Search Results (55)

Objectives: This study aimed to evaluate the effects of a combined treatment consisting of O₂O₃ injections and McKenzie-based physiotherapy exercises, compared to a Control group treated with O₂O₃ injections and a Back School physiotherapy program, in reducing pain and disability in individuals with chronic non-specific neck pain. Methods: In this prospective double-arm pilot study, patients with chronic non-specific neck pain and a Numerical Rating Scale (NRS) > 4 were enrolled. All patients received eight weekly sessions of O₂O₃ injections (10 μg/mL, 10 mL total, and 2 mL bilaterally into the cervical paravertebral muscles). Patients were then randomly assigned (1:1 ratio) to either an experimental group receiving McKenzie physiotherapy or a Control group undergoing Back School techniques, with five sessions per week over two weeks. Outcome measures included the Neck Disability Index (NDI), NRS, EuroQol-5D-3L (EQ5D3L), and EuroQol Visual Analog Scale (EQ-VAS). Results: A total of 41 patients were included and divided into two groups: Back School (n = 21; mean age: 63.9 ± 13.4 years) and McKenzie (n = 20; mean age: of 57.3 ± 12.9 years). Both groups showed significant improvement in NDI, NRS, EQ5D3L, and EQ-VAS following the O₂O₃ injection cycle (∆T0–T1 p < 0.001). The subsequent addition of physical therapy led to further improvements across all outcomes in both groups (∆T1–T2 p < 0.001), with the McKenzie group showing slightly greater benefits, despite the lack of significant differences. Conclusions: This study demonstrated the effects of combining O₂O₃ injections with either McKenzie or Back School therapy in improving pain, disability, and quality of life in patients with chronic non-specific neck pain. Full article

In this Reply, we address the criticisms raised by Franzini, Valdenassi, and Chirumbolo concerning our study on the effects of ozonized saline solution (O3SS) on microglial polarization and endothelial responses in vitro. We clarify that the primary aim of the original work was mechanistic, relying on rigorously controlled cellular models that are universally recognized as essential preclinical tools in translational medicine. We reaffirm the validity of our experimental approach, including the preparation and characterization of O3SS based on empirically validated methodologies, direct ozone quantification, and standardized protocols consistent with the existing literature and clinical practice. Concerns regarding ozone chemistry, dose relevance, and hypochlorite formation are addressed through analytical validation, biological threshold considerations, and the use of certified assays. We further justify the choice of BV2 microglia and HUVEC cells as established and widely used models for investigating inflammatory and vascular pathways under reproducible conditions. Statistical analyses, gene expression interpretation, and the absence of comparative pharmacological agents are discussed in the context of the study’s focused objectives. Finally, we place our findings within the established framework of ozone as an indirect pro-oxidant that elicits adaptive redox signaling (“oxidative eustress”), emphasizing the translational relevance of in vitro systems for elucidating early mechanistic events. Overall, we maintain that our study provides a robust, balanced, and evidence-based contribution to the understanding of ozone-derived redox biology. Full article

Background and Objectives: Diabetes mellitus (DM) is a major global health concern, with diabetes-related foot disease (DFD) representing one of its most severe complications, often resulting in chronic infection, osteomyelitis, and limb amputation. Conventional therapies frequently fail in refractory cases, necessitating novel adjunctive strategies. Ozone therapy (OT) possesses antimicrobial, immunomodulatory, and oxygen-enhancing properties, while negative pressure wound therapy (NPWT) facilitates granulation, exudate removal, and tissue perfusion. This study explored the combined efficacy of OT and NPWT in advanced DFD. Materials and Methods: An exploratory, retrospective, observational cohort study was conducted at a specialized wound care center in Gdańsk, Poland, between 2019 and 2022. The study included 30 patients (n = 30) with refractory DFD involving both soft tissue and bone infection who had not responded to previous conventional treatment. The analyzed treatment approach consisted of surgical debridement, application of topical ozonated preparations, and (NPWT) with instillation of ozonated saline administered over a six-week period. Clinical outcomes included wound healing assessed using the Wagner classification and wound volume reduction, pain intensity measured using the Numeric Rating Scale (NRS), inflammatory biomarkers (C-reactive protein [CRP] and procalcitonin [PCT]), and microbiological characteristics of wound cultures. Statistical analyses were performed using the Wilcoxon signed-rank test and the chi-square test, and regression modeling was applied to identify potential predictors of therapeutic response. Statistical significance was defined as p < 0.05. Results: By week six, 100% of ulcers improved to Wagner stage ≤1, with 26.7% achieving stage 0. Median wound volume decreased from 5.5 cm³ to 0 cm³ (p < 0.001). Pain scores declined from 7.2 ± 0.96 points to 0.2 ± 0.5 points (p < 0.001). CRP and PCT levels decreased significantly (p < 0.001), and microbiological clearance was observed in all cases. Higher body mass index (BMI) was associated with poorer pain reduction. Conclusions: The combination of standard wound care with OT and NPWT was associated with clinically relevant improvements in wound healing, infection control, systemic inflammation, and pain reduction in patients with refractory DFD. Although limited by a non-controlled design and small cohort size, these findings support further randomized controlled trials to define the role of this combined approach in integrated diabetic foot care. Full article

