THE EFFECTIVENESS OF THE INFLUENCE OF OZONE IN THE DURATION OF PERITONITIS AND ADHESION PROCESS IN THE EXPERIMENT

Sh.A. Yusupov

Samarkand State Medical Institute, Republic of Uzbekistan

Introduction. In spite of the big success in the abdominal surgery, problems of peritonitis and peritoneal commissures are still actual [1, 2, 3, 9]. In recent years, medicine has become, widely applied ozone [4, 5, 6]. However, there is practically no work devoted on the effects of ozone for the duration of peritonitis and adhesions process.

The aim of our investigation was to study the effects of ozone on the development of peritonitis and adhesions in the experiment.

Materials and methods. Experimental investigations were carried out on 38 white Wistar rats weighing 140-160 g. Modeling of acute peritonitis was conducted by the method of I.M. Baybekov and V.A. Horoshaeva (1991) [7, 8].

The rats were divided into 2 groups of 19 each. Animals 1 (control) group, after the development of their peritonitis, midline laparotomy was performed and abdominal cavity was dried from the purulent masses with the use of sterile wipes. In the lower right corner of the wound it was left the polychlorvinile drainage and the abdominal cavity was sutured.

In the second 2 group of animals, after drainage of the abdominal cavity from the purulent masses, it was blown by the dry ozone-oxygen mixture for 3 minutes. The ozone-oxygen mixture is prepared by the use of OTRE-01 apparatus with an ozone concentration 5.8 mg/l. They also left the tree-changes require the tube and sutured the abdominal cavity. Also in the investigated patients it has been left drainage tube and abdominal cavity has been sutured.

At 2 and 3 days after surgical operation rats 2 groups through the drainage tube into the peritoneal cavity it was injected 10 sm3 of dry ozone-oxygen mixture of the same concentration of ozone, after which both groups were removed drainage tubes.

At 3, 7 and 14 days after surgical operation, the animals were taken out of the experiment by momentary lekapitation. The samples of omentum, diaphragmatic parts of peritoneum, small bowels with mesentery and adhesions were investigated by the use of light-optic and electron microscopy.

The drugs for light-optic microscopy were prepared by the accepted method.

The results of investigation and discussion. For transmission electron microscopy (TEM), the material is fixed in 2.5% glutaraldehyde solution of 0.1 M phosphate buffer (pH – 7,2) and in 1% solution of osmium tetroxide, after dehydration and impregnation was poured into a mixture of epon and araldita. Semithin and ultrathin sections are manufactured by use of "Reichert Jung" ultramicrotome (Reichert, Austria) and it was respectively colored by methylene blue and basic fooksin or by uranylacetate and lead citrate (Karupu V.J., 1986). Ultrathin sections were investigated by the use of electron microscope H-600 (Hitachi, Japan).

For scanning electron microscopy (SEM) samples after dehydration it was dried by the use of critical point method in the “NSP-2” apparatus (Hitachi, Japan). From the part of the material, flooded in resin it was made sections thickness of 6,8 mm. They are pasted on the cover glasses or on the aluminum foil, and after removal of the resin in the saturated solution of sodium hydroxide in absolute ethanol it was dried and sprayed with gold and examined in the SEM.

In 3 days after surgical operation in the animals of control group bowel loops were macroscopically swollen and there were turbid exudates, peritoneal surface was dull with purulent-fibrinous overlays. It was noted the presence of the significant adhesion process. Adhesions are easily torn and are located mainly between loops of small intestine.

During the histological examination in the greater omentum it was determined swelling and significant polymorphocellular infiltration with predominance of polymorpho-nuclear neutrophilic leucocytes. Vessels are dilated; there were many clots in the lumen of them. The similar situation occurs in the mesentery of the small intestine and of the diaphragmatic part of peritoneum.

Scanning electron microscopy (SEM) of the peritoneum has been showed a significant enlargement of intercellular gaps, and even the discrepancies of mesotheliocyte. There were strands of fibrin, peritoneal macrophages and mast cells on the surface of the mesothelium. The surface of many mesotheliocytes was with small erosions.

Transmission electron microscopy (TEM) has been showed a significant dilatation of the lumen of micro-vessels. Cytoplasm of endotheliocytes was made very thin and vascularized. Under mesotheliocytes it was determined congestion transsudate.

During histological investigation of the adhesions it was determined that in the bases of them was connective tissue which was consisting of the tender collagenous fibers between them there were fibroblasts. Also there have been determined macrophages and lymph cells.

During the electronic-microscopic investigation of the adhesions’ surface it was determined mesotheliocytes of the oblong forms, apical surface of them were bulged in the lumen of the abdominal cavity. There was a big amount of microvillus on the surface of cytoplasmatic membrane.

In 7 days after surgical operation in the abdominal cavity the amount of exudate was decreased. Peritoneal edematous was preserved and interintestinal abscesses were observed. Adhesion process was extended through the whole abdominal cavity. Adhesions mostly localized between loops of the small bowel and they were mostly rough.

The light-optical investigation of peritoneum showed that inflammatory changes were less expressed and rarely there has been found the dilatation blood vessels with thrombosis. The parts of peritoneum were found with impaired mesothelial lining.

SEM and TEM investigations have been showed that in areas with impaired mesothelial lining, mesotheliocytes have flattened form with a fairly large nuclei. In the cytoplasm of the cells it has been found a few mitochondria and profiles of endoplasmic reticulum.

