physiological tonicity improves human chondrogenic marker expression through nuclear factor of activated t-cells 5 in vitro生理紧张性改善人类chondrogenic标记表达式通过核转录因子的激活t细胞5体外.pdf

van der Windt et al. Arthritis Research Therapy 2010, 12:R100 /content/12/3/R100 R E S E A R C H A R T I C L E Open Access Research article Physiological tonicity improves human chondrogenic marker expression through nuclear factor of activated T-cells 5 in vitro 1 1 1 1 2 2 Anna E van der Windt , Esther Haak , Ruud HJ Das , Nicole Kops , Tim JM Welting , Marjolein MJ Caron , Niek P van 3 1 1 1 Til , Jan AN Verhaar , Harrie Weinans and Holger Jahr* Abstract Introduction: Chondrocytes experience a hypertonic environment compared with plasma (280 mOsm) due to the high fixed negative charge density of cartilage. Standard isolation of chondrocytes removes their hypertonic matrix, exposing them to nonphysiological conditions. During in vitro expansion, chondrocytes quickly lose their specialized phenotype, making them inappropriate for cell-based regenerative strategies. We aimed to elucidate the effects of tonicity during isolation and in vitro expansion on chondrocyte phenotype. Methods: Human articular chondrocytes were isolated and subsequently expanded at control tonicity (280 mOsm) or at moderately elevated, physiological tonicity (380 mOsm). The effects of physiological tonicity on chondrocyte proliferation and chondrogenic marker expression were evaluated. The role of Tonicity-responsive Enhancer Binding Protein in response to physiological tonicity was investigated using nuclear factor of activated T-cells 5 (NFAT5) RNA interference. Results: Moderately elevated, physiological tonicity (380 mOsm) did not affect chondrocyte proliferation, while higher tonicities inhibited proliferation and diminished cell viability. Physiological to