Objective: To compare the efficacy and safety of gas-based therapies for chronic wounds using a systematic review and network meta-analysis (NMA). Methods: Following PRISMA 2020, we systematically searched PubMed, Embase, Web of Science, Cochrane CENTRAL, and CBM from inception to 1 October 2025, screened studies in duplicate, and resolved disagreements by arbitration (κ = 0.87). Randomized controlled trials (RCTs) enrolling adults with chronic wounds were eligible; the primary endpoint was complete wound healing. Pairwise meta-analysis used risk ratios (RRs) with 95% CIs; heterogeneity was assessed with Q/I² and random-effects models were applied when appropriate. A frequentist NMA synthesized direct and indirect evidence, and treatments were ranked with SUCRA. Publication bias (Egger/Begg) and evidence certainty (GRADE) were evaluated. Results: Twenty-seven RCTs comprising 1673 participants were included. In pairwise pooling, gas therapies significantly increased complete healing versus standard care (random-effects RR = 2.17, 95% CI 1.61–2.94), with substantial heterogeneity (I² = 75.7%); results were directionally consistent and robust to sensitivity analyses. Prespecified subgroup analyses suggested effect modification by intervention type and wound etiology. In the NMA, most gas modalities showed beneficial trends versus standard care; however, SUCRA ranking placed standard care highest (93.9%), a finding attributed by the authors to network structure and between-study variability. Ozone therapy and topical oxygen ranked next, whereas HBOT and cold atmospheric plasma ranked mid-range; CO₂ therapy ranked lowest due to sparse evidence. Small-study effects were likely (Egger p < 0.001; Begg p = 0.013), and overall certainty was graded as moderate, limited primarily by heterogeneity, imprecision, and potential publication bias. Conclusions: Across RCTs, gas therapies as a class improve the probability of complete healing in chronic wounds relative to standard care, but effect sizes vary by modality and wound type. Given heterogeneity, possible publication bias, and inconsistencies within the evidence network, these findings should be applied with caution. HBOT remains the modality supported by the broadest evidence base, while large, high-quality, multicenter RCTs are needed to refine comparative effectiveness and safety rankings across gas therapies. Full article

This commentary critically evaluates the translational relevance of a recent study investigating the effects of ozonated saline solution (O₃SS) on microglial and endothelial cell models. While the original research proposes potential antioxidant and anti-inflammatory benefits of low-dose ozone exposure, we identify significant methodological and conceptual flaws that undermine its conclusions. Key concerns include the unjustified assumption that ozone behaves similarly in microwell cultures and clinical infusion settings, despite known physicochemical differences affecting ozone stability and reactivity. The use of immortalized BV2 and HUVEC cells, which lack the complexity of in vivo systems, further limits the study’s applicability. The absence of accurate ozone quantification, proper controls, protein-level validation, and kinetic modeling exacerbates these weaknesses. Our analysis also demonstrates, through differential equation modeling, that ozone rapidly decays in saline solutions, making systemic delivery via infusion chemically implausible as a therapeutic approach. Moreover, the extrapolation of in vitro gene expression data to systemic therapeutic claims lacks scientific justification. We conclude that while the observed cellular responses in vitro are of academic interest, they do not support the efficacy or safety of O₃SS in clinical settings. A more rigorous approach is necessary to substantiate the biomedical potential of ozonated solutions. Full article

