A Simple and Efficient Method for Analyzing Murine Gingival Immune Cells Using Flow Cytometry: An Animal Study.

Abstract

BACKGROUND: Elucidating the immune cell network is of great significance for understanding the pathogenesis of periodontitis. Flow cytometry is the preferred tool for quantifying the composition of immune cells in the pathological process of inflammation. Perfusion prior to tissue harvesting is necessary for flow cytometric analysis of various tissues. However, whether perfusion is necessary for flow cytometric analysis of murine gingival tissues has never been reported. METHOD: Forty-eight mice were randomly divided into four groups: control + non-perfused group, control + perfused group, periodontitis + non-perfused group, and periodontitis + perfused group. Ligation-induced experimental periodontitis was established in periodontitis mice. The circulating blood was perfused out in the perfused groups before harvesting tissue samples. Flow cytometry was used to analyze the composition of immune cells in murine palatal gingival tissues. RESULTS: Experimental periodontitis induced alveolar bone resorption and inflammatory cell infiltration. Perfusion can successfully remove blood from the vessels of murine periodontal tissues. Flow cytometric analysis showed that there was no significant difference in the proportion of various immune cells in murine gingival tissues between the perfusion groups and the non-perfusion groups. CONCLUSION: The present study was the first to use flow cytometry to compare perfused mouse gingiva with non-perfused mouse gingiva and found no significant difference in immune cell composition between the two groups. In addition, this study provides a simple and efficient protocol for the collection and processing of murine gingival tissue, which could provide facilities for understanding the role and mechanism of the immune cell network in periodontal diseases.