22.07.2003 RV 1

The Use of Ozone in MedicineMechanisms of Action

Munich May 23 –25, 2003Renate Viebahn-Hänsler

22.07.2003 RV 2

Ozone Treatment in Surgery

1915 A. Wolff1935 E. Payr1937 P. Aubourg

1977 O. Rokitansky1981 H. Werkmeister,1987 H.G. Knoch,2002 Calderon, N.A. Kauffmann, T

Calderon, Kauffmann 2002

22.07.2003 RV 3

Mechanism of Action in Topical Applications

1. Microbicidal effectsie bactericidal, fungicidalvirustatic

2. Wound cleansing effect3. Wound healing effect

22.07.2003 RV 4

Mechanisms of Action of Ozone in Systemic Applications

1. Activation of Red Blood Cell Metabolism

2. Activation of Immunocompetent Cells

3. Activation and Induction of Biological Antioxydants and Radical Scavengers

22.07.2003 RV 5

Indications of Ozonetherapy Today and the Pharmacological Background

Disinfection, wound healingDental Medicine

Activation of antioxidantsand radical scavengers

Inflammatory conditions

Activation of immunocompetent cells

Immunodeficiency and immunodisbalance

Activation of RBC-metabolism

Arterial circulatory disorders

Disinfection, wound cleansing

External ulcers and skin lesions

Mechanism of Action

Indications

22.07.2003 RV 6

Pharmacological Effects 1: Red Blood Cells

1975 Buckley et al.1979 Freeman, Miller et al.1977-1986 Washüttl et al.2001 Lell, Viebahn et al.2002 Hoffmann, Viebahn

Peroxide formation andactivation of the RBC-metabolism via the glutathion system with improvement of oxygen releaseOxygen partial pressure pO2 in mm Hg

Oxygen saturation

without DPG

with DPG

22.07.2003 RV 7

Mechanisms of Ozone Reactions

a. structure of the ozone-molecule

b. mechanisms ofozone-reactions

c. „ozone-peroxides“

a.

b.

c.

22.07.2003 RV 8

The Action of Ozone on the Red Blood Cell Metabolism

22.07.2003 RV 9

2,3-DPG ( in % ) in vivo:Arterial Circulatory Disorders, stage III +IV. (Rokitansky et al. 1981)

0

50

100

150

200

250

control ozone

22.07.2003 RV 10

Mechanism of Action 1Activation of Red Blood Cell Metabolism

increase in ATP and 2,3-DPG

Shift of HbO2/Hb-balance to the right and

Improvement of oxygen-release

22.07.2003 RV 11

2,3-DPG in Blood Preserves in mg/L Serum (N = 5)

0

10

20

30

40

50

60

70

0 24 h 48 h

22.07.2003 RV 12

ATP and 2,3-DPG in % as Measure for Activation of Cell

Metabolism in Elderlies

020406080

100120140160

0 3 5 8

2,3-DPG ATPWeeks of treatment

22.07.2003 RV 13

Pharmacological Effects 2: immunocompetent cells

1990-2002 Bocci et al.

Induction of cytokinssuch as interferons,interleukins and growth factors

Cytokin induction in % dep.on O3-conc.

0

50

100

150

200

250

IL-1 IL-2 GM-CSF

oxygen 42 µg 78 µg

22.07.2003 RV 14

Pharmacological Effects 3: Radical Scavengers

1998 León et al.

1999 Peralta et al.

Activation ofantioxidative enzymesand radical scavengers as protective effect indamage by free radicals

0

10

20

30

40

50

60

70

SOD GSH

Kontrolle Ischaemia/Reperfusion Ozone+I/R

H2O2

Protective effect of ozone in

Hepatic Ischaemia - Reperfusion

22.07.2003 RV 15

The Action of Ozone on the Red Blood Cell Metabolism

22.07.2003 RV 16

Hepatic Ischemia/Reperfusion Damage(Peralta et al 1999)

0

10

20

30

40

50

60

70

SOD GSH

control I/R I/R+Ozone

H2O2

H2O2 ( µmol/g); SOD (u/mg Protein); GSH ( nmol/mg Protein)

