Saliva as a medium to detect and measure biomarkers related to pain

Hajer Jasim, Anders Carlsson, Britt Hedenberg-Magnusson, Bijar Ghafouri, Malin Ernberg, Hajer Jasim, Anders Carlsson, Britt Hedenberg-Magnusson, Bijar Ghafouri, Malin Ernberg

Abstract

Saliva is often neglected as a body fluid of diagnostic or prognostic value, even though generally well accepted by the patients. This is due to lack of a standardized collection procedure. The aim of this study was to identify the ideal saliva collection technique and develop new sensitive methods to detect and analyse markers related to pain in healthy pain-free subjects. Plasma and five different saliva collection approached was evaluated during strictly controlled conditions. Levels of nerve growth factor (NGF), calcitonin gene-related peptide (CGRP) and brain derived neurotropic factor (BDNF) were determined using novel western blotting based technology. Glutamate and substance P (SP) was determined using commercial available methods. Several new isoforms were found for NGF, CGRP and BDNF in saliva. The isoform pattern showed significant variation in both expression and chemiluminescence levels between different collection methods. New sensitive methods to study pain related markers in saliva were developed in this study. Furthermore, we are first to demonstrate a correlation between the Glutamate concentration in stimulated whole saliva and blood. However, the fundamental conclusion drawn is the importance of consistency in the collection method.

Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Illustrative overview of the main salivary glands and different collection approaches used in the study. The colours are associated with the different collection methods explained in the diagrams.
Figure 2
Figure 2
Salivary and plasma NGF, CGRP, BDNF, glutamate and substance P concentration in 20 healthy individuals matched for age and gender (Fig. 2A–H). Large variations were observed between different collection methods. Several isoforms was detected for NGF, CGRP and BDNF. The isoform pattern showed significant variation in both expression and chemiluminescence levels between different collection methods (Friedman; P 