Background: Oxygen–ozone (O₂–O₃) therapy is a minimally invasive treatment for discogenic lumbar pain. Although rare, spinal infections—specifically spondylodiscitis—have been reported following intradiscal injections. To date, Lactobacillus iners has not been described as a causative agent in this context. Case Presentation: A 55-year-old immunocompetent woman presented with progressive lumbosciatica and elevated inflammatory markers three months after intradiscal O₂–O₃ therapy. MRI revealed L4–L5 spondylodiscitis with paravertebral involvement. Surgical biopsy confirmed L. iners as the pathogen. She underwent decompression and received targeted intravenous antibiotics, achieving full clinical and radiological recovery. Methods: A systematic literature review was performed using PubMed, MEDLINE, and Scopus to identify reports of spondylodiscitis following oxygen–ozone therapy. Six cases were included based on predefined inclusion criteria. Results: The 8 identified cases involved a range of pathogens, including Staphylococcus aureus, Streptococcus beta-haemolyticus, Escherichia coli, Achromobacter xylosoxidans, Mycobacterium abscessus, and Streptococcus intermedius, and one culture-negative infection. Clinical presentations varied from radiculopathy to sepsis. Management strategies encompassed both conservative (antibiotics alone) and surgical approaches, depending on neurological status and abscess formation. Outcomes were favorable in all cases except one fatality. Conclusions: This report is the first to describe L. iners spondylodiscitis in an immunocompetent patient following O₂–O₃ therapy. Clinicians should vigilantly evaluate post-infiltration spinal infections, maintain a low threshold for imaging and biopsy, and implement pathogen-targeted antibiotic regimens, with surgical intervention as needed. Full article

Background/Objectives: Mild cognitive impairment (MCI) is accompanied by olfactory dysfunction, and few interventions target shared chemosensory–cognitive mechanisms. We retrospectively examined whether a 5-week oxygen–ozone major autohemotherapy (MAH) cycle is associated with coupled improvements in olfactory and cognitive performance in adults with MCI. Methods: We analyzed 81 individuals with MCI who completed 10 MAH sessions (twice weekly) and 93 matched healthy controls. In the MCI group, olfactory function was measured before and after MAH using Sniffin’ Sticks^® threshold–discrimination–identification (TDI) scores; global cognition was assessed with the Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA). We evaluated between-group and pre–post changes and used Spearman correlations to assess olfactory–cognitive coupling. Results: At baseline, MCI participants showed lower TDI and MoCA scores than controls and more hyposmia/anosmia. Following MAH, the proportion of normosmic patients increased from 32.1% to 50.6%, with fewer anosmic cases. TDI scores improved but remained lower than in controls. MMSE scores were unchanged, whereas MoCA total scores increased, with domain-level gains and a significant improvement in Language Repetition. TDI gains were modestly correlated with MoCA total and selected domain changes. Conclusions: In this retrospective cohort, MAH was associated with partial restoration improvements of olfactory function and improved cognitive performance. Correlated olfactory–cognitive changes were observed within the treated MCI group; however, causal attribution to O₂–O₃ MAH cannot be established without randomized, double-blind, sham-controlled trials with coupled olfactory–cognitive gains consistent with a shared, potentially modifiable substrate. Prospective randomized trials are needed to confirm efficacy and clinical utility. Full article

