Morphometric analysis of collagen and inflammatory cells in periodontal disease

Ranko Golijanin; Bojan Kujundžić; Zoran Milosavljević; Dragan R Milovanović; Zlatibor Andjelković; Miroslav Obrenović; Radivoje Nikolić

doi:10.2298/4098

Ranko Golijanin Department of Dentistry, Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia; Faculty of Dental Medicine, University of East Sarajevo, Foča, Republic of Srpska, Bosnia and Herzegovina
Bojan Kujundžić Faculty of Dental Medicine, University of East Sarajevo, Foča, Republic of Srpska, Bosnia and Herzegovina
Zoran Milosavljević Department of Histology and Embriology, Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia
Dragan R Milovanović Department of Pharmacology and Toxicology,Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia; Clinical Center “Kragujevac”, Kragujevac, Serbia
Zlatibor Andjelković Department oh Histology, Medical Faculty University of Pristina, Kosovska Mitrovica, Serbia
Miroslav Obrenović Faculty of Medicine, University of East Sarajevo, Foča, Republic of Srpska, Bosnia and Herzegovina
Radivoje Nikolić Department of Surgery, Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia; Clinical Center “Kragujevac”, Kragujevac, Serbia

DOI: https://doi.org/10.2298/4098

Keywords: peridontal diseases, gingiva, histological techniques, collagen, macrophages, plasma cells,

Abstract

Background/Aim. Periodontal disease affects gingival tissue and supporting apparatus of the teeth leading to its decay. The aim of this study was to highlight and precisely determine histological changes in the gum tissue. Methods. Gingival biopsy samples from 53 healthy and parodontopathy-affected patients were used. Clinical staging of the disease was performed. Tissue specimens were fixed and routinely processed. Sections, 5 μm thin, were stained with hematoxylin and eosin, histochemical Van-Gieson for the collagen content, Spicer method for mast-cells and immunochemical method with anti-CD68 and anti-CD38 for the labelling of the macrophages and plasma-cells. Morphometric analysis was performed by a M42 test system. Results. While the disease advanced, collagen and fibroblast volume density decreased almost twice in the severe cases compared to the control ones, but a significant variation was observed within the investigated groups. The mast-cell number increased nearly two times, while the macrophage content was up to three times higher in severe parodontopathy than in healthy gingival tissue. However, the relative proportion of these cells stayed around 6% in all cases. Plasma-cells had the most prominent increase in the number (over 8 times) compared to the control, but again, a variation within investigated groups was very high. Conclusion. Gingival tissue destruction caused by inflammatory process leads to significant changes in collagen density and population of resident connective tissue cells. Although inflammatory cells dominated with the disease advancing, a high variation within the same investigated groups suggests fluctuation of the pathological process.

References

Page RC. Gingivitis. J Clin Periodontol 1986; 13(5): 345−55.

Ohlrich EJ, Cullinan MP, Seymour GJ. The immunopathogenesis of periodontal disease. Aust Dent J 2009; 54(Suppl 1): 2−10.

Seymour GJ, Greenspan JS. The phenotypic characterization of lymphocyte subpopulations in established human periodontal disease. J Periodontal Res 1979; 14(1): 39−46.

Séguier S, Godeau G, Brousse N. Collagen Fibers and Inflamma-tory Cells in Healthy and Diseased Human Gingival Tissues: A Comparative and Quantitative Study by Immunohistochemi-stry and Automated Image Analysis. J Periodontol 2000; 71(7): 1079−85.

Ejeil A, Gaultier F, Igondjo-Tchen S, Senni K, Pellat B, Godeau G, et al. Are Cytokines Linked to Collagen Breakdown During Peri-odontal Disease Progression. J Periodontol 2003; 74(2): 196−201.

Lappin DF, MacLeod CP, Kerr A, Mitchell T, Kinane DF. Anti-inflammatory cytokine IL-10 and T cell cytokine profile in pe-riodontitis granulation tissue. Clin Exp Immunol 2001; 123(2): 294−300.

Séguier S, Gogly B, Bodineau A, Godeau G, Brousse N. Is Collagen Breakdown During Periodontitis Linked to Inflammatory Cells and Expression of Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases in Human Gingival Tissue. J Periodontol 2001; 72(10): 1398−406.

Younes R, Ghorra C, Khalife S, Igondjo-Tchen-Changotade S, Yousfi M, Willig C, et al.. Pertinent cell population to characterize pe-riodontal disease. Tissue Cell 2009; 41(2): 141−50.

