Osteochondritis Dissecans (OCD)-Derived Chondrocytes Display Increased Senescence, Oxidative Stress, Chaperone-Mediated Autophagy and, in Co-Culture with Adipose-Derived Stem Cells (ASCs), Enhanced Expression of MMP-13.
Abstract: Osteochondritis dissecans (OCD) in equids, especially in sport horses, has become a growing issue as it contributes to the occurrence of lameness. Thus the aim of this study was to investigate the cytophysiological properties of OCD chondrocytes including expression of chondrogenic genes, apoptosis, mitochondria dynamics and autophagy. Horse chondrocytes were isolated from healthy (HE) and OCD cartilages. Properties of cells were evaluated using multiple assays e.g., polymerase chain reaction (PCR), immunofluorescence, Western blot. OCD chondrocytes were characterized by increased apoptosis and senescence. Expression of chondrogenic genes (vimentin, aggrecan) was decreased while mRNA levels of matrix metalloproteinase 13 significantly upregulated in comparison to HE cells. Moreover, OCD cells displayed increased mitochondrial fusion while fission events were diminished. Interestingly, chaperone mediated autophagy was triggered in those cells and it predominated over macroautophagy. Furthermore, co-culture of LPS-treated chondrocytes with adipose-derived stem cells (ASC) decreased p62/sequestosome 1 (SQSTM) and increases MMP-13 expression in OCD cells. Our results suggest that OCD affected horse chondrocytes are characterized by senescent phenotype due to endoplasmic reticulum stress and mitochondria dynamics deterioration. Expression of chondrogenic markers is decreased in those cells while expression of chaperone mediated autophagy (CMA)-related genes increased. Increased malfunctioning of cells leads to loss of their functionality and capacity to maintain tissue homeostasis.
Publication Date: 2019-03-08 PubMed ID: 30857162PubMed Central: PMC6462951DOI: 10.3390/jcm8030328Google Scholar: Lookup
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Summary
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This research studied the behavior of cells affected by Osteochondritis dissecans (OCD) in horses, finding that these cells display increased senescence and oxidative stress, and when paired with adipose-derived stem cells, the expression of matrix metalloproteinase 13 is enhanced.
Objective of the Study
- The main focus of the study was the investigation of the physiological behavior of chondrocytes (cartilage cells) sourced from horses suffering from Osteochondritis dissecans (OCD), a joint disorder prevalent in athletic horses leading to lameness.
- Specific objectives included studying the expression of cartilage-specific genes, cell death mechanisms, dynamics of mitochondria (the energy factory of cells), and cell self-digestion process called autophagy.
Methodology
- Horse chondrocytes were extracted from both healthy and OCD-affected cartilages.
- These cells were analyzed using various techniques such as polymerase chain reaction (PCR) and immunofluorescence to study gene expression and visualize proteins of interest, and Western blotting to detect specific proteins.
- Certain cells were also co-cultured alongside adipose-derived stem cells (ASC), and the effects were observed.
Findings
- The study found that cells from OCD-affected cartilage showed higher rates of apoptosis (programmed cell death) and cellular aging (senescence).
- There was a decrease in the expression of key cartilage-related genes such as vimentin and aggrecan, while the level of matrix metalloproteinase 13, a molecule involved in the degrading of extracellular components, was significantly increased.
- The study also revealed increased merger of mitochondria in OCD cells while the splitting events were reduced, implying disruptive energy metabolism within these cells.
- Furthermore, increased chaperone-mediated autophagy was observed, a process wherein specific proteins are targeted and transported into lysosomes for degradation, indicating enhanced catabolic activity.
- Upon co-culturing with ASCs, a decline in the p62/ SQSTM protein, but an increase in the matrix metalloproteinase 13 expression was noticed in the OCD cells.
Conclusion
- The research concludes that cells affected by OCD in horses show an accelerated aging phenotype, attributed to stress in the endoplasmic reticulum (organism in cells that assists in protein folding and lipid metabolism) and disruptions in mitochondria dynamics.
- Such cells display diminished expression of vital cartilage markers while simultaneously showing increased expression of genes linked to chaperone-mediated autophagy.
- These dysfunctional features may lead to the impairment of the cells’ functionality and their capacity to maintain balance in tissue functionality, thereby potentially contributing to the progression of OCD.
Cite This Article
APA
Kornicka K, Al Naem M, Röcken M, Zmiertka M, Marycz K.
(2019).
Osteochondritis Dissecans (OCD)-Derived Chondrocytes Display Increased Senescence, Oxidative Stress, Chaperone-Mediated Autophagy and, in Co-Culture with Adipose-Derived Stem Cells (ASCs), Enhanced Expression of MMP-13.
J Clin Med, 8(3), 328.
https://doi.org/10.3390/jcm8030328 Publication
Researcher Affiliations
- International Institute of Translational Medicine, Jesionowa, 11, Malin, 55-114 Wisznia Mała, Poland. kornicka.katarzyna@gmail.com.
- Department of Experimental Biology, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland. kornicka.katarzyna@gmail.com.
- Faculty of Veterinary Medicine, Equine Clinic-Equine Surgery, Justus-Liebig-University, 35392 Gießen, Germany. mohamad.al-naem@vetmed.uni-giessen.de.
- Faculty of Veterinary Medicine, Equine Clinic-Equine Surgery, Justus-Liebig-University, 35392 Gießen, Germany. Michael.Roecken@vetmed.uni-giessen.de.
- International Institute of Translational Medicine, Jesionowa, 11, Malin, 55-114 Wisznia Mała, Poland. zmiertka.marta@gmail.com.
- International Institute of Translational Medicine, Jesionowa, 11, Malin, 55-114 Wisznia Mała, Poland. krzysztofmarycz@interia.pl.
- Department of Experimental Biology, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland. krzysztofmarycz@interia.pl.
- Faculty of Veterinary Medicine, Equine Clinic-Equine Surgery, Justus-Liebig-University, 35392 Gießen, Germany. krzysztofmarycz@interia.pl.
Conflict of Interest Statement
The authors declare no conflict of interest.
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