Photo-crosslinked chitosan-gelatin xerogel-like coating onto "cold" plasma functionalized poly(lactic acid) film as cell culture support.

This paper reports on biofunctionalisation of a poly(lactic acid) (PLA) film by surface activation through cold plasma treatment followed by coating with a chitosan-gelatin xerogel. The UV cross-linking of the xerogel precursor was simultaneously performed with the fixation onto the PLA support. This has a strong effect on surface properties, in terms of wettability, surface free energy, morphology and micromechanical features. The hydrophilic - hydrophobic character of the surface, determined by contact angle measurements, was tuned along the process, passing from moderate hydrophobic PLA to enhanced hydrophilic plasma activated surface, which favors coating adhesion, then to moderate hydrophobic chitosan-gelatin coating. The coating has a Lewis amphoteric surface, with a porous xerogel-like morphology, as revealed by scanning electron microscopy images. By riboflavin mediated UV cross-linking the chitosan-gelatin coating becomes high adhesive and with a more pronounced plasticity, as shown by AFM force-distance spectroscopy. Thus prepared surface-coated PLA supports were successfully tested for growth of dermal fibroblasts, which are known for their induction potential of chondrogenic cells, which is very important in cartilage tissue engineering.

Skin treatment with non-thermal plasma modulates the immune system through miR-223-3p and its target genes.

Non-thermal plasma, a partially ionized gas, holds significant potential for clinical applications, including wound-healing support, oral therapies, and anti-tumour treatments. While its applications showed promising outcomes, the underlying molecular mechanisms remain incompletely understood. We thus apply non-thermal plasma to mouse auricular skin and conducted non-coding RNA sequencing, as well as single-cell blood sequencing. In a time-series analysis (five timepoints spanning 2 hours), we compare the expression of microRNAs in the plasma-treated left ears to the unexposed right ears of the same mice as well as to the ears of unexposed control mice. Our findings indicate specific effects in the treated ears for a set of five miRNAs: mmu-miR-144-5p, mmu-miR-144-3p, mmu-miR-142a-5p, mmu-miR-223-3p, and mmu-miR-451a. Interestingly, mmu-miR-223-3p also exhibits an increase over time in the right non-treated ear of the exposed mice, suggesting systemic effects. Notably, this miRNA, along with mmu-miR-142a-5p and mmu-miR-144-3p, regulates genes and pathways associated with wound healing and tissue regeneration (namely ErbB, FoxO, Hippo, and PI3K-Akt signalling). This co-regulation is particularly remarkable considering the significant seed dissimilarities among the miRNAs. Finally, single-cell sequencing of PBMCs reveals the downregulation of 12 from 15 target genes in B-cells, Cd4+ and Cd8+ T-cells. Collectively, our data provide evidence for a systemic effect of non-thermal plasma.

Accelerating electrostatic particle-in-cell simulation: A novel FPGA-based approach for efficient plasma investigations.

Particle-in-cell (PIC) simulation serves as a widely employed method for investigating plasma, a prevalent state of matter in the universe. This simulation approach is instrumental in exploring characteristics such as particle acceleration by turbulence and fluid, as well as delving into the properties of plasma at both the kinetic scale and macroscopic processes. However, the simulation itself imposes a significant computational burden. This research proposes a novel implementation approach to address the computationally intensive phase of the electrostatic PIC simulation, specifically the Particle-to-Interpolation phase. This is achieved by utilizing a high-speed Field Programmable Gate Array (FPGA) computation platform. The suggested approach incorporates various optimization techniques and diminishes memory access latency by leveraging the flexibility and performance attributes of the Intel FPGA device. The results obtained from our study highlight the effectiveness of the proposed design, showcasing the capability to execute hundreds of functional operations in each clock cycle. This stands in contrast to the limited operations performed in a general-purpose single-core computation platform (CPU). The suggested hardware approach is also scalable and can be deployed on more advanced FPGAs with higher capabilities, resulting in a significant improvement in performance.

Training in the use of the water jet and cold atmospheric plasma jet for the decontamination of dental implants.

