Modelling condensation and the initiation of chondrogenesis in chick wing bud mesenchymal cells levitated in an ultrasound trap

Edwards, Gareth O.; Coakley, W. Terence; Ralphs, J. R.; Archer, Charles W.

doi:10.22203/ecm.v019a01

Cited by 14 publications

(9 citation statements)

References 68 publications

(67 reference statements)

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“…7). Once entrapped, MSCs could be stimulated to undergo cellular proliferation and chondrogenic differentiation by growth factor stimulation as well as cell-cell interactions [62,63], even in the absence of potential cell-scaffold interactions. This cellular proliferation enabled MSCs to overgrow the bottom of the scaffold and produce a smooth shiny surface (Fig.…”

Section: Discussionmentioning

confidence: 99%

The effects of crosslinking of scaffolds engineered from cartilage ECM on the chondrogenic differentiation of MSCs

et al. 2013

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Scaffolds fabricated from cartilage extracellular matrix provide a chondroinductive environment that stimulates cartilaginous matrix synthesis in a variety of cell types. A limitation of these cartilage-derived matrix (CDM) scaffolds is that they contract during in vitro culture, which unpredictably alters their shape. The current study examined the hypothesis that collagen crosslinking techniques could inhibit cell-mediated contraction of CDM scaffolds. We analyzed the effects of dehydrothermal (DHT) treatment, ultraviolet light irradiation (UV), and the chemical crosslinker carbodiimide (CAR) on scaffold contraction and chondrogenic differentiation of adult human bone marrow-derived stem cells (MSCs). Both physical and chemical crosslinking treatments retained the original scaffold dimensions. DHT and UV treatments produced significantly higher glycosaminoglycan and collagen contents than CAR crosslinked and non-crosslinked constructs. Crosslinking treatments influenced the composition of newly synthesized matrix, and DHT treatment best matched the composition of native cartilage. DHT, UV, and non-crosslinked CDM films supported cell attachment, while CAR crosslinking inhibited cell adhesion. These results affirm that collagen crosslinking treatments can prevent cell-mediated contraction of CDM scaffolds. Interestingly, crosslinking treatments influence chondrogenic differentiation. These effects seem to be mediated by modifications to cell-matrix interactions between MSCs and the CDM; however, further work is necessary to elucidate a specific mechanism.

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Section: Discussionmentioning

confidence: 99%

The effects of crosslinking of scaffolds engineered from cartilage ECM on the chondrogenic differentiation of MSCs

et al. 2013

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“…42 Additionally, gap junctional intercellular communication may play a major role in cartilaginous differentiation thereby influencing linked processes such as altered Ca 2+ or second messenger exchange. 43 Adult articular cartilage chondrocytes exist as individual cells embedded in the ECM, and direct intercellular communication via gap junctions occurs mainly among the flattened chondrocytes facing the outer cartilage layer. 40,44 However, chondrocytes extracted from adult articular cartilage and grown in primary culture express connexin 43 (Cx43) and form functional gap junctions capable of sustaining the propagation of intercellular calcium waves.…”

Section: Discussionmentioning

confidence: 99%

Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes

et al. 2013

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Cartilage regeneration based on isolated and culture-expanded chondrocytes has been studied in various in vitro models, but the quality varies with respect to the morphology and the physiology of the synthesized tissues. The aim of our study was to promote in vitro chondrogenesis of human articular chondrocytes using a novel three-dimensional (3-D) cultivation system in combination with the chondrogenic differentiation factors transforming growth factor beta 2 (TGF-β2) and L-ascorbic acid. Articular chondrocytes isolated from six elderly patients were expanded in monolayer culture. A single-cell suspension of the dedifferentiated chondrocytes was then added to agar-coated dishes without using any scaffold material, in the presence, or absence of TGF-β2 and/or L-ascorbic acid. Three-dimensional cartilage-like constructs, called single spheroids, and microtissues consisting of several spheroids fused together, named as fusions, were formed. Generated tissues were mainly characterized using histological and immunohistochemical techniques. The morphology of the in vitro tissues shared some similarities to native hyaline cartilage in regard to differentiated S100-positive chondrocytes within a cartilaginous matrix, with strong collagen type II expression and increased synthesis of proteoglycans. Finally, our innovative scaffold-free fusion culture technique supported enhanced chondrogenesis of human articular chondrocytes in vitro. These 3-D hyaline cartilage-like microtissues will be useful for in vitro studies of cartilage differentiation and regeneration, enabling optimization of functional tissue engineering and possibly contributing to the development of new approaches to treat traumatic cartilage defects or osteoarthritis.

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“…The developmental relevance of these rapidly forming aggregates is suggested by a recent study in which cells identical to those used here were permitted to recover from trypsinization and levitated in an ultrasound trap. Such cells formed aggregates via their native attachment moieties, also within two minutes, and contained detectable amounts of the chondrogenic transcription factor Sox9 in their nuclei within three hours [67]. …”

Section: Discussionmentioning

confidence: 99%

A regulatory network of two galectins mediates the earliest steps of avian limb skeletal morphogenesis

et al. 2011

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BackgroundThe skeletal elements of vertebrate embryonic limbs are prefigured by rod- and spot-like condensations of precartilage mesenchymal cells. The formation of these condensations depends on cell-matrix and cell-cell interactions, but how they are initiated and patterned is as yet unresolved.ResultsHere we provide evidence that galectins, β-galactoside-binding lectins with β-sandwich folding, play fundamental roles in these processes. We show that among the five chicken galectin (CG) genes, two, CG-1A, and CG-8, are markedly elevated in expression at prospective sites of condensation in vitro and in vivo, with their protein products appearing earlier in development than any previously described marker. The two molecules enhance one another's gene expression but have opposite effects on condensation formation and cartilage development in vivo and in vitro: CG-1A, a non-covalent homodimer, promotes this process, while the tandem-repeat-type CG-8 antagonizes it. Correspondingly, knockdown of CG-1A inhibits the formation of skeletal elements while knockdown of CG-8 enhances it. The apparent paradox of mutual activation at the gene expression level coupled with antagonistic roles in skeletogenesis is resolved by analysis of the direct effect of the proteins on precartilage cells. Specifically, CG-1A causes their aggregation, whereas CG-8, which has no adhesive function of its own, blocks this effect. The developmental appearance and regulation of the unknown cell surface moieties ("ligands") to which CG-1A and CG-8 bind were indicative of specific cognate- and cross-regulatory interactions.ConclusionOur findings indicate that CG-1A and CG-8 constitute a multiscale network that is a major mediator, earlier-acting than any previously described, of the formation and patterning of precartilage mesenchymal condensations in the developing limb. This network functions autonomously of limb bud signaling centers or other limb bud positional cues.

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Modelling condensation and the initiation of chondrogenesis in chick wing bud mesenchymal cells levitated in an ultrasound trap

Cited by 14 publications

References 68 publications

The effects of crosslinking of scaffolds engineered from cartilage ECM on the chondrogenic differentiation of MSCs

The effects of crosslinking of scaffolds engineered from cartilage ECM on the chondrogenic differentiation of MSCs

Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes

A regulatory network of two galectins mediates the earliest steps of avian limb skeletal morphogenesis

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