Vitamin C attenuates the cytotoxic effects of Porphyromonas gingivalis on human gingival fibroblasts
Introduction
Epidemiological studies indicate a low vitamin C concentration in plasma as a risk factor for periodontitis.1, 2, 3, 4 Our group recently reported that patients with periodontitis – in contrast to healthy subjects – had decreased vitamin C levels in their plasma.5 Although the specific role of vitamin C deficiency is widely unknown, this vitamin has long been a candidate for modifying periodontal diseases.6 This could be due to the various biological functions which are also connected with the periodontal tissues. For example, studies have shown that vitamin C increases the number of collagen bundles in the regenerating periodontal tissue,7 detoxifies histamine in gingival inflammation,8, 9 and reduces gingival oxidative stress.10 Particularly, the role of oxidative stress in periodontal disease is well established.11 The investigation of oxidative stress factors in the saliva by our group resulted in increased levels of reactive oxygen species in periodontitis patients corresponding with decreased anti-oxidative capacities.12 In this connection the antioxidant properties of vitamin C produced interest. As a water-soluble vitamin it protects especially aqueous environments against oxidative attacks. For example, leucocytes accumulate vitamin C to protect themselves during phagocytosis against oxidative species.13 Fibroblasts also require vitamin C for normal cell growth in the context of collagen synthesis14 and as an antioxidant.15 Gingival fibroblasts play a key role in the regulation of the physiological turnover of connective tissues by depleting collagen and remodelling new collagen fibrils in the periodontal tissue.16 Contact with periodontal-pathogenic bacteria disturbs this sensitive balance resulting in collagen degradation and consequently in pocket formation.17 One of the invasive bacteria in the plaque biofilm is Porphyromonas gingivalis, a black-pigmented, anaerobic gram-negative species.18 It produces a broad array of potential virulence factors, including lipopolysaccharides, fimbriae and proteases that affect the host response and damage oral cells.19, 20, 21 In periodontal fibroblasts it suppresses the proliferation and induces a reduction in collagen content.22 Because vitamin C stimulate the collagen synthesis in fibroblasts it might prevent the negative effects of P. gingivalis. A current study demonstrated that antibodies to P. gingivalis were negatively correlated with the plasma vitamin C concentration,3 which confirmed the hypothesis of a special interaction between P. gingivalis and vitamin C. The purpose of the present study was to investigate the influence of vitamin C on the cytotoxic effects of P. gingivalis on human gingival fibroblasts in vitro.
Section snippets
Culture of human gingival fibroblasts
Primary cultures of human gingival fibroblasts were obtained from a gingival biopsy of a donator with clinically healthy periodontium23 after the positive vote of the ethical committee of the Friedrich Schiller University Jena (B1881-10/06). Briefly, cells were cultured in Dulbecco's modified Eagle medium (DMEM; PAA Laboratories, Linz, Austria) supplemented with 10% fetal calf serum (FCS; PAA Laboratories, Linz, Austria) and antibiotics (10 U ml−1 penicillin, 10 mg/ml streptomycin, 25 μg/ml
Measurement of cell viability
Co-incubation with vitamin C significantly increased the viability rate of HGF (Fig. 1). The highest viability rate was observed with a vitamin C concentration of 12 μg/ml and exceeded the values of untreated HGF by 29.2 ± 6.4% (p < 0.05). The viability rates were lower with concentrations about 12 μg/ml, but the rates increased also the level of untreated controls.
Fig. 2 shows the viability rates varying with exposure to viable P. gingivalis ATCC 33277 and exposure time. In comparison to controls
Discussion
The present results demonstrate that vitamin C decreases apoptosis and prevents viability loss in HGF exposed to viable P. gingivalis.
Several studies have investigated the effect of antioxidants on bacteria-induced apoptosis in different cell lines. For example, researchers found that the treatment with tocopherol (vitamin E) increased the survival of Pseudomonas aeruginosa-infected cells.24 Before this investigation, there were no studies describing the effect of antioxidants in a context with
References (43)
Plasma vitamin C is inversely associated with periodontitis
J Evid Based Dent Pract
(2008)- et al.
The role of ascorbic acid deficiency in human gingivitis—a new hypothesis
J Theor Biol
(1984) - et al.
Effect of ascorbic acid nutriture on blood histamine and neutrophil chemotaxis in guinea pigs
J Nutr
(1991) - et al.
Effects of vitamin C intake on gingival oxidative stress in rat periodontitis
Free Radic Biol Med
(2009) - et al.
Activated polymorphonuclear leucocytes consume vitamin C
FEBS Lett
(1984) - et al.
Purification and characterization of Porphyromonas gingivalis outer membrane antigens
Arch Oral Biol
(1995) - et al.
Inhibition of polymorphonuclear leucocyte phagocytosis by Porphyromonas gingivalis culture products in patients with adult periodontitis
Arch Oral Biol
(1993) - et al.
Pseudomonas aeruginosa induces apoptosis in human endothelial cells
Microb Pathog
(2000) - et al.
Etiology and pathogenesis of periodontal disease
Dent Clin North Am
(2005) - et al.
respiratory infections and the immune system
Trends Immunol
(2003)
l-Ascorbic acid induces apoptosis in acute myeloid leukaemia cells via hydrogen peroxide-mediated mechanism
Int J Biochem Cell Biol
Staphylococcal enterotoxin C injection in combination with ascorbic acid promotes the differentiation of bone marrow-derived mesenchymal stem cells into osteoblasts in vitro
Biochem Biophys Res Commun
Effects of ascorbic acid on proliferation and collagen synthesis in relation to the donor age of human dermal fibroblasts
J Invest Dermatol
Regulation of apoptosis by vitamin C
J Biol Chem
Java project on periodontal diseases: the relationship between vitamin C and the severity of periodontitis
J Clin Periodontol
Periodontitis is associated with a low concentration of vitamin C in plasma
Clin Diagn Lab Immunol
Periodontal health related to plasma ascorbic acid
Proc Finn Dent Soc
Grapefruit consumption improves vitamin C status in periodontitis patients
Br Dent J
Oral scurvy and periodontal disease
J Can Dent Assoc
Role of ascorbic acid in periodontal ligament cell differentiation
J Periodontol
The role of reactive oxygen and antioxidant species in periodontal tissue destruction
Periodontol 2000
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