OBJECTIVES: Clinical trials testing new devices require prior training on dummies to minimize the "learning curve" for patients. Dentists were trained using a novel water jet device for mechanical cleaning of dental implants and with a novel cold plasma device for surface functionalisation during a simulated open flap peri-implantitis therapy. The hypothesis was that there would be a learning curve for both devices. MATERIALS AND METHODS: 11 dentists instrumented 44 implants in a dummy-fixed jaw model. The effect of the water jet treatment was assessed as stain removal and the effect of cold plasma treatment as surface wettability. Both results were analysed using photographs. To improve treatment skills, each dentist treated four implants and checked the results immediately after the treatment as feedback. RESULTS: Water jet treatment significantly improved from the first to the second implant from 62.7% to 75.3% stain removal, with no further improvement up to the fourth implant. The wettability with cold plasma application reached immediately a high level at the first implant and was unchanged to the 4th implant (mean scores 2.7 out of 3). CONCLUSION: A moderate learning curve was found for handling of the water jet but none for handling of the cold plasma. CLINICAL RELEVANCE: Scientific rational for study: Two new devices were developed for peri-implantitis treatment (Dental water jet, cold plasma). Dentists were trained in the use of these devices prior to the trial to minimize learning effects. PRINCIPAL FINDINGS: Experienced dentists learn the handling of the water jet very rapidly and for cold plasma they do not need much training. PRACTICAL IMPLICATIONS: A clinical study is in process. When the planned clinical study will be finished, we will find out, if this dummy head exercise really minimised the learning curve for these devices.

Human coronavirus 229E inactivation on porous and nonporous materials using dielectric barrier discharge (DBD) plasma.

Viruses can spread through contaminated aerosols and contaminated surface materials, and effective disinfection techniques are essential for virus inactivation. Nonthermal plasma-generated reactive oxygen and nitrogen species can effectively inactivate the coronavirus. We aim to interpret the coronavirus inactivation level and mechanism of surface interaction with materials with and without dielectric barrier discharge (DBD) plasma treatment. Nonthermal plasma, particularly surface-type DBD plasma, can inactivate human coronavirus 229E (HCoV-229E) on porous (paper, wood, mask) and nonporous (plastic, stainless steel, glass, Cu) materials. Virus inactivation was analyzed using a 50% tissue culture infectivity dose (TCID50) using cell line, flow cytometry, and immunofluorescence. Surfaces contaminated with HCoV-229E were treated at different time intervals (0-5 h) with and without plasma exposure (natural decay in ambient air conditions). HCoV-229E persistence conformed to the following order: plastic > cover glass > stainless steel > mask > wood > paper > Cu with and without plasma exposure. HCoV-229E was more stable in plastic, cover glass, and stainless steel in 5 h, and the viable virus titer gradually decreased from its initial log10 order of 6.892 to 1.72, 1.53, and 1.32 TCID50/mL, respectively, under plasma exposure. No virus was observed in Cu after treatment for 5 h. The use of airflow, ambient nitrogen, and argon did not promote virus inactivation. Flow cytometry and immunofluorescence analysis demonstrated a low expression level of spike protein (fluorescence intensity) during plasma treatment and in E and M genes expression compared with the virus control.

Machine learning-aided evaluation of oxidative strength of cold atmospheric plasma-treated water.

Plasma medicine is gaining attraction in the medical field, particularly the use of cold atmospheric plasma (CAP) in biomedicine. The chemistry of the plasma is complex, and the reactive oxygen species (ROS) within it are the basis for the biological effect of CAP on the target. Understanding how the oxidative power of ROS responds to diverse plasma parameters is vital for standardizing the effective application of CAP. The proven applicability of machine learning (ML) in the field of medicine is encouraging, as it can also be applied in the field of plasma medicine to correlate the oxidative strength of plasma-treated water (PTW) according to different parameters. In this study, plasma-treated water was mixed with potassium iodide-starch reagent for color formation that could be linked to the oxidative capacity of PTW. Corresponding images were captured resulting from the exposure of the color-forming agent to water treated with plasma for different time points. Several ML models were trained to distinguish the color changes sourced by the oxidative strength of ROS. The AdaBoost Classifier (ABC) algorithm demonstrated better performance among the classification models used by extracting color-based features from the images. Our results, with a test accuracy of 63.5%, might carry a potential for future standardization in the field of plasma medicine with an automated system that can be created to interpret the oxidative properties of ROS in different plasma treatment parameters via ML.

Modeling the growth of Aspergillus brasiliensis affected by a nonthermal plasma.

