In Vitro and In Vivo Effects of Melatonin-Containing Combinations in Human Pancreatic Ductal Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) has poor prognosis and high mortality rates. Therefore, it is necessary to identify new targets and therapeutic strategies to improve the prognosis of patients with PDAC. Integrative therapies are increasingly being used to boost the efficacy of the known anticancer therapeutic approaches. Hence, this study aimed to evaluate the effects of a novel combination of different potential anticancer molecules, melatonin (MLT), cannabidiol (CBD), and oxygen-ozone (O2/O3) to treat PDAC using in vitro and in vivo models of human PDAC. The effect of this combination was investigated in combination with gemcitabine (GEM), the most common chemotherapeutic drug used for PDAC treatment. The combination of MLT + CBD + O2/O3 was more effective than the individual treatments in inhibiting PDAC cell viability and proliferation, inducing cell death, and modulating the RAS pathway protein levels. Moreover, different combinations of treatments reduced tumor mass in the PDAC mouse model, thus promoting the effect of GEM. In conclusion, a mixture of MLT + CBD + O2/O3 could serve as a potential adjuvant therapeutic strategy for PDAC.

Oxygen-Ozone Therapy for Reducing Pro-Inflammatory Cytokines Serum Levels in Musculoskeletal and Temporomandibular Disorders: A Comprehensive Review.

To date, the application of oxygen-ozone (O2O3) therapy has significantly increased in the common clinical practice in several pathological conditions. However, beyond the favorable clinical effects, the biochemical effects of O2O3 are still far from being understood. This comprehensive review aimed at investigating the state of the art about the effects of O2O3 therapy on pro-inflammatory cytokines serum levels as a modulator of oxidative stress in patients with musculoskeletal and temporomandibular disorders (TMD). The efficacy of O2O3 therapy could be related to the moderate oxidative stress modulation produced by the interaction of ozone with biological components. More in detail, O2O3 therapy is widely used as an adjuvant therapeutic option in several pathological conditions characterized by chronic inflammatory processes and immune overactivation. In this context, most musculoskeletal and temporomandibular disorders (TMD) share these two pathophysiological processes. Despite the paucity of in vivo studies, this comprehensive review suggests that O2O3 therapy might reduce serum levels of interleukin 6 in patients with TMD, low back pain, knee osteoarthritis and rheumatic diseases with a concrete and measurable interaction with the inflammatory pathway. However, to date, further studies are needed to clarify the effects of this promising therapy on inflammatory mediators and their clinical implications.

Intradiscal oxygen-ozone therapy for the treatment of symptomatic lumbar disc herniation: A preliminary study.

PURPOSE: To assess safety and effectiveness of computed tomography (CT)-guided intradiscal oxygen-ozone therapy (O2-O3 therapy) for the treatment of symptomatic lumbar disc herniation and radiological changes. MATERIALS AND METHODS: This study was conducted in twenty patients presenting lumbar disc herniation with resistant lumbar or lumbar radicular pain They underwent intradiscal oxygen-ozone therapy under CT guidance. They were treated at one- or two-disc levels, representing a total of 24 discs treated. MR imaging examinations were obtained before treatment and 2 months post-procedure to analyse treatment-related disc modifications including modification of the surfaces of the disc and of the herniated disc, and the variations in disc height according to the disc height index. Clinical outcomes were assessed using the visual analogue scale (VAS) to evaluate the severity of pain before the procedure, at primary (2 months) and at secondary (12 months) follow-ups. RESULTS: All the procedures were technically successful. The median VAS scores were 7.95 before the procedure, 3.9 at 2 months and 2.95 at 12 months. MRI analysis showed a significant decrease in herniation size at 2 months (-20%, p = 0.008). No immediate or late complications were observed. Only three patients (13.6%) underwent lumbar spine microdiscectomy in the year following ozone therapy. The treatment appeared to be more effective in cases of nerve root symptomatology. CONCLUSION: This study suggests that intradiscal O2-O3 therapy is safe and effective for the treatment of lumbar disc herniation associated with resistant lumbar or lumbar radicular pain.

The effects of oxygen-ozone therapy on regulatory T-cell responses in multiple sclerosis patients.

