Culture of Chondrocytes in Alginate Beads

Ceuninck, Frédéric De; Lesur, C.; Pastoureau, Philippe; Caliez, Audrey; Sabatini, Massimo

doi:10.1385/1-59259-810-2:015

Cited by 45 publications

(54 citation statements)

References 18 publications

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“…13,49,50 Scaffolds composed of natural polymers such as, polysaccharide-based hydrogels (alginate beads, chitosan, or agarose) or protein-based hydrogels (collagen or fibrin) are now commonly used in cartilage tissue engineering. [51][52][53] However, because of the ease with which synthetic polymer hydrogels can be functionalized and modified, polyethyleneglycol (PEG) and PEG-derived hydrogels or -fibrins are also making their way into cartilage tissue engineering. 49,54,55 Unfortunately, both types of scaffolds yield products that fall short of the biochemical and mechanical properties of the native tissue.…”

Section: Cartilage Tissue Engineeringmentioning

confidence: 99%

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

Kotecha

Klatt

Magin

2013

Tissue Engineering Part B: Reviews

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A key technical challenge in cartilage tissue engineering is the development of a noninvasive method for monitoring the composition, structure, and function of the tissue at different growth stages. Due to its noninvasive, three-dimensional imaging capabilities and the breadth of available contrast mechanisms, magnetic resonance imaging (MRI) techniques can be expected to play a leading role in assessing engineered cartilage. In this review, we describe the new MR-based tools (spectroscopy, imaging, and elastography) that can provide quantitative biomarkers for cartilage tissue development both in vitro and in vivo. Magnetic resonance spectroscopy can identify the changing molecular structure and alternations in the conformation of major macromolecules (collagen and proteoglycans) using parameters such as chemical shift, relaxation rates, and magnetic spin couplings. MRI provides high-resolution images whose contrast reflects developing tissue microstructure and porosity through changes in local relaxation times and the apparent diffusion coefficient. Magnetic resonance elastography uses low-frequency mechanical vibrations in conjunction with MRI to measure soft tissue mechanical properties (shear modulus and viscosity). When combined, these three techniques provide a noninvasive, multiscale window for characterizing cartilage tissue growth at all stages of tissue development, from the initial cell seeding of scaffolds to the development of the extracellular matrix during construct incubation, and finally, to the postimplantation assessment of tissue integration in animals and patients.

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Section: Cartilage Tissue Engineeringmentioning

confidence: 99%

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

Kotecha

Klatt

Magin

2013

Tissue Engineering Part B: Reviews

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“…The incubation of articular chondrocytes suspended in alginate beads is a common tissue growth strategy in the field of cartilage tissue engineering [38][39][40][41][42]. Bovine chondrocytes (4 millions cells/ml) were cultured using chondrogenic growth media in alginate beads according to the published protocol [39].…”

Section: Chondrocytes Based Cartilage Tissue Engineering Constructsmentioning

confidence: 99%

Monitoring Tissue Engineering and Regeneration by Magnetic Resonance Imaging and Spectroscopy

Kotecha¹

2012

J Tissue Sci Eng

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In this article, based on the invited talk at the "Tissue Science 2012" meeting in Chicago on October 1-3, 2012, we describe some examples of characterization of engineered cartilage and bone tissue using magnetic resonance spectroscopy and imaging. Two different models of engineered cartilage and engineered bone tissue constructs were used for these studies: 1) chondrocyte based cartilage tissue engineering constructs: human and bovine chondrocytes seeded in alginate beads (Hydrogel scaffold model) or bovine chondrocytes grown as pellets (scaffold free model); 2) mesenchymal stem cell (MSC) based cartilage and bone tissue engineering constructs: human mesenchymal stem cell (HMSCs) seeded in cartilage biomimetic scaffolds (collagen/chitosan scaffold integrated with extracellular matrix of cartilage) or HMSCs seeded in collagen/chitosan scaffolds. Magnetic resonance spectroscopy and imaging experiments using 9.4 T (400 MHz proton frequency), 11.7 T (500 MHz proton frequency) or 14.1 T (600 MHz proton frequency) MR spectrometers/Imager were performed on these constructs over two to four weeks of tissue culture time. Specifically, water suppressed proton NMR spectroscopy; proton and sodium multi-quantum coherence spectroscopy and proton T1, T2 and ADC parametric MRI were used to study the chondrogenesis and osteogenesis of these tissues. We found that the change in MR relaxation and diffusion coefficient parameters correlate well with the growth of engineered tissues. We found that the MR parameters and the change in these parameters in growing tissue are strongly influenced by the choice of scaffolds. We also found as expected that the tissue-engineered cartilage lacked order or preference in collagen orientation. Further work is underway to elucidate these findings. We anticipate that in future, MRI will augment histological and immunohistochemical techniques by providing a complimentary and real time quantitative assessment of engineered tissue growth at all growth stages: (i) cell seeding to pre implantation; (ii) preclinical validation studies post implantation in small and large animal models; (iii) clinical studies of performance of engineered tissues.

