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Abstract
Objective. Tissue inhibitor of metalloproteinases 1 (TIMP‐1) has been identified as a potential biomarker in diseases such as cancer, cardiovascular diseases and diabetes. Since TIMP‐1 resides in most tissues and bodily fluids, we evaluated the potential of using saliva to obtain reproducible TIMP‐1 measurements in a non‐invasive manner. Material and methods. Samples of unstimulated and stimulated whole saliva and saliva collected from individual glands were analysed for TIMP‐1 content. A TIMP‐1 ELISA was validated for use in saliva testing and the most optimal sampling and handling procedures for reproducible measurements identified. Western blotting and MALDI‐TOF mass spectrometry were used for confirmatory analyses. Results. The TIMP‐1 ELISA was found suitable for saliva measurements. All saliva secretions contained TIMP‐1, but in different concentrations ranging from 2.81 ng/mL in submandibular/sublingual saliva to 173.88 ng/mL in parotid saliva. TIMP‐1 concentrations were influenced to a varying degree by fluctuations in flow. We found the lowest output in submandibular/sublingual saliva stimulated with 0.5 % citric acid (3.56 ng/min) and highest output in chewing‐stimulated whole saliva (267.01 ng/min). Conclusion. This study shows that saliva contains authentic TIMP‐1, the concentration of which was found to depend on gland type and salivary flow. Stimulated whole saliva is suggested as a reliable and easily accessible source for TIMP‐1 determinations in bodily fluids.
Acknowledgements
This work was supported by The Kornerup Fund, The Aase and Ejnar Danielsen Fund, The Danish Cancer Society, The IMK Foundation, The Danish Medical Research Counsel, Familien Hede Nielsens Fund, The Danish Cancer Research Foundation, Ib Henriksens Foundation, Kathrine og Vigo Skovgaard Foundation, Redaktør Kaaresens Foundation, and The Obel Family Foundation. HJN is Danish Cancer Society Professor of Surgical Oncology.