Multiple sclerosis (MS) is a common degenerative disorder of the central nervous system. The decreased frequency and dysfunction of Treg cells cause inflammation and disease progression. Ozone autohemotherapy can be used as a potential therapeutic approach to regulate the immune system responses and inflammation in MS. For this purpose, 20 relapsing-remitting multiple sclerosis patients were under treatment with ozone twice weekly for 6 months. The frequency of Treg cell, the expression levels of the Treg cell-related factors (FoxP3, IL-10, TGF-ß, miR-17, miR-27, and miR-146A), and the secretion levels of IL-10 and TGF-ß were assessed. We found a significant increase in the number of Treg cells, expression levels of FoxP3, miRNAs (miR-17 and miR-27), IL-10, and TGF-ß factors in patients after oxygen-ozone (O2 -O3 ) therapy compared to before treatment. In contrast, oxygen-ozone therapy notably decreased the expression level of miR-146a in treated patients. Interestingly, the secretion levels of both IL-10 and TGF-ß cytokines were considerably increased in both serum and supernatant of cultured peripheral blood mononuclear cells in posttreatment condition compared to pretreatment condition. According to results, oxygen-ozone therapy raised the frequency of Treg cell and its relevant factors in treated MS patients. Oxygen-ozone therapy would contribute to improving the MS patients by elevating the Treg cell responses.

Low Ozone Concentrations Differentially Affect the Structural and Functional Features of Non-Activated and Activated Fibroblasts In Vitro.

Oxygen-ozone (O2-O3) therapy is increasingly applied as a complementary/adjuvant treatment for several diseases; however, the biological mechanisms accounting for the efficacy of low O3 concentrations need further investigations to understand the possibly multiple effects on the different cell types. In this work, we focused our attention on fibroblasts as ubiquitous connective cells playing roles in the body architecture, in the homeostasis of tissue-resident cells, and in many physiological and pathological processes. Using an established human fibroblast cell line as an in vitro model, we adopted a multimodal approach to explore a panel of cell structural and functional features, combining light and electron microscopy, Western blot analysis, real-time quantitative polymerase chain reaction, and multiplex assays for cytokines. The administration of O2-O3 gas mixtures induced multiple effects on fibroblasts, depending on their activation state: in non-activated fibroblasts, O3 stimulated proliferation, formation of cell surface protrusions, antioxidant response, and IL-6 and TGF-ß1 secretion, while in LPS-activated fibroblasts, O3 stimulated only antioxidant response and cytokines secretion. Therefore, the low O3 concentrations used in this study induced activation-like responses in non-activated fibroblasts, whereas in already activated fibroblasts, the cell protective capability was potentiated.

[The use of the functional state of the joints for the estimation of the effectiveness of the application of oxygen/ozone therapy for the rehabilitative treatment of the patients suffering from knee arthritis]. / Funktsional'noe sostoianie sustavov kak kriterii otsenki éffektivnosti ispol'zovaniia ozonoterapii v vosstanovitel'nom lechenii bol'nykh gonartrozom.

BACKGROUND: In the recent years, the ever increasing tendency toward the growth in the prevalence of degenerative-dystrophic articular diseases has been observed especially among the young subjects and the middle-aged people. The enhanced economic burden of treating the patients with this condition on the public health care system frequently comes not only from chronic pathology of the joints but also from the development of concomitant disorders which dictates the necessity of the search for the new methods for the management of such patients. AIM: The objective of the present study was the optimization of the treatment of the patients presenting with stage I-II gonarthrosis by the para-articular application of oxygen/ozone therapy at the early stages of knee arthritis and the evaluation of the clinical effectiveness of this approach. MATERIAL AND METHODS: A total of 89 patients suffering from I-II sg gonarthrosis were recruited for the participation in the study. They included 67.2% of men and 32.8% of women. The age of the patients varied from 33 to 54 (mean 42±5) years, the duration of the disease averaged 4±1.2 years. All the patients were randomly divided into two groups: matched for the age, gender, the duration and severity of the disease, and the methods of its treatment. The main group was comprised of 45 patients whereas the group of comparison consisted of 44 patients. The generally accepted therapeutic modalities were used to treat the patients of both groups. Those included in the main group (n=45) additionally received para-articular oxygen/ozone therapy applied to the affected knee at a permanent ozone concentration of 8 gr/l (the injection area 15-20 sq.cm), main group n=45). The results of the treatment were evaluated within 1 and 6 months after the onset of therapy. RESULTS: The application of the up-to-date methods of oxygen/ozone therapy in the combination with the universally-accepted drug-based treatments for the patients of the main group resulted in the significant (p<0.01) improvement of the indicators of the biomechanical activity of the knee joints in comparison with that achieved by means of the generic drug-based treatment alone. DISCUSSION: The results of the present study give evidence of the feasibility of application of oxygen/ozone therapy for the treatment of degenerative-dystrophic articular diseases of the knees. The positive dynamics of the indicators of the functional activity of the knee joints, intensity and duration of the pain syndrome as well as the reduction of the period of the temporary incapacity for work associated with exacerbation of knee arthrosis suggests the high clinical and economic effectiveness of the para-articular application of oxygen/ozone therapy. CONCLUSIONS: The present study has demonstrated the expediency and effectiveness of the application of oxygen/ozone therapy during exacerbation of stage І-ІІ knee arthritis as evaluated based on the WOMAC index, VAS, and Lyshom scale within one and six months after the onset of the treatment. The resulting reduction in the number of days of temporary incapacity for work in the patients suffering from knee arthritis gives evidence of the clinical and economic validity of the proposed method of the combined treatment of this condition based on the application of oxygen/ozone therapy.