Aging is accompanied by progressive gastrointestinal structural and functional decline, increased intestinal permeability, dysbiosis, and impaired mucosal immunity, collectively elevating susceptibility to infections, chronic inflammation, and multimorbidity. These age-related changes are further exacerbated by polypharmacy, metabolic disorders, and lifestyle factors, positioning the gastrointestinal tract as a central driver of systemic physiological decline. Gut-centered interventions have emerged as critical strategies to mitigate these vulnerabilities and support healthy aging. Dietary modulation, prebiotic and probiotic supplementation, and microbiota-targeted approaches have demonstrated efficacy in improving gut microbial diversity, enhancing short-chain fatty acid production, restoring epithelial integrity, and modulating immune signaling in older adults. Beyond nutritional strategies, non-nutritional interventions such as molecular hydrogen and medical ozone offer complementary mechanisms by selectively neutralizing reactive oxygen species, reducing pro-inflammatory signaling, modulating gut microbiota, and promoting mucosal repair. Hydrogen-based therapies, administered via hydrogen-rich water or inhalation, confer antioxidant, anti-inflammatory, and cytoprotective effects, while ozone therapy exhibits broad-spectrum antimicrobial activity, enhances tissue oxygenation, and stimulates epithelial and vascular repair. Economic considerations further differentiate these modalities, with hydrogenated water positioned as a premium wellness product and ozonated water representing a cost-effective, scalable option for geriatric gastrointestinal care. Although preclinical and early clinical studies are promising, evidence in older adults remains limited, emphasizing the need for well-designed, age-specific trials to establish safety, dosing, and efficacy. Integrating dietary, microbiota-targeted, and emerging non-nutritional gut-centered interventions offers a multimodal framework to preserve gut integrity, immune competence, and functional health, potentially mitigating age-related decline and supporting overall health span in older populations. Full article

Background and Clinical Significance: Breast cancer is the most frequent malignancy in women. Metastatic breast cancer is considered a treatable but incurable condition, with a median overall survival of only 2–3 years. Among its subtypes, triple-negative breast cancer (TNBC) accounts for a high proportion of breast cancer-related deaths. It is characterized by an aggressive clinical course, early recurrence, and a strong propensity for visceral and brain metastases. Case Presentation: We report the case of a Caucasian woman who developed systemic disease recurrence with lung and mediastinal lymph node metastases, occurring two years after her primary diagnosis and treatment for TNBC. The patient received three months of chemotherapy combined with an adjuvant integrative protocol consisting of melatonin, cannabidiol, and oxygen–ozone therapy. This combined approach led to the complete disappearance of the lung nodules. Subsequently, stereotactic radiotherapy was performed and, in association with the ongoing integrative treatment, resulted in a significant reduction in mediastinal adenopathy. Introduction of immunotherapy, supported continuously by the same adjuvant strategy, achieved a complete and durable remission. Strikingly, the patient remained disease-free five years after the diagnosis of lung and mediastinal metastases. Conclusions: This clinical case highlights the potential benefit of using melatonin, cannabidiol, and oxygen–ozone therapy as part of an integrative approach in patients with aggressive metastatic TNBC. While it is not possible to establish causality from a single case, the sustained remission observed suggests that such unconventional adjuvant strategies could play a supportive role in enhancing the efficacy of standard oncologic therapies. Full article

Background/Aim: This systematic review and meta-analysis aimed to evaluate the clinical efficacy of oxygen-based adjunct therapies in patients with periodontitis, including ozone therapy, hyperbaric oxygen therapy, and local oxygen delivery, as adjuncts to subgingival instrumentation. These interventions have been proposed to counteract tissue hypoxia and inflammation, which sustain an environment favorable to anaerobic pathogens in periodontitis. Methods: An electronic search was conducted in MEDLINE PubMed, the Cochrane Library, the Cochrane Central Register of Controlled Trials, and SciELO. Risk of bias was assessed using the Cochrane Risk of Bias Tool 2. Standardized mean difference was calculated for gains in clinical attachment level, and a random effects model was applied due to high variability. Results: The meta-analysis of adjunct ozone therapies presented a pooled standardized mean difference of 0.53 (95% CI [−0.14, 1.19]), indicating a clinically relevant medium effect in favor of ozone therapies, though this effect was not statistically significant and substantial heterogeneity was observed (I² = 70%, p < 0.01). Meta-analysis was restricted to adjunct ozone therapies due to the limited availability of qualifying studies for hyperbaric oxygen therapy and local oxygen therapies. Conclusions: While the medium effect size in favor of ozone therapies could be clinically relevant, the statistical non-significance underscores the need for more evidence before widespread adoption. Individual studies reported significant benefits for adjunct HBOT and ozonated olive oil, but comparison between oxygen delivery modes was not possible due to heterogeneous protocols. Full article

