Anticariogenic activity of acanthoic acid isolated from Acanthopanax koreanum $N_{AKAI}$ against oral pathogens

김홍준,정승일,이화정,주영승,Kim, Hong-Jun,Jeong, Seung-Il,Lee, Hwa-Jung,Ju, Young-Sung 한국한의학연구원 2006 한국한의학연구원논문집 Vol.12 No.1

치면세균막내의 미생물 중 Streptococcus mutans $N_{AKAI}$는 치면세균막형성에 가장 중요한 역할을 하는 원인균으로 알려져 있으며, Streptococcus mutans $N_{AKAI}$는 치아우식을 일으키는 주요 원인균으로 주목을 받고 있다. Streptococcus mutans $N_{AKAI}$균에 항균효과를 나타내는 화합물 acanthoic acid (AA, $C_{20}H_{30}O_2$)을 Acanthopanax koreanum $N_{AKAI}$의 클로로포롬 추출물에서 각종 컬럼크로마토그래피법으로 추출분리하여 ms, 1H-NMR 그리고 13C-NMR 등을 이용하여 구조를 동정하였다. AA의 Streptococcus mutans $N_{AKAI}$에 대한 최소억제농도(MIC)와 최소살균농도(MBC)를 조사하였으며, 이 때 각각 2와 $4\:{\mu}g/mL$을 나타냈었다. 또한 다른 천연물에서 분리된 화합물인 tanshinone IIA는 8/16\;${\mu}g/mL$, 녹차의 추출물과 eucalyptoi은 $250/500\;{\mu}g/mL$이었다. 그리고 또 다른 치주질환의 원균인 Streptococcus sobrinus $N_{AKAI}$, Streptococcus sanguis $N_{AKAI}$ 및 Streptococcus gordonii $N_{AKAI}$의 최소억제 농도의 범위는 $4{\sim}32\;{\mu}g/mL$이었다. 우리의 연구결과에서 Acanthopanax koreanum $N_{AKAI}$에서 분리한 acanthoic acid 치주질환 원인균들을 억제하는 항균제로의 가능성을 확인하였다. This research was aimed to search for natural antimicrobial agents that are sefe for humans and specific for oral pathogens. Acanthoic was isolated from the chloroform fraction of methanol extract of Acanthopanax koreanum $N_{AKAI}$ and its structure were elucidated by 13-NMR, 1H-NNR and ESIMS. Antibacterial activity of acanthoic acid was investigated by the minimum inhibitory (MIC) and minimun bactericidal (MBC) concentration. MIC/MBC of acanthoic acid against Streptococcus mutans $N_{AKAI}$ causing dental caries was determined to be $2/4\;{\mu}g/mL$, which was much lower than these of other natural antimicrobial agents such as $8/16\;{\mu}g/mL$ of sangurinarine and $250/500\;{\mu}/mL$ of green tea extract, $500/600\;{\mu}g/mL$ of thymol and borneol. Acanthoic and significantly inhibited the growth of other cariogenic bacteria such as Streptococcus sobrinus $N_{AKAI}$ and Streptococcus sanguis $N_{AKAI}$, and Streptococcus gordonii $N_{AKAI}$ in the MIC range of $4{\sim}32\;{\mu}g/mL$. My finding suggests that acanthoic acid could be employed as a potential antibacterial agent for preventing dental caries.

The Effect of Chlorhexidine on the formation of bone nodules by Periodontal ligament Cells in Vitro

최희준,지숙,국중기,장현선,박주철,김흥중,김종관,김병옥,Choi, Hui-Jun,Ji, Suk,Kook, Joong-Ki,Jang, Hyun-Seon,Park, Joo-Cheol,Kim, Heung-Joong,Kim, Chong-Gwan,Kim, Byung-Ock The Korean Academy of Periodontoloy 2006 Journal of Periodontal & Implant Science Vol.36 No.2