AIM: The main objective of the study was to develop and validate a model for the growth of Aspergillus brasiliensis on surfaces, specifically on agar culture medium. An additional aim was to determine conditions for complete growth inhibition of this micromycete using two different nonthermal plasma (NTP) sources. METHODS AND RESULTS: The developed model uses two key parameters, namely the growth rate and growth delay, which depend on the cultivation temperature and the amount of inoculum. These parameters well describe the growth of A. brasiliensis and the effect of NTP on it. For complete fungus inactivation, a single 10-minute exposure to a diffuse coplanar surface barrier discharge was sufficient, while a point-to-ring corona discharge required several repeated 10-minute exposures at 24-h intervals. CONCLUSIONS: The article presents a model for simulating the surface growth of A. brasiliensis and evaluates the effectiveness of two NTP sources in deactivating fungi on agar media.

Predictive modeling of molds effective elimination by external inactivation sources.

Presented paper deals with a novel application of the (nonlinear) logistic equation to model an elimination of microscopic filaments types of fungi-molds from affected materials via different external inactivation techniques. It is shown that if the inactivation rate of the external source is greater than the maximum natural growth rate of mycelium, the mold colony becomes destroyed after a finite time. Otherwise, the mycelium may survive the external attack only at a sufficiently large initial concentration of the inoculum. Theoretically determined growth curves are compared with the experimental data for Aspergillus brasiliensis mold inactivated by using both cold atmospheric plasma (CAP) and UV-germicidal lamp. Model presented in the article may be applied also to other classes of microorganisms (e.g. bacteria).

Rheological and gelling properties of atmospheric pressure cold plasma treated finger millet (Eleusine coracana) starch.

Interest in exploring alternative starch sources like finger millet is rising due to wide starch applications. However, native starch often lacks desired qualities, including rheological properties. Modification is thus necessary for specific end uses. Plasma treatment as a greener and sustainable method for starch modification was therefore, studied for its ability to impact rheological properties of finger millet starch (FMS). Considerable changes in the rheological properties on FMS was noted, a significant decrease and increase (p < 0.05) in the peak viscosity (from 3.35 to 0.553 Pa.s) and paste clarity respectively was observed, indicating occurrence of depolymerization. However, intermediate plasma-treated samples (200 V) observed a decrease in paste clarity attributed to aggregate formation and cross-linking. Cross-linking was also confirmed by findings of frequency sweep where a continuous decrease in G' values of plasma treated FMS gel was interrupted by sudden increase. Despite depolymerization causing alteration of rheological behaviour such as decrease in shear thinning properties, gel strength observed a contradictory increase. This was attributed to incorporation of functional group and absence of shear responsible for network formation giving higher gel strength to FMS gels. This is elaborated in detail in the study. The study thus concluded that cold plasma significantly impacted all the rheological properties of the FMS and hence can prove to be beneficial for modification of starch rheological parameters.

Bilateral Thermal Keratopathy Due to Plasma Skin Regeneration.

A 40-year-old woman underwent periocular plasma skin regeneration, a cosmetic treatment for periorbital rejuvenation. She subsequently developed bilateral thermal keratitis, manifesting as blurred vision, irritation, and redness, with a vision decrease to 20/60 and 20/50 in her OD and OS, respectively. Examination demonstrated bilateral large, irregular corneal epithelial defects and edema, necessitating treatment with amniotic membrane grafts, bandage contact lenses, and hypertonic saline. One year posttreatment, her visual acuity improved to 20/20 and 20/25, albeit with ongoing symptomatic dryness and bilateral anterior stromal haze. This case, as only the second reported instance of ocular damage from periocular plasma skin regeneration, underscores the need for heightened awareness of potential ocular complications following plasma skin regeneration and reinforces the importance of protective measures during periocular procedures.

Cold atmospheric plasma enhances morphological and biochemical attributes of tomato seedlings.

Cold atmospheric plasma (CAP) is a physical technology with notable effects on living organisms. In the present study, tomato seeds (Solanum lycopersicum var. Bassimo Mill.) were exposed to CAP for various time intervals, ranging from 1 to 5 min, in both continuous and intermittent periods, and were compared with a control group that received no CAP treatment. Seedlings grown from treated seeds exhibited improvements in levels of growth traits, photosynthetic pigments, and metabolite contents when compared to the control group. Seedlings from seeds treated with S04 displayed significant increases in shoot and root lengths, by 32.45% and 20.60% respectively, compared to the control group. Moreover, seedlings from seeds treated with S01 showed a 101.90% increase in total protein, whereas those treated with S02 experienced a 119.52% increase in carbohydrate content. These findings highlight the substantial improvements in growth characteristics, photosynthetic pigments, and metabolite levels in seedlings from treated seeds relative to controls. Total antioxidant capacity was boosted by CAP exposure. The activities of enzymes including superoxide dismutase, catalase, and peroxidases were stimulated by S02 and exceeded control treatment by (177.48%, 137.41%, and 103.32%), respectively. Additionally, exposure to S04 increased the levels of non-enzymatic antioxidants like flavonoids, phenolics, saponins, and tannins over the control group (38.08%, 30.10%, 117.19%, and 94.44%), respectively. Our results indicate that CAP-seed priming is an innovative and cost-effective approach to enhance the growth, bioactive components, and yield of tomato seedlings.

