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

Abstract

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.

Combination of Polysaccharide Nanofibers Derived from Cellulose and Chitin Promotes the Adhesion, Migration and Proliferation of Mouse Fibroblast Cells

“…At the beginning, intact collagen from animal sources was used to promote cell attachment and angiogenesis in tissue engineering [ 4 ]. In a similar way, natural hyaluronan was also effective for wound healing when cultured with chondrocytes around damaged human cartilages [ 5 ]. However, such ECM components are often derived from animal origins, resulting in a risk of various infectious diseases and immune responses [ 6 ].…”

Extracellular matrix production in vitro in cartilage tissue engineering

“…Besides dynamic environment mentioned above, cryopreservation has a negative effect on the chondrogenic capacity of chondrocytes. Human chondrocytes showed significantly decreased collagen type II expression after cryopreservation [ 86 ]. Although new methods and scaffolds are being under investigation for the preservation and revitalization of articular cartilage or chondrocytes [ 87 - 89 ], it is better to use freshly cultured chondrocytes for clinical application if it is possible.…”