사람치주인대섬유모세포(human periodontal ligament fibroblast, PDLF)의 기능 손상과 클로르헥시딘(Chlorhexidine, CHX)의 세포독성에 관한 분자적인 기전은 최근까지도 불명확하다. 이 연구의 목적은 PDLF에 의한 골결절 형성에 있어서 CHX의 효과를 평가하고, 치주수술후에 치주병원균의 최소억제농도(minimal inhibitory concentration, MIC)를 평가하고자 하였다. CHX의 세포독성을 평가하기 위해서 MTT assay법을 실시하였다. CHX은 0.12%에서 0.00012%까지, 즉 10-1000배로 희석시킨 후 30, 60, 120초 동안 PDLF에 적용되었고, 석회화된 결절은 alizarin red 용엑에 염색되었다. 치주병원균에 대한 CHX의 MIC가 평가되었다. 이 연구 결과, 세포생존율 검사에서는, 단지 0.12% CHX 에 노출되었던 세포들만 세포 증식 소견을 다소 나타내었다. 모든 CHX 농도(0.12%-0.00012%)에서 PDLF에 의한 골결절 형성은 의미있는 감소를 나타내었다. 또한 치주병원균에 대한 CHX의 MIC는 0.0012%로 나타났다. PDLF의 골결절 형성에 영향을 주는 농도(0.00012%)는 세포독성을 나타내는 농도(0.12%)보다 더 낮은 농도를 보였고, 치주병원균의 최소억제에 필요한 농도는 0.0012%로 나타났다. 이런한 결과들은 통상적으로 상용되는CHX이 PDLF에 의한 골결절 형성에 있어서 영향을 미칠 수 있음을 시사하였다.

구강 내 세균에 대한 Essential oil의 항균효과에 관한 연구

백병주,양연미,이경열,이용훈,김미아,김재곤,이선영 大韓小兒齒科學會 2009 大韓小兒齒科學會誌 Vol.36 No.1

Essential oils are mixture of volatile, lipophilic compounds originating from plants. Essential oils have potential biological effects, i.e., antibacterial, antifungal, spasmolytic and antiplasmodial activities and insect-repellent property. In this study, five essential oils, namely R, LG, FR, O, and NM, extracted from various aromatic plants were used to test their antimicrobial activity against the oral microorganisms. The effects of essential oils were investigated against eight important bacteria, Streptococcus mutans (S. mutans), Staphylococcus aureus (S. aureus), Streptococcus sanguis (S. sanguis), Streptococcus anginosus (S. anginosus), Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans), Streptococcus sobrinus (S. sobrinus), Staphylococcus epidermidis (S. epidermidis), and Escherichia coli (E. coli). Essential oils, except NM, effectively inhibited the growth of tested oral pathogenic microorganisms dose-dependently. However, the essential oils didn’t show a significant inhibitory effect against E. coli and S. epidermidis. Consequently, these results represented that essential oil-mediated anti-microbial activity was prominent against the oral pathogenic bacteria. For example, minimum bactericidal concentration(MBC) of R, LG, FR oil against A. actinomycetemcomitans was very low as 0.078 mg/mL. In addition, minimum inhibitory concentration (MIC) of R, LG, FR, O oil against S. mutans was low as 0.156 mg/mL in vitro. Essential oil은 식물로부터 추출한 휘발성의 호지방성 화합물로서 essential oil은 잠재적으로 항균, 항진균, 진경련, 항말라리아성, 곤충퇴치능력 등의 생물학적 효과를 가지고 있다. 본 연구에서는 다양한 방향식물에서 추출한 Citral, Pineole, Linalool, Eugenol, Limonene, Pinene 등으로 구성된 다섯종류의 essential oil을 사용하였으며, 여덟 군의 중요한 병원성 세균인 Streptococcus mutans(S. mutans), Staphylococcus aureus(S. aureus), Streptococcus sanguis(S. sanguis), Streptococcus anginosus(S. anginosus), Actinobacillus actinomycetemcomitans(A. actinomycetem- comitans), Streptococcus sobrinus(S. sobrinus), Staphylococcus epidermidis(S. epidermidis), Esherichia coli (E. coli)에 대한 저항성으로서 항균능력을 평가하였다. Essential oil의 항균능력은 용액희석방법을 사용하여 검사하였으며, essential oil 농도범위는 10 mg/mL, 5 mg/mL, 2.5 mg/mL, 1.25 mg/mL, 0.625 mg/mL, 0.516 mg/mL, 0.3125 mg/mL, 0.078 mg/mL, 0.039 mg/mL, 그리고 0.015 mg/mL였다. Essential oil은 용량 의존적으로 시험 균주의 성장을 효과적으로 억제하였으며, NM을 제외하고 R, LG, FR, O oil은 저농도에서 구강 병원성 미생물의 성장을 억제하였다. 또한 사용된 essential oil은 E.coli와 S. epidermidis의 성장에는 탁월한 억제효과를 보이지는 못했으나 구강 병원성 미생물에 대해서는 특별한 성장 억제효과를 나타냈다.