Cold Atmospheric Pressure Plasma-Activated Medium Modulates Cellular Functions of Human Mesenchymal Stem/Stromal Cells In Vitro.

Cold atmospheric pressure plasma (CAP) offers a variety of therapeutic possibilities and induces the formation of reactive chemical species associated with oxidative stress. Mesenchymal stem/stromal cells (MSCs) play a central role in tissue regeneration, partly because of their antioxidant properties and ability to migrate into regenerating areas. During the therapeutic application, MSCs are directly exposed to the reactive species of CAP. Therefore, the investigation of CAP-induced effects on MSCs is essential. In this study, we quantified the amount of ROS due to the CAP activation of the culture medium. In addition, cell number, metabolic activity, stress signals, and migration were analyzed after the treatment of MSCs with a CAP-activated medium. CAP-activated media induced a significant increase in ROS but did not cause cytotoxic effects on MSCs when the treatment was singular and short-term (one day). This single treatment led to increased cell migration, an essential process in wound healing. In parallel, there was an increase in various cell stress proteins, indicating an adaptation to oxidative stress. Repeated treatments with the CAP-activated medium impaired the viability of the MSCs. The results shown here provide information on the influence of treatment frequency and intensity, which could be necessary for the therapeutic application of CAP.

Antimicrobial efficacy of direct air gas soft jet plasma for the in vitro reduction of oral bacterial biofilms.

The aim of this study was to evaluate the antimicrobial efficacy of an air gas soft jet CAP for its potential use in removing oral biofilms, given that plasma-based technologies have emerged as promising methods in periodontology. Two types of biofilms were developed, one by Streptococcus mutans UA 159 bacterial strain and the other by a complex mixture of saliva microorganisms isolated from a patient with periodontitis. This latter biofilm was characterized via Next Generation Sequencing to determine the main bacterial phyla. The CAP source was applied at a distance of 6 mm for different time points. A statistically significant reduction of both CFU count and XTT was already detected after 60 s of CAP treatment. CLSM analysis supported CAP effectiveness in killing the microorganisms inside the biofilm and in reducing the thickness of the biofilm matrix. Cytotoxicity tests demonstrated the possible use of CAP without important side effects towards human gingival fibroblasts cell line. The current study showed that CAP treatment was able to significantly reduce preformed biofilms developed by both S. mutans and microorganisms isolated by a saliva sample. Further studies should be conducted on biofilms developed by additional saliva donors to support the potential of this innovative strategy to counteract oral pathogens responsible for periodontal diseases.

In-vitro biofilm removal from TiUnite® implant surface with an air polishing and two different plasma devices.

BACKGROUND: We investigated the efficacy of two different cold atmospheric pressure jet plasma devices (CAP09 and CAPmed) and an air polishing device with glycine powder (AP) either applied as monotherapies or combined therapies (AP + CAP09; AP + CAPmed), in microbial biofilm removal from discs with anodised titanium surface. METHODS: Discs covered with 7-day-old microbial biofilm were treated either with CAP09, CAPmed, AP, AP + CAP09 or AP + CAPmed and compared with negative and positive controls. Biofilm removal was assessed with flourescence and electron microscopy immediately after treatment and after 5 days of reincubation of the treated discs. RESULTS: Treatment with CAP09 or CAPmed did not lead to an effective biofilm removal, whereas treatment with AP detached the complete biofilm, which however regrew to baseline magnitude after 5 days of reincubation. Both combination therapies (AP + CAP09 and AP + CAPmed) achieved a complete biofilm removal immediately after cleaning. However, biofilm regrew after 5 days on 50% of the discs treated with the combination therapy. CONCLUSION: AP treatment alone can remove gross biofilm immediately from anodised titanium surfaces. However, it did not impede regrowth after 5 days, because microorganisms were probably hidden in holes and troughs, from which they could regrow, and which were inaccessible to AP. The combination of AP and plasma treatment probably removed or inactivated microorganisms also from these hard to access spots. These results were independent of the choice of plasma device.

