University of Twente Student Theses


The Influence of Hydrogel, Temperature, Oxygen, and Cytokines on Human Chondrocytes using in vitro Culture-Based Models

Hengel, Eline van (2021) The Influence of Hydrogel, Temperature, Oxygen, and Cytokines on Human Chondrocytes using in vitro Culture-Based Models.

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Abstract:Osteoarthritis is one of the most common knee cartilage diseases affecting over 1,4 million people in the Netherlands in 2018 and results in cartilage destruction and impaired mobility. Due to the unique structure and characteristics of articular cartilage, there is currently no appropriate in vitro model available that fully mimics the in vivo situation, limiting the research on disease progress and drug development. Important requirements for a culture model are the low physiological temperature (32,6 °C) and oxygen levels (2,5% O2) of cartilage, and the ability to culture in a dynamic three-dimensional environment, enabling both mechanical and biochemical stimuli. In this study, the aim was to establish a chondrocyte in vitro culture system which comprises all required factors. To this end, the project was split into two sections: the first part to study hydrogels for a dynamic culture, the second part to study a physiological culture environment. Current protocols rely on agarose as a matrix for chondrocyte culture, but this gel lacks various features, such as cell attachment sites and the ability for cells to proliferate. Therefore, the use of Dex-TA/HA-TA hydrogel was evaluated for a three-dimensional dynamic culture system. Dex/HA chondrocytes showed viability, maintenance of the chondrogenic phenotype, motility, and enhanced production of degradative factors, suggesting the potential as culture matrix for a better representation of in vivo cartilage, as an alternative to agarose. Secondly, we determined the effect of temperature 33 °C (intra-articular), 37 °C (body core), and 39 °C (hyperthermic cartilage) under hypoxic circumstances for monolayer and pellet cultures. A culture temperature between 33 °C and 37 °C under hypoxic circumstances was optimal for chondrocyte culture; positively influencing the chondrocyte-specific genes; cartilage development; and the maintenance of the differentiated phenotype. Lastly, under the same temperature and oxygen conditions, an inflammatory response was triggered by pro-inflammatory cytokine stimulation. The effects of cytokine stimulation were enhanced under hypoxic conditions, resulting in a stress response. In the future, a reliable platform for chondrocyte culture could be established by combining the Dex/HA hydrogel culture and physiological culture circumstances, this could be applied as a disease model for Osteoarthritis by hyper physiological mechanical and cytokine stimulation.
Item Type:Essay (Master)
University of Twente, Nederland
Faculty:TNW: Science and Technology
Subject:42 biology
Programme:Biomedical Engineering MSc (66226)
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