開發新型抗菌塗層於骨科移植材料上之應用

Development of new antibacterial coating for orthopedic transplantation materials

詹皓群(HAO-CHUN CHAN)

指導教授 ： 吳瑞璋

共同指導教授 ： 黃郁慈(Yu-Tzu Huang)

中原大學/工學院/化學工程研究所/碩士(2021年)

https://doi.org/10.6840/cycu202100408

摘要

本實驗檢測多種抗菌胜肽（Antimicrobial peptides, AMPs）對金黃色葡萄球菌（革蘭氏陽性菌）以及大腸桿菌（革蘭氏陰性菌）的抗菌能力，並探討其作用機制與是否適合應用於移植材料的塗層。透過OD值、AlamarBlue螢光測試的結果，確認各AMPs的IC50濃度（half maximal inhibitory concentration：將細菌生長量抑制50 ％所需的濃度）與MIC濃度（minimum inhibitory concentration：將細菌生長量抑制到接近0 ％所需的濃度）。選出了兩種對陽性菌與陰性菌皆有效的AMPs：D-GL13K（對大腸桿菌之IC50：3 μg/ml，MIC：3.5 μg/ml。對金黃色葡萄球菌之IC50：1.7 μg/ml，MIC：2.5 μg/ml。）和MSI-594（對大腸桿菌之IC50：1 μg/ml，MIC：4 μg/ml。對金黃色葡萄球菌之IC50：6 μg/ml，MIC：7 μg/ml），並透過場發式電子顯微鏡（FESEM）觀察細菌受損情況，確認AMPs的作用機制為膜破壞。此外，透過FESEM檢測EDC改質PEEK的表面變化，結果得到EDC會將PEEK表面變得粗糙，並初步測試了改質PEEK的抗菌能力，結果顯示MSI-594與EDC混合塗層於PEEK表面後依然保持對大腸桿菌的抗菌能力。
根據上述結果，確認了D-GL13K與MSI-594的抗菌潛力，以及利用EDC改質材料的可行性，未來可以進行AMPs塗層改質PEEK，並利用EDC增強塗層穩定性的相關測試，以增加移植材料的抗菌能力，降低細菌感染的風險。

關鍵字

抗菌胜肽 ； 大腸桿菌 ； 金黃色葡萄球菌 ； 膜破壞 ； PEEK改質

並列摘要

This study reports the antibacterial capability of several types of Antimicrobial peptides (AMPs) against Staphylococcus aureus (S. aureus, a gram positive bacterium) and Escherichia coli (E. coli, a gram negative bacterium) and also investigates if AMPs-coating films are applicable on the materials for transplantation.
The IC50 (half maximum inhibitory concentration) and MIC (minimum inhibitory concentration) of all AMPs candidates were calculated using the results of their visible light OD values and AlamarBlue fluorescence readings. Two AMPs candidates, D-GL13K and MSI-594, were concluded effective in inhibiting both gram positive and negative bacteria. The data obtained for D-GL13K against E. coli were IC50 3 μg/ml and MIC 3.5 μg/ml and against S. aureus IC50 1.7 μg/ml and MIC 2.5 μg/ml. The data obtained for MSI-594 against E. coli were IC50 1 μg/ml and MIC 4 μg/ml and against S. aureus IC50 6 μg/ml and MIC 7 μg/ml.The damage mechanism of bacteria surface was observed by FE-SEM. It was concluded that bacteria in tests were inhibited by membrane destruction. The surface modification of PEEK（Polyetheretherketone）by EDC (1-ethyl-(3-dimethylaminopropyl) carbondimimide) was also investigated. Its surface became rough by FE-SEM observation. The antibacterial activity of modified PEEK was tested. The results showed that the mixed coating of MSI-594 and EDC still maintained the antibacterial activity against E. coli.
This study concludes the inhibiting effectiveness of D-GL13K and MSI-594 against test antibacterial and also the potential feasibility of surface modification of PEEK for transplantation. This study could be extended to the surface modification of PEEK using an AMPs coating enhanced its stability by EDC. That modification could improve the antibacterial capabilities of materials for implantation from the risks of infection.

並列關鍵字

AMPs ； E. coli ； S. aureus ； Membrane destruction ； Modified PEEK

參考文獻

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