Essential oils의 구강내 세균에 대한 항세균 및 성장억제 효과

이영수 ( Young Soo Lee ),김신규 ( Sin Gyu Kim ),양태철 ( Tae Cheol Yang ),김광석 ( Gwang Seok Kim ),전재규 ( Jae Gyu Jeon ),장기완 ( Kee Wan Chang ) 대한예방치과·구강보건학회 2006 大韓口腔保健學會誌 Vol.30 No.4

Sodium benzoate의 최소억제농도 측정을 위한 새로운 방법

차문석,김미림,박찬성 慶山大學校 1998 論文集 Vol.16 No.1

The sensitivity of various pathogenic bacteria (Aeromonas hydrophila.Escherichia coli 0157 :H7, Listeria monocytogenes, Staphylococcus aureus 196E, Salmonella typhimurium) to the sodium benzoate was tested. Tryptic soy broth(TSB) containing 0-2% (w/v) of sodium benzoate adjusted pH 5.5,6.0, 6.5 and 7.0 was inoculated with 10^(5)∼10^(6) CFU/ml of pathogenic bacteria and incubated at 35℃ for 24 hours. Bacterial growth was determined by turbidity at 660 nm and viable cell counts of bacterial culture at each pH. Minimum inhibitoy concentration (MIC) of each bacteria increased with increasing pH of sodium benzoate. S. aureus and A. hydrophila were very resistant to sodium benzoate, the MICs were 0.5%, 1.0%, 2.0% at pH 5.5, 6.0,6.5 respectively. The MICs of S. typhimurium and L. monocytogenes to sodiumbenzoate were 0.3%, 0.5%, 0.7%, 1.5% at pH 5.5, 6.0, 6.5, 7.0 respectively.The MICs of E. coli were 0.5% at pH 5.5 and 6.0, 1.0% at pH6.5 and 7.0. The color of bacterial culture by resazurin test was changed to blue → violet → pink→ pale pink →colorless with increasing viable cell counts of bacteria. Turbidity was accorded with viable cell counts and the color of resazurin test. MICs of sodium benzoate against each pathogenic bacteria were 97% turbidometric inhibition of bacteria and were blue color by resazurin test. Resazurin test was a new, simple and rapid method to determine the MIC of sodium benzoate.

Potassium Sorbate의 최소억제농도 측정을 위한 Resazurin반응 이용

박찬성 慶山大學校 1999 論文集 Vol.17 No.1

The sensitivity of various pathogenic bacteria(Aeromonas hydrophila. Escherichia coli 0157:H7. Listeria monocytogenes. Staphylococcus aureus 196E, Salmonella typhimurium) to the potassium sorbate was tested. Tryptic soy broth(TSB) containing 0∼2%(w/v) of potassium sorbate adjusted pH 5.5, 6.0, 6.5 and 7.0 was inoculated with 10의5승∼10의6승 CFU/ml of pathogenic bacteria and incubated at 35℃ for 24 hours. Bacterial growth was determined by turbidity at 660 nm and viable cell counts of bacterial culture at each pH. Minimum inhibitoy concentration(MIC) of each bacteria increased with increasing pH of culture broth. The MICs of S. aureus and A. hydrophila were 0.5%, 1.0%, 2.0% at pH 5.5, 6.0, 6.5respectively. S. aureus and A. hydrophila were not inhibited at pH 7.0and they were very resistant to potassium sorbate. The MICs of S. typhimurium and L. monocytogenes to potassium sorbate were 0.3%,0.5%, 0.7%, 1.5% at pH 5.5. 6.0, 6.5, 7.0 respectively. The MICs of E. coli were 0.3%. 0.5%, 0.7%, 0.8% at pH 5.5, 6.0, 6.5, 7.0 respectively and it was the most sensitive strain to potassium sorbate. The color of bacterial culture by resazurin test was changed to blue → violet → pink → pale pink → colorless with increasing viable cell counts of bacteria. Turbidity was accorded with viable cell counts and the color of resazurin test MICs of potassium sorbate against each pathogenic bacteria were 97% turbidometric inhibition of bacteria and were blue color by resazurin test. Resazurin test was a new, simple and rapid method to determine the MIC of potassium sorbate.

