TGF-β1 and IGF-1 influence the re-differentiation capacity of human chondrocytes in 3D pellet cultures in relation to different oxygen concentrations

Jonitz, Anika; Lochner, Katrin; Tischer, Thomas; Hansmann, Doris; Bader, Rainer

doi:10.3892/ijmm.2012.1042

Cited by 33 publications

(40 citation statements)

References 25 publications

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“…This was observed for both oxygen conditions, but was more pronounced under hypoxia. This finding suggests an additive effect of both growth factors, which was also observed in previous studies (23) as well as by other research groups (18,32,33). Additionally, it was previously reported that intracellular signalling pathways of TGF-β1 and IGF-1 are partially linked (7,34).…”

Section: Discussionsupporting

confidence: 91%

Gene expression analysis of growth factor receptors in human chondrocytes in monolayer and 3D pellet cultures

Witt

Salamon

Boy

et al. 2017

International Journal of Molecular Medicine

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The main goal of cartilage repair is to create functional tissue by enhancing the in vitro conditions to more physiological in vivo conditions. Chondrogenic growth factors play an important role in influencing cartilage homeostasis. Insulin-like growth factor (IGF)-1 and transforming growth factor (TGF)-β1 affect the expression of collagen type II (Col2) and glycosaminoglycans (GAGs) and, therefore, the targeted use of growth factors could make chondrogenic redifferentiation more efficient. In the present study, human chondrocytes were postmortally isolated from healthy articular cartilage and cultivated as monolayer or 3D pellet cultures either under normoxia or hypoxia and stimulated with IGF-1 and/or TGF-β1 to compare the impact of the different growth factors. The mRNA levels of the specific receptors (IGF1R, TGFBR1, TGFBR2) were analyzed at different time points. Moreover, gene expression rates of collagen type 1 and 2 in pellet cultures were observed over a period of 5 weeks. Additionally, hyaline-like Col2 protein and sulphated GAG (sGAG) levels were quantified. Stimulation with IGF-1 resulted in an enhanced expression of IGF1R and TGFBR2 whereas TGF-β1 stimulated TGFBR1 in the monolayer and pellet cultures. In monolayer, the differences reached levels of significance. This effect was more pronounced under hypoxic culture conditions. In pellet cultures, increased amounts of Col2 protein and sGAGs after incubation with TGF-β1 and/or IGF-1 were validated. In summary, constructing a gene expression profile regarding mRNA levels of specific growth factor receptors in monolayer cultures could be helpful for a targeted application of growth factors in cartilage tissue engineering.

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Section: Discussionsupporting

confidence: 91%

Gene expression analysis of growth factor receptors in human chondrocytes in monolayer and 3D pellet cultures

Witt

Salamon

Boy

et al. 2017

International Journal of Molecular Medicine

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“…As a preliminary prove of the protocols’ efficiency, we considered gene expression analysis as the best option, since it is affordable and the required instrumentation (PCR, inverted optical microscope) is most likely available in most cell biology laboratories. The set of analysed genes was carefully chosen considering the available literature to monitor cartilage phenotype alterations (Caterson et al, 2001; Diekman et al, 2010; Grogan et al, 2014; Jonitz et al, 2012; Seda Tigli et al, 2009; Shi et al, 2014). Based on the mentioned, the correlation between COL2 and COL1 in addition to ACAN, seemed to be the most suitable.…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Naranda

Gradišnik

Gorenjak

et al. 2017

PeerJ

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BackgroundCartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue.MethodsIsolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production.ResultsCartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well.DiscussionIn this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very frequently performed orthopaedic surgery during which both femoral condyles are removed. The latter present the ideal source for a simple and relatively cheap isolation of chondrocytes as was confirmed in our study.

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“…3 Recent in vitro studies indicate that the most efficient chondrogenic differentiation of chondrocyte-like progenitor cells and mesenchymal stem cells is achieved by using standard cell culture medium supplemented with growth factors like insulin-like growth factor (IGF) 1, transforming growth factor (TGF) β1 and 2, fibroblast growth factor (FGF) 2, and bone morphogenetic protein (BMP) 2. [4][5][6] It is known that IGF-1 enhances the synthesis of proteoglycans and directly decreases the degradation of proteoglycans in cartilage. 7 Moreover, it was shown that IGF-1 helps to maintain the survival of chondrocytes.…”

Section: Introductionmentioning

confidence: 99%

“…With regard to the clinical application for cartilage repair, cells have been incubated in three-dimensional culture systems, like pellets and microspheres, 4,14 as well as hydrogels or biomaterials. [15][16][17][18] Superficial cartilage defects have mainly been treated with hyaluronic acid (HA), while chondral and osteochondral lesions have been medicated by means of HA and autologous cartilage transplantation (ACT) since 1994.…”

Section: Introductionmentioning

confidence: 99%

Positive impact of IGF-1-coupled nanoparticles on the differentiation potential of human chondrocytes cultured on collagen scaffolds

Pasold

Zander

Heskamp

et al. 2015

IJN

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Purpose In the present study, silica nanoparticles (sNP) coupled with insulin-like growth factor 1 (IGF-1) were loaded on a collagen-based scaffold intended for cartilage repair, and the influence on the viability, proliferation, and differentiation potential of human primary articular chondrocytes was examined. Methods Human chondrocytes were isolated from the hyaline cartilage of patients (n=4, female, mean age: 73±5.1 years) undergoing primary total knee joint replacement. Cells were dedifferentiated and then cultivated on a bioresorbable collagen matrix supplemented with fluorescent sNP coupled with IGF-1 (sNP–IGF-1). After 3, 7, and 14 days of cultivation, cell viability and integrity into the collagen scaffold as well as metabolic cell activity and synthesis rate of matrix proteins (collagen type I and II) were analyzed. Results The number of vital cells increased over 14 days of cultivation, and the cells were able to infiltrate the collagen matrix (up to 120 μm by day 7). Chondrocytes cultured on the collagen scaffold supplemented with sNP–IGF-1 showed an increase in metabolic activity (5.98-fold), and reduced collagen type I (1.58-fold), but significantly increased collagen type II expression levels (1.53-fold; P =0.02) after 7 days of cultivation compared to 3 days. In contrast, chondrocytes grown in a monolayer on plastic supplemented with sNP-IGF-1 had significantly lower metabolic activity (1.32-fold; P =0.007), a consistent amount of collagen type I, and significantly reduced collagen type II protein expression (1.86-fold; P =0.001) after 7 days compared to 3 days. Conclusion Collagen-based scaffolds enriched with growth factors, such as IGF-1 coupled to nanoparticles, represent an improved therapeutic intervention for the targeted and controlled treatment of articular cartilage lesions.

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TGF-β1 and IGF-1 influence the re-differentiation capacity of human chondrocytes in 3D pellet cultures in relation to different oxygen concentrations

Cited by 33 publications

References 25 publications

Gene expression analysis of growth factor receptors in human chondrocytes in monolayer and 3D pellet cultures

Gene expression analysis of growth factor receptors in human chondrocytes in monolayer and 3D pellet cultures

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Positive impact of IGF-1-coupled nanoparticles on the differentiation potential of human chondrocytes cultured on collagen scaffolds

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