Tissue and Cell 41 (2009) 141–150 Contents lists available at ScienceDirect Tissue and Cell journal homepage: www.elsevier.com/locate/tice Pertinent cell population to characterize periodontal disease R. Younes a,b,∗,1 , C. Ghorra c,1 , S. Khalife d,1 , S. Igondjo-Tchen-Changotade b , M. Yousfi b , C. Willig b , K. Senni b , G. Godeau b,∗∗ , N. Naaman e a Department of Oral Surgery, Faculty of Dental Medicine, St Joseph University, P.O. Box 17-5208, Beirut 1104-2020, Lebanon b Laboratoire de Physiopathologie des Tissus non minéralisés, Faculté de Chirurgie Dentaire, Université Paris Descartes, 1, rue Maurice Arnoux 92120 Montrouge, France c Department of Anatomy and Pathology, Hôtel-Dieu de France, CHU St Joseph University, P.O. Box 17-5208, Beirut 1104-2020, Lebanon d Department of Biostatistics Faculty of medicine, St Joseph University, P.O. Box 17-5208, Beirut 1104-2020, Lebanon e Department of Periodontology, Faculty of Dental Medicine, St Joseph University, P.O. Box 17-5208, Beirut 1104-2020, Lebanon article info Article history: Received 30 June 2008 Received in revised form 26 September 2008 Accepted 30 September 2008 Available online 29 November 2008 Keywords: Periodontal disease Collagen Extracellular matrix Inflammatory cells/plasma cells abstract The purpose of this in situ study is to quantify the inflammatory cell subsets and the area fraction (AA%) occupied by collagen fibers in human healthy and diseased (four different stages) gingival connective tissue in order to establish a possible correlation between periodontal disease resulting in collagen breakdown and specific inflammatory cell subsets. Paraffin gingival tissue sections from eight healthy controls (group 0), 10 patients with gingivitis (group 1), 10 patients with moderate periodontitis (group 2) and 10 patients with severe periodontitis (group 3) were immunohistochemically investigated using antibodies against CD-45+, CD-3+, CD-8+, CD-20+, CD-68+, and EMA+ (plasma cells). The AA% occupied by gingival collagen fibers significantly decreased from 54.12% in group (0) to 38.58% in group (1), to 31.87% in group (2), and to 25.46% in group (3). In progressive lesions of periodontal dis- ease, CD-3+ and CD-8+ cell numbers were increased in early stages within the connective tissue, while CD-20+ cell numbers were increased only in late stages. On the other hand, EMA+, CD-68+ and CD-45+ cell numbers were progressively increased from group (0) to group (3). We demonstrated that CD-68+ mono- cyte/macrophages, CD-45+ leukocyte common antigen and notably EMA+ plasma cells are pertinently correlated with the severity of periodontal disease and related collagen breakdown. © 2008 Elsevier Ltd. All rights reserved. 1. Introduction The periodontal disease is an inflammatory process initiated and maintained by bacterial plaque and its metabolic products that trigger local infiltration of inflammatory cells associated with breakdown of the extracellular matrix macromolecules (Page and Schroeder, 1976). Among these, collagen quantitatively constitutes the major component of the gingival connective tissue and plays a key role in its architecture and is therefore implicated in patho- logical states such as periodontitis (Séguier et al., 2000). The composition of the inflammatory cells which infiltrate different tis- sue compartments in periodontal lesions may indicate which arm ∗ Corresponding author at: Department of Oral Surgery, Faculty of Dental Medicine, St Joseph University, P.O. Box 17-5208, Beirut 1104-2020, Lebanon. Tel.: +961 3360033; fax: +961 1396196. ∗∗ Corresponding author. Tel.: +33 1 58076789; fax: +33 1 58076787. E-mail addresses: ronald.younes@usj.edu.lb (R. Younes), godeau g@yahoo.fr (G. Godeau). 1 These authors contributed equally to this study. of the immune system most effectively deploys locally to protect the periodontal tissues. Bandtzaeg and Kraus (1965) showed the presence of immunoglobulin producing plasma cells in the gingi- val tissues of patients with periodontal disease. This was the first evidence which demonstrated that adaptive immune mechanisms play a role in the pathogenesis of periodontal inflammation. In 1970, Ivanyi and Lehner (1970) using peripheral blood lymphocyte trans- formation assays, highlighted a role for cell-mediated immunity in periodontal disease. Since then, immunohistological studies have provided evidence for the immunological nature of the response to plaque bacteria (Listgarten, 1986). Thus, in the gingival connective tissue, many studies have shown important changes in cellular populations during the periodontal disease. It is clear from histological studies that the predominant cell type of the chronic gingival lesion is the lymphocyte (Page and Schroeder, 1976; Seymour et al., 1979a,b). In the early stages of gingival inflammation, pro-inflammatory cytokines secreted by activated monocytes, macrophages and other cells (e.g. fibroblasts, epithelial and endothelial cells) predominate (Page, 1986). The macrophages are key cells of the innate immune system involved in the progression of periodontal disease from an acute inflam- 0040-8166/$ – see front matter © 2008 Elsevier Ltd. All rights reserved. doi:10.1016/j.tice.2008.09.003