The efficacy of ultrasonography-guided oxygen-ozone therapy versus corticosteroids in patients with knee osteoarthritis: A multicenter randomized controlled trial.

BACKGROUND: Osteoarthritis (OA) is a widely-known disease distinguished by the breakdown of joint cartilage, leading to pain and morning stiffness. In this context, the role of corticosteroids is well known, but there is still a gap of knowledge on the duty of oxygen-ozone therapy (O2-O3). OBJECTIVE: To evaluate for effectiveness of ultrasound-guided O2-O3 injections compared with corticosteroid injections among patients diagnosed with knee OA. METHODS: This randomized controlled clinical trial was conducted on participants with knee OA who were randomly sorted into two groups: group A, undergoing corticosteroid group (n= 47) and group B, undergoing O2-O3 (n= 49) were injected within the knee joint under ultrasound guidance. The primary outcome measure was the change in the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) score between baseline and 12-weeks post-injection. Secondary outcome measures included visual analog scale scores, joint effusion and a knee flexion ROM. Assessments were recorded at baseline and 4-weeks and 12-weeks post-injection. For the examination of intra- and inter-group variations at various time points, a repeated-measure analysis of variance (two-way ANOVA) was employed. RESULTS: Ninety-six participants completed this study. Based on repeated measurement analysis of variance, a significant effect of time was found for all outcome measures in both groups. Both groups showed clinically significant improvements in knee pain, quality of life and, function. Baseline, 4-week post-injection and 12-week post-injection WOMAC scores (mean ± standard deviation) were 72.54 ± 18.89, 45,95 ± 13.30 and 37.10 ± 19.87 (p= 0.00, p= 0.00, p= 0.00; respectively) in the corticosteroid group, respectively and 68.23 ± 20.18, 42.99 ± 18.67, and 33.43 ± 18.24 (p= 0.00, p= 0.00, p= 0.00; respectively) in the ozone group, respectively. However, no significant group × time interaction was determined regarding all outcome measures. CONCLUSION: The study demonstrates the efficacy of O2-O3 compared to steroid injections regarding functioning and pain relief among patients with diagnosed knee OA.

Chemical Variations Induced by Ozonization of 5% Glucose Solution and Evaluation of Generated Compounds.

The most commonly studied method of administering ozone therapy is systemic ozone therapy. However, there may be situations where this method is not feasible due to technical issues, such as poor vein condition or anemia. As an alternative method, pre-ozonized solutions, such as 0.9% saline solution, have been investigated for their ease of preparation and administration. However, concerns have been raised regarding the formation of chlorine compounds. Currently, there is no available literature on the treatment potential of pre-ozonized glucose solution. The objective of this study is to compare and evaluate the chemical changes induced by ozonization of a 5% glucose solution and determine if any toxic compounds are produced. Our findings indicate that the chemical alterations following ozone infusion are quantitatively minimal and pose a negligible risk in terms of safety.

Oxygen-ozone therapy for pain relief in patients with trapeziometacarpal osteoarthritis: a proof-of-concept study.

