About 700 types of microorganisms live in the oral cavity. Because the supply of nutrients necessary for the growth of microorganisms is continuous, the proliferated bacteria form a dental plaque, which is the mechanism that causes bacterial oral diseases . Therefore, the best way to prevent this is to remove the dental plaque, representatively using Tetracyclines, Metrodinidazole, Penicillin, Clindamycin, and Ciprofloxacin out of the various types of antibacterial agents . Chemical antibacterial agents inhibit bone loss and are effective for periodontitis . But their use is limited due to side effects such as the generation of resistant bacteria and mycosis .
Therefore, this study verified O’Leary index, PI, and GI as periodontal-related indicators and antibacterial effects on related pathogens that cause periodontal diseases to develop a natural mouthwash using Sambucus williamsii var. coreana extract, which has a natural antibiotic effect preventing periodontal disease. As a result of this study, O'Leary index, PI, and GI all showed a marked decrease from Treatment to After 5 days compared to the saline solution gargle group when applying mouthwash containing Sambucus williamsii var. coreana extract. Also, as the application time of the mouthwash containing Sambucus williamsii var. coreana extract increased, the values of periodontal-related indicators were effectively lowered, verifying that there was an effect of improving the oral environment. According to a previous study, Sambucus williamsii var. coreana extract had antioxidant and anti-inflammatory effects and had no cytotoxicity . Therefore, it is considered appropriate to apply it in the oral cavity as a mouthwash to suppress and prevent occurring periodontal disease.
P. micra, which is known as a bacterium associated with periodontal disease, is a gram-positive, non-motile, anaerobic coccus  that is more common in patients with severe or moderate periodontitis than those with gingivitis and mild periodontitis . In this study, a mouthwash containing Sambucus williamsii var. coreana extract was effective in the mandible. The effect appeared immediately after application, and the antibacterial effect showed up over time until After 5 days. P. micra is known to act as a core pathogen in developing the pathogenesis of the periodontal disease , and signaling molecules derived from P. micra stimulate the expression of the P. gingivalis proteolytic gingipain enzyme thirteen times more to enhance the growth and toxicity of Porphyromonas gingivalis, a major periodontal pathogen . P. gingivalis, a gram-negative anaerobic bacterium, is associated with periodontal destruction and increased risk of disease recurrence  and is the foremost periopathogen for periodontal disease progression . After applying the mouthwash containing Sambucus williamsii var. coreana extract to P. gingivalis, it decreased sharply in the maxilla and the mandible compared to the saline solution gargle group. The mouthwash was verified to be very effective against P. gingivalis as it decreased continuously from the immediate application. Therefore, a mouthwash using Sambucus williamsii var. coreana extract can inhibit and block the growth of core pathogens of periodontal disease. P. gingivalis is also considered the major pathogen because of its ability to change the normal microflora into a dysbiotic community . Among dysbiotic species, the increase in P. gingivalis, a major periodontal disease-related pathogen, is accompanied by various bacterial species including P. intermedia and F. nucleatum . Since the antibacterial effect was shown by marked reduction of P. gingivalis in this study, the use of mouthwash containing Sambucus williamsii var. coreana extract will play a huge role in reducing various anaerobic bacteria. P. intermedia, one of the anaerobic black bacteria, is known to be one of the powerful causative agents of adult-type periodontitis and various types of periodontal disease . In addition, since the expression frequency of P. nigrenscens was reported to be more than double that of P. intermedia in the inflammatory site of patients with periodontal disease , dealing with P. nigrenscens is also necessary. Comparing with the saline solution gargle group, P. intermedia was effectively reduced in the mandible immediately after applying a mouthwash containing Sambucus williamsii var. coreana extract. There was no effect on P. nigrenscens immediately after using a mouthwash containing Sambucus williamsii var. coreana extract, but it was found to be effective in both maxilla and mandible after using the mouthwash for 5 days. Regarding the change according to the application time, both P. intermedia and P. nigrenscens in the mandible were more effectively reduced over time by a mouthwash containing Sambucus williamsii var. coreana extract. As a result, a mouthwash containing Sambucus williamsii var. coreana extract reduced P. gingivalis immediately after application. P. nigrenscens and P. intermedia also decreased as the application time increased. F. nocleatum is one of the oral microflora and also a causative agent of periodontitis. It is also related to preterm birth, colorectal cancer, inflammatory bowel disease, respiratory tract infections, cardiovascular disease, rheumatoid arthritis, and Alzheimer’s disease . T. denticola, a bacterium that causes progressive periodontitis, was reported that be associated with loss of attached gingiva and bleeding when the depth of the periodontal pocket and the periodontal pocket were measured . In the results of this study, F. nocleatum was reduced compared to the saline solution gargle group in After 5 days in the maxilla upon using a mouthwash containing Sambucus williamsii var. coreana extract. On the other hand, in the case of T. denticola, the mouthwash containing Sambucus williamsii var. coreana extract was found to be effective until After 5 days as the bacteria completely disappeared from Treatment in the mandible. In addition, E. corrodens is a bacterium that causes acute periodontitis and promotes complex infection. C. rectus is known as a bacterium involved in root canal infection and periodontal disease . Both bacteria appear in patients with severe periodontal disease. When comparing the saline solution gargle group and the Sambucus williamsii coreana var. extract gargle group, there was no significant difference between the two groups regarding E. corrodens. In the case of the saline solution gargle group, there was a significant difference in the mandible in After 5 days when the application time increased. It means that if the application time increases, the reduction of acute periodontitis-causing bacteria can occur even with a saline solution. The number of C. rectus was reduced in Sambucus williamsii var. coreana extract gargle group showing the antibacterial effect. In both the maxilla and the mandible, there were continuous differences immediately after applying a mouthwash containing Sambucus williamsii var. coreana extract. Based on these oral clinical results, Sambucus williamsii var. coreana extract was found to be an excellent natural substance with antibacterial effects for improving periodontal disease.
As periodontal disease progresses, bacterial species diversify , so continuous management of anaerobic bacteria related to periodontal disease is required. Based on the results of the study, a mouthwash containing Sambucus williamsii var. coreana extract reduced bacteria in the oral cavity such as P. gingivalis, C. rectus, T. denticola, P. intermedia, and E. corrodens immediately after application. As the application time of a mouthwash containing Sambucus williamsii var. coreana extract extended, the more marked antibacterial effect appeared, which means it is excellent in improving periodontal disease. According to the results of this study, mouthwash was more effective in the mandible. It seems that the teeth and periodontal tissues of the mandible were in contact with and stored more in the mouthwash when keeping the mouthwash in the mouth. Therefore, to maximize the effect of mouthwash, it will be more effective to gargle mouthwash enough to cover the entire oral cavity rather than simply holding it.
Based on this study verifying the practicality and development of oral health care products using Sambucus williamsii var. coreana extract, a mouthwash containing Sambucus williamsii var. coreana extract can inhibit and prevent periodontal disease. As a natural ingredient with sufficiently excellent effects, Sambucus williamsii var. coreana extract can be used for oral health by improving periodontal disease.