Electron Microscopic Studying the Impact of Ozone on Chronic Hepatitis C Patients as Antifibrotic
Ahmed Sadek1
, Soheir Mansy2, Ahmed M Abd El Hady3, Olfat Hammam3, Afaf A Abdel Hady4, Wafaa Wafy5, Eman R. Youness6*1Department of Hepatology and Gastroenterology, Theodor Bilharz Research Institute. Imbaba Giza, Egypt.
2Department of Electron microiscopy, Theodor Bilharz Research Institute. Imbaba Giza, Egypt.
3Department of Pathology, Theodor Bilharz Research Institute. Imbaba Giza, Egypt.
4 Department of Clinical Chemistry, Theodor Bilharz Research Institute. Imbaba Giza, Egypt.
5 Department of Public Health, Theodor Bilharz Research Institute, Imbaba Giza, Egypt.
6 Department of Medical Biochemistry, Medical Researches and Clinical Studies Institute, National Research Centre Cairo, Egypt.
Corresponding Author E-mail:hoctober2000@yahoo.com
Abstract: We aimed to evaluate the histopathological and ultrastructural changes detected by light and electron microscopy, induced in patients having chronic hepatitis C receiving ozone/oxygen gas treatment. Twenty six patients with chronic hepatitis C who were at different stages of liver fibrosis, non-responders to interferon plus ribavirin therapy (n = 9), had contraindications (n = 2), or were not compliant (n = 15) were included. At baseline and 12 weeks after administering the ozone/oxygen gas mixture, liver biopsies were carried out utilizing both the major rectal insufflations and auto-hemotherapy along with clinical evaluation, kidney and liver function assessments, liver biopsies and abdominal ultrasonography. Before and 12 weeks after the treatment of ozone, quantitative PCR was performed. Two pathologists evaluated the histological activity index (HAI) using Ishak's score while working in blind settings, taking into account the degree of inflammation and the stage of fibrosis. Electron microscopy was done for all cases before and after treatment. Significant improvements in liver enzymes in hepatic fibrosis and inflammatory activity based on Ishak scoring system were detected. The mean grade of inflammation dropped from 10.3 to 8.4 and the mean stage of fibrosis dropped from 2.3 to 2.0 both with P value < 0.001 and < 0.05 respectively. Mean PCR values showed significant increase after 12 weeks of treatment from 17059 to 218736 with P value <0.05. As regards electron microscopy findings, the ultra-structural manifestations of HCV infection were disclosed in liver specimens exposed or not to ozone treatment. Stellate cells were often encountered in unexposed samples to ozone, meanwhile they disappeared after ozone exposure. Apoptotic hepatocytes which were frequently encountered before treatment are rarely seen in specimens after treatment with ozone. Signs of cellular regeneration in the form of binucleated cells, RER enveloping mitochondria, hepatocyte progenitors insinuating between the cells at the sinusoid pole are seen after treatment. Circulating inflammatory cells in the sinusoids and infilterating the lobule were decreased after treatment. Peroxisomes were increased after exposure to ozone with longitudinal orientation of mitochondrial cristae thus increasing the antioxidative activity of hepatocytes. No single significant complication was recorded in a total of >1000 settings of ozone therapy. In conclusion, ozone oxygen gas mixture is a direct anti-fibrotic and anti-inflammatory agent in treatment of chronic hepatitis C patients without improving viral PCR as evidenced by histopathology, electron microscopy and quantitative PCR.
Keywords: Chronic liver disease; Electron microscopy; Hepatitis C; Liver fibrosis; Ozone/oxygen gas mixture Back to TOC