Bendeck MP. Macrophage Matrix Metalloproteinase-9 Regulates Angiogenesis in Ischemic Muscle. Circ Res 2004; 94(2): 138−9.

Kessenbrock K, Brown M, Werb Z. Measuring matrix metallopro-teinase activity in macrophages and polymorphonuclear leuko-cytes. Curr Protoc Immunol 2011; 14: 14−24.

Ainamo J, Löe H. Anatomical Characteristics of Gingiva: A Clinical and Microscopic Study of the Free and Attached Gin-giva. J Periodontol 1966; 37(1): 5−13.

Mühlemann HR, Son S. Gingival sulcus bleeding: a leading symptom in initial gingivitis. Helv Odontol Acta 1971; 15(2): 107−13.

Armitage GC. Periodontal diagnoses and classification of peri-odontal diseases. Periodontol 2000 2004; 34(1): 9−21.

Hillmann G, Krause S, Ozdemir A, Dogan S, Geurtsen W. Immu-nohistological and morphometric analysis of inflammatory cells in rapidly progressive periodontitis and adult periodonti-tis. Clin Oral Investig 2001; 5(4): 227−35.

Lorencini M, Silva JA, Almeida CA, Bruni-Cardoso A, Carvalho HF, Stach-Machado DR. A new paradigm in the periodontal disease progression: Gingival connective tissue remodeling with simultaneous collagen degradation and fibers thickening. Tis-sue Cell 2009; 41(1): 43−50.

Daniel A, Dupont M. Stereological analysis of human gingival connective tissues. Clinically healthy gingiva. J Biol Buccale 1980; 8(2): 141−53.

Vardar-Sengul S, Arora S, Baylas H, Mercola D. Expression Pro-file of Human Gingival Fibroblasts Induced by Interleukin-1β Reveals Central Role of Nuclear Factor-Kappa B in Stabilizing Human Gingival Fibroblasts During Inflammation. J Peri-odontol 2009; 80(5): 833−49.

Oyarzún A, Arancibia R, Hidalgo R, Peñafiel C, Cáceres M, González MJ, et al. Involvement of MT1-MMP and TIMP-2 in human periodontal disease. Oral Dis 2010; 16(4): 388−95.

Seppälä B, Sorsa T, Ainamo J. Morphometric analysis of cellular and vascular changes in gingival connective tissue in long-term insulin-dependent diabetes. J Periodontol 1997; 68(12): 1237−45.

Havemose-Poulsen A, Holmstrup P. Factors Affecting IL-1-Mediated Collagen Metabolism By Fibroblasts and the Patho-genesis of Periodontal Disease: A Review of the Literature. Crit Rev Oral Biol Med 1997; 8(2): 217−36.

Batista AC, Rodini CO, Lara VS. Quantification of mast cells in different stages of human periodontal disease. Oral Dis 2005; 11(4): 249−54.

Huang S, Lu F, Chen Y, Huang B, Liu M. Mast Cell Degranula-tion in Human Periodontitis. J Periodontol 2013; 84(2): 248−55.

Gemmell E, Carter CL, Seymour GJ. Mast Cells in Human Peri-odontal Disease. J Dent Res 2004; 83(5): 384−7.

Beklen A, Ainola M, Hukkanen M, Gurgan C, Sorsa T, Konttinen YT. MMPs, IL-1, and TNF are Regulated by IL-17 in Peri-odontitis. J Dent Res 2007; 86(4): 347−51.

Wallace AM, Sandford AJ, English JC, Burkett KM, Li H, Finley RJ, et al. Matrix metalloproteinase expression by human alveolar macrophages in relation to emphysema. COPD 2008; 5(1): 13−23.

Kocher T, Schwahn C, Gesch D, Bernhardt O, John U, Meisel P, et al. Risk determinants of periodontal disease - an analysis of the Study of Health in Pomerania (SHIP 0). J Clin Periodontol 2005; 32(1): 59−67.

Lins RD, Figueiredo CR, Queiroz LM, da Silveira EJ, Freitas RA. Immunohistochemical evaluation of the inflammatory re-sponse in periodontal disease. Braz Dent J 2008; 19(1): 9−14.

Kim YC, Ko Y, Hong SD, Kim KY, Lee YH, Chae C, et al. Pres-ence of Porphyromonas gingivalis and plasma cell dominance in gingival tissues with periodontitis. Oral Dis 2010; 16(4): 375−81.