Oxygen–ozone (O₂–O₃) therapy is widely used for treating lumbar disc herniation. However, controversy remains regarding the safest and most effective route of administration. While intradiscal injection is purported to show clinical efficacy, it has also been associated with serious complications. In contrast, the intramuscular route can exhibit a more favourable safety profile and comparable pain outcomes, suggesting its potential as a safer alternative in selected patient populations. This mixed-method study combined computed tomography (CT) imaging, biophysical diffusion modelling, and a meta-analysis of clinical trials to evaluate whether intramuscular O₂–O₃ therapy can achieve disc penetration and therapeutic efficacy comparable to intradiscal nucleolysis, while minimizing procedural risk. Literature searches across PubMed, Scopus, and Cochrane databases identified seven eligible studies (four randomized controlled trials and three cohort studies), encompassing a total of 120 patients. Statistical analyses included Hedges’ g, odds ratios, and number needed to harm (NNH). CT imaging demonstrated gas migration into the intervertebral disc within minutes after intramuscular injection, confirming the plausibility of diffusion through annular micro-fissures. The meta-analysis revealed substantial pain reduction with intramuscular therapy (Hedges’ g = −1.55) and very high efficacy with intradiscal treatment (g = 2.87), though the latter was associated with significantly greater heterogeneity and higher complication rates. The relative risk of severe adverse events was 6.57 times higher for intradiscal procedures (NNH ≈ 1180). O₂–O₃ therapy offers a biologically plausible, safer, and effective alternative to intradiscal injection, supporting its adoption as a first-line, minimally invasive strategy for managing lumbar disc herniation. Full article

Ozone (O₃) has re-emerged in periodontology for its antimicrobial, oxygenating, and immunomodulatory actions, yet its role in regeneration remains contentious. This narrative review synthesizes current evidence on adjunctive ozone use in periodontal therapy, delineates cellular constraints—especially in periodontal ligament fibroblasts (PDLFs)—and explores mitigation strategies using bioactive compounds and advanced delivery platforms. Two recent meta-analyses indicate that adjunctive ozone with scaling and root planing yields statistically significant reductions in probing depth and gingival inflammation, with no significant effects on bleeding on probing, plaque control, or clinical attachment level; interpretation is limited by heterogeneity of formulations, concentrations, and delivery methods. Mechanistically, ozone imposes a dose-dependent oxidative burden that depletes glutathione and inhibits glutathione peroxidase and superoxide dismutase, precipitating lipid peroxidation, mitochondrial dysfunction, ATP depletion, and PDLF apoptosis. Concurrent activation of NF-κB and upregulation of IL-6/TNF-α, together with matrix metalloproteinase-mediated extracellular matrix degradation and tissue dehydration (notably with gaseous applications), further impairs fibroblast migration, adhesion, and ECM remodeling, constraining regenerative potential. Emerging countermeasures include co-administration of polyphenols (epigallocatechin-3-gallate, resveratrol, curcumin, quercetin), coenzyme Q10, vitamin C, and hyaluronic acid to restore redox balance, stabilize mitochondria, down-modulate inflammatory cascades, and preserve ECM integrity. Nanocarrier-based platforms (nanoemulsions, polymeric nanoparticles, liposomes, hydrogels, bioadhesive films) offer controlled ozone release and co-delivery of protectants, potentially widening the therapeutic window while minimizing cytotoxicity. Overall, current evidence supports ozone as an experimental adjunct rather than a routine regenerative modality. Priority research needs include protocol standardization, dose–response definition, long-term safety, and rigorously powered randomized trials evaluating bioactive-ozone combinations and nanocarrier systems in clinically relevant periodontal endpoints. Full article