22.07.2003 RV 17

OZONE and Prevention (León et al. 2002)Histological results correspond completely to biochemical measurements

Rectal Ozone Insufflationin an animal model. Glycogen Depletion in Liver Cells.

a) Control

b) CCl4-inducedGlycogen-Depletion

c) 15 preventive Ozone Application

a

b

c

22.07.2003 RV 18

Protection Against Hepatic Cellular Damage inducedby Carbontetrachloride in an Animal Model through 10 rectal Insufflations before CCl4 –Application. (León et al. 1998)

0

20.000

40.000

60.000

80.000

100.000

120.000

control

SOD (u/g)

CCl4 O3+CCl4 O2+CCl4

22.07.2003 RV 19

Reperfusion Damage in Renal Cells (Calunga et al.2001)

Control Isch/Reperf.

O2 + I/R O3 + I/R

22.07.2003 RV 20

Reperfusion Damage in Kidneys in an Animal Model following rectal ozone application compared to pure oxygenHistological results: % without lesion, N=10/group. (Calunga et al. 2002)

0

20

40

60

80

100

Kontrolle I/R Oxygen+I/R Ozon+I/R

5 d 10 d 15 d

22.07.2003 RV 21

Mechanism of Action 3Enzymatic Antioxydants and Radical Scavengers

-such as SOD, GSH-peroxidase and GSH - reductase ...-

are induced and activated by Ozone-formed peroxides,

thus increasing the organism`s antioxydative capacity.

22.07.2003 RV 22

Pharmacological Effects 4: Ozone and Prevention

1999 Schulz et al.- Improvement of survival

rate in septic peritonitis in an animal model by pretreatment with ozone.

- Synergistic effect with antibiotics.

2001 Lell, Viebahnet al.

- Growth inhibitory effect ofplasmodium in infectedRBC´s by pretreatment with ozone.

0%

20%

40%

60%

80%

100%

ozone III IV V

anibiotic O3+antib. control

0

5

10

15

20

25

30

35

day 1 day 2 day 3 day 4 day 5 day 6

Parasites Ozone+Par.

Survival rate in lethal septic peritonitis

Parasitemia in %

22.07.2003 RV 23

Peritonitis Model in Rats. Survival Rate withOzone in % (Schulz et al.1999)

0%

10%

20%

30%

40%

50%

60%

70%

control oxygen 100 50 10µg/ml

22.07.2003 RV 24

Peritonitis Model in Rats. Survival Rate in %Treatment: III Claforan. IV Tavanic. V Tazobac.

0%

20%

40%

60%

80%

100%

ozone III IV V

antibiotic O3+antib.

22.07.2003 RV 25

a. Ring Shaped Plasmodia and Mature Schizontsb. Female Plasmodia falciparum Gametes

(acc. Lieske et al. 1991)

a b

22.07.2003 RV 26

Plasmodium falciparum infected Red Blood Cells + Ozone. Parasitemia in % (Lell, Viebahn, Kremsner 2001)

05

10152025303540

day 2 day 4 day 5 day 6

Parasites Par.+Ozone

22.07.2003 RV 27

P.falciparum infected Red Blood Cells+Ozone :pre and post infection. Parasitemia in %

0

5

10

15

20

25

30

35

day 1 day 2 day 3 day 4 day 5 day 6

Parasites Par+Ozone Ozone+Par. Oz+Par+Oz

22.07.2003 RV 28

Ozone and PreventionIndications , Applications and Dosages

Herpes (post Zoster Neuralgia)Susceptibility to InfectionsGeneral Immunoactivationand RevitalizationPresurgical measurePreventive Application in Chemotherapie and RadiationAllergies

Major Autohemotherapywith 800 to 2000µg Ozone per treatment (50 ml with 16-40 µg/ml)

Rectal Insufflation with 10-20µg/ml and a Volume of 150-300 ml, in children 30-50 ml

Minor Autohemotherapywith 220 µg Ozone in 3-5 ml Patient´s Own Blood