Biological freezing of human articular chondrocytes

Almqvist, Karl; Wang, L; Broddelez, C; Em, Veys; Verbruggen, Gust

doi:10.1053/joca.2000.0394

Cited by 30 publications

(18 citation statements)

References 30 publications

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“…Despite low cell viability observed after thawing, cryopreservation allowed for storage of chondrocytes for up to six months with little functional alterations, similar to that found by Almqvist et al (2001) who froze chondrocytes in a culture medium containing DMSO at the same concentration used here.…”

Section: Discussionsupporting

confidence: 78%

Platelet rich plasma associated with heterologous fresh and thawed chondrocytes on osteochondral lesions of rabbits

Filgueiras

Carlo

Alves³

et al. 2014

Arq. Bras. Med. Vet. Zootec.

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Chondrocytes obtained from stifle joint of New Zealand White rabbits were cultivated. Half of cells were maintained in culture for later implantation and the others frozen during six months to evaluate viability. A circular osteochondral defect was created in the right stifle of other twenty seven rabbits. The control group (CG) received no treatment. The thawed (TH) and fresh (FH) heterologous groups received, respectively, an implant of cultivated thawed or fresh heterologous chondrocytes associated with platelet rich plasma (PRP). The CG group showed greatest pain and lameness compared to the other groups seven days after the implantation. Microscopically, at 45 and 90 days, the TH and FH groups showed filling with cartilaginous tissue containing chondrocytes surrounded by a dense matrix of glycosaminoglycans. In the CG group, healing occurred with vascularized fibrous connective tissue without integration to the subchondral bone. Cryopreserved heterologous chondrocytes were viable for implantation and healing of osteochondral lesions; the association with PRP allows the fixation of cells in the lesion and offers growth factors which accelerates repair with tissue similar to articular hyaline cartilage.Keywords: rabbits, articular cartilage, chondrocyte implantation, platelet rich plasma, type II collagen RESUMO Cultivaram-se condrócitos obtidos da articulação do joelho de coelhos. Metade das células foi mantida em cultura para posterior implantação, e a outra metade foi congelada durante seis meses com a finalidade de avaliar a viabilidade. Criou-se um defeito circular osteocondral no joelho direito de outros vinte e sete coelhos. O grupo controle (GC) não recebeu tratamento. Os grupos descongelado (TH) e fresco (FH) receberam, respectivamente, implantes heterólogos de condrócitos cultivados descongelados e frescos, associados com PRP. O grupo GC apresentou maior dor e claudicação em comparação com os

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Section: Discussionsupporting

confidence: 78%

Platelet rich plasma associated with heterologous fresh and thawed chondrocytes on osteochondral lesions of rabbits

Filgueiras

Carlo

Alves³

et al. 2014

Arq. Bras. Med. Vet. Zootec.

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“…Only a few studies have been published that study the affect of cryopreservation on chondrocyte phenotype, with somewhat conflicting results. It has been reported that total aggrecan synthesis was not affected by cryopreservation of chondrocytes [20]. Further, COL2 production has also been demonstrated in cryopreserved chondrocytes [21].…”

Section: Discussionmentioning

confidence: 94%

Influence of cryopreservation, cultivation time and patient’s age on gene expression in Hyalograft® C cartilage transplants

Albrecht

Tichy

Nürnberger

et al. 2013

International Orthopaedics (SICOT)

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Purpose Our aim was to evaluate the impact of cryopreservation, cultivation time and patient's age on the expression of specific chondrogenic markers in Hyalograft® C transplants. Methods Gene expression of chondrocyte markers [collagen type I (COL1A1), COL2A1, aggrecan, versican, melanoma inhibitory activity (MIA) and interleukin (IL)-1β] was analysed in cartilage biopsies (n=17) and Hyalograft® C transplant samples (non-cryopreserved=78, cryopreserved=13) by quantitative real-time polymerase chain reaction (PCR). Correlation analyses were performed to evaluate the influence of the above-named parameters on the level of gene expression. Results Cryopreservation of cells was found to decrease COL2A1 and MIA significantly (4.6-fold, p<0.01 and 2-fold, p<0.045, respectively). The duration of cryopreservation had no further influence on the expression of these factors. No correlation was detected between cultivation time (75±31 days) and the expression level of any gene. Cartilage transplants from older patients (>35 years) exhibited a significantly higher IL-1β expression (3.7-fold, p<0.039) than transplants from younger patients (≤35 years). Conclusions Our data demonstrate that cryopreservation has a profound impact on chondrocyte metabolic activity by decreasing the expression of COL2A1 and MIA in Hyalograft® C transplants, independent of the duration of cryopreservation.

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“…Almqvist et al (2001) used the same concentration for freezing human chondrocytes for 24 hours and did not observe alterations in cellular viability or matrix production. Nevertheless, in this study, freezing time was six months, which may have caused the low viability immediately after thawing all samples.…”

Section: Discussionmentioning

confidence: 99%

“…However, during freezing, there is a possibility of intracellular crystals formation, and they may break the cell membrane. The addition of a cryoprotector to the culture medium, such as dimethylsulfoxide (DMSO) and serum proteins from fetal bovine serum (FBS), protect the cells from thermal and osmotic shock (Almqvist et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Effect of freezing on rabbit cultured chondrocytes

Filgueiras¹,

Carlo²,

Alves³

et al. 2011

Arq. Bras. Med. Vet. Zootec.

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This work evaluated the effect of freezing on chondrocytes maintained in culture, aiming the establishment of a cell bank for future application as heterologous implant. Chondrocytes extracted from joint cartilage of nine healthy New Zealand White rabbits were cultivated and frozen with the cryoprotector 5% dimethylsulfoxide for six months. Phenotypic and scanning electron microscopy analyses were carried out to identify morphological and functional differences between fresh and thawed cells. After enzymatic digestion, a total of 4.8x10 5 cells per rabbit were obtained. Fresh chondrocytes showed a high mitotic rate and abundant matrix was present up to 60 days of culture. Loss of phenotypic stability was notable in the thawed chondrocytes, with a low labeling of proteoglycans and weak immunostaining of type II collagen. The present study showed important loss of chondrocyte viability under the freezing conditions. For future in vivo studies of heterologous implant, these results suggests that a high number of cells should be implanted in the host site in order to achieve an adequate number of viable cells. Furthermore, the chondrocytes should be implanted after two weeks of culture, when the highest viability rate is found.

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Biological freezing of human articular chondrocytes

Cited by 30 publications

References 30 publications

Platelet rich plasma associated with heterologous fresh and thawed chondrocytes on osteochondral lesions of rabbits

Platelet rich plasma associated with heterologous fresh and thawed chondrocytes on osteochondral lesions of rabbits

Influence of cryopreservation, cultivation time and patient’s age on gene expression in Hyalograft® C cartilage transplants

Effect of freezing on rabbit cultured chondrocytes

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