Immunocytochemical indicators of apoptosis in gingival tissues of patients with chronic periodontitis

  • Aldona Kasprzak Chair and Department of Histology and Embryology, Poznan University of Medical Sciences, Poland
  • Miłosz Hausmann Chair and Department of Histology and Embryology, Poznan University of Medical Sciences, Poland
  • Agata Małkowska-Lanzafame Department and Clinics of Conservative Dentistry and Periodontology, Poznan University of Medical Sciences, Poland
  • Joanna Surdyk-Zasada Chair and Department of Histology and Embryology, Poznan University of Medical Sciences, Poland
  • Wiesława Przybyszewska Chair and Department of Histology and Embryology, Poznan University of Medical Sciences, Poland
  • Elżbieta Siodła Chair and Department of Histology and Embryology, Poznan University of Medical Sciences, Poland
  • Anna Surdacka Department and Clinics of Conservative Dentistry and Periodontology, Poznan University of Medical Sciences, Poland
Keywords: chronic periodontitis, apoptosis, immunocytochemistry


Introduction. Inflammatory mechanisms of chronic periodontitis (CP) may be linked to various forms of disturbances in apoptosis.Aim. The study aimed at comparison of tissue expression of anti-apoptotic protein (Bcl-2) and proapoptotic proteins (p53, caspase-3) in gingival tissues of 30 patients with CP and of 15 with healthy periodontium.Material and methods. Gingival samples (n = 68) were obtained during the open curettage procedure with gingivectomy of adult patients (18 women and 12 men) with CP. Classical immunocytochemical (IHC) method was used to detect apoptotic proteins, and the obtained expression was evaluated using semi-quantitative IRS scale.Results. No differences could be revealed in expression intensity or reciprocal correlations between apoptotic proteins within the group of patients with CP. Greater expression of the two apoptotic proteins (Bcl-2 and p53) were detected in patients with CP than in control individuals. Moreover, a more pronounced expression of Bcl-2 was demonstrated in gingival samples of patients with localised form as compared to generalised form of CP. Expression of caspase-3 (effector phase of apoptosis) manifested no differences between CP and control individuals. Greater expression of the anti-apoptotic protein Bcl-2 and caspase-3 was detected in cells of inflammatory infiltrates in lamina propria than in keratinocytes.Conclusions. In CP significant alterations developed in expression of indicators of apoptosis, with prevalence of Bcl-2 and p53 expression, as compared to the control. The localised form of CP was linked to higher proportion of Bcl-2-positive cells of inflammatory infiltrates, suggesting that apoptosis was inhibited mainly in this form of CP. The comparable expression of caspase-3 in gingival cells with CP and in control and absence of correlation with clinical data suggested that the process of apoptosis did not play a significant role in destruction of periodontium tissues in CP.


Seymur GJ, Gemmel E, Reinhardt RA et al. Immunopathogenesis of chronic inflammatory periodontal disease: cellular and molecular mechanisms. J Periodont Res. 1993 Nov;28(6 Pt 2):478–86.

Loesche WJ, Grossman NS. Periodontal disease as specific, albeit chronic, infection: diagnosis and treatment. Clin Microbiol Rev. 2001 Oct;14(4):727–52.

Slots J, Kamma JJ, Sugar C. The herpesvirus-Porphyromonas gingivalis-periodontitis axis. J Periodontal Res. 2003 Jun;38(3):318–23.

Graves DT, Jiang Y, Genco C. Periodontal disease: bacterial virulence factors, host response and impact on systemic health. Curr Opin Infect Dis. 2000 Jun;13(3):227–32.

Wassenaar A, Reinhardus C, Thepen T et al. Cloning, characterization, and antigen specificity of T-lymphocyte subsets extracted from gingival tissue of chronic adult periodontitis patients. Infect Immun. 1995 Jun;63(6):2147–53.

Arakawa S, Nakajima T, Ishikura H et al. Novel apoptosis-inducing activity in Bacteroides forsynthus: a comparative study with three serotypes of Actinobacillus actinomycetemcomitans. Infect Immun. 2000 Aug;68(8):4611–5.

Berker E, Kantarci A, Hasturk H et al. Effect of neutrophil apoptosis on monocytic cytokine response to Porphyromonas gingivalis lipopolysaccharide. J Periodontol. 2005 Jun;76(6):964–71.

Gamonal J. Apoptosis in chronic adult periodontitis analyzed by in situ DNA breaks, electron microscopy, and immunohistochemistry. J Periodontol. 2001 Apr;72(4):517–25.

Jarnbring F, Somogyi E, Dalton J et al. Quantitative assessment of apoptotic and proliferative gingival keratinocytes in oral and sulcular epithelium in patients with gingivitis and periodontitis. J Clin Periodontol. 2002 Dec;29(12):1065–71.

Bantel H, Beikler T, Flemmig TF et al. Caspase activation is involved in chronic periodontitis. FEBS Lett. 2005 Oct;579(25):5559–64.

Kato S, Muro M, Akifusa S et al. Evidence of apoptosis of murine macrophages by Actinobacillus actinomycetemcomitans infection. Infect Immun. 1995 Oct;63(10):3914–19.

Kato S, Nakashima K, Inoue M et al. Human epithelial cell death caused by Actinobacillus actinomycetemcomitans infection. J Med Microbiol. 2000 Aug;49(8):739–45.

Geatch DR, Harris JI, Heasman PA et al. In vitro studies of lymphocyte apoptosis induced by the periodontal pathogen Porphyromonas gingivalis. J Periodontal Res. 1999 Feb;34(2):70–8.

Wang PL, Shirasu S, Shinohara M et al. Induction of apoptosis in human gingival fibroblasts by a Porphyromonas gingivalis protease preparation. Arch Oral Biol. 1999 Apr;44(4):337–42.

