Haemolysis Generated with the Use of Glass Bottles Instead of PVC, DEHP-Free Blood Collection Bags in the Oxygen-Ozone Therapy
Franzini M, Chirumbolo S, Valdenassi L. Maedica, 2025.
- Study type
- Other
- Population
- No patients: modelling of haemolysis in blood collected for major autohaemotherapy in glass bottles versus DEHP-free PVC bags, drawing on 25 laboratory experiments
- Published
- 2025, Maedica
- Identifiers
- PMID 40880716, DOI 10.26574/maedica.2025.20.2.257
What the authors reported
The authors modelled haemolysis, methaemoglobin formation, and inflammation with differential equations, drawing on 25 laboratory experiments (12 DEHP-free PVC bags, 13 glass bottles) and the literature. They report faster red-cell rupture in glass (p = 0.03433) and a higher safety index with plastic bags, and conclude that glass bottles should be phased out of major autohaemotherapy.
What limits this result
A self-described commentary based on modelling; no adverse events in patients are reported. The equations and tables appear only as images, the 25 experiments are not described in detail, and the text and a figure caption disagree on how the safety index changes with ozone dose. Two of the three authors are from SIOOT, and the products compared are named brands.
Harms reported after ozone therapy, and studies of how ozone exposure affects health. One study rarely settles a medical question; our guides weigh each paper against the rest of the evidence.
How the study was done
- Design
- Mathematical modelling with ordinary differential equations, drawing on laboratory experiments and published data; the text calls the paper a commentary
The authors applied ordinary differential equations (ODEs) to model haemolysis, biochemical stability and inflammatory responses
- Setting
- Italian Scientific Society of Oxygen Ozone Therapy (SIOOT) and High Master School of Oxygen Ozone Therapy, University of Pavia, Italy (two authors); University of Verona (one author)
- Read from
- Read from the full text of the paper. Read on October 2, 2026.
Source: Abstract on PubMed
Study arms
| Group | Participants | What they received |
|---|---|---|
| DEHP-free PVC bags (laboratory experiments) | 12 | Blood collected for oxygen-ozone major autohaemotherapy in SANO 3 DEHP-free PVC bags (Haemopharm, Milan) |
| Glass bottles (laboratory experiments) | 13 | Blood collected in Ozonosan glass bottles (Hänsler Medical, Germany) |
Source: Full text on PubMed Central
What it measured
- Red-cell integrity and biochemical stability over time (model)
- Better preserved in plastic than in glass
The results indicate that plastic storage is superior, preserving RBC integrity and biochemical stability better than glass over time.
- Haemolysis progression, glass versus plastic
- Statistical difference, p = 0.03433
Therefore, on the basis of these estimations, Figure 2 shows the haemolysis progression over one hour, with minutes on the X-axis, showing a statistical difference p = 0.03433.
- Foam formation by vacuum strength
- About 30% foam at about 400 mm Hg and about 5% at about 100 mm Hg
Obviously, where ΔP is higher (~400 mm Hg), there is more turbulence, so more foam (~30%), whereas where ΔP is lower (~100 mm Hg), there is less turbulence, so less foam (~5%).
- Modelled patient safety index by container
- Higher with plastic bags than with glass bottles
The plastic bag storage method consistently results in a higher SI than glass bottles.
- Modelled treatment failure index
- Lowest with plastic bags across all ozone doses
Plastic bags remain the best choice, showing the lowest failure rate across all ozone doses.
- Safety index and ozone dose, as stated in the text
- Decreases rapidly with increasing ozone dose for plastic bags; the trend is dampened in glass bottles
- Safety index and ozone dose, as stated in a figure caption
- All safety indices rise with ozone dose, at different rates
- Authors' conclusion
- Glass bottles should be phased out to minimize haemolysis-related risks
Further research should continue refining the methodology, but current evidence suggests that glass bottles should be unequivocally phased out to minimize haemolysis-related risks.
Source: Full text on PubMed Central
Safety as reported
The authors argue that red-cell rupture releases lactate dehydrogenase and methaemoglobin, which contribute to inflammatory responses; no adverse events in patients are reportedIncreased red blood cell rupture leads to the release of lactate dehydrogenase and meta-haemoglobin, both of which contribute to inflammatory responses, potentially compromising the intended therapeutic effects of major autohaemotherapy
Source: Full text on PubMed Central
Funding and conflicts
Conflicts of interest: none declared; financial support: none declared
Source: Full text on PubMed Central
Where we cite this study
Does EBO2 (EBOO) Work? The Evidence, Study by Study
Every study in our EBO2 (EBOO) library in one place: design, size, what was measured and found, its limits, and a check for any study a clinic cites.
Ozone Therapy Adverse Events by Route: What Case Reports Show, and Where EBO2 (EBOO) Fits
Every ozone therapy adverse event we could read in case reports and series, grouped by route, plus how to search FDA's MAUDE data and report to MedWatch.