RESUMEN
As the risk of antibiotic-resistant bacteria increases, interest in non-antibiotic treatment is also increasing. Among the methods used in non-antibiotic therapy, natural antibiotics such as essential oils have disadvantages such as low efficiency. In the case of phototherapy, the light used for antibacterial activities has low penetration into the human body because of its short wavelength, making it of low medical utility. To solve this problem, this study aimed to determine conditions for enhancing the antibacterial activity of natural phytochemicals and visible light. Four natural phytochemical extracts that showed high antibacterial properties in previous studies were analyzed. Synergistic effects on antibacterial activity and cytotoxicity were determined when natural phytochemical extracts and visible light were simultaneously used. As a result, it was confirmed that the antibacterial activity increased by four times when Sanguisorba officinalis L. was irradiated with 465 nm for 10 min and 520 nm for 40 min, and Uncaria gambir Roxb. was irradiated with 465 nm for 10 min and 520 nm for 60 min compared to when Sanguisorba officinalis L. and Uncaria gambir Roxb. were used alone. The synergistic effect on antibacterial activity was independent of the absorption peak of the natural phytochemical extracts. In addition, in the case of natural phytochemical extracts with improved antibacterial activity, it was confirmed that the improvement of antibacterial activity was increased in inverse proportion to the light irradiation wavelength and in proportion to the light irradiation time. The antibacterial activity was enhanced regardless of antibiotic resistance. In the case of cytotoxicity, it was confirmed that there was no toxicity to A549 cells when treated with 465 nm, the shortest wavelength among the natural phytochemical extracts. These results show how to replace blue light, which has been underutilized due to its low transmittance and cytotoxicity. They also demonstrate the high medical potential of using natural phytochemical and visible light as a combination therapy.
RESUMEN
BACKGROUND: Periodontal disease is a highly prevalent illness that affects many dogs, reaching up to 85 % prevalence in individuals over the age of 4 years. Currently the drug of choice for combating the formation of dental plaque in these animals, the etiologic agent of the disease, is chlorhexidine, which has several side effects reported. Thus, surveys are conducted throughout the world in order to identify potential substitutes for antimicrobial therapy and prevention of periodontal disease. The objective of the work was to evaluate the antimicrobial activity of ß-caryophyllene against bacteria from dog's dental plaque in vitro and in vivo. The minimum inhibitory concentration was evaluated by agar microdilution assay, the induction or inhibition of bacterial adherence by sub-inhibitory concentrations in 96-well plates, and reduction of dental plaque formation in mongrel dogs subjected to topical solution with ß-caryophyllene for 15 days. RESULTS: Results showed minimum inhibitory concentrations above 100 mg/mL for 25 % of the isolates, 100 mg/mL for 3 %, 50 mg/mL for 25 %, 25 mg/mL for 12 %, 12.5 mg/mL for 19 % and 6.25 mg/mL for 16 %. Bacterial adherences of three Enterococcus sp., one Streptococcus sp., one Haemophilus sp., one Aerococcus sp., one Bacillus sp. and one Lactococcus sp. isolates were inhibited by subinhibitory concentration. One Lactococcus sp., one Bacillus sp. and one Streptococcus sp. were stimulated to adhere by concentrations of 0.19, 1.56 and 0.78 mg/mL, respectively. In vivo assay showed reduction in dental plaque formation by ß-caryophyllene, with final plaque coverage of 23.3 ± 2.6 % of the total area of the teeth, with significant difference compared with chlorhexidine group (37.5 ± 3.7 % - p < 0.05) and negative control group (65.5 ± 2.5 % - p < 0.001). CONCLUSIONS: The results showed that ß-caryophyllene has antimicrobial activity against the proliferation of dog's dental plaque-forming bacteria representing a suitable alternative to the use of chlorhexidine in prophylaxis and treatment of periodontal disease of dogs.