Non-invasive Physical Plasma Modulates Macrophage Polarization: A Potential Strategy for Tumor Microenvironment Remodeling.

BACKGROUND/AIM: Non-invasive physical plasma (NIPP) has shown promise in the treatment of cancer. However, conflicting results have been reported regarding the effect of NIPP on macrophage polarization. As tumor-associated macrophages (TAMs) are essential in the regulation of cancer development, this study aimed to determine the role of NIPP treatment in macrophage polarization and tumor-microenvironment (TME) remodeling. MATERIALS AND METHODS: A portable NIPP device, Plasma Care (Terraplasma Medical, Garching, Germany), was employed as the source of NIPP. The human monocytic cell line THP-1 was adopted as the cell model for macrophage differentiation and polarization. The effects of NIPP treatment on temperature, pH value, and oxidative stress induction of the culture medium were examined to validate the feasibility of applying the NIPP device in subsequent cell treatment. The changes in morphology, viability, and proliferation of THP-1 cells after NIPP treatment were determined. The expression of M1/M2 macrophage markers was examined by real-time quantitative polymerase chain reaction. RESULTS: No significant changes were observed in temperature and pH value after NIPP treatment, while the formation of hydrogen peroxide was promoted in a time-dependent manner. Cell morphology, viability, and proliferation were not affected by up to 6 minutes of NIPP treatment. In monocytes, 6 minutes of NIPP treatment significantly increased the expression of M1 markers (TNF-α and IL-6) and suppressed the M2 marker (CD206), findings which were consistent in the monocyte-derived macrophages. Furthermore, NIPP treatment also significantly promoted M1 polarization in the monocyte-derived macrophages induced by phorbol 12-myristate 13-acetate. CONCLUSION: NIPP is a safe and robust oxidative stress inducer and showed potential in TAM regulation by promoting M1 macrophage polarization.

Enhancing high-efficiency breeding and microbial microdroplet cultivation techniques for Ganoderma lucidum.

Ganoderma lucidum is known for its bioactive compounds, such as polysaccharides and triterpenoids, which are crucial in food and medicine. However, liquid fermentation encounters challenges in terms of strain differentiation and stability. In this research, we employed atmospheric room temperature plasma mutation and a microbial microdroplet culture system to identify strains with enhanced biomass and triterpenoid production. The three mutant strains, YB05, YB09, and YB18, exhibited accelerated growth rates and antagonized the initial strain G0023 more effectively than the controls. Notably, YB18 displayed the fastest growth, with a 17.25% increase in colony radius. Shake flask cultivation demonstrated that, compared with the initial strain, YB05 and YB18 had 26.33% and 17.85% greater biomass, respectively. Moreover, the triterpenoid production of YB05 and YB18 surpassed that of the control by 32.10% and 15.72%, respectively, as confirmed by colorimetric detection. Importantly, these mutant strains remained stable for five generations. This study revealed a comprehensive screening system utilizing atmospheric pressure, room temperature plasma mutation technology and microbial droplet cultivation. This innovative approach offers a promising pathway for obtaining advantageous Ganoderma strains for liquid fermentation. The methodology of atmospheric room temperature plasma mutation and microbial microdroplet culture systems is detailed for better comprehension.

Dielectric barrier discharge plasma pre-treatment to facilitate the acetylation process of corn starch under heating/cooling cycles.

The objective of the current work was to evaluate the impacts of dielectric barrier discharge plasma and repeated dry-heat treatments on the acetylation process of corn starch. The combined modification resulted in a higher substitution degree of acetate groups on starch chains compared to the acetylation treatment alone. This outcome was linked to the increase in surface area and structural organization level of granules achieved through the application of plasma and heating/cooling cycles, respectively. The successful esterification of starch structure was verified through FTIR (1710 cm-1) and 1H NMR (2 ppm). With the increase in plasma treatment duration up to 20 min, gelatinization enthalpy increased (10.81 J/g) due to the cross-linking reaction. Starch acetate produced through the combined treatment could find the application in the development of low-calorie food formulations due to its high resistant starch (70.5 g/100 g) and low viscosity (43 mPa s).

Overcoming antibiotic resistance: non-thermal plasma and antibiotics combination inhibits important pathogens.

