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Abstract
Summary and Conclusion
Diabetes mellitus is one of the most prevalent metabolic diseases. It affects every cell and every biochemical process in the body resulting in a major health problem; it is characterized by chronic hyperglycemia with disturbances of carbohydrate, fat and protein metabolism resulting from defects in insulin secretion, insulin action, or both.
Ozone therapy has been used as an alternative medical approach for the last few decades and it has encountered some objections in spite of the finding that neither acute nor chronic side effects have ensued in millions of patients treated with ozone.
The aim of the present investigation was to study the effect of systemic ozone therapy on the submandibular and sublingual salivary glands of diabetic rats.
Materials and methods:
Forty adult male albino rats with an average body weight 140 ± 10.5 grams were used in this investigation.
The animals were divided into 4 equal groups, 10 animals each as follows:
Group I: animals served as non diabetic controls.
Group II: animals received a daily intraperitoneal injection of one cm3 of ozone – oxygen (O3O2) gas mixture with ozone concentration 70 microgram /1 cm3.
Group III: animals (diabetic group) received alloxan (ureide of mesoxallic acid) supplied in powder form, dissolved in distilled water to be given as a single intraperitoneal dose of 150 mg/kg body weight for the induction of diabetes mellitus.
Group IV: animals were treated with alloxan to induce diabetes mellitus as in group III and couple of days later, they received O3O2 gas mixture daily in the same dose and route of administration as group II animals.
The animals were caged, five animals per cage. They were fed natural diet and supplied drinking tap water adlibitum throughout the experimental period which lasted for two months.
At the end of the experiment, the animals of the different groups were sacrificed by cervical dislocation, their submandibular and sublingual salivary glands were dissected free, Fixed in 10% neutral buffered formalin for 24 hours, washed, dehydrated in ascending grades of ethyl alcohol, cleared in zylene and embedded in paraffin.
5-6 micron thick sections were cut and stained with:-
6. Hematoxylin and eosin for histological examination.
7. Mercuric brome phenol blue for total proteins.
8. Periodic acid Schiff (PAS) method (before and after diastase digestion) for demonstration of glycogen and neutral mucopolysaccharides respectively.
9. Alcian blue PH 1 and PH 2.5 for sulfated and nonsulfated acid mucopolysaccharides respectively. .
10. Immunoperoxidase staining procedure using monoclonal mouse antibody LP 34 clone (IgG-1 kappa isotope ) at a dilution of 1:50 in phosphate buffer saline ( PBS) for immunohistochemical localization of cytokeratin 5,6 & 18.
Specimens from the pancreas were taken from the animals of the different groups, fixed in 10% neutral buffered formalin, washed, processed, embedded in paraffin and sectioned to be stained with Hematoxylin and eosin for histological evaluation of the islets ofIV ( treated with ozone ).
II)Histological results:
A) .