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Analytical methods : advancing methods and applications2022; 14(37); 3661-3670; doi: 10.1039/d2ay00779g

Optical photothermal infrared spectroscopy can differentiate equine osteoarthritic plasma extracellular vesicles from healthy controls.

Abstract: Equine osteoarthritis is a chronic degenerative disease of the articular joint, characterised by cartilage degradation resulting in pain and reduced mobility and thus is a prominent equine welfare concern. Diagnosis is usually at a late stage through clinical examination and radiographic imaging, whilst treatment is symptomatic not curative. Extracellular vesicles are nanoparticles that are involved in intercellular communication. The objective of this study was to investigate the feasibility of Raman and Optical Photothermal Infrared Spectroscopies to detect osteoarthritis using plasma-derived extracellular vesicles, specifically differentiating extracellular vesicles in diseased and healthy controls within the parameters of the techniques used. Plasma samples were derived from thoroughbred racehorses. A total of 14 samples were selected (control; = 6 and diseased; = 8). Extracellular vesicles were isolated using differential ultracentrifugation and characterised using nanoparticle tracking analysis, transmission electron microscopy, and human tetraspanin chips. Samples were then analysed using combined Raman and Optical Photothermal Infrared Spectroscopies. Infrared spectra were collected between 950-1800 cm. Raman spectra had bands between the wavelengths of 900-1800 cm analysed. Spectral data for both Raman and Optical Photothermal Infrared Spectroscopy were used to generate clustering principal components analysis and classification models were generated using partial least squared discriminant analysis in order to characterize the techniques' ability to distinguish diseased samples. Optical Photothermal Infrared Spectroscopy could differentiate osteoarthritic extracellular vesicles from healthy with good classification (93.4% correct classification rate) whereas Raman displayed poor classification (correct classification rate = -64.3%). Inspection of the infrared spectra indicated that plasma-derived extracellular vesicles from osteoarthritic horses contained increased signal for proteins, lipids and nucleic acids. For the first time we demonstrated the ability to use optical photothermal infrared spectroscopy combined with Raman spectroscopy to interrogate extracellular vesicles and osteoarthritis-related samples. Optical Photothermal Infrared Spectroscopy was superior to Raman in this study, and could distinguish osteoarthritis samples, suggestive of its potential use diagnostically to identify osteoarthritis in equine patients. This study demonstrates the potential of Raman and Optical Photothermal Infrared Spectroscopy to be used as a future diagnostic tool in clinical practice, with the capacity to detect changes in extracellular vesicles from clinically derived samples.
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Publication Date: 2022-09-29 PubMed ID: 36066093PubMed Central: PMC9521322DOI: 10.1039/d2ay00779gGoogle 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 research investigates the use of Raman and Optical Photothermal Infrared Spectroscopies to detect equine osteoarthritis in plasma-derived extracellular vesicles. The study showed that Optical Photothermal Infrared Spectroscopy could effectively differentiate osteoarthritic extracellular vesicles from healthy ones.

Objective and Methodology

  • The primary aim of this research was to study the feasibility of using Raman and Optical Photothermal Infrared Spectroscopies for the detection of osteoarthritis in horses through analysis of plasma-derived extracellular vesicles. The team aimed to categorize and distinguish vesicles in diseased samples and healthy controls, using certain parameters of the techniques.
  • The team chose samples from thoroughbred racehorses, with a total of 14 samples (6 control and 8 diseased). Plasma samples were analyzed after isolation using differential ultracentrifugation and characterization through nanoparticle tracking analysis, transmission electron microscopy, and human tetraspanin chips.

Data Analysis and Results

  • The team analyzed the samples using combined Raman and Optical Photothermal Infrared Spectroscopies. Infrared spectra were collected between 950-1800 cm, while the Raman spectra took into account bands between the wavelengths of 900-1800 cm.
  • Data from this spectral analysis were used in generating principal components analysis clusters and classification models. These were created using partial least squared discriminant analysis, which facilitated the characterization of the techniques’ ability to distinguish diseased samples.
  • The results showed that Optical Photothermal Infrared Spectroscopy could distinguish osteoarthritic extracellular vesicles from healthy ones quite accurately (93.4% correct classification rate). However, Raman displayed poor classification, with a negative correct classification rate.

Implications and Conclusions

  • The signal increase for proteins, lipids, and nucleic acids in the infrared spectra showed that plasma-derived extracellular vesicles from osteoarthritic horses had increased contents of these physical constituents.
  • The study proved the potential of Optical Photothermal Infrared Spectroscopy, combined with Raman spectroscopy, in analyzing extracellular vesicles and osteoarthritis-related samples. The former was superior in distinguishing osteoarthritis samples, indicating its potential diagnostic use for identifying osteoarthritis in equine patients.
  • The study’s findings suggest that these spectral analysis techniques could become valuable tools for future diagnostics in clinical practice, having been demonstrated to effectively detect changes in clinically derived samples of extracellular vesicles.

Cite This Article

APA
Clarke EJ, Lima C, Anderson JR, Castanheira C, Beckett A, James V, Hyett J, Goodacre R, Peffers MJ. (2022). Optical photothermal infrared spectroscopy can differentiate equine osteoarthritic plasma extracellular vesicles from healthy controls. Anal Methods, 14(37), 3661-3670. https://doi.org/10.1039/d2ay00779g

Publication

Analytical methods : advancing methods and applications
ISSN: 1759-9679
NlmUniqueID: 101519733
Country: England
Language: English
Volume: 14
Issue: 37
Pages: 3661-3670

Researcher Affiliations

Clarke, Emily J
  • Department of Musculoskeletal Biology and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, William Henry Duncan Building, 6 W Derby St, Liverpool L7 8TX, UK. eclarke@liverpool.ac.uk.
Lima, Cassio
  • Centre for Metabolomics Research, Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool, L69 7BE, UK.
Anderson, James R
  • Department of Musculoskeletal Biology and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, William Henry Duncan Building, 6 W Derby St, Liverpool L7 8TX, UK. eclarke@liverpool.ac.uk.
Castanheira, Catarina
  • Department of Musculoskeletal Biology and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, William Henry Duncan Building, 6 W Derby St, Liverpool L7 8TX, UK. eclarke@liverpool.ac.uk.
Beckett, Alison
  • Biomedical Electron Microscopy Unit, University of Liverpool, UK.
James, Victoria
  • School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Loughborough LE12 5RD, UK.
Hyett, Jacob
  • Department of Musculoskeletal Biology and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, William Henry Duncan Building, 6 W Derby St, Liverpool L7 8TX, UK. eclarke@liverpool.ac.uk.
Goodacre, Royston
  • Centre for Metabolomics Research, Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool, L69 7BE, UK.
Peffers, Mandy J
  • Department of Musculoskeletal Biology and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, William Henry Duncan Building, 6 W Derby St, Liverpool L7 8TX, UK. eclarke@liverpool.ac.uk.

MeSH Terms

  • Animals
  • Extracellular Vesicles
  • Horses
  • Humans
  • Lipids
  • Nucleic Acids
  • Osteoarthritis / diagnosis
  • Osteoarthritis / veterinary
  • Spectroscopy, Near-Infrared / methods

Grant Funding

  • MR/P020941/1 / Medical Research Council
  • 107471/Z/15/Z / Wellcome Trust
  • Versus Arthritis

Conflict of Interest Statement

The authors declare no conflict of interest.

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