Antibiotic resistance (ATBR) is increasing every year as the overuse of antibiotics (ATBs) and the lack of newly emerging antimicrobial agents lead to an efficient pathogen escape from ATBs action. This trend is alarming and the World Health Organization warned in 2021 that ATBR could become the leading cause of death worldwide by 2050. The development of novel ATBs is not fast enough considering the situation, and alternative strategies are therefore urgently required. One such alternative may be the use of non-thermal plasma (NTP), a well-established antimicrobial agent actively used in a growing number of medical fields. Despite its efficiency, NTP alone is not always sufficient to completely eliminate pathogens. However, NTP combined with ATBs is more potent and evidence has been emerging over the last few years proving this is a robust and highly effective strategy to fight resistant pathogens. This minireview summarizes experimental research addressing the potential of the NTP-ATBs combination, particularly for inhibiting planktonic and biofilm growth and treating infections in mouse models caused by methicillin-resistant Staphylococcus aureus or Pseudomonas aeruginosa. The published studies highlight this combination as a promising solution to emerging ATBR, and further research is therefore highly desirable.

Evaluation of degradation efficacy and toxicity mitigation for 4-nitrophenol using argon and air-mixed argon plasma jets.

This paper investigates the effects of argon (Ar) and that of Ar mixed with ambient air (Ar-Air) cold plasma jets (CPJs) on 4-nitrophenol (4-NP) degradation using low input power. The introduction of ambient air into the Ar-Air plasma jet enhances ionization-driven processes during high-voltage discharge by utilizing nitrogen and oxygen molecules from ambient air, resulting in increased reactive oxygen and nitrogen species (RONS) production, which synergistically interacts with argon. This substantial generation of RONS establishes Ar-Air plasma jet as an effective method for treating 4-NP contamination in deionized water (DW). Notably, the Ar-Air plasma jet treatment outperforms that of the Ar jet. It achieves a higher degradation rate of 97.2% and a maximum energy efficiency of 57.3 gkW-1h-1, following a 6-min (min) treatment with 100 mgL-1 4-NP in DW. In contrast, Ar jet treatment yielded a lower degradation rate and an energy efficiency of 75.6% and 47.8 gkW-1h-1, respectively, under identical conditions. Furthermore, the first-order rate coefficient for 4-NP degradation was measured at 0.23 min-1 for the Ar plasma jet and significantly higher at 0.56 min-1 for the Ar-Air plasma jet. Reactive oxygen species, such as hydroxyl radical and ozone, along with energy from excited species and plasma-generated electron transfers, are responsible for CPJ-assisted 4-NP breakdown. In summary, this study examines RONS production from Ar and Ar-Air plasma jets, evaluates their 4-NP removal efficacy, and investigates the biocompatibility of 4-NP that has been degraded after plasma treatment.

Plasma catalysis. A study of the influence of oxysulfur, SOx• species through the degradation of sulfonated dyes by non-thermal plasma.

This work studied the degradation reaction of sulfonated dyes, indigo carmine, phenol red, and their mixtures by non-thermal plasma (NTP). Interestingly, the degradation rate constant showed a faster process and lower activation energy (Ea) for the dye mixtures than for the degradation reaction of the individual dyes. This unexpected result opened up new opportunities for understanding plasma chemistry and the interaction between reactive species formed by the plasma and the target molecule. As no catalyst or chemical additive was added to the reactor, the decrease in Ea came from a self-synergistic effect (SSE), through the dye molecules fragmentation, which resulted in plasma catalysis. The hypothesis proposed in this work is that oxysulfur (SOx) species are formed by the desulfonation reaction of dyes. The sulfonic groups (SO3) present in the chemical structures of dyes can function as precursors for forming several SOx•- species. Studies based on oxygenated sulfonated species such as SO3•-, SO4•- and SO5•- have been widely applied in advanced oxidative and reductive processes due to their satisfactory efficiency and low cost. Among them, SO4•- is the key reactive species with the best performance in the degradation of pollutants due to its high oxidation potential (E° = 2.60 V). In addition, it is an alternative source of HO• in aqueous media, improving the oxidation reaction. In order to elucidate the SSE, the kinetic process was followed by UV-Vis analysis, and the reactive species, such as alkyl, hydroxyl, and oxy-sulfur radicals were identified by Electron Paramagnetic Resonance. The by-products of the NTP degradation reaction were analyzed by ultrafast liquid chromatography coupled with a mass spectrometer, and a fragmentation route was proposed.