Effects of cold atmospheric-pressure plasma in combination with doxorubicin drug against breast cancer cells in vitro and invivo.

Cold atmospheric plasma (CAP) has been suggested for medical applications that can be applied indirectly through plasma-activated medium (PAM) and recently it has been introduced as an innovative therapeutic approach for all cancer types. Studies have exhibited that ROS/RNS are key factors in CAP-dependent apoptosis; nevertheless, ROS/RNS stability are weak. Combination therapy is considered an effective strategy to overcome these problems. In the present research, we revealed that the combination of CAP and doxorubicin (DOX) significantly induces the apoptosis of breast cancer cells both in vitro and in vivo. Our results indicated that both Ar and He/O2 CAP treatment as well as DOX drug alone reduced cell growth. CAP/PAM treatment in combination with DOX induced apoptosis in MCF-7 breast cancer cells and 4T1-implanted BALB/c mice, resulting in a significant increase in antitumor activity. The apoptotic effects of CAP-DOX on MCF-7 cells were inferred from altered expression of BAX and cleaved-caspase-3 which mechanistically take place through the mitochondrial pathway mediated by Bcl-2 family members. Besides, the BAX/BCL-2 ratio is significantly higher in the simultaneous treatment of CAP and DOX. This ratio was equal to 2.82 ± 0.24, 2.54 ± 0.30, and 11.27 ± 0.31 for treatment with DOX, He/O2 plasma, and combination treatment, respectively. Additionally, the tumor growth rate of He/O2-PAM + DOX and Ar-PAM + DOX treatments was significantly inhibited by PAM-injection, and the tumor growth rate of PAM alone or DOX alone was slightly reduced. It can be concluded that the effect of PAM + DOX may increase the anticancer activity and decrease the dose required for the chemotherapeutic treatment.

Antimicrobial activity of cold atmospheric-pressure argon plasma combined with chicory (Cichorium intybus L.) extract against P. aeruginosa and E. coli biofilms.

The present study reports a significant combined antibacterial activity of Cichorium intybus L. (known as Chicory) natural extract with cold atmospheric-pressure argon plasma treatment against multi-drug resistant (MDR) Gram-negative bacteria. To detect reactive species that are generated in the argon plasma, optical emission spectra were recorded. The molecular bands were allocated to the hydroxyl radicals (OH) and neutral nitrogen molecules (N2). Moreover, the atomic lines form the emitted spectra were determined to argon atoms (Ar) and the oxygen atoms (O), respectively. The results revealed that Chicory extract treatment at a concentration of 0.043 g/ml reduced the metabolic activity of P. aeruginosa cells by 42%, while, a reduced metabolic activity of 50.6% was found for E. coli biofilms. Moreover, the combination of Chicory extract with 3 min Ar-plasma introduced a synergistic effect, so that it exhibited a significantly reduced metabolic activity of P. aeruginosa to 84.1%, and E. coli ones to 86.7%, respectively. The relationship between cell viability and membrane integrity of P. aeruginosa and E. coli biofilms treated with Chicory extract and argon plasma jet were also analyzed by CLSM. It was found that after the combined treatment, a noticeable membrane disruption was formed. Besides, it was concluded that E. coli biofilms showed a higher sensitivity to Ar-plasma than P. aeruginosa biofilm at longer plasma exposure times. This study suggests that the anti-biofilm therapy based on a combined effect of Chicory extract and cold argon plasma treatment can serve as a considerable green method for treatment of antimicrobial MDR bacteria.

Inactivation of Staphylococcus aureus and Escherichia coli Biofilms by Air-Based Atmospheric-Pressure DBD Plasma.

Air-based atmospheric-pressure plasma is an effective non-thermal method in deactivating various kinds of microbial biofilms with several advantages, including high bactericidal efficiency and low treatment costs. Bacterial biofilm formation is a major determinant in establishment of bacterial infection and also resistance to antibacterial chemotherapy. This study aims to assess the anti-biofilm potential of air-based atmospheric-pressure DBD plasma against Staphylococcus aureus and Escherichia coli biofilms. The biofilms of Staphylococcus aureus and Escherichia coli were exposed to air-based atmospheric-pressure DBD plasma for up to 4 min (control, 30 s, 90 s, 3 min, and 4 min) and their biofilm formation level, viability, and membrane integrity were determined. Based on the results, plasma exposure caused disruption up to 70% and 85% for S. aureus and E. coli biofilms, respectively. The biofilm disruption potential of air-based atmospheric-pressure DBD plasma was confirmed using the scanning electron microscopy (SEM). Besides, based on confocal laser scanning microscopy (CLSM), plasma exposure caused a significant bacterial inactivation and E. coli was found as more susceptible strain than S. aureus. In conclusion, atmospheric-pressure DBD plasma could be considered an efficient non-thermal approach against bacterial pathogenicity by biofilm disruption and thus prevention of infection establishment.