PURPOSE: Trapeziometacarpal osteoarthritis (TMC-OA) is a prevalent hand disorder affecting a growing number of people worldwide. While a multidisciplinary approach might provide additional advantages, the analgesic and anti-inflammatory role of intra-articular oxygen-ozone (O2O3) injections combined with physical therapy is still unknown. To assess the impact of a multimodal therapeutic approach combining O2O3 injections with physical therapy in patients with TMC-OA. MATERIALS AND METHODS: A prospective open-label study conducted in the Physical and Rehabilitation Medicine Unit of the "Renato Dulbecco" University Hospital of Catanzaro. We assessed patients with TMC-OA who had not responded to standard medical therapy. Participants received O2O3 therapy and targeted physical therapy for 4 weeks. Pain relief, muscle strength, and physical functioning were assessed at baseline and after 4, 12 and 24 weeks (respectively T0, T1, T2, and T3). RESULTS: Seventeen patients with a mean age of 67.1 ± 6.1 years were included in the study. Short-term improvements in pain intensity were observed (T0: 6.221 ± 1.514; T1: 3.172 ± 1.1451; p < .001) and were maintained over a 24-week follow-up period (T0: 6.221 ± 1.514; T3: 4.393 ± 1.438; p: 0.006). Significant changes were reported also in terms of muscle strength and physical functioning. O2O3 therapy was well-tolerated with no adverse effects. CONCLUSIONS: A combination of O2O3 injections and physical therapy might be considered in patients with TMC-OA. Further investigation is warranted to assess the effectiveness of O2O3 therapy in managing TMC-OA.

The addition of intra-articular trapeziometacarpal O₂O₃ injections to physical therapy is safe and reliable for thumb osteoarthritisO₂O₃ injection could be considered a second-line mini-invasive approach option when simple analgesic and non-pharmacologic interventions have failed, and surgical treatment is not yet indicatedO₂O₃ injections in combination with physical therapy may provide benefits in terms of pain relief in patients with TMC joint OA in whom previous conventional medical therapy has been unsuccessful.

Intramuscular paravertebral oxygen-ozone therapy for chronic neck pain and low back pain: evaluation of 6-month clinical outcomes.

Spinal pain is recognized as the most common cause of disability, work absenteeism and need of healthcare services worldwide. Although many strategies have been developed for conservative treatment of spinal pain, its increasing prevalence diagnosis highlights the need for new treatments. Oxygen-ozone (O2-O3) therapy is considered to be an alternative therapy due to its analgesic and anti-inflammatory effects. This retrospective study evaluated the effects of O2-O3 intramuscular paravertebral injections in 76 patients with chronic neck pain or low back pain, in terms of pain and disability reduction, quality of life improvement, and analgesic drug intake. Patients were evaluated before, at the end of the treatment, and at 1, 3 and 6 months after the last treatment, using Numeric Rating Scale, Neck Disability Index or Oswestry Disability Index, and Short Form-12 Health Survey. There were significant beneficial effects of O2-O3 therapy in reducing pain and disability reduction and improving quality of life during the 6-month follow-up period. O2-O3 therapy was associated with a reduction in analgesic drug intake at each assessment. Our results allow us not only to support treatment with O2-O3 intramuscular paravertebral injections as a safe and beneficial treatment for chronic low back pain, but also to consider it as a valuable conservative therapy for patients with chronic neck pain.

Systemic Ozone Therapy May Improve the Perception of Quality of Life: Ozonized Glucose Solution 5% Extends Reach to Patients Ineligible for Blood Infusion.

Ozone therapy is a complementary treatment that has gained popularity due to its safety and wide range of applications. Systemic ozone therapy involves withdrawing 100 to 200 ml of blood, treating it with an oxygen-ozone mixture, and then reinfusing it. This process requires large-caliber venous access, which can be a limitation. To overcome this, alternative administration methods have been explored, including the use of ozonized solutions. The aim of this study is to evaluate the clinical effects of systemic ozone therapy on the perception of quality of life and to analyze the outcomes of different administration methods. Three groups of patients were treated: one group received classical systemic ozone therapy, another received ozone therapy via intravenous infusion of a 5% glucose solution, and the third group alternated between the two methods. The results showed an improvement in perceived quality of life in all groups, regardless of the method used. Thus, systemic ozone therapy showed efficacy in improving the perception of quality of life in our group. Moreover, intravenous infusion of a 5% glucose solution has made it possible to treat patients who could not be treated with the classical method, achieving similar results.

Cognitive, Neuropsychological and Biological Effects of Oxygen-Ozone Therapy on Frailty: A Study Protocol for a 5-Week, Randomized, Placebo-Controlled Trial.