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“…Alginic acid, the pivotal hydrogel component, is considered to be suitable due to biodegradability and biocompatibility Stevens et al, 2004]. Alginates are well understood in their variability caused by monomer composition, sequential arrangement, G-block length, molecular size and concentrations [De Ceuninck et al, 2004]. New modifi cations, e.g.…”

Section: Hydrogel As Biomaterialsmentioning

confidence: 99%

“…They can be used in many compositions for 3-D cell encapsulation and culture due to variable properties defi ned by monomer composition, sequential arrangement, G-block length, molecular size, Ca 2+ and polymer concentration [Smidsrod and Skjak-Braek, 1990]. Established protocols and thorough characterization make this a favorable material [Draget et al, 1997;De Ceuninck et al, 2004]. Thus, alginates can serve as temporary conductive structures for anchorage-independent spatial cell arrangement assuring cell and tissue differentiation.…”

Section: Introductionmentioning

confidence: 99%

Morphology and Function of Ovine Articular Cartilage Chondrocytes in 3-D Hydrogel Culture

Schagemann

Mrosek

Landers

et al. 2006

Cells Tissues Organs

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Different cell- and biomaterial-based tissue engineering techniques are under investigation to restore damaged tissue. Strategies that use chondrogenic cells or tissues in combination with bioresorbable delivery materials are considered to be suitable to regenerate bio-artificial cartilage. Three-dimensional (3-D) cell embedding techniques can provide anchorage-independent cell growth and homogenous spatial cell arrangement, which play a key role in the maintenance of the characteristic phenotype and thus the formation of differentiated tissue. We developed a new injectable high water content (90%) hydrogel formulation with 5% sodium alginic acid and 5% gelatin as a temporary supportive intercellular matrix for 3-D cell culture. The objective was to determine whether the in vitro hydrogel culture of chondrocytes could preserve hyaline characteristics and thus could provide cartilage regeneration in vitro. Chondrocytes harvested from knee joints of skeletally mature sheep were cultured 3-D in hydrogel (7 × 10⁶ cells/ml, 2.8-µl beads) for up to 10 weeks. Cell morphology and viability were evaluated with light microscopy, and proliferative activity was assessed with antibromodeoxyuridine immunofluorescence. Expression of collagens type I (COL1) and II (COL2), cartilage proteoglycans (PG) and hyaluronan synthases (HAS) were studied immunohistochemically. We observed that up to 36% of chondrocytes proliferated, while almost 100% presented a differentiated spheroidal phenotype. After an initial decrease at 2 weeks, cell density recovered to 85% of the initial absolute value at 10 weeks. Expression of hyaline matrix molecules resembled the in vivo pattern with increasing spatial deposition of PG and COL2. The proportion of PG-positive cells increased from initially 13 to 53% after 10 weeks, in contrast to consistently 100% COL2-positive cells. We conclude that 3-D hydrogel culture, even without mechanical stimulation or growth factor application, can keep chondrocytes in a differentiated state and provides a chondrogenic cell environment for in vitro cartilage regeneration for at least 10 weeks. Moreover, this hydrogel appears to be a suitable cell delivery material for subsequent in vivo implantation.

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Culture of Chondrocytes in Alginate Beads

Cited by 45 publications

References 18 publications

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

Monitoring Tissue Engineering and Regeneration by Magnetic Resonance Imaging and Spectroscopy

Morphology and Function of Ovine Articular Cartilage Chondrocytes in 3-D Hydrogel Culture

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