-signalling was essential for MSC-chondrogenesis and its prochondrogenic rather than prohypertrophic role may explain why inhibition of canonical BMP-signalling could not uncouple cartilage matrix production from hypertrophy as this was achieved with pulsed PTHrP-application.

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“…β-catenin accumulation activates transcription in conjunction with co-transcription factors, LEFs/TCFs [32]. It was previously reported that LiCl (5–10 mM) can effectively induce the canonical Wnt signaling pathway and mediate cell differentiation of MSCs [16,17,33] and articular chondrocytes [34]. SB216763 has been shown to regulate cell proliferation and survival and to induce the transcription of β-catenin-dependent genes [24].…”

Section: Discussionmentioning

confidence: 99%

Enhanced Chondrogenic Differentiation of Human Umbilical Cord Wharton's Jelly Derived Mesenchymal Stem Cells by GSK-3 Inhibitors

et al. 2017

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Articular cartilage is an avascular, alymphatic, and aneural system with very low regeneration potential because of its limited capacity for self-repair. Mesenchymal stem cells (MSCs) are the preferred choice for cell-based therapies. Glycogen synthase kinase 3 (GSK-3) inhibitors are compounds that can induce the Wnt signaling pathway, which is involved in chondrogenesis and cartilage development. Here, we investigated the influence of lithium chloride (LiCl) and SB216763 synergistically with TGF-β3 on chondrogenic differentiation in human mesenchymal stem cells derived from Wharton’s jelly tissue (hWJ-MSCs). hWJ-MSCs were cultured and chondrogenic differentiation was induced in monolayer and pellet experiments using chondrogenic medium, chondrogenic medium supplemented with LiCl, or SB216763 for 4 weeks. After in vitro differentiation, cultured cells were examined for the expression of Sox9, ACAN, Col2a1, and β-catenin markers. Glycosaminoglycan (GAG) accumulation was also examined by Alcian blue staining. The results indicated that SB216763 was more effective than LiCl as evidenced by a higher up-regulation of the expression of cartilage-specific markers, including Sox9, ACAN, Col2a1 as well as GAG accumulation. Moreover, collagen type II expression was strongly observed in cells cultured in the chondrogenic medium + SB216763 as evidenced by western blot analysis. Both treatments appeared to mediate the Wnt signaling pathway by up-regulating β-catenin gene expression. Further analyses showed that all treatments suppressed the progression of chondrocyte hypertrophy, determined by decreased expression of Col10a1 and Runx2. These results indicate that LiCl and SB216763 are potential candidates for further in vivo therapeutic trials and would be of great importance for cartilage regeneration.

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BMP activation and Wnt-signalling affect biochemistry and functional biomechanical properties of cartilage tissue engineering constructs

Abstract: While Wnt-activation is less effective, BMP-4/-7 heterodimer stimulation approximated native cartilage features in less than 50% of standard culture time representing a promising strategy for functional cartilage TE to improve biomechanical parameters of engineered cartilage.

Cited by 31 publications

References 47 publications

Mechanical properties and structure‐function relationships of human chondrocyte‐seeded cartilage constructs after in vitro culture

Mechanical properties and structure‐function relationships of human chondrocyte‐seeded cartilage constructs after in vitro culture

Differential expression of TGF-β superfamily members and role of Smad1/5/9-signalling in chondral versus endochondral chondrocyte differentiation

Enhanced Chondrogenic Differentiation of Human Umbilical Cord Wharton's Jelly Derived Mesenchymal Stem Cells by GSK-3 Inhibitors

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