During histological examination it has been determined that the basis of adhesions is loose connective tissue consisting of thin bundles of collagen fibers, between which it has been localized fibroblasts and a little amount of blood capillaries. Fibroblasts have an oblong shape; their nuclei are large, hyperchrom. Blood vessels were lined by endothelial cells with oval nuclei. By the visceral peritoneum in the thick of the adhesions it was grown smooth muscle cells, the source of which was mesocolon. Besides structures which were described the above in the thick of the adhesions we observed a few macrophages, lymphocytes and neutrophils leucocytes. The surface of the adhesion has been lined by mesothelial cells.

On the 14th day in the abdominal cavity there was a small amount exudate, in some cases it was found encapsulated interintestinal abscesses. Microscopic peritoneal mesothelial lining has been restored its integrity. However, the cells were left swollen, dome-like changed. Endoplasmic reticulum was granular and vacuolated. Dilatation of capillaries, the phenomenon of stasis and sludging of erythrocytes were less expressed.

Adhesion process in the abdominal cavity has been significant. Adhesions were rough, in different forms; in some places they clamped intestinal loops. During histological investigation it was noted a big amount of collagenous fibers, bunches of them were thick and rough and they had less amount of fibroblasts. The last ones had stretched form with prolonged nuclei. It was determined macrophages, a few lymphocytes and smooth muscle cells. The surface of the adhesions was covered by flattened mesotheliocytes.

In the control group the amount of dead animals was 52,6%.

In the second group of animals which has been used ozone in the 3-d day after surgical operation in the abdominal cavity macroscopical localization of bowel loops was not disordered and we have found a little amount of half-transparent exudate. Peritoneum was slightly thickened, smooth, on the surface of it we determined few strands of fibrin, and it was noted the presence of a single thin short adhesions that were easily torn and deformed intestine.

The light-optic investigations have been showed that inflammatory changes were less significantly expressed. The integrity of mesothelial lining was less disordered than in the animals of the control group. Analogous formed adhesions have been presented by the smooth bunches of collagenous fibers and fibroblasts. The single formed adhesions were presented by the smooth bunches of collagenous fibers and fibroblasts. We have practically no determined smooth muscle cells.

On the 7-th day peritoneum was macroscopically clean, bright with no exudates in it. It has adhesions which easily torn and it has not deformed intestine.

During the light-optic investigation it was noted a small infiltration by the polymorph cell elements with the predominance of lymphocytes. It was determined a small amount of perivascular infiltration and moderate thickening of the blood vessels without thrombus. In the wall of the small intestine moderate infiltration was determined only in the stroma of fibers and between them. Intestinal fibers were in the right form without cell desquamation. There was predominance of prismatic cells in the epithelial lining of the intestinal fibers.

Serous membrane of the small intestine was without any signs of inflammation and injures. Blood vessels have been had the usual structure.

The results of SEM and TEM also have been testified that ozonotherapy led to the significant production of the pathological changes of ultra-structures caused by experimental peritonitis. There were determined the long and thin microvillus on the surface of mesotheliocytes. In the cytoplasm of mesotheliocytes it was determined a slightly big amount of mitochondria and the single profiles of endoplasmic reticulum. Underlying blood vessels was moderately enlarged. Endotheliocytes were with the strait surface of lumen and with narrow cytoplasm.

The light connective tissue was a basis of the adhesions presented by thin, lightly localized collagenous fibers, fibroblasts and blood capillaries. In the comparison with analogous period in the control group the amount of fibroblasts and blood capillaries was significantly low.

On the 14 day in the abdominal cavity exudate was not macroscopically determined, peritoneum was clear, intestinal loops were loosely localized. The determined single adhesions were short, thin and easily tear. There was no determined any adhesions in 78,9% of animals. Light-optic, SEM and TEM investigations have been showed that peritoneum has the usual structure.

The structure of the adhesions was differing from the same ones in the control group; we examined the signs pointing to the decreasing of synthetic activities of fibroblasts: the size and the amount of the cells were decreased, also it was decreased the amount of collagenous fibers.

Conclusion. Our investigations have been showed that the use of ozone is significantly effective in the experimental peritonitis and it reduces the intensiveness of adhesion process. Ozone could be recommended in the clinical practice for treatment and prevention of the same pathology.

References

1. Bairov G.A. Adhesion intestinal obstruction: In the book “Emergency Pediatric Surgery”. - St.Petersburg, 1997. - P. 189-200

2. Bairov G.A., Roshal L.M. Purulent surgery in children. – Moscow, 1991 – P. 41-48

3. Jenchevskiy V.A. Adhesion Disease. – Moscow: Medicine, 1989. – P. 10-15.

4. Peretagin S.P. Ozonotherapy: Methodic recommendation. – N-Novgorod, – 1996. – P. 14.

5. Seppo А. Oxidation drugs and ozone as a disinfected and antibacterial medications. – Tallin, 1996. – Vol. 1 – P. 73-78.

6. Salimov Sh.G. Ozonotherapy in the complex treatment of purulent-inflammatory diseases in children // Actual Problems of Pediatric Surgery. Republican Collection of the Scientific Works. – Andijan, 1997. – P. 157-160.

7. Khoroshaev V.A., Baybekov I.M., Vorogeykin I.M. Morphology of mesotheliocytes of the liver capsule in the normal condition and in the experimental peritonitis // Bulletin of experiments in biology and medicine. – 1988. – №9. – P. 374-377.

8. Khoroshaev V.A. Pathomorphological features of the peritoneum in the acute expressed peritonitis //In the book “Peritonitis”. – Novosibirsk, 1994. – Р. 34-36.

9. Tsuman V.G., Mashkov A.E., Kosarev V.A. //Surgery. – 1994. – № 9. – P. 23-25.