Proteoglycan synthesis by cultured human chondrocytes

Aulthouse, Amy Lynn; Becker, Konrad B.; Beck, Michael

doi:10.1002/jemt.1070280607

Cited by 6 publications

(2 citation statements)

References 26 publications

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“…Besides TGF-β 2 , ascorbic acid was used as an inducing factor for chondrogenesis in several studies, resulting in conflicting outcomes. 32,33 In our study, ascorbic acid did not have any influence on the PG, collagen type II, or S100 contents in microtissues.…”

Section: Discussionsupporting

confidence: 43%

Featured Article:In vitrodevelopment of personalized cartilage microtissues uncovers an individualized differentiation capacity of human chondrocytes

Martin

Lehmann

Sack

et al. 2017

Exp Biol Med (Maywood)

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Personalized features in the treatment of knee injuries and articular replacement therapies play an important role in modern life with increasing demand. Therefore, cell-based therapeutic approaches for the regeneration of traumatic defects of cartilage tissue were developed. However, great variations in the quality of repair tissue or therapeutic outcome were observed. The aim of the study was to capture and visualize individual differentiation capacities of chondrocytes derived from different donors with regard to a possible personal regeneration capacity using a cell-based therapy. The redifferentiation potential of monolayer cultured cells was analyzed in a scaffold-free three-dimensional tissue model. Furthermore, stimulating options using cartilage maturation factors such as L-ascorbic acid and transforming growth factor beta 2 (TGF-β) on this process were of special interest. Cells and tissues were analyzed via histological and immunohistochemical methods. Gene expression was measured by quantitative real-time polymerase chain reaction. In monolayer culture, cells from all donors showed an almost identical differentiation profile. In contrast, the differentiation state of cartilage-like three-dimensional microtissues revealed clear differences with respect to individual donors. Analyses at the protein and mRNA levels showed high variations regarding cartilage-typical matrix components (e.g. proteoglycans, collagen type II) and intracellular proteins (e.g. S100). Interestingly, only donor chondrocytes with a basic tendency to re-differentiate in a three-dimensional environment were able to increase this tissue-specific maturation when exposed to L-ascorbic acid and/or TGF-β2. Our approach revealed clear-cut possibilities for classification of individual donors into responders or non-responders. On the basis of these results an in vitro platform could be designed to discriminate responders from non-responders. This in vitro three-dimensional test system may be a suitable basis to establish a "personalized diagnostic tool" with the opportunity to assess the capacity of expanded chondrocytes to respond to an autologous cell-based therapy. Impact statement A challenge in cell-based cartilage regeneration therapies is the identification of a "personalized diagnostic tool" to predict the chondrogenic potency of cells from patients who are going to be treated with autologous cells. Comparing the phenotype of isolated chondrocytes from different donors in vitro revealed an individual cartilage-specific differentiation capacity. These personalized features are not detectable in vitro until the monolayer cells have the possibility to rearrange in 3D tissues. Cells from articular cartilage in monolayer culture may not be a suitable basis to discriminate responders from non-responders with respect to a personalized cell-based therapy to treat cartilage defects. A more physiological 3D (micro-)environment enable the cells to present their individual differentiation capacity. The here described microtissue model might be...

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Section: Discussionsupporting

confidence: 43%

Featured Article:In vitrodevelopment of personalized cartilage microtissues uncovers an individualized differentiation capacity of human chondrocytes

Martin

Lehmann

Sack

et al. 2017

Exp Biol Med (Maywood)

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“…The supplementation with L-ascorbic acid is frequently reported to mediate pro-differentiating effects towards the chondrogenic lineage, 34 but the effect of L-ascorbic acid on proteoglycan synthesis by chondrocytes is debatable. 35,36 Our results show that L-ascorbic acid alone could not stimulate proteoglycan synthesis (Figure 5 B1). GAG expression was detected in fusions cultivated with and without L-ascorbic acid, indicating that the fusion culture itself stimulated GAG expression rather than L-ascorbic acid (compare Figure 5 A2,A3 with B2,B3).…”

Section: Discussionmentioning

confidence: 71%

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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Human Intervertebral Disc Cells from the Annulus: Three-Dimensional Culture in Agarose or Alginate and Responsiveness to TGF-β1

Desai

et al. 1997

Experimental Cell Research

282

164

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Proteoglycan synthesis by cultured human chondrocytes

Cited by 6 publications

References 26 publications

Featured Article:In vitrodevelopment of personalized cartilage microtissues uncovers an individualized differentiation capacity of human chondrocytes

Featured Article:In vitrodevelopment of personalized cartilage microtissues uncovers an individualized differentiation capacity of human chondrocytes

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

Human Intervertebral Disc Cells from the Annulus: Three-Dimensional Culture in Agarose or Alginate and Responsiveness to TGF-β1

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