Cognitive frailty (CF) is a heterogeneous syndrome that is becoming one of the most serious health problems as the world's population age is increasing. Elucidating its biological mechanisms as well as prevention and treatments is becoming increasingly significant, particularly in view of the associated health costs. We presented the study protocol of a research project funded by the Italian Ministry of Health (grant number RF-2016-02363298) aiming to investigate the cognitive and neuropsychological effects of a 5-week treatment with therapy based on the regenerative properties of ozone (O3) in a cohort of subjects stratified according to CF scores. We also studied the potential effects of O3 on blood-based biomarkers indicative of specific biological systems that may be altered in CF. Seventy-five older persons were recruited and randomly assigned to receive the active treatment (150 cc of oxygen-O2-O3 mixture at the concentration of 30 µg of O3 per cc of O2), O2, or the placebo (air) for 5 weeks. The main endpoints were the change in the scores of clinical scales from baseline (T0) to weeks 3 (T3), 9 (T9), and 15 (T15) after treatment and the change in biomarker levels resulting from transcriptomics, proteomics, and metabolomic patterns at the same times. The positive results from this study could have important clinical implications.

Alpha-Lipoic Acid, Palmitoylethanolamide, Myrrh, and Oxygen-Ozone Therapy Improve Pharmacological Therapy in Acute Painful Lumbosacral Radiculopathy due to Herniated Disc.

BACKGROUND: Neuropathic mechanisms largely contribute to radicular Low Back Pain (LBP) and an increase in oxidative stress is recognized as one of the possible causes of nerve damage, inducing axonal degeneration and myelin degradation of nerve fibers. OBJECTIVES: We investigated whether a combination of nutraceutical supplements and oxygen-ozone (O2-O3) therapy might reduce disability and improve clinical effects of pharmacological therapy in patients with acute radicular LBP. STUDY DESIGN: This is a prospective, open-label, comparative observational study approved by the Institutional Review Board of the Sapienza University of Rome (RS 6285/2021). SETTING: Physical Medicine and Rehabilitation Unit of Sant'Andrea Hospital. METHODS: Within the scope of this study, 62 patients with acute radicular LBP diagnosed with disc herniation were assigned into 4 groups. The first group was assigned pharmacological therapy (n = 16), the second group was assigned pharmacological therapy and nutraceutical supplements (n = 15), the third group was assigned pharmacological therapy and O2-O3 therapy (n = 15), and the fourth group was assigned pharmacological therapy, nutraceutical supplements, and O2-O3therapy (n = 16). All patients who participated in the study were evaluated at the beginning of the study, 2 weeks, and 4 weeks (T2) after the beginning of treatment using the Numeric Rating Scale (NRS-11), Oswestry Disability Index (ODI), and 12-item Short-Form Health Survey. Opioid analgesic intake was noted from baseline to end of treatment (T2). RESULTS: In each group was observed a statistically significant difference for all measures compared to the baseline. At the T2 evaluation time between groups for the Mann-Whitney U test, a statistically significant difference was found: in the ODI scale between groups B and A (P = 0.004), groups C and A (P < 0.001), and groups D and A (P < 0.001); in the NRS-11 between groups B and A (P = 0.017), groups C and A (P = 0.002), and groups D and A (P < 0.001); in the 12-item Physical Component Summary score between groups B and A (P = 0.003), groups C and A (P = 0.002), and groups D and A (P < 0.001), while no significant differences between groups were observed in the 12-item Mental Component Summary score. The average days of opioid usage were similar in the 4 groups (8.33 in group A, 8.33 in group B, 8.33 in group C, and 8.75 in group D). However, the percentage of patients requiring adjuvant opioid therapy differed remarkably: 60% in group A, 40% in group B, 20% in group C, and 25% in group D. LIMITATIONS: A small number of patients were recruited, and we did not perform long-term follow-up. CONCLUSIONS: This study supports a multimodal approach combining nutraceutical supplements and O2-O3 therapy with pharmacological therapy in the treatment of acute radicular LBP secondary to disc herniation. The combination of neurotrophic and antioxidant therapies represents an etiopathogenetic approach, not purely symptomatic, that reduces symptomatology and avoids progression of the nerve damage.

Percutaneous Computed Tomography-Guided Oxygen-Ozone (O2O3) Injection Therapy in Patients with Lower Back Pain-An Interventional Two-Year Follow-Up Study of 321 Patients.

