Misaki Yagi , Kentaro Endo, Keiichiro Komori and Ichiro Sekiya. Sci. Rep. 13, 7697 (2023).
https://doi.org/10.1038/s41598-023-34825-1
Copyright: © Yagi, M., Endo, K., Komori, K. and Sekiya, I. 2023
This article is licensed under a Creative Commons Attribution 4.0 International License (CC BY).
Background
Osteoarthritis (OA) is a condition characterized by degeneration and deformity of joint tissues, such as bones and in joint cartilage and has been associated with the impairing the development of cartilage. It is an age-related disease that can significantly impact quality of life. The loss of homeostasis of OA, and it has been found that chondrocyte senescence is an important factor in the onset and progression of the disease. In this study, effects of oxidative and inflammatory stresses on rat chondrocyte senescence have been investigated.
Research Achievements
From this study, it showed that oxidative and inflammatory stresses increased the SA‑β‑gal activity in rat chondrocytes. Oxidative stress impaired ECM‑producing ability and inhibited the proliferation of rat chondrocytes, while inflammatory stress increased the expression of SASP* factors and the degradation of ECM as well. SASP is the most important senescence phenotype in OA progression because it triggers an imbalance in ECM production and degradation. These findings suggest that inflammatory stress, rather than oxidative stress, is the dominant driver of SASP in OA.
Use of PrimeSurface in this Study
Rat chondrocyte 3D spheroids were formed using PrimeSurface® 96U plates. Oxidative (H2O2) and inflammatory (IL/TNF) stresses were added before or after spheroid formation and effects (production and degradation) on cartilage ECM were evaluated.
- Cell density: 2.5×104 cells/well
- Medium: chondrogenic medium
- Culture period: 2 weeks
- Stress treatment: 5 days
- Analysis method: histological staining and Glycosaminoglycan (GAG) & DNA Quantification
Results：
Fig. 5, 6
(A) Experimental schema
(B) Immunostaining
(C) Quantification of GAG and DNA
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