Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2021, 165(4):367-374 | DOI: 10.5507/bp.2021.020

Alpha-defensins determination in different types of synovial fluid and parallel collected serum samples by ELISA

Pavlina Kusnierovaa, Iveta Bystronovaa, b, Pavel Walderc, Rudolf Hlubekc, Frantisek Vsianskya, David Stejskala
a Institute of Laboratory Diagnostics, Department of Clinical Biochemistry, University Hospital Ostrava, Czech Republic
b Department of Epidemiology and Public Health, Faculty of Medicine, University of Ostrava, Czech Republic
c Orthopedic Department, University Hospital Ostrava, Czech Republic

Aims: The study aims were to verify the serum (S) and synovial fluid (SF) reference intervals (RIs) for human neutrophil defensins (HNP1-3); measure S and SF defensin concentrations in different types of SF, including non-inflammatory, inflammatory non-pyogenic, inflammatory pyogenic, and hemorrhagic; and to compare the HNP1-3 concentrations in SF and S with those of other inflammatory biomarkers.

Methods: SF and S samples were collected from 92 patients. HNP1-3 concentrations were determined using enzyme-linked immunosorbent assays; glucose, lactate, interleukin-6, and procalcitonin using an automatic analyzer; and presepsin using a Pathfast system. There were 61 non-inflammatory, 11 inflammatory non-pyogenic, 11 inflammatory pyogenic, and 9 hemorrhagic SF. Non-inflammatory SF was divided into non-inflammatory normal and non-inflammatory osteoarthritis. The former was used to estimate the HNP1-3 RI in SF and S.

Results: The estimated HNP1-3 RIs of SF and S were 12.47-437.42 mg/L and 5.45-44.75 µg/L, respectively. HNP1-3 differed significantly between S and SF and individual groups of SF (P<0.001 and P=0.001, respectively). There were significant relationships between SF HNP1-3 and S HNP1-3 (P<0.001), S C-reactive protein (P<0.001), and S interleukin-6 (P=0.007), and between SF HNP1-3 and SF C-reactive protein (P=0.004) and SF interleukin-6 (P<0.001). The highest kappa coefficient was between SF HNP1-3 and SF interleukin-6 (κ=0.507).

Conclusions: We validated the SF HNP1-3 diagnostic kit and demonstrated that SF and S HNP1-3 are promising biomarkers for distinguishing inflammatory from non-inflammatory joint diseases.

Keywords: alpha-defensins, synovial fluid, inflammatory joint disease, enzyme-linked immunosorbent assay

Received: June 10, 2020; Revised: April 6, 2021; Accepted: April 7, 2021; Prepublished online: April 19, 2021; Published: November 23, 2021  Show citation

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Kusnierova, P., Bystronova, I., Walder, P., Hlubek, R., Vsiansky, F., & Stejskal, D. (2021). Alpha-defensins determination in different types of synovial fluid and parallel collected serum samples by ELISA. Biomedical papers, 165(4), 367-374. doi: 10.5507/bp.2021.020
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    References

