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Bone reports2018; 9; 19-26; doi: 10.1016/j.bonr.2018.06.003

Expression of pro-apoptotic markers is increased along the osteochondral junction in naturally occurring osteochondrosis.

Abstract: Osteochondrosis (OC) is a naturally occurring disease of the articular-epiphyseal cartilage and subchondral bone layers, leading to pain and decreased mobility. The objective of this study was to characterize gene and protein expression of apoptotic markers in chondrocytes surrounding cartilage canals and along the osteochondral junction of osteochondrosis (OC)-affected and normal cartilage, using naturally occurring disease in horses. Paraffin-embedded osteochondral samples (6 OC, 8 normal controls) and cDNA from chondrocytes captured with laser capture microdissection (4 OC, 6 normal controls) were obtained from the lateral trochlear ridge of femoropatellar joints in 14 immature horses (1-6 months of age). Equine-specific caspase-3, caspase-8, caspase-10, Fas, Bcl-2, BAG-1, TNFα, cytochrome , thymosin-β10, and 18S mRNA expression levels were evaluated by two-step real-time quantitative PCR. Percentage of cell death was determined using the TUNEL method. Protein expression of caspase-10, Fas, cytochrome , and thymosin-β10 was determined following immunohistochemistry. Statistical analysis was performed using the Wilcoxon rank sum test or two-sample -test ( < 0.05). In OC samples, there was significantly increased gene expression of caspase-10, Fas, cytochrome , and thymosin-β10 in chondrocytes along the osteochondral junction and increased Fas gene expression in chondrocytes adjacent to cartilage canals, compared to controls. In OC samples, higher matrix Fas and cytochrome protein expression, lower mitochondrial cytochrome protein expression, and a trend for higher cytoplasmic caspase-10 protein expression were found. Collectively, these results suggest that both extrinsic and intrinsic apoptotic pathways are activated in OC cartilage. Increased apoptosis of osteochondral junction chondrocytes may play a role in OC, based on increased gene expression of several pro-apoptotic markers in this location.
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Publication Date: 2018-06-21 PubMed ID: 29998174PubMed Central: PMC6038796DOI: 10.1016/j.bonr.2018.06.003Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study investigates the increased expression of pro-apoptotic markers, markers indicative of programmed cell death, near the osteochondral junction in cases of osteochondrosis (OC), a disease affecting the joints. The research utilized naturally occurring instances of OC in horses and controlled comparisons to evaluate gene and protein expressions by employing a quantitative approach and techniques like immunohistochemistry.

Research Objective and Procedures

  • The primary objective of the study was to compare gene and protein expressions of pro-apoptotic markers in chondrocytes, which are cells found in healthy cartilage, located near cartilage canals and along the osteochondral junction in normal and OC-affected cartilage. This analysis was conducted using naturally occurring cases of OC in horses.
  • Examination and comparison were made using samples from the femoropatellar joints of 14 immature horses (aged one to six months). The samples included 6 from OC-affected horses and 8 from healthy controls at the mRNA level. Protein expressions were evaluated using laser-captured microdissection from 4 OC-affected and 6 control chondrocytes.

Gathering and Analyzing Data

  • The researchers measured the expression levels of several equine specific mRNA, including caspase-3, caspase-8, caspase-10, Fas, Bcl-2, BAG-1, TNFα, cytochrome, thymosin-β10, and 18S by employing a two-step real-time quantitative PCR method.
  • Cell death percentage was determined using the TUNEL method. Protein expressions of caspase-10, Fas, cytochrome, and thymosin-β10 were observed through immunohistochemistry. To determine statistical relevance, they used the Wilcoxon rank sum test or two-sample t-test.

Results and Conclusion

  • The results showed that in OC samples, gene expressions of caspase-10, Fas, cytochrome, and thymosin-β10 were significantly increased in chondrocytes along the osteochondral junction compared to controls. Plus, Fas gene expression was increased in chondrocytes near cartilage canals.
  • Upon examining protein expression, it was found that OC samples had higher matrix Fas and cytochrome protein expressions, lower mitochondrial cytochrome protein expression, and a trend towards higher cytoplasmic caspase-10 protein expression.
  • The overall findings suggest a role of chondrocyte apoptosis or programmed cell death in osteochondrosis, both through intrinsic and extrinsic pathways. This conclusion was based on the observed increased expression of several pro-apoptotic markers in these specific locations within the osteochondral junction.

Cite This Article

APA
Semevolos SA, Duesterdieck-Zellmer KF, Larson M, Kinsley MA. (2018). Expression of pro-apoptotic markers is increased along the osteochondral junction in naturally occurring osteochondrosis. Bone Rep, 9, 19-26. https://doi.org/10.1016/j.bonr.2018.06.003

Publication

Bone reports
ISSN: 2352-1872
NlmUniqueID: 101646176
Country: United States
Language: English
Volume: 9
Pages: 19-26

Researcher Affiliations

Semevolos, Stacy A
  • Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
Duesterdieck-Zellmer, Katja F
  • Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
Larson, Maureen
  • Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
Kinsley, Marc A
  • Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.

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Citations

This article has been cited 3 times.
  1. Grissom SK, Semevolos SA, Duesterdieck-Zellmer K. Role of cartilage and bone matrix regulation in early equine osteochondrosis. Bone Rep 2023 Jun;18:101653.
    doi: 10.1016/j.bonr.2023.101653pubmed: 36632355google scholar: lookup
  2. Chiaradia E, Pepe M, Orvietani PL, Renzone G, Magini A, Sforna M, Emiliani C, Di Meo A, Scaloni A. Proteome Alterations in Equine Osteochondrotic Chondrocytes. Int J Mol Sci 2019 Dec 7;20(24).
    doi: 10.3390/ijms20246179pubmed: 31817880google scholar: lookup
  3. Kornicka K, Al Naem M, Röcken M, Zmiertka M, Marycz K. 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 2019 Mar 8;8(3).
    doi: 10.3390/jcm8030328pubmed: 30857162google scholar: lookup
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