Tissue-specific gene expression in chondrocytes grown on three-dimensional hyaluronic acid scaffolds

Girotto, Davide; Urbani, Serena; Brun, Paola; Renier, Davide; Barbucci, Rolando; Abatangelo, Giovanni

doi:10.1016/s0142-9612(03)00160-1

Cited by 89 publications

(56 citation statements)

References 39 publications

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“…Following the study published by Girotto et al (2003), we selected the genes of the transcription factor SOX9 and of the collagen gene COL3A1 as reliable markers for in vitro studies of chondrocytes with respect to cartilage differentiation. We found that the transcript level of SOX9 changed during the 21 d incubation only if the cells were incubated onto the HA-Mg 2+ -Na-polyP matrix (Fig.…”

Section: Expression Of Sox9 and Col3a1 In Response To Polyp Exposurementioning

confidence: 99%

Artificial cartilage bio-matrix formed of hyaluronic acid and Mg2+-polyphosphate

Wang¹,

Ackermann²,

Tolba³

et al. 2016

eCM

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Here we show that inorganic polyphosphate (polyP), a polyanionic metabolic regulator consisting of multiple phosphate residues linked by energy-rich phosphoanhydride bonds, is present in the synovial fluid. In a biomimetic approach, to enhance cartilage synthesis and regeneration, we prepared amorphous polyP microparticles with Mg 2+ as counterions. The particles were characterised by X-ray diffraction (XRD), energy-dispersive X-ray (EDX) and Fourier transformed infrared spectroscopic (FTIR) analyses. Similar particles were obtained after addition of Mg 2+ ions to a solution containing hyaluronic acid, as a major component of the synovial fluid, and soluble NapolyP. The viscous paste-like material formed, composed of globular microparticles with diameter of 400 nm, strongly promoted the adhesion of chondrocytes and caused a significant upregulation of the expression of the genes encoding collagen type 3A1, as a marker for chondrocyte differentiation, and SOX9, a transcription factor that regulates chondrocyte differentiation and proliferation. The expression level of the collagen type 3A1 gene was also enhanced by exposure of chondrocytes to synovial fluid that was found to contain polyP with a size of about 80 phosphate residues. This stimulatory effect was abolished after pre-incubation of the synovial fluid with the polyP degrading alkaline phosphatase. We propose a strategy for treatment of joint dysfunctions caused by osteoarthritis based on the application of amorphous Mg 2+ -polyP microparticles that prevent calcium crystal formation in the synovial fluid using scavenging Ca -polyP to hyaluronic acid at the cartilage surface.

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Section: Expression Of Sox9 and Col3a1 In Response To Polyp Exposurementioning

confidence: 99%

Artificial cartilage bio-matrix formed of hyaluronic acid and Mg2+-polyphosphate

Wang¹,

Ackermann²,

Tolba³

et al. 2016

eCM

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“…In all tested scaffolds, transcript levels of cartilage-related genes, like Col 2, Agr, Sox 9, were upregulated. [27][28][29][30][31][32] Two different types of hydrogels were examined in the present study: alginate and alginate/agarose. A 14-day incubation period was selected to allow for sufficient redifferentiation.…”

Section: Human Chondrocyte Gene Expression In Vitromentioning

confidence: 99%

Quantitative analysis of gene expression in human articular chondrocytes assigned for autologous implantation

Barlič

Drobnič

Maličev

et al. 2008

Journal Orthopaedic Research

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Autologous chondrocyte implantation (ACI) relies on the implantation of in vitro expanded cells. The aim was to study the dedifferentiation of human articular chondrocytes under different cultivating conditions [days 0-10 in the primary culture (P0); passages in a monolayer from P0 to P3; monolayer vs. alginate and monolayer vs. alginate/agarose hydrogels] using real-time PCR analysis. The relative gene expressions for collagen type I and II, aggrecan and versican were quantified and the corresponding differentiation indexes (Col2/Col1, Agr/Ver) were calculated. The values of both differentiation indexes decreased exponentially with time in the P0 monolayer culture, and continued with a significant decrease over the subsequent monolayer passages. On the contrary, the chondrocytes seeded in either of the hydrogels significantly increased the indexes compared to their parallel monolayer cultures. These results indicate that alginate and alginate/agarose hydrogels offer an appropriate environment for human articular chondrocytes to redifferentiate after being expanded in vitro. Therefore the three-dimensional (3D) hydrogel chondrocyte cultures present not only surgical, but also biological advantage over the classic suspension-periosteum chondrocyte implantation. ß

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“…Many applications that take advantage of the exceptionally high viscosity characteristics to facilitate a thickened gel-like barrier were first found almost indispensable for surgery procedures in ophthalmology, such as intra-ocular lenses implantation. Other uses of hyaluronan were reported for the treatment of degenerative joint dysfunction in horses and gradually expanded to the experimental treatment to humans [9][10][11][12].…”

Section: Introductionmentioning

confidence: 99%

Influences of hyaluronan on type II collagen fibrillogenesis in vitro

Kuo

Wang²,

Niu

et al. 2007

J Mater Sci: Mater Med

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The effect to the kinetics of type II collagen fibrillogenesis with the addition of hyaluronan (HA), (M w of 1.8 · 10 6 Da), at various concentrations of HA (0.01, 0.05 and 0.1 wt.%) for a series of fibril formation systems was examined in this study. Evidences deduced from the turbidity-time curves revealed that the inclusion of HA had minor or no impact to the fibrillogenesis of type II collagen (collagen conc. at 0.2 mg/mL). The apparent rate constants, k lag (lag phase) increased slightly but k g (growth phase) decreased not very significantly with addition of HA, as compared to the case of pure collagen. This leads us to believe tentatively that, with the addition of HA to collagen solutions, the nucleation process of the fibril formation might have been sped up slightly whereas the growth process slowed up slightly. However, data from TEM observations on the resulting fibrils indicated that the presence of HA did not significantly affect the diameters and the characteristic D-banding periods of the collagen fiber formed. And, from the statistical analyses, we found only insignificant difference (P > 0.05) between the specimens from the various experimental groups. It seems to indicate that the ultimate packing of collagen monomers was probably not interfered or affected significantly by the presence of HA in vitro.

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Tissue-specific gene expression in chondrocytes grown on three-dimensional hyaluronic acid scaffolds

Cited by 89 publications

References 39 publications

Artificial cartilage bio-matrix formed of hyaluronic acid and Mg2+-polyphosphate

Artificial cartilage bio-matrix formed of hyaluronic acid and Mg2+-polyphosphate

Quantitative analysis of gene expression in human articular chondrocytes assigned for autologous implantation

Influences of hyaluronan on type II collagen fibrillogenesis in vitro

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