Experimental Analysis of Tear Fluid and Its Processing for the Diagnosis of Multiple Sclerosis

Vladimíra Tomečková; Soňa Tkáčiková; Ivan Talian; Gabriela Fabriciová; Andrej Hovan; Daria Kondrakhova; Katarína Zakutanská; Miriama Skirková; Vladimír Komanický; Natália Tomašovičová

doi:10.3390/s23115251

Experimental Analysis of Tear Fluid and Its Processing for the Diagnosis of Multiple Sclerosis

Sensors (Basel). 2023 Jun 1;23(11):5251. doi: 10.3390/s23115251.

Affiliations

¹ Department of Medical and Clinical Biochemistry, Faculty of Medicine, Pavol Jozef Šafárik University in Košice, Trieda SNP 1, 040 11 Košice, Slovakia.
² Department of Medical and Clinical Biophysics, Faculty of Medicine, Pavol Jozef Šafárik University in Košice, Trieda SNP 1, 040 11 Košice, Slovakia.
³ Department of Biophysics, Institute of Physics, Faculty of Science, Pavol Jozef Šafárik University in Košice, Jesenná 5, 041 54 Košice, Slovakia.
⁴ Department of Condensed Matter Physics, Institute of Physics, Faculty of Science, Pavol Jozef Šafárik University in Košice, Park Angelinum 9, 041 54 Košice, Slovakia.
⁵ Department of Magnetism, Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 040 01 Košice, Slovakia.
⁶ Department of Opthalmology, Faculty of Medicine, Pavol Jozef Šafárik University in Košice, Trieda SNP 1, 040 11 Košice, Slovakia.

Abstract

A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic-force microscopy. Infrared spectroscopy found no significant difference between the tear fluid of MS patients and the control spectra; all three significant peaks were located at around the same positions. Raman analysis showed differences between the spectra of the tear fluid of MS patients and the spectra of healthy subjects, which indicated a decrease in tryptophan and phenylalanine content and changes in the relative contributions of the secondary structures of the polypeptide chains of tear proteins. Atomic-force microscopy exhibited a surface fern-shaped dendrite morphology of the tear fluid of patients with MS, with less roughness on both oriented silicon (100) and glass substrates compared to the tear fluid of control subjects. The results of liquid chromatography-mass spectrometry showed downregulation of glycosphingolipid metabolism, sphingolipid metabolism, and lipid metabolism. Proteomic analysis identified upregulated proteins in the tear fluid of patients with MS such as cystatine, phospholipid transfer protein, transcobalamin-1, immunoglobulin lambda variable 1-47, lactoperoxidase, and ferroptosis suppressor protein 1; and downregulated proteins such as haptoglobin, prosaposin, cytoskeletal keratin type I pre-mRNA-processing factor 17, neutrophil gelatinase-associated lipocalin, and phospholipase A2. This study showed that the tear proteome in patients with MS is modified and can reflect inflammation. Tear fluid is not a commonly used biological material in clinico-biochemical laboratories. Experimental proteomics has the potential to become a promising contemporary tool for personalized medicine, and it might be applied in clinical practice by providing a detailed analysis of the tear-fluid proteomic profile of patients with MS.

Keywords: Raman spectroscopy; atomic force microscopy; infrared spectroscopy; multiple sclerosis; proteomics; tear fluid.

MeSH terms

Chromatography, Liquid
Humans
Mass Spectrometry
Multiple Sclerosis* / diagnosis
Proteomics* / methods
Tears / chemistry

Grants and funding

This research was funded by a grant of the Slovak Research and Development Agency under the contract APVV-20-0324 and by a grant of the Research Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic under Grant: Integrative strategy in the development of personalized medicine of selected cancers and its impact on quality of life, ITMS2014+: 313011V446.