OBJECTIVES: To assess the effect of oxygen-ozone therapy guided by percutaneous Computed Tomography (CT) compared to corticosteroids in individuals experiencing lower back pain (LBP) not attributed to underlying bone-related issues. METHODS: A total of 321 patients (192 males and 129 females, mean age: 51.5 ± 15.1 years) with LBP were assigned to three treatment groups: group A) oxygen-ozone only, group B) corticosteroids only, group C) oxygen-ozone and corticosteroids. Treatment was administered via CT-guided injections to the intervertebral disc (i.e., intradiscal location). Clinical improvement of pain and functionality was assessed via self-reported pain scales and magnetic resonance (MR) and CT imaging. RESULTS: At all follow-up times, the mean score of the numeric rating scale and the total global pain scale (GPS) of study groups receiving oxygen-ozone (groups A and C) were statistically significantly lower than the study group receiving corticosteroids only (group B), with p < 0.001. There was a statistically significant difference between groups A and C at 30 days for the numeric rating scale. CONCLUSIONS: The percutaneous application of oxygen-ozone in patients with LBP due to degeneration of the lumbosacral spine showed long-lasting significant pain reduction of up to two years post-treatment when compared to corticosteroids alone. Combination therapy of oxygen-ozone and corticosteroids can be useful as corticosteroids showed statistically significant improvement in LBP earlier than the oxygen-ozone-only treatment.

Efficacy of Oxygen-Ozone Therapy and Platelet-Rich Plasma for the Treatment of Knee Osteoarthritis: A Meta-analysis and Systematic Review.

Context: This systematic review and meta-analysis evaluated the effect of the intra-articular injection of platelet-rich plasma (PRP) and oxygen-ozone therapy and provided an evidence-based methodology to treat KOA. Method: Databases, including Cochrane Library, PubMed, and EMBASE, were searched. The retrieval period was before 2021. Two reviewers performed the process of screening and data extraction. Mean differences were calculated [95% confidence interval (CI)] with an inverse-variance method and fixed effect model. Meta-analysis was performed using the latest version of STATA version 16. Results: A total of 12 studies out of 769 articles were evaluated. The mean difference of visual analog scale score between ozone and control groups in the first month after injection was -0.02 (MD, -0.02; 95% CI: -0.32, 0.28; P < 0.05). Mean differences of WOMAC pain, stiffness, and physical function score between baseline and after PRP were -3.53 (MD: -3.53; 95% CI: -4.04, -3.02; P = 0.00), -0.60 (MD: -0.60; 95% CI: -4.0 - 0.864, -0.34; P = 0.00), and -5.96 (MD: -5.96; 95% CI: -7.83, -4.09; P = 0.00). Conclusions: Our results showed that to treat knee osteoarthritis, using PRP for a longer period of 6 - 12 months after the intervention shows better clinical results, while oxygen-ozone therapy has short-term results.

Ozone at low concentration modulates microglial activity in vitro: A multimodal microscopy and biomolecular study.

Oxygen-ozone (O2 -O3 ) therapy is an adjuvant/complementary treatment based on the activation of antioxidant and cytoprotective pathways driven by the nuclear factor erythroid 2-related factor 2 (Nrf2). Many drugs, including dimethyl fumarate (DMF), that are used to reduce inflammation in oxidative-stress-related neurodegenerative diseases, act through the Nrf2-pathway. The scope of the present investigation was to get a deeper insight into the mechanisms responsible for the beneficial result of O2 -O3 treatment in some neurodegenerative diseases. To do this, we used an integrated approach of multimodal microscopy (bright-field and fluorescence microscopy, transmission and scanning electron microscopy) and biomolecular techniques to investigate the effects of the low O3 concentrations currently used in clinical practice in lipopolysaccharide (LPS)-activated microglial cells human microglial clone 3 (HMC3) and in DMF-treated LPS-activated (LPS + DMF) HMC3 cells. The results at light and electron microscopy showed that LPS-activation induced morphological modifications of HMC3 cells from elongated/branched to larger roundish shape, cytoplasmic accumulation of lipid droplets, decreased electron density of the cytoplasm and mitochondria, decreased amount of Nrf2 and increased migration rate, while biomolecular data demonstrated that Heme oxygenase 1 gene expression and the secretion of the pro-inflammatory cytokines, Interleukin-6, and tumor necrosis factor-α augmented. O3 treatment did not affect cell viability, proliferation, and morphological features of both LPS-activated and LPS + DMF cells, whereas the cell motility and the secretion of pro-inflammatory cytokines were significantly decreased. This evidence suggests that modulation of microglia activity may contribute to the beneficial effects of the O2 -O3 therapy in patients with neurodegenerative disorders characterized by chronic inflammation. HIGHLIGHTS: Low-dose ozone (O3 ) does not damage activated microglial cells in vitro Low-dose O3 decreases cell motility and pro-inflammatory cytokine secretion in activated microglial cells in vitro Low-dose O3 potentiates the effect of an anti-inflammatory drug on activated microglial cells.