References

    1. Fischer HP, Eich W, Russell IJ. A possible role for saliva as a diagnostic fluid in patients with chronic pain. Seminars in arthritis and rheumatism. 1998;27:348–359. doi: 10.1016/S0049-0172(98)80014-0.
    1. Castrillon EE, et al. Interstitial glutamate concentration is elevated in the masseter muscle of myofascial temporomandibular disorder patients. Journal of orofacial pain. 2010;24:350–360.
    1. Rosendal L, et al. Increase in muscle nociceptive substances and anaerobic metabolism in patients with trapezius myalgia: microdialysis in rest and during exercise. Pain. 2004;112:324–334. doi: 10.1016/j.pain.2004.09.017.
    1. Shimada, A. et al. Increased pain and muscle glutamate concentration after single ingestion of monosodium glutamate by myofascial temporomandibular disorders patients. European journal of pain (London, England), 10.1002/ejp.874 (2016).
    1. Nam JW, Chung JW, Kho HS, Chung SC, Kim YK. Nerve growth factor concentration in human saliva. Oral diseases. 2007;13:187–192. doi: 10.1111/j.1601-0825.2006.01265.x.
    1. Ashina M, Bendtsen L, Jensen R, Schifter S, Olesen J. Evidence for increased plasma levels of calcitonin gene-related peptide in migraine outside of attacks. Pain. 2000;86:133–138. doi: 10.1016/S0304-3959(00)00232-3.
    1. Jang MU, Park JW, Kho HS, Chung SC, Chung JW. Plasma and saliva levels of nerve growth factor and neuropeptides in chronic migraine patients. Oral diseases. 2011;17:187–193. doi: 10.1111/j.1601-0825.2010.01717.x.
    1. Zidverc-Trajkovic J, et al. Calcitonin gene-related peptide levels in saliva of patients with burning mouth syndrome. Journal of oral pathology & medicine: official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology. 2009;38:29–33. doi: 10.1111/j.1600-0714.2008.00721.x.
    1. Fischer M, et al. Brain-derived neurotrophic factor in primary headaches. The journal of headache and pain. 2012;13:469–475. doi: 10.1007/s10194-012-0454-5.
    1. Greco R, et al. Role of calcitonin gene-related peptide and substance P in different models of pain. Cephalalgia: an international journal of headache. 2008;28:114–126.
    1. Mandel AL, Ozdener H, Utermohlen V. Identification of pro- and mature brain-derived neurotrophic factor in human saliva. Archives of oral biology. 2009;54:689–695. doi: 10.1016/j.archoralbio.2009.04.005.
    1. Drobitch RK, Svensson CK. Therapeutic drug monitoring in saliva. An update. Clinical pharmacokinetics. 1992;23:365–379. doi: 10.2165/00003088-199223050-00003.
    1. Jasim H, Olausson P, Hedenberg-Magnusson B, Ernberg M, Ghafouri B. The proteomic profile of whole and glandular saliva in healthy pain-free subjects. Scientific reports. 2016;6:39073. doi: 10.1038/srep39073.
    1. Segal A, Wong DT. Salivary diagnostics: enhancing disease detection and making medicine better. European journal of dental education: official journal of the Association for Dental Education in Europe. 2008;12(Suppl 1):22–29.
    1. Lee YH, Wong DT. Saliva: an emerging biofluid for early detection of diseases. American journal of dentistry. 2009;22:241–248.
    1. Al Kawas S, Rahim ZH, Ferguson DB. Potential uses of human salivary protein and peptide analysis in the diagnosis of disease. Archives of oral biology. 2012;57:1–9. doi: 10.1016/j.archoralbio.2011.06.013.
    1. Helmerhorst EJ, Oppenheim FG. Saliva: a dynamic proteome. Journal of dental research. 2007;86:680–693. doi: 10.1177/154405910708600802.
    1. Schiffman E, et al. Diagnostic Criteria for Temporomandibular Disorders (DC/TMD) for Clinical and Research Applications: recommendations of the International RDC/TMD Consortium Network* and Orofacial Pain Special Interest Groupdagger. Journal of oral & facial pain and headache. 2014;28:6–27. doi: 10.11607/jop.1151.
    1. Gerdle B, et al. Chronic widespread pain: increased glutamate and lactate concentrations in the trapezius muscle and plasma. The Clinical journal of pain. 2014;30:409–420.
    1. Wozniak KM, Rojas C, Wu Y, Slusher BS. The role of glutamate signaling in pain processes and its regulation by GCP II inhibition. Current medicinal chemistry. 2012;19:1323–1334. doi: 10.2174/092986712799462630.
    1. Gerdle B, Ghafouri B, Ernberg M, Larsson B. Chronic musculoskeletal pain: review of mechanisms and biochemical biomarkers as assessed by the microdialysis technique. Journal of pain research. 2014;7:313–326. doi: 10.2147/JPR.S59144.
    1. McNearney T, Speegle D, Lawand N, Lisse J, Westlund KN. Excitatory amino acid profiles of synovial fluid from patients with arthritis. The Journal of rheumatology. 2000;27:739–745.
    1. Mandel AL, Ozdener H, Utermohlen V. Brain-derived neurotrophic factor in human saliva: ELISA optimization and biological correlates. Journal of immunoassay & immunochemistry. 2011;32:18–30. doi: 10.1080/15321819.2011.538625.
    1. Parris WC, Kambam JR, Naukam RJ, Rama Sastry BV. Immunoreactive substance P is decreased in saliva of patients with chronic back pain syndromes. Anesthesia and analgesia. 1990;70:63–67. doi: 10.1213/00000539-199001000-00010.
    1. Humphrey SP, Williamson RT. A review of saliva: normal composition, flow, and function. The Journal of prosthetic dentistry. 2001;85:162–169. doi: 10.1067/mpr.2001.113778.
    1. Nieuw Amerongen AV, Ligtenberg AJ, Veerman EC. Implications for diagnostics in the biochemistry and physiology of saliva. Annals of the New York Academy of Sciences. 2007;1098:1–6. doi: 10.1196/annals.1384.033.
    1. Goedert M, Nagy JI, Emson PC. The origin of substance P in the rat submandibular gland and its major duct. Brain research. 1982;252:327–333. doi: 10.1016/0006-8993(82)90400-0.
    1. Cruz-Almeida Y, et al. Age differences in salivary markers of inflammation in response to experimental pain: does venipuncture matter? Journal of pain research. 2017;10:2365–2372. doi: 10.2147/JPR.S138460.
    1. Komatsu K, Hasegawa H, Honda T, Yabashi A, Kawasaki T. Nerve growth factor in saliva stimulated by mastication. Oral Science International. 2008;5:78–84. doi: 10.1016/S1348-8643(08)80011-0.
    1. Olausson P, Gerdle B, Ghafouri N, Larsson B, Ghafouri B. Identification of proteins from interstitium of trapezius muscle in women with chronic myalgia using microdialysis in combination with proteomics. PloS one. 2012;7:e52560. doi: 10.1371/journal.pone.0052560.
    1. Golatowski C, et al. Comparative evaluation of saliva collection methods for proteome analysis. Clinica chimica acta; international journal of clinical chemistry. 2013;419:42–46. doi: 10.1016/j.cca.2013.01.013.
    1. Topkas E, Keith P, Dimeski G, Cooper-White J, Punyadeera C. Evaluation of saliva collection devices for the analysis of proteins. Clinica chimica acta; international journal of clinical chemistry. 2012;413:1066–1070. doi: 10.1016/j.cca.2012.02.020.
    1. Baum BJ. Evaluation of stimulated parotid saliva flow rate in different age groups. Journal of dental research. 1981;60:1292–1296. doi: 10.1177/00220345810600070101.
    1. Hu S, et al. Differentially expressed protein markers in human submandibular and sublingual secretions. International journal of oncology. 2004;25:1423–1430.
    1. Fleissig Y, et al. Comparative proteomic analysis of human oral fluids according to gender and age. Oral diseases. 2010;16:831–838. doi: 10.1111/j.1601-0825.2010.01696.x.

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