The application of atelocollagen gel in combination with porous scaffolds for cartilage tissue engineering and its suitable conditions

Yamaoka, Hisayo; Tanaka, Yoko; Nishizawa, Satoru; Asawa, Yukiyo; Takato, Tsuyoshi; Hoshi, Kazuto

doi:10.1002/jbm.a.32509

Cited by 26 publications

(30 citation statements)

References 36 publications

(46 reference statements)

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“…The COL2 evaluated in this study is a specific cartilage component. COL2 forms fibrils that permit cartilage to entrap proteoglycan aggregates and im- the surgical invasiveness of the biopsy and the physical strain on patients (22,29,31,32). Minimal invasiveness is one of the major goals of biomedical techniques, and our method may increase the advantages of this approach (33).…”

Section: Resultsmentioning

confidence: 99%

Application of floating cells for improved harvest in human chondrocyte culture

Yonenaga¹,

Nishizawa²,

Fujihara³

et al. 2012

Biomed. Res.

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Cell culture medium, which must be discarded during medium change, may contain many cells that do not attach to culture plates. In the present study, we focused on these floating cells and attempted to determine their usefulness for cartilage regeneration. We counted the number of floating cells discarded during medium change and compared the proliferation and differentiation between floating cells and their adherent counterparts. Chondrocyte monolayer culture at a density of 5 × 10 3 cells/cm 2 produced viable floating cells at a rate of 2.7-3.2 × 10 3 cells/cm 2 per primary culture. When only the floating cells from one dish were harvested and replated in another dish, the number of cells was 2. ). The floating and adherent cells showed similar proliferation and differentiation properties. The recovery of floating cells from the culture medium could provide an approximately 1.5-fold increase in cell number over conventional monolayer culture. Thus, the collection of floating cells may be regarded as a simple, easy, and reliable method to increase the cell harvest for chondrocytes.

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Section: Resultsmentioning

confidence: 99%

Application of floating cells for improved harvest in human chondrocyte culture

Yonenaga¹,

Nishizawa²,

Fujihara³

et al. 2012

Biomed. Res.

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show abstract

“…To increase the area of cartilage regeneration and apply it to osteoarthritis, we have to make total number of cultured chondrocytes higher compared with that obtained by conventional culture, and should establish three-dimensional mass culture technique for chondrocytes. Although in the present culture using the hollow fiber module, we enzymatically dissolved collagen gel to collect cells, this collagen gel may serve as a scaffold for chondrocytes [18,19]. We prepared the hollow fibers using a non-absorbable material inappropriate for transplantation.…”

Section: Discussionmentioning

confidence: 99%

Hollow Fiber Module Applied for Effective Proliferation and Harvest of Cultured Chondrocytes

Mori¹,

Watanabe²,

Nakagawa³

et al. 2013

MSA

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Steady and useful culture for chondrocytes is essential for cartilage regenerative medicine. However, in conventional plate culture, the chondrocytes become dedifferentiated and lose their ability to make cartilage matrices. Three-dimensional culture mimicking the physiological environment in native chondrocytes is useful to maintain the chondrocyte properties during the proliferation culture. However, the three-dimensional culture is practically a hard task due to difficult harvest of the cells. Thus, we attempted to apply porous materials, hollow fibers for the three-dimensional culture, and developed their module to realize the effective harvest of the cells. Polyethersulfone-based hollow fibers, whose safety and cell affinity were confirmed by the experiment of the coculture with human chondrocytes, were collected to fabricate a module. The hollow fiber module was installed with screw ends, and enabled the easy removal of chondrocytes from the inner unit. Cultured human chondrocytes embedded within collagen hydrogel were put into the outer lumen of the hollow fiber module, while chondrocyte prolfieration medium was perfused through the inner lumen at 0 to 30 mL/min. After 2 weeks' culture, the flow rate of 3 to 10 mL/min effectively supported the chondrocyte proliferation. Then, long-term culture using the hollow fiber module at flow rate of 5 mL/min was performed, revealing that the cell growth in this module at 3 weeks was approximately twice larger than that in static culture. The numbers of viable cells could be maintained by week 7. The hollow fiber module installed with screw ends can effectively culture and harvest the chondrocytes.

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“…Human auricular chondrocytes and 3% atelocollagen gel (KOKEN, Tokyo, Japan) were mixed, adjusting the cell concentration to 1.0 × 10 8 cells/ml and gel concentration to 1% as regenerative cartilage [4,13].…”

Section: Isolation and Analysis Of Chondrocytesmentioning

confidence: 99%

“…However, its application is limited to local articular defects or silicon removal after cosmetic surgery of the nose. We introduced a porous material of scaffold and succeeded in the development of tissue-engineered cartilage with a three-dimensional shape and mechanical strength [4,5]. Briefly, we regenerate cartilage with atelocollagen hydrogel and a PLLA porous scaffold used as a composite scaffold and apply it for treatment of nasal deformation in patients with cleft lip and palate [6].…”

Section: Introductionmentioning

confidence: 99%

Usefulness of Agarose Mold as a Storage Container for Three-Dimensional Tissue-Engineered Cartilage

Mori¹,

Kanazawa²,

Watanabe³

et al. 2013

MSA

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The efficiency of substance exchange may be decreased when the thickness and volume of such a tissue-engineered cartilage that is composed of cultured cells and porous scaffold increase. Moreover, during the transport of this construct with complicated shapes, excessive and focal mechanical loading may cause deformation. The establishment of incubation and transport methods is necessary for the three-dimensional tissue-engineered cartilage. Therefore, we investigated the preparation of an agarose mold with a concavity similar to the shape of 3-dimensional tissue-engineered cartilage to prevent excessive and focal concentration of stress, while avoiding interference with substance exchange as much as possible. Firstly, we investigated the preparation at 1% -4% agarose concentrations. Since the mechanical strength was insufficient at 1%, 2% was regarded as appropriate. Using 2% agarose, we prepared a mold with a 5 × 5 × 5 mm concavity to accommodate tissue-engineered cartilage (5 × 5 × 5 mm mixture of 1.5 × 10 7 cells and collagen gel), and stored the regenerative cartilage in it for 2 and 24 hours. On comparison with storage in a plastic mold with the same shape in which substance exchanged from side and bottom was impossible, although no significant differences were noted in the number or viability of cells after 2 hours, these were markedly reduced in the plastic mold after 24 hours. It was confirmed that favorable cell numbers and viability were maintained by immediately retaining the regenerative cartilage in the culture medium in the agarose mold and keeping the temperature at 37˚C. Since this agarose mold also buffers against mechanical forces loaded on the three-dimensional regenerative tissue, it may be useful as a container for storage and transport of large-sized three-dimensional regenerative tissue.

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The application of atelocollagen gel in combination with porous scaffolds for cartilage tissue engineering and its suitable conditions

Cited by 26 publications

References 36 publications

Application of floating cells for improved harvest in human chondrocyte culture

Application of floating cells for improved harvest in human chondrocyte culture

Hollow Fiber Module Applied for Effective Proliferation and Harvest of Cultured Chondrocytes

Usefulness of Agarose Mold as a Storage Container for Three-Dimensional Tissue-Engineered Cartilage

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