米諾四環素加入亞甲基藍作為局部緩釋性藥物之牙周病原菌感受性及細胞毒性之研究

In vitro cytotoxicity and susceptibility tests to periodontol pathogens with a combination of minocycline and methylene blue as a LDSRS

錢弘恩(Hung-En Chien)

指導教授 ： 陳志豪

中山醫學大學/醫學院/微生物免疫研究所/碩士(2012年)

https://doi.org/10.6834/CSMU.2012.00014

摘要

防止牙菌斑的再堆積是治療牙周病復發的主要措施，目前已知最有效的方法是機械性清除牙菌斑及結石。但某些類型之牙周病還需要使用藥物治療作為輔助治療，近年來國內外對使用抗菌藥物治療牙周炎進行了許多研究，並已研製出多種牙周病治療藥物，但仍有缺點，像是無法有效治療某些重症及慢性牙周病。本研究之目的為設計出一種理想之牙周病藥物治療局部緩釋劑配方，以期能更有效治療牙周病，其理論基礎主要是配方以minocycline (MNC)為主要成份，加入亞甲基藍 (methylene blue ) 利用其氧化還原作用 (oxidation-reduction) 中為氧原子接受者產生還原作用，使原來MNC 易因曝光而氧化破壞產生細胞毒性之缺點因產生保護作用而改善，並且亞甲基藍之吸收峰值為 (609-668 nm) 之藍色光是MNC (黃色) 之互補色，因其互補作用可防止MNC被光破壞，因而產生雙重保護作用之效果。表一至表四為MNC對E. coli組、Staphylococcus aureus組、Porphyromonas gingivalis及Prevotella intermedia組之紙錠擴散法抑菌實驗，結果顯示其抑菌圈之平均直徑分別為5.3 ± 0.6 mm、10.0 ± 1.0 mm、4.8 mm ± 0.3 mm及12.8 ± 0.8 mm，證實MNC對革蘭氏陽性、陰性菌及牙周病原菌均有明顯的抑菌效果。若經光照破壞後其抑菌圈之平均直徑分別降低為2.5 ± 0.5 mm、7.6 ± 0.8 mm、2.5 ± 0.0 mm及9.8 ± 0.3 mm。但MNC加甲基藍經光照後其抑菌圈之平均直徑分別為3.5 ± 0.5 mm、8.3 ± 1.0 mm、3.4 ± 0.2 mm及12.0 ± 0.5 mm。結果顯示MNC加入亞甲基藍可以避免因光照破壞，其保護效果平均提升達58.5%，而在細胞毒性試驗中，可以發現亞甲基藍加入MNC被光照後約減少60%之光毒性產生。
結論：經以上實驗證實，本研究設計之複方局部緩釋性藥物應用於牙周病治療，對於藥物保存之方便性以及藥物使用之安全性都有改善，且更有效抑制牙周囊袋中病原菌之生長，對牙周病的化學療法之改善應具有正面之意義。

關鍵字

牙周病 ； 米諾四環素 ； 亞甲基藍 ； 局部緩釋性藥物

並列摘要

The recurrence of periodontitis after treatment is the most troublesome problem in clinical dentistry. How to avoid the re-accumulation of biofilm in periodontal pocket is the main approach for resolving this problem. Studies shown that mechanical debridement is effective for the majority of periodontitis patients, but there still have some cases need meditation for adjunctive therapy. Many studies investigated systemic antibiotics in the treatment of periodontal diseases but were not indicated in the management of cases of chronic refractory periodontitis. To solve this problem, local delivery slow-released systems (LDSRS) have been developed. Although many studies had been done recently and showed good effect, other studies showed conflicting data. For this reason, improvement of LDSRS was necessary.
The aim of this study was to develop a new formula to improve LDSRS. The basic of the theory was to use a new component, methylene blue (MB), and combine it with the main ingredient, minocycline (MNC), which is easily oxidized to produce cytotoxicity, especially under light exposure. Due to the blue color of MB is the complementary color of yellow of MNC. Mixing the two components may reduce oxidation and degradation of MNC from light. The results showed that MNC had significant antimicrobial effect to testing strains: Escherichia coli, Staphylococcus aureus, Porphyromonas gingivalis and Prevotella intermedia. The diameters of the inhibition zone of the antimicrobial tests were 5.3 ± 0.6 mm, 10.0 ± 1.0 mm, 4.8 mm ± 0.3 mm and 12.8 ± 0.8 respectively, and after light wrecked, the diameters were decreased to 2.5mm ± 0.5 mm, 7.7 ± 0.8 mm, 2.5 ± 0.0 mm and 9.8 ± 0.3 mm respectively. In group MNC plus MB, the diameters increased to 3.5 ± 0.5 mm, 8.3 ± 1.0 mm, 3.4 ± 0.2 mm and 12.0 ± 0.5 mm after light wrecked, which showed that MB plus MNC can avoid light wrecking and increased the antibacterial effect up to 58.5 %. To evaluate the safety of MB along and the new combination of LDSRS, the cytotoxicity test was performed. The results showed that MB could effectively reduce MNC producing cytotoxicity about 60% due to light degradation.
In conclusion the combination of MB and MNC as a new LDSRS drug can inhibit pathogens growth more efficiency, prevent from light wrecking, and increase the duration for the drug storage and may have the potential valuable in clinical usage.

並列關鍵字

periodontol diease ； minocycline ； methelen blue ； LSDRS

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