    1. Dungl P. Orthopedics, Grada Publishing, a.s. 2014; 23-25. ISBN 978-80-247-4357-8.
    2. Selsted ME, Harwig SS, Ganz T, Schilling JW, Lehrer RI. Primary structures of three human neutrophil defensins. J Clin Invest 1985;76:1436-39. Go to original source... Go to PubMed...
    3. Ganz T, Selsted ME, Szklarek D, Harwig SS, Daher K, Bainton DF, Lehrer RI. Defensins. Natural peptide antibiotics of human neutrophils. J Clin Invest 1985;76:1427-35. Go to original source... Go to PubMed...
    4. Zhang Y, Lu W, Hong M. The membrane-bound structure and topology of a human α-defensin indicate a dimer pore mechanism for membrane disruption. Biochemistry 2010;49(45):9770-82. Go to original source... Go to PubMed...
    5. Human HNP1-3 ELISA Kit, Product information and manual, HycultBiotech, Edition 08-16. [online]. In: https://www.hycultbiotech.com/hk317-01 [cit. 2020-10-06].].
    6. McHugh ML. Interrater reliability: the kappa statistic. Biochemia Medica 2012;22(3):276-82. Go to original source...
    7. Stata Version 13, Stata Press Publication, Texas, 2013. ISBN-13:978-1-59718-129-7.
    8. Clinical and Laboratory Standards Institute (CLSI). Defining, establishing, and verifying reference intervals in the clinical laboratory; approved guideline. CLSI document C28 - A3. 3rd ed. Wayne: PA: Clinical and Laboratory Standards Institute; 2008.
    9. CRP Latex, Beckman Coulter, Inc. 250S. Kraemer Blvd., Brea, CA 92821 USA. Version: 2013-07.
    10. Interleukin 6 (IL6), Siemens Healthare Diagnostics Inc. Version: RPBL1223/R4_CS Rev. D, 2019-10.
    11. Kemik O, Kemik AS, Sumer A, Begenik H, Purisa S, Tuzun S. Human neutrophil peptides 1, 2 and 3 (HNP 1-3): elevated serum levels in colorectal cancer and novel marker of lymphatic and hepatic metastasis. Hum Exp Toxicol 2013;32(2):167-71. Go to original source... Go to PubMed...
    12. Qi YY, Zhou XJ, Cheng FJ, Zhang H. Elevated plasma α-Defensins (HNP1-3) levels correlated with IgA1 glycosylation and susceptibility to IgA nephropathy. Dis Markers 2016;2016:8123138. Go to original source... Go to PubMed...
    13. Melicherčík P, Klapková E, Kotaška K, Jahoda D, Landor I, Čeřovský V. High-performance liquid chromatography as a novel method for the determination of α-Defensins in synovial fluid for diagnosis of orthopedic infections. Diagnostics (Basel) 2020;10(1):33. Go to original source... Go to PubMed...
    14. Saleh A, George J, Faour M, Klika AK, Higuera CA. Serum biomarkers in periprosthetic joint infections. Bone Joint Re. 2018;7(1):85-93. Go to original source... Go to PubMed...
    15. Cipriano CA, Brown NM, Michael AM, Moric M, Sporer SM, Della Valle CJ. Serum and synovial fluid analysis for diagnosing chronic periprosthetic infection in patients with inflammatory arthritis. J Bone Joint Surg [Am] 2012;94-A:594-600. Go to original source... Go to PubMed...
    16. Berbari E, Mabry T, Tsaras G, Spangehl M, Erwin PJ, Murad MH, Steckelberg J, Osmon D. Inflammatory blood laboratory levels as markers of prosthetic joint infection: a systematic review and meta-analysis. J Bone Joint Surg [Am] 2010;92-A:2102-9. Go to original source... Go to PubMed...
    17. Glehr M, Friesenbichler J, Hofmann G, Bernhardt GA, Zacherl M, Avian A, Windhager R, Leithner A. Novel biomarkers to detect infection in revision hip and knee arthroplasties. Clin Orthop Relat Res 2013;471(8):2621-28. Go to original source... Go to PubMed...
    18. Ettinger M, Calliess T, Kielstein JT, Sibai J, Brückner T, Lichtinghagen R, Windhagen H, Lukasz A. Circulating biomarkers for discrimination between aseptic joint failure, low-grade infection, and high-grade septic failure. Clin Infect Dis 2015;61:332-41. Go to original source... Go to PubMed...
    19. Gollwitzer H, Dombrowski Y, Prodinger PM, Peric M, Summer B, Hapfelmeier A, Saldamli B, Pankow F, von Eisenhart-Rothe R, Imhoff AB, Schauber J, Thomas P, Burgkart R, Banke IJ. Antimicrobial peptides and proinflammatory cytokines in periprosthetic joint infection. J Bone Joint Surg Am 2013;95(7):644-51. Go to original source... Go to PubMed...
    20. Randau TM, Friedrich MJ, Wimmer MD, Reichert B, Kuberra D, Stoffel-Wagner B, Limmer A, Wirtz DC, Gravius S. Interleukin-6 in serum and in synovial fluid enhances the differentiation between periprosthetic joint infection and aseptic loosening. PLoS One 2014;9:e89045. Go to original source... Go to PubMed...
    21. Prucha M, Bellingan G, Zazula R. Sepsis biomarkers. Clin Chim Acta 2015;440:97-103. Go to original source... Go to PubMed...
    22. Xie K, Qu X, Yan M. Procalcitonin and α-defensin for diagnosis of periprosthetic joint infections. J Arthroplasty 2017;32:1387-94. Go to original source... Go to PubMed...
    23. Agilli M, Sener I, Yesildal F, Honca T, Aydin I, Akgul EO, Yaman H. A new marker for the diagnosis of sepsis: Presepsin. J Investig Biochem 2012;1(1):55-7. Go to original source...
    24. Faix JD. Presepsin - the new kid on the sepsis block. Clin Biochem 2014;47(7-8):503-4. Go to original source... Go to PubMed...
    25. Kim H, Hur M, Moon HW, Yun YM, Di Somma S; GREAT Network. Multi-marker approach using procalcitonin, presepsin, galectin-3, and soluble suppression of tumorigenicity 2 for the prediction of mortality in sepsis. Ann Intensive Care 2017;7(1):27. Go to original source... Go to PubMed...
    26. Masson S, Caironi P, Spanuth E, Thomae R, Panigada M, Sangiorgi G, Fumagalli R, Mauri T, Isgrò S, Fanizza C, Romero M, Tognoni G, Latini R, Gattinoni L; ALBIOS Study Investigators. Presepsin (soluble CD14 subtype) and procalcitonin levels for mortality prediction in sepsis: data from the Albumin Italian Outcome Sepsis trial. Crit Care 2014;18(1):R6. Go to original source... Go to PubMed...
    27. Sparacino M, Russo E, Vitali S, Torri A, Pedna M F, Di Viesti M, Gagliardi M, Crepaldi E, Castiglioni M, Agnoletti V, Sambri V. Plasma concentration of presepsin and its relationship to the diagnosis of infections in multiple trauma patients admitted to intensive care. Microbiologia Medica 2017;32(2). Go to original source...
    28. Imagama T, Tokushige A, Seki K, Seki T, Nakashima D, Ogasa H, Sakai T, Taguchi T. Early diagnosis of septic arthritis using synovial fluid presepsin: A preliminary study. J Infect Chemother 2019;25(3):170-4. Go to original source... Go to PubMed...
    29. Marazzi MG, Randelli F, Brioschi M, Drago L, Romanò CL, Banfi G, Massaccesi L, Crapanzano C, Morelli F, Corsi Romanelli MM, Galliera E. Presepsin: A potential biomarker of PJI? A comparative analysis with known and new infection biomarkers. Int J Immunopathol Pharmacol 2018;31:394632017749356. Go to original source... Go to PubMed...

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