Bacterial steroidogenesis by periodontal pathogens and the effect of bacterial enzymes on steroid conversions by human gingival fibroblasts in culture.
Studies were performed to investigate the effect of microbial culture supernatants of periodontal pathogens on the metabolism of radiolabelled testosterone in the presence or absence of human gingival fibroblasts. Subgingival plaque samples were obtained on paper points from 3 sites with probing depth values of 6-8 mm. Samples were incubated with 14C-testosterone for 24 h in brain heart infusion (BHI) broth. Similar incubations were also carried out with strains of A. actinomycetemcomitans, P. Intermedius and P. gingivalis to study the metabolism of radiolabelled testosterone by these periodontal pathogens. At the end of a 24 h incubation period with fibroblasts and supernatants or sonicates, the radioactive metabolites were extracted with ethyl acetate, evaporated and subjected to thin layer chromatography. The separated metabolites were quantified by scanning the radioactive plates using a Berthold linear analyser. When three sub-gingival plaque samples were incubated with radiolabelled testosterone there were 50-fold, 10-12-fold and 15-17-fold increases in 5 alpha-dihydrotestosterone (DHT) synthesis over 4-androstenedione production in these mixed microbial cultures. The two strains of P. intermedius produced 3- and 20-fold increases in 4-androstenedione production and DHT synthesis respectively. Both strains of A. actinomycetemcomitans and P. gingivalis showed 3-4-fold and 12-28-fold increases respectively in 4-androstenedione synthesis over that of DHT. Culture supernatants of P. intermedius and P. gingivalis caused 3-fold and 2-fold increases in DHT synthesis by fibroblasts over controls. There was little change in the case of the third pathogen. Since DHT has implications on matrix synthesis by fibroblasts in the environment of plaque associated inflammatory periodontal disease, bacterial metabolism and the effect of bacterial supernatants on human gingival fibroblasts can influence the degree of inflammatory repair.
Soory
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Department of Periodontology, King's College School of Medicine and Dentistry, London, England.
D016976:Aggregatibacter actinomycetemcomitans / Q000201:enzymology / Q000378:metabolism; D000735:Androstenedione / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D002478:Cells, Cultured; D042944:Cholestenone 5 alpha-Reductase; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D003773:Dental Plaque / Q000378:metabolism; D013196:Dihydrotestosterone / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism / Q000382:microbiology; D005881:Gingiva / Q000166:cytology / Q000378:metabolism / Q000382:microbiology; D006801:Humans; D010088:Oxidoreductases / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000378:metabolism*; D018713:Prevotella intermedia / Q000201:enzymology / Q000378:metabolism*; D013739:Testosterone / Q000378:metabolism*
D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D013196:Dihydrotestosterone; D013739:Testosterone; D000735:Androstenedione; D010088:Oxidoreductases; D042944:Cholestenone 5 alpha-Reductase