Objective: To evaluate the in vitro antimicrobial and antibiofilm properties and the immune modulatory activity of cannabidiol (CBD) and cannabigerol (CBG) on oral bacteria and periodontal ligament fibroblasts (PLF). Methods: Cytotoxicity was assessed by propidium iodide flow cytometry on fibroblasts derived from the periodontal ligament. The minimum inhibitory concentration (MIC) of CBD and CBG for S. mutans and C. albicans and the metabolic activity of a subgingival 33-species biofilm under CBD and CBG treatments were determined. The Quantification of cytokines was performed using the LEGENDplex kit (BioLegend, Ref 740930, San Diego, CA, USA). Results: CBD-treated cell viability was greater than 95%, and for CBG, it was higher than 88%. MIC for S. mutans with CBD was 20 µM, and 10 µM for CBG. For C. albicans, no inhibitory effect was observed. Multispecies biofilm metabolic activity was reduced by 50.38% with CBD at 125 µg/mL (p = 0.03) and 39.9% with CBG at 62 µg/mL (p = 0.023). CBD exposure at 500 µg/mL reduced the metabolic activity of the formed biofilm by 15.41%, but CBG did not have an effect. CBG at 10 µM caused considerable production of anti-inflammatory mediators such as TGF-β and IL-4 at 12 h. CBD at 10 µM to 20 µM produced the highest amount of IFN-γ. Conclusion: Both CBG and CBD inhibit S. mutans; they also moderately lower the metabolic activity of multispecies biofilms that form; however, CBD had an effect on biofilms that had already developed. This, together with the production of anti-inflammatory mediators and the maintenance of the viability of mammalian cells from the oral cavity, make these substances promising for clinical use and should be taken into account for future studies.
Background. In health, microorganisms have been associated with the disease, although the current knowledge shows that the microbiota present in various anatomical sites is associated with multiple benefits. Objective. This study aimed to evaluate and compare the genitourinary microbiota of chronic prostatitis symptoms patients and fertile men. Materials and Methods. In this preliminary study, ten volunteers have included 5 volunteers with symptoms of chronic prostatitis (prostatitis group) and five fertile volunteers, asymptomatic for urogenital infections (control group) matched by age. Bacterial diversity analysis was performed using the 16S molecular marker to compare the microbiota present in urine and semen samples from chronic prostatitis symptoms and fertile volunteers. Seminal quality, nitric oxide levels, and seminal and serum concentration of proinflammatory cytokines were quantified. Results. Fertile men present a greater variety of operational taxonomical units-OTUs in semen (67.5%) and urine (17.6%) samples than chronic prostatitis symptoms men. Chronic prostatitis symptoms men presented a higher concentration of IL-12p70 in seminal plasma. No statistically significant differences were observed in conventional and functional seminal parameters. The species diversity in semen samples was similar in healthy men than prostatitis patients, inverted Simpson index median 5.3 (5.0–10.7) vs. 4.5 (2.1–7.8, ). Nevertheless, the microbiota present in the semen and urine samples of fertile men presents more OTUs. Less microbial diversity could be associated with chronic prostatitis symptoms. The presence of bacteria in the genitourinary tract is not always associated with the disease. Understanding the factors that affect the microbiota can implement lifestyle habits that prevent chronic prostatitis. Conclusion. Chronic prostatitis does not seem to affect male fertility; however, studies with a larger sample size are required. Our preliminary results strengthen the potential role; the greater bacterial diversity is a protective factor for chronic prostatitis.
To detect DNA of different microorganisms, in semen samples from apparently healthy men and correlate their presence with seminal quality. Semen samples from 81 healthy volunteers were collected, and semen parameters were analyzed. DNA extraction was performed using the phenol-chloroform technique, and the microorganisms were detected by the amplification of specific primers using polymerase chain reaction. DNA from at least one of the microorganisms was detected in 78 samples. The most frequent microorganism found in semen were: Lactobacillus spp. (70%), Neisseria gonorrhoeae (N. gonorrhoeae) (36%), Streptococcus epidermidis (64%), Klebsiella pneumoniae (56%), Staphylococcus aureus (32%), Chlamydia trachomatis (C. trachomatis) (28%), Pseudomonas aeruginosa (27%). The seminal parameters of all semen samples were over the lower reference values for normal semen analysis. To compare with negative samples, seminal volume was higher for the Escherichia coli positive samples and lower for Pseudomonas aeruginosa positive samples. Semen samples positive for Staphylococcus aureus had worse sperm morphology. The frequency of progressive motility was higher in positive samples for N. gonorrhoeae and C. trachomatis. Positive semen samples for C. trachomatis had a higher concentration per milliliter. It is common to find microorganisms in semen of asymptomatic men, including those responsible for sexually transmitted infections. Antimicrobial treatment is recommended only in those individuals with a sexually transmitted infection (C. trachomatis and N. gonorrhoeae) and always promote condom use.
This work evaluated the Lippia origanoides derivatives in vitro effect on polymicrobial biofilms of Streptococcus mutans, Lactobacillus rhamnosus and Candida albicans. Additionally, the cytotoxic effect of the oils on human skin keratinocytes (HaCaT) and fibroblasts of the periodontal ligament (FLP) cell lines was evaluated. The minimum inhibitory concentration, the inhibitory activity on monomicrobial (S. mutans) and polymicrobial biofilm (S. mutans, L. rhamnosus and C. albicans) of L. origanoides four essential oils and terpenes (thymol and carvacrol) were evaluated. The cytotoxic effect of each one of the compounds was measured, and all the tests were compared against chlorhexidine. All the evaluated compounds reached an inhibition percentage of S. mutans monomicrobial biofilms formation of 100 % at 600 µg/mL (p < 0.0001). The highest concentration (2 MIC) eradicated 100 % of S. mutans-preformed biofilms after 5 min L. origanoides carvacrol + thymol and thymol chemotypes showed marked reductions in topography, the number of microbial cells and extracellular matrix on polymicrobial biofilm. The cytotoxic effect of the compounds was very similar to chlorhexidine. L. origanoides essential oils have an inhibitory effect on mono and polymicrobial biofilms. The oils present a similar cytotoxic effect to chlorhexidine on HaCaT and FLP cell lines. However, including these compounds in formulations for clinical use is an exciting proposal yet to be investigated.
1 July 2017 in Clínica E Investigación En Ginecología Y Obstetricia