Patients with Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) Greatly Improved Fatigue Symptoms When Treated with Oxygen-Ozone Autohemotherapy.

(1) Background: Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a chronic syndrome characterized by fatigue as its major and most outstanding symptom. Previous evidence has supported the ability of ozone to relief ME/CFS related fatigue in affected patients (2) Methods: A number of 200 ME/CFS previously diagnosed patients, (mean age 33 ± 13 SD years) were consecutively treated with oxygen-ozone autohemotherapy (O2-O3-AHT). Fatigue was evaluated via an FSS 7-scoring questionnaire before and following 30 days after treatment. (3) Results: Almost half (43.5%) of the treated patients evolved their FSS scale from the worst (7) to the best (1) score, assessing the highest improvement from being treated with O2-O3-AHT. Furthermore 77.5% of patients experienced significant ameliorations of fatigue, of 4-6 delta score. No patient showed side effects, yet experienced long lasting fatigue disappearance, by three months follow up (4) Conclusions: Treatment with O2-O3-AHT greatly improves ME/CFS related fatigue, aside from sex and age distribution.

Ozone therapy for patients with COVID-19 pneumonia: Preliminary report of a prospective case-control study.

BACKGROUND: There is still no specific treatment strategies for COVID-19 other than supportive management. DESIGN: A prospective case-control study determined by admittance to the hospital based on bed availability. PARTICIPANTS: Eighteen patients with COVID-19 infection (laboratory confirmed) severe pneumonia admitted to hospital between 20th March and 19th April 2020. Patients admitted to the hospital during the study period were assigned to different beds based on bed availability. Depending on the bed the patient was admitted, the treatment was ozone autohemotherapy or standard treatment. Patients in the case group received ozonated blood twice daily starting on the day of admission for a median of four days. Each treatment involved administration of 200 mL autologous whole blood enriched with 200 mL of oxygen-ozone mixture with a 40 µg/mL ozone concentration. MAIN OUTCOMES: The primary outcome was time from hospital admission to clinical improvement. RESULTS: Nine patients (50%) received ozonated autohemotherapy beginning on the day of admission. Ozonated autohemotherapy was associated with shorter time to clinical improvement (median [IQR]), 7 days [6-10] vs 28 days [8-31], p = 0.04) and better outcomes at 14-days (88.8% vs 33.3%, p = 0.01). In risk-adjusted analyses, ozonated autohemotherapy was associated with a shorter mean time to clinical improvement (-11.3 days, p = 0.04, 95% CI -22.25 to -0.42). CONCLUSION: Ozonated autohemotherapy was associated with a significantly shorter time to clinical improvement in this prospective case-control study. Given the small sample size and study design, these results require evaluation in larger randomized controlled trials. CLINICAL TRIAL REGISTRATION NUMBER: NCT04444531.

Male vs. Female Differences in Responding to Oxygen-Ozone Autohemotherapy (O2-O3-AHT) in Patients with Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS).

(1) Background: Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a syndrome that has fatigue as its major symptom. Evidence suggests that ozone is able to relieve ME/CFS-related fatigue in affected patients. (2) Objective: To evaluate whether differences exist between males and females in ozone therapy outputs in ME/CFS. (3) Methods: In total, 200 patients previously diagnosed with ME/CFS (mean age 33 ± 13 SD years) underwent treatment with oxygen-ozone autohemotherapy (O2-O3-AHT). Fatigue was investigated via an FSS 7-scoring questionnaire before and following 1 month after treatment. (4) Results: The Mann-Whitney test (MW test) assessed the significance of this difference (H = 13.8041, p = 0.0002), and female patients showed better outcomes than males. This difference was particularly striking in the youngest age cohort (14-29 years), and a KW test resulted in H = 7.1609, p = 0.007 for the Δ = 28.3% (males = 3.8, females = 5.3). (5) Conclusions: When treated with O2-O3-AHT, females respond better than males.