Ozone (O₃) therapy has demonstrated antioxidant and anti-inflammatory properties, but the systemic administration of ozonated saline solution (O₃SS) remains underexplored. This study evaluates the cytotoxicity, antioxidant response, and immunomodulatory effects of O₃SS on murine microglial (BV2) and human endothelial (HUVEC) cells. Cells were exposed to increasing doses of O₃ (1, 5, or 10 μg/NmL) dissolved in saline. Viability assays showed that low doses (1 and 5 μg/NmL) enhanced cell proliferation without cytotoxicity, while the highest dose (10 μg/NmL) reduced viability and increased cell death. O₃SS treatment upregulated antioxidant genes, including Nrf2 and SOD1, and decreased reactive oxygen species in lipopolysaccharide (LPS)-stimulated microglia. Additionally, O₃SS modulated microglial phenotype by reducing pro-inflammatory markers (iNOS, IL-1β) and increasing anti-inflammatory markers (Arg-1, IL-10). Immunofluorescence confirmed enhanced Arg-1 protein expression, indicating a shift toward an anti-inflammatory state. These results suggest that low-dose O₃SS activates cellular antioxidant defenses and promotes an anti-inflammatory microglial phenotype, supporting its potential as a safe systemic O₃ therapy. Further studies are warranted to confirm in vivo efficacy and optimize clinical protocols. Full article

Background: Tendinopathy is a degenerative condition caused by mechanical overload, accounting for approximately 30% of musculoskeletal healthcare cases. It progresses through a process characterized by collagen disorganization, altered vascularization, and neuronal ingrowth. Traditional conservative treatments, such as therapeutic exercises, non-steroidal anti-inflammatory drugs, and physical therapies, are useful, but their effectiveness is sometimes only partial and there is a need to search for new potential solutions. Recent interest in oxygen–ozone (O₂-O₃) therapy stems from preliminary observations suggesting potential anti-inflammatory and regenerative effects. Nevertheless, its clinical role remains speculative and warrants thorough investigation beyond anecdotal evidence. Considering the heterogeneity of clinical presentations and treatment responses among patients, O₂-O₃ therapy has been proposed as a promising tool for tailoring personalized treatment strategies for tendinopathy. This review critically appraises the available literature concerning the mechanistic rationale and clinical applications of O₂-O₃ therapy in tendinopathy, with attention to both its theoretical underpinnings and the quality of empirical evidence. Methods: A literature search was conducted on O₂-O₃ therapy for tendinopathy using PubMed, Cochrane, and Embase, filtering for full-text articles published between 2004 and 2024. Recent clinical trials were included irrespective of evidence level, while excluding systematic reviews, duplicates, and irrelevant studies. Results: Ozone has been shown to modulate oxidative stress, promote neovascularization, and suppress pro-inflammatory cytokines. Both clinical and in vivo studies indicate that O₂-O₃ therapy relieves pain, enhances tendon healing, and improves biomechanical properties. Some comparative studies suggest that O₂-O₃ therapy might provide more sustained symptoms control than corticosteroids, but the heterogeneity of follow-up durations and outcome measures prevents definitive conclusions. Conclusions: O₂-O₃ therapy emerges as a potentially valuable adjunct in the management of chronic tendinopathy, particularly in cases unresponsive to conventional treatments. However, its clinical role remains to be clearly defined and its possible role in personalized medicine needs further exploration, particularly in relation to patient stratification and individualized treatment protocols. Further high-quality randomized controlled trials are warranted to validate its efficacy, determine long-term outcomes, and standardize treatment protocols to ensure safety and reproducibility. Full article

Non-thermal plasma has attracted strong interest in medicine and agriculture due to its ability to generate reactive oxygen and nitrogen species (RONS). These species can stimulate wound healing and seed germination, but at higher levels they induce DNA damage—useful in cancer therapy but harmful when healthy cells must be preserved. Direct study of DNA damage in cells is difficult because of repair processes and protective barriers. To address this, we applied a dual-model system combining plasmid DNA and human lymphocytes exposed to plasma from the RPS40 device. Using selective scavengers, we identified hydroxyl radicals, ozone, and reactive nitrogen species as key mediators of DNA strand breaks and structural changes. Our results support a mechanistic model in which long-lived plasma-derived species (NOx, ozone, acids) dissolve in water and subsequently generate short-lived radicals such as hydroxyl radicals and peroxynitrite. These reactive molecules then directly attack DNA. This integrated approach—linking plasmid and cellular assays with scavenger-based identification of RONS—offers a novel and cost-effective method for dissecting plasma–DNA interactions. The findings provide mechanistic insight into how plasma-activated water damages DNA, guiding the safer and more effective application of plasma technologies in biomedical and agricultural contexts. Full article