A novel method of sampling gingival crevicular fluid from a mouse model of periodontitis

Using a mouse model of silk ligature-induced periodontal disease (PD), we report a novel method of sampling mouse gingival crevicular fluid (GCF) to evaluate the time-dependent secretion patterns of bone resorption-related cytokines. GCF is a serum transudate containing host-derived biomarkers which can represent cellular response in the periodontium. As such, human clinical evaluations of PD status rely on sampling this critical secretion. At the same time, a method of sampling GCF from mice is absent, hindering the translational value of mouse models of PD. Therefore, we herein report a novel method of sampling GCF from a mouse model of periodontitis, involving a series of easy steps. First, the original ligature used for induction of PD was removed, and a fresh ligature for sampling GCF was placed in the gingival crevice for ten minutes. Immediately afterwards, the volume of GCF collected in the sampling ligature was measured using a high precision weighing balance. The sampling ligature containing GCF was then immersed in a solution of PBS-Tween 20 and subjected to ELISA. This enabled us to monitor the volume of GCF and detect time-dependent changes in the expression of such cytokines as IL-1b, TNF-α, IL-6, RANKL, and OPG associated with the levels of alveolar bone loss, as reflected in GCF collected from a mouse model of PD. Therefore, this novel GCF sampling method can be used to measure various cytokines in GCF relative to the dynamic changes in periodontal bone loss induced in a mouse model of PD. Correspondence: Toshihisa Kawai, DDS, PhD, Department of Immunology and Infectious diseases, The Forsyth Institute, 245 First Street, Cambridge, MA 02142, Tel: 617-892-8317, Fax: 617-892-8437, [email protected]. # Contributed equally to this work HHS Public Access Author manuscript J Immunol Methods. Author manuscript; available in PMC 2017 November 01. Published in final edited form as: J Immunol Methods. 2016 November ; 438: 21–25. doi:10.1016/j.jim.2016.08.008. Author Manuscript Author Manuscript Author Manuscript Autho