การศึกษาพลาสมาเจ็ทแบบไดอิเล็คทริกแบร์ริเออร์ดิสชาจ์นเพื่อการฆ่าเชื้อแบคทีเรียในแผลติดเชื้อ

Abstract

The cold atmospheric pressure plasma (CAPP) technique has been recognized in medical fields to bring a new innovative approach in biomedical applications such a contaminated wound healing enhancement as well as relieving the patient’s pain without side effects. Contaminated wounds are a major health problem in many countries. Factors of patient health problems are such as diabetes, contaminated wounds, bacteria as well as others. The patients endures pain and requires long treatment periods using antibiotics or other therapies. Moreover, using antibiotics for a long period of time bacterial resistance occurs, such as methicillin-resistant Staphylococcus aureus (MRSA). Bacteria have the ability to develop resistance to antibiotics faster than one can develop new drugs. This being the main problem in contaminated wound healing. In this study, the dielectric barrier discharge plasma jet (DBDJ) was used for bactericidal and treated the Primary Human Dermal Fibroblasts Adult (HDFa) cells to study the side effect of DBDJ. This DBDJ is driven by high voltage dc pulse at 20 kHz and using 1 L/min of helium (He) as plasma gas. The DBDJ plasma varied the plasma dissipated power from 0.27 W to 0.50 W and the exposure time 15 s to 60 s. Plasma radical species were utilized by using an optical emission spectroscopy (OES). The results of the OES study found NO and OH radical groups, which play an important role in bactericidal and contaminated wound healing. The increase of radical plasma density depends on the plasma dissipated power. The Colony Forming unit (CFU) method was used to monitor the efficiency of bacteria killing. Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) were used in the vitro bacteria killing test. The results showed that the plasma dissipated power and exposure time were major factors in bactericidal. When increasing the plasma dissipated power to 0.50 W and exposure time to 60 s, the effect of bacteria killing increased to 100%. Therefore, this condition was choosing to study the effect of plasma on bacteria biofilm and HDFa cells. The result from fluorescent images by live/dead assay showed that DBDJ had high efficiency to removed biofilm. Studying the effects of DBDJ on HDFa cells using optical microscopes, live/dead assay and Muse Cell Analyzer with the Muse Count & Viability Assay Kit and Muse Annexin V and Dead Cell Assay Kit, the result showed the DBDJ being highly efficient for bactericidal as well as not having any side effects on HDFa cells under the same conditions. Therefore, the DBDJ at plasma dissipated power 0.50 W and exposure time 60 s has a high efficiency to kill bacteria, destroy bacteria biofilm by without damage cell and also without side effect to cell viability, apoptosis and death of HDFa cells as in vitro is essential for clinical use.