Hydrostatic pressure-generated reactive oxygen species induce osteoarthritic conditions in cartilage pellet cultures

Bernhard Rieder,Sylvia Nürnberger,Andreas H Teuschl,Dominik Rünzler,Anna M Weihs,Carina Huber-Gries,Dorota Szwarc,Adelheid Weidinger,Heinz Redl

doi:10.1038/s41598-018-34718-8

Abstract

Osteoarthritis (OA) is one of the most common causes of disability and represents a major socio-economic burden. Despite intensive research, the molecular mechanisms responsible for the initiation and progression of OA remain inconclusive. In recent years experimental findings revealed elevated levels of reactive oxygen species (ROS) as a major factor contributing to the onset and progression of OA. Hence, we designed a hydrostatic pressure bioreactor system that is capable of stimulating cartilage cell cultures with elevated ROS levels. Increased ROS levels in the media did not only lead to an inhibition of glycosaminoglycans and collagen II formation but also to a reduction of already formed glycosaminoglycans and collagen II in chondrogenic mesenchymal stem cell pellet cultures. These effects were associated with the elevated activity of matrix metalloproteinases as well as the increased expression of several inflammatory cytokines. ROS activated different signaling pathways including PI3K/Akt and MAPK/ERK which are known to be involved in OA initiation and progression. Utilizing the presented bioreactor system, an OA in vitro model based on the generation of ROS was developed that enables the further investigation of ROS effects on cartilage degradation but can also be used as a versatile tool for anti-oxidative drug testing.

Highlights

Osteoarthritis (OA) is the most common type of arthritis, affecting 25% of the adult population
The addition of the iron-chelator led to significantly lower reactive oxygen species (ROS) generation, whereas the addition of superoxide dismutase (SOD) resulted in high levels of ROS
The conversion of SOD was driving the main reaction of Fe2+ and O2 to Fe3+ and superoxide, which resulted in high ROS levels (Fig. 3B)

Summary

Introduction

Osteoarthritis (OA) is the most common type of arthritis, affecting 25% of the adult population. It was believed that the loss of articular cartilage subsequently results in altered biomechanics combined with cellular changes which over time led to severe changes of the subchondral bone, synovium, menisci, ligaments, periarticular muscles and nerves[5]. This hypothesis is supported by results of in vivo models in which mechanical instability of the knee joint was induced, e.g. by transection of the anterior cruciate ligament[6,7] to promote excessive wear of cartilage structures. Different in vivo and in vitro OA models have been established to decipher the roles of specific factors contributing to the disease

Methods

Results

Conclusion