Ansai T, Yu W, Urnowey S et al. Contruction of a pepO gene-deficient mutant of Porphyromonas gingivalis: potential role of endopeptidase O in the invasion of host cells. Oral Microbiol Immunol. 2003 Dec;18(6):398–400.

Baba A, Abe N, Kadowaki T et al. Arg-gingipain is responsible for the degradation of cell adhesion molecules of human gingival fibroblasts and their death induced by Porphyromonas gingivalis. Biol Chem. 2001 May;382(5):817–24.

Sheets SM, Potempa J, Travis J et al. Gingipains from Porphyromonas gingivalis W83 induce Cell Adhesion Molecule cleavage and apoptosis in endothelial cells. Infect Immun. 2005 Mar;73(3):1543–52.

Urnowey S, Ansai T, Bitko V et al. Temporal activation of anti- and pro-apoptotic factors in human gingival fibroblasts infected with the periodontal pathogen, Porphyromonas gingivalis: potential role of bacterial proteases in host signalling. BMC Microbiol. 2006 Mar;6(1):26, DOI. 10.1186/1471–2180–6-26.

Mezyk-Kopec R, Bzowska M, Potemba J et al. Inactivation of membrane tumor necrosis factor alpha by gingipains from Porphyromonas gingivalis. Infect Immun. 2005 Mar;73(3):1506–14.

Yamamoto M, Kawabata K, Fujihashi K et al. Absence of exogenous interleukin-4-induced apoptosis of gingival macrophages may contribute to chronic inflammation in periodontal diseases. Am J Pathol. 1996 Jan;148(1):331–9.

Sawa T, Nishimura F, Ohyama H et al. In vitro induction of activation-induced cell death in lymphocytes from chronic periodontal lesions by exogenous Fas Ligand. Infect Immun. 1999 Mar;67(3):1450–54.

Nakhjiri SF, Park Y, Yilmaz O et al. Inhibition of epithelial cell apoptosis by Porphyromonas gingivalis. FEMS Microbiol Lett. 2001 Jun;200(2):145–9.

Yilmaz O, Jungas T, Verbeke P et al. Activation of the phosphatidylinositol 3-kinase/Akt pathway contributes to survival of primary epithelial cells infected with the periodontal pathogen Porphyromonas gingivalis. Infect Immun. 2004 Jul;72(7):3743–51.

Koulouri O, Lappin DF, Radvar M et al. Cell division, synthetic capacity and apoptosis in periodontal lesions analysed by in situ hybridization and immunohistochemistry. J Clin Periodontol. 1999 Aug;26(8):552–9.

Szmyt M, Kasprzak A, Malkowski W et al. Tissue expression of S100 proteins in gallbladder mucosa of the patients with calculous cholecystitis. Folia Histochem Cytobiol. 2013, 51(2):1–8, DOI: 10.5603/FHC.2013.0022

Remmele W, Stegner HE. Recomendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue. Pathologe. 1987 May;8(3):138–40.

Hockenbery DM, Zutter M, Hickey W et al. Bcl-2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci USA. 1991 Aug;88(16):6961–65.

Bulut S, Uslu H, Ozdemir BH et al. Expression of caspase-3, p53 and Bcl-2 in generalized aggressive periodontitis. Head Face Med. 2006 Jun;2:17, DOI:10.1 1186/1746-160X-2-17.

Monaghan P, Robertson D, Amos TA et al. Ultrastructural localisation of BCL-2 protein. J Histochem Cytochem. 1992 Dec;40(12):1819–25.

Carvalho-Filho PC, Trindade SC, Olczak T et al. Porphyromonas gingivalis HmuY stimulates expression of Bcl-2 and Fas by human CD3+ T cells. BMC Microbiol. 2013 Sep 11;13:206. DOI: 10.1186/171-2180-13-206.

Jacks T, Weinberg RA. Cell cycle control and its watchman. Nature. 1996 Jun;381(6584):643–4.

Ghosh A, Joo NE, Chen TC et al. Proapoptotic fibronectin fragment induces the degradation of ubiquitinated p53 via proteasomes in periodontal ligament cells. J Periodontal Res. 2010 Aug;45(4):481–7.

Fischer U, Janicke RU, Schulze-Osthoff K. Many cuts to ruin: a comprehensive update of caspase substrates. Cell Death Differ. 2003 Jan;10(1):76–100.

Krajewska M, Wang HG, Krajewski Z et al. Immunocytochemical analysis of in vivo patterns of expression of CPP32 (caspase-3), a cell death protease. Cancer Res. 1997 Apr;57(8):1605–13.

Lucas H, Bartold PM, Dharmapatni AA et al. Inhibition of apoptosis in periodontitis. J Dent Res. 2010 Jan;89(1): 29–33.

Tonetti MS, Cortellini D, Lang NP. In situ detection of apoptosis at sites of chronic bacterially induced inflammation in human gingival. Infect Immun. 1998 Nov;66(11):5190–95.

Kang J, de Brito Bezerra B, Pacios S et al. Aggregatibacter actinomycetemcomitans infection enhances apoptosis in vivo through a caspase-3-dependent mechanism in experimental periodontitis. Infect Immun. 2012 Jun;80(6):2247–56.

How to Cite
Kasprzak, A., Hausmann, M., Małkowska-Lanzafame, A., Surdyk-Zasada, J., Przybyszewska, W., Siodła, E., & Surdacka, A. (2016). Immunocytochemical indicators of apoptosis in gingival tissues of patients with chronic periodontitis. Journal of Medical Science, 83(1), 37-46. Retrieved from
Original Papers