Ozone In-Vitro Examination

An Electron Microscopical Examination Of Cellular Constituents Of Human Whole Blood After In-Vitro Exposure To Ozone Gas

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Abstract

A transmission and scanning electron microscopical examination was undertaken of four main cell types in human whole blood after exposure to ozone gas in-vitro. Cellular morphology of erythrocytes, lymphocytes, neutrophilic and basophilic leukocytes and thrombocytes was analyzed following ozonization of human whole blood with 44, 65, 80, 90, and 110 ug/ml oxygen/ozone mixture under normo- and hyperbaric (1 bar) conditions with a high total gas mass.

Introduction

Over 7000 physicians and naturopathic practitioners in Europe use intravenous ozone auto-hemotherapy for a wide variety of medical conditions including gastrointestinal, cardiologic, peripheral and central vascular, cardiologic disease, immune deficiency illnesses (hiv/aids), and others. Controversy continues to exist regarding the maximum gas concentration per amount of blood as well as the total mass of ozone and pressure conditions during major auto-hemotherapy for a variety of indications with respect to the effects on human cell morphology. Due to the many thousands of patients per year treated with this therapeutic modality, it was therefore of dire interest to observe cellular morphological reactions under extreme ozone gas conditions.

Methodology

Five milliliters of human whole blood was drawn into a 20ml syringe and immediately exposed to 15ml of 44, 55, 65, 80, 90, and 110ug/ml of oxygen/ozone gas mixture. Thereafter the blood was either compromised to 1/2 of the volume of the syringe representing 1 bar (hyperbaric) or left at normobaric conditions and shaken for 45 seconds. Five to 10 drops of the ozonized blood was then introduced into an Ito-Forssmann perfusion fixation solution and processed for observation under a Zeiss EM 10 transmission or JEOL scanning electron microscope.

Results

Thrombocytes, neutrophilic and basophilic leukocytes, lymphocytes and erythrocytes exhibited normal cytological morphology under normo- and hyperbaric conditions up to and including 80 ug/ml (fig. 1,2,3,4,). Lymphocytes, erythrocytes and neutrophils demonstrated evident cyto-pathology under normo- and hyperbaric conditions from 90 ug/ml. Basophilic leukocytes, and thrombocytes were irrevocably damaged at 110 ug/ml (fig. 5,6,7,8).

Conclusion

Extreme conditions were selected in this study in order to provoke cyto-pathological alterations after exposure to ozone gas to human whole blood. That is, 5ml of blood and 15ml of an oxygen/ozone mixture which in normal auto-hemotherapy treatment with 150ml of blood, would extrapolate to a total mass of 19,800 ug/ml ozone, is greatly beyond physiological therapeutic doses administered. Transmission and scanning electron microscopical morphological alterations of cell membranes, organelles and inclusions are clearly observed from 90 ug/ml under normo- and hyperbaric (1bar) conditions and onwards. We therefore conclude and suggest that practitioners do not exceed 80 ug/ml concentrations under normo- or hyperbaric pressure.

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