近年來全球高齡化與抗藥性問題日趨嚴重，其所衍生之問題已成為公共衛生重要議題，就慢性傷口而言，以綠膿桿菌(Pseudomonas aeruginosa)感染最為常見，因多重抗藥性P.aeruginosa的增加，提高了該菌感染治療的困難度，而不適當的治療，更促使死亡率增加。因此未來於治療P.aeruginosa時，可能須找出新的治療方式，以解決P.aeruginosa多重抗藥性增加之問題。本研究探討微電流應用於綠膿桿菌體外菌落之抗菌作用。其特定目標為；(I) 確立電抗菌之體外實驗模型。(II) 探討電流對綠膿桿菌生長之影響，並利用重複性試驗找出其相關式。電抗菌原理是使用電化學反應，加速銀針釋放銀離子和電極吸附細菌之能力，且因銀對細胞作用產生細胞毒性，抑制細菌於患處生長，進而達到降低菌種數量的目的，進而有利於日後感染性傷口之應用。

本研究首先考量電抗菌的臨床需求與功能性設計，並依照臨床感染綠膿桿菌之慢性傷口特性，設計電抗菌體外菌落實驗模型，並根據敏感性測試與菌種培養成長速率校正結果，採用菌液濃度為7.5*104及1.5*104 CFU/mL。最後，以電源供應模組來模擬預設電流刺激參數(4、0.4及0.04mA)對P.aeruginosa的抑菌效果，並利用重複性實驗來探討微電流應用於綠膿桿菌體外菌落之抗菌作用，並找出其相互關係式。根據重複性試驗結果可知，其抗菌效果與電流強度及電流刺激時間成正相關，且位於正極附近抗菌效果較佳，並出現綠膿桿菌附著物，而附著物內含活性菌量低且無法進行二次培養。當菌液濃度為7.5*104 CFU/mL，以4、0.4及0.04mA電流強度進行60分鐘實驗，其抗菌效果可達為79%、66%及59%；當菌液濃度為1.5*104 CFU/mL，以4、0.4及0.04mA電流強度進行60分鐘實驗，其抗菌效果分別為74%、43%、29%。因此本研究確立新式的體外菌落模型運用於電流抑菌效應，且可於體外模擬綠膿桿菌抗藥性產生之過程，與自體免疫之相互關係，而未來應用於治療感染綠膿桿菌慢性傷口，可提供相關參數以供參考，並且對於全球性抗藥性問題，提供一個新的治療概念，以解決P.aeruginosa多重抗藥性增加之問題。

In the recent years, the problem of drug resistance becomes severe, and the topics of treatments for drug resistant bacteria become top sanitaria issues. Among them, treatments for Pseudomonas aeruginosa (PA) infection raise the greatest attention. The growing number of multi-drug resistance PA increases the difficulties to treat the infection, which raises the fatal rate. To solve the problem of multi-drug resistance of PA, we need an innovative solution. In this study, we will discuss how the current controls the growth of PA. Our goal is (1) to develop the model of Electro-bacterial inhibition and (2) to study how the current treatment restraints the growth of Pseudomonas. Due to the characters of having no efficient treatment and being easy to become drug resistance, Pseudomonas infection is difficult to control and costs a great amount of medical resources. Based on Electro-Chemical reaction, the Electro-bacterial inhibition accelerates the speed the silver needle emits silver ions which react with body cell to generate cell toxicity and restraint bacteria from growing around the wound, and reaches the goal of reducing the number of bacteria species. Hence, we propose the theory of electro-bacterial inhibition, which is expected to use the current to inhibit the wound to heal and to increase the strength of wound tension. Based on the theory of Electrophysiology, Microbiology, Wound healing, Electrochemistry and Biostatics, we will quantitatively analyze the relation between the strength of the current and the growth of Pseudomonas, which is expected to be adopted on the clinical practice and to ease the problem of medical resource overuse.

The in vitro model is designed based on the clinical demands after taking into account the characteristics of the P. aeruginosa infection. In addition, the density of bacteria liquid is set at the level of 7.5*104 and 1.5*104 CFU/mL according to the sensitivity analysis and calibration of growth rate. The anti-bacteria effects of current to P. aeruginosa infection are then confirmed by adopting power supplying module and adjusting the strength of current (4, 0.4, 0.04mA). According to the repeated trials conducted, there’s a positive correlation between the current strength and the anti-bacteria effect and a positive correlation between the time to stimulate and. The anti-bacteria effect of current is notably great around the positive electrode. From the attachment found around where, the activated bacterium contains is lower and could not proceed secondary culture. In conditions where the density of bacteria liquid is 7.5*104 CFU/mL, the anti-bacteria effects of the 60-minutes trials are 79%, 66%, 59% with current levels of 4, 0.4, 0.04mA respectively. In conditions where the density of bacteria liquid is 1.5*104 CFU/mL, the anti-bacteria effect of the 60-minutes trials are 74%, 43%, 29% with current levels of 4, 0.4, 0.04mA.

The study establishes a new standard of in vitro model used to explain how current restrains the bacteria development around wounds and to simulate the process that drug-resistance of P. aeruginosa is formed, which could be adopted to the treatment of the P. aeruginosa infection in the future and helps to resolve the problem of multi-drug resistance.

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