원저 : 예방치과학 분야 ; 사람 치면세균막에서 분리 및 동정된 Fusobacterium nucleatum의 수종 항생제에 대한 내성 검사

최재흥 ( Jae Heung Choi ),국중기 ( Joong Ki Kook ),김진수 ( Jin Soo Kim ),김화숙 ( Hwa Soon Kim ),이병진 ( Byoung Jin Lee ),김동기 ( Dong Kie Kim ) 대한구강보건학회 2008 大韓口腔保健學會誌 Vol.32 No.4

구강 미생물 7종에 대한 멘톨과 불소치약의 항균 활성 비교 연구

조세희(Cho, Sehee),안태규(Ahn, Tae-Kyu),윤찬건(Yun, Chan-Gun),김지우(Kim, Jiwoo),전예담(Jeon, Yedam),황진서(Hwang, Jinseo),김지인(Kim, Jee-In),이현경(Lee, Hyeongyeong),임재환(Lim, Jae-Hwan) 한국과학영재교육학회 2020 과학영재교육 Vol.12 No.3

구강에는 600종 이상의 미생물이 존재하는 것으로 보고되고 있다. 알코올과 항생제 제품을 대체하여 천연물을 사용한 구강 위생 제제의 개발연구가 보고되고 있으나, 특정 성분들의 균주별 항균 활성 등에 관한 연구는 부족하다. 본 연구에서는 멘톨, 녹차 추출물, 불소치약, 플루오린화나트륨을 사용하여 구강 미생물 7종 -S. mutans, E. coli, B. subtilis, S. aureus, B. cereus, P. aeruginosa, L. brevis- 에 대한 항균효능을 확인하고자 하였다. 디스크 확산법을 통해 멘톨(200 mg/mL)은 구강 미생물 7종, 치약의 경우는 2종의 균주 -S. mutans, B. cereus- 에서 10.4~ 21.9 mm의 clear zone이 형성되는 것을 확인하였다. 불소치약과 멘톨의 복합 처리 시에는 구강 미생물 4종에서 증가된 항균 활성을 확인하였다. 또한 MIC 분석을 통해, 멘톨은 200~500 ㎍/mL의 농도에서 모든 균주에 대한 최소억제농도를 확인할 수 있었고 플루오린화나트륨의 경우 E. coli에서 40 ㎍/mL의 억제농도를 확인할 수 있었다. 결과적으로, 구강청결제 및 구강질환 항생물질의 대체소재로서 멘톨의 활용 가능성을 제시하고자 한다. More than 600 kinds of microorganisms exist in the oral cavity. Studies on the development of oral hygiene preparations using natural products as a substitute for alcohol and antibiotic products have been reported, but studies on the antibacterial activity of specific ingredients by strain are insufficient. In this study, menthol, green tea extract, fluoride toothpaste, and sodium fluoride were used against 7 kinds of oral microorganisms, S. mutans, E. coli, B. subtilis, S. aureus, B. cereus, P. aeruginosa, L. brevis. Using the disk diffusion method, It was confirmed that clear zones of 10.4 to 21.9 mm are formed in 7 types of oral microorganisms when menthol (200 mg/mL) is treated and 2 strains, S. mutans and B. cereus, when treated with fluoride toothpaste. When combined treatment with fluoride toothpaste and menthol, increased antibacterial activity was observed in four oral microbes. Through the MIC analysis, the minimum inhibitory concentration of menthol for all strains was confirmed at a concentration of 200-500 ㎍/mL, and the inhibitory concentration of sodium fluoride was confirmed at 40 ㎍/mL in E. coli. As a result, we suggest that the possibility of using menthol as a substitute for oral cleansers and antibiotics for oral diseases.

실험용 항균성 불소바니쉬의 최소억제농도와 최소살균농도

손주리(Ju-Lee Son),김아진(Ah-Jin Kim),오승한(Seunghan Oh),배지명(Ji-Myung Bae) 대한치과재료학회 2018 대한치과재료학회지 Vol.45 No.2

농산물에서 분리한 무름병균의 항생제 최소 억제농도 조사

김다운(Dawoon Kim),김나예슬(Nayeseul Kim),김채린(Chaerin Kim),정명인(Myung-In Jeong),오광교(Kwang Kyo Oh),김보은(Bo-Eun Kim),류재기(Jae-Gee Ryu),정지은(Jieun Jung),지삼녀(Samnyu Jee),류경열(Kyoung-Yul Ryu) 한국농약과학회 2021 농약과학회지 Vol.25 No.4