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

Int Orthop. 2013 Nov;37(11):2297-303. doi: 10.1007/s00264-013-2009-z. Epub 2013 Jul 17.

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.

Publication types

  • Evaluation Study

MeSH terms

  • Adult
  • Age Factors
  • Aggrecans / genetics
  • Aggrecans / metabolism
  • Bone Transplantation / methods*
  • Cartilage, Articular / cytology
  • Cartilage, Articular / metabolism*
  • Chondrocytes / cytology
  • Chondrocytes / metabolism
  • Chondrocytes / transplantation*
  • Chondrogenesis / genetics*
  • Chondrogenesis / physiology*
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • Cryopreservation / methods*
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / metabolism
  • Female
  • Gene Expression / genetics
  • Gene Expression / physiology*
  • Humans
  • Interleukin-1beta / genetics
  • Interleukin-1beta / metabolism
  • Male
  • Middle Aged
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Time Factors
  • Tissue Engineering
  • Versicans / genetics
  • Versicans / metabolism

Substances

  • Aggrecans
  • Collagen Type I
  • Extracellular Matrix Proteins
  • Interleukin-1beta
  • MIA protein, human
  • Neoplasm Proteins
  • Versicans