Effects of Ammonia and Volatile Fatty Acids Produced by Oral Bacteria on Tissue Culture Cells

Abstract

Culture filtrates of several bacterial species isolated from the oral cavity were tested for their effects on two types of tissue culture cells: Vero cells, the continuous cell line of African green monkey kidney cells; and chondrocytes, isolated from 15-day-old chick embryo tibiae. Only a limited number of bacterial species — i.e., the asaccharolytic black-pigmented Bacteroides species and Fusobacterium species - affected the two cell types. The effect on Vero cells, detected by the rounding of the cells, correlated with the butyric acid concentration in the bacterial supernatant, which confirms previous findings. A small enhancement of this effect was found with propionic acid and ammonium ions. The same strains which affected Vero cells also affected chondrocytes, detected by a vacuolization of the cells. However, volatile fatty acids on their own had no visible effect on these cells. Instead, ammonium ion in the culture filtrate, when present in concentrations of 20 to 60 mmollL, proved to be responsible for vacuolization of the chondrocytes. The volatile fatty acids (butyric and propionic) had a limited additive effect. No effects were visible with cell extracts of the bacteria.

Get full access to this article

View all access options for this article.

References

Bickel, M. and Cimasoni, G. (1985): The pH of Human Crevicular Fluid Measured by a New Microanalytical Technique, J Periodont Res 20:35-40.

Dean, R.T. ; Jessup, W. ; and Roberts, C.R. (1984): Effects of Exogenous Amines on Mammalian Cells, with Particular Reference to Membrane Flow, Biochem J 217:27-40.

Duguid, R. ; Al-Makadsi, F. ; and Cowley, G.C. (1980): Cytotoxic Material Extracted from Human Dental Plaque and Oral Streptococci, Arch Oral Biol 25:349-352.

Grenier, D. and Mayrand, D. (1985): Cytotoxic Effects of Culture Supernatants of Oral Bacteria and Various Organic Acids on Vero Cells, Can J Microbiol 31:302-304.

Helgeland, K. (1984): Inhibitory Effect of NH4 Cl on Secretion of Collagen in Human Gingival Fibroblasts, Scand J Dent Res 92:419-435.

Helgeland, K. (1985): pH and the Effect of NH4 Cl on Human Gingival Fibroblasts, Scand J Dent Res 93:39-45.

Holdeman, L.V. ; Cato, E.P. ; and Moore, W.E.C. (1977): Anaerobic Laboratory Manual, 4th ed. Blacksburg, VA: Anaerobe Laboratory, Virginia Polytechnic Institute and State University.

Kamen, P.R. (1981): The Effects of Bacterial Sonicates on Human Keratinizing Stratified Squamous Epithelium in vitro, J Periodont Res 16:323-330.

Larjava, H. ; Jalkanen, M. ; Penttinen, R. ; and Paunio, K. (1983): Enhanced Synthesis of Hyaluronic Acid by Human Gingival Fibroblasts Exposed to Human Dental Bacterial Extract, J Periodont Res 18:31-39.

10.

Levine, M. (1985): The Role for Butyrate and Propionate in Mediating HeLa-cells Growth Inhibition by Human Dental Plaque Fluid from Adult Periodontal Disease, Arch Oral Biol 30 :155-159.

11.

Ng, W. and Tonzetich, J. (1984): Effects of Hydrogen Sulfide and Methyl Mercaptan on the Permeability of Oral Mucosa, J Dent Res 63:994-997.

12.

Shenker, B.J. ; Kushner, M.E. ; and Tsai, C.C. (1982): Inhibition of Fibroblast Proliferation by Actinobacillus actinomycetemcomitans, Infect Immun 28:986-992.

13.

Singer, R.E. and Buckner, B.A. (1981): Butyrate and Propionate : Important Components of Toxic Dental Plaque Extracts, Infect Immun 32:458-463.

14.

Slots, J. (1982): Importance of Black-pigmented Bacteroides in Human Periodontal Disease. In: Host Parasite Interactions in Periodontal Diseases, R.J. Genco and S.E. Mergenhagen , Eds., Washington, D.C. : American Society for Microbiology, pp. 27-45.

15.

Slots, J. and Genco, R.J. (1984): Black-pigmented Bacteroides Species, Capnocytophaga Species, and Actinobacillus actinomycetemcomitans in Human Periodontal Disease: Virulence Factors in Colonization, Survival, and Tissue Destruction, J Dent Res 63:412-421.

16.

Taichman, N.S. ; Klass, J.E. ; Shenker, B.J. ; Macarak, E.J. ; Boehringer, H. ; and Tsai, C.C. (1984): Suspected Periodontopathic Organisms Alter in vitro Proliferation of Endothelial Cells, J Periodont Res 19:583-586.

17.

Tonzetich, J. and McBride, B.C. (1981): Characterization of Volatile Sulphur Production by Pathogenic and Non-pathogenic Strains of Oral Bacteroides, Arch Oral Biol 26:963-969.

18.

Touw, J.J.A. ; van Kampen, G.P.J. ; van Steenbergen, T.J.M. ; Veldhuuzen, J.P. ; and de Graaff, J. (1982a): The Effect of Culture Filtrates of Oral Strains of Black-pigmented Bacteroides on the Matrix Production of Chick Embryo Cartilage Cells in vitro, J Periodont Res 17:351-357.

19.

Touw, J.J.A. ; van Steenbergen, T.J.M. ; and de Graaff, J. (1982b): Butyrate: A Cytotoxin for Vero Cells Produced by Bacteroides gingivalis and Bacteroides asaccharolyticus , Antonie van Leeuwenhoek 48:315-325.

20.

Tsai, C.C. ; McArthur, W.P. ; Baehni, P.C. ; Hammond, B.F. ; and Taichman, N.S. (1979): Extraction and Partial Characterization of a Leukotoxin from a Plaque-derived Gram-negative Microorganism, Infect Immun 25:427-439.

21.

Van Kampen, G.P.J. ; Van Steenbergen, T.J.M. ; Schip-Per, C.A. ; DE Graaff, J. ; and Veldhuijzen, J.P. (1984): Proteoglycan Production by Chick Embryonic Chondrocytes is Inhibited by Culture Filtrate of Bacteroides gingivalis, J Periodont Res 19:483-488.

22.

Van Steenbergen, T.J.M. ; Den Ouden, M.D. ; Touw, J.J.A. ; and De Graaff, J. (1982): Cytotoxic Activity of Bacteroides gingivalis and Bacteroides asaccharolyticus, J Med Microbiol 5:253-258.

23.

Van Steenbergen, T.J.M. ; Van Winkelhoff, A.J. ; and De Graaff, J. (1984): Pathogenic Synergy: Mixed Infections in the Oral Cavity, Antonie van Leeuwenhoek 50:789-798.

24.

Van Winkelhoff, A.J. ; Carlee, A.W. ; and De Graaff, J. (1985): Bacteroides endodontalis and Other Black-pigmented Bacteroides in Odontogenic Abscesses, Infect Immun 49:494-497.

25.

Yessup, W. ; Shirazi, M.T. ; and Dean, R.T. (1983): Inhibition of Some Spontaneous Secretory Processes in Macrophages and Fibroblasts by Ammonium Chloride, Biochem Pharmacol 32 :2703-2710.