The Use of Biotechnology in the Prevention of Failure in the Imposition of Small Intestinal Anastomoses in the Experiment

Abstract

Summary. The aimof the study is to develop a method of sealing the sutured of the small intestine anastomosis in conditions of high risk anastomosis leakege by stopping the inflammation’ cascade in experiment.
 Materials and methods.The work was experimental in nature, was performed in the SI «ZIGUS NAMSU» in collaboration with the Scientific Research Institute of Biology V. N. Karazin KhNU in the period from 2018 to 2019. To increase the tightness of the intestinal anastomoses applied under conditions of laparatomy, experimental studies were conducted on laboratory animals. The studies were carried out after the permission of the ethics committee SI «ZIGUS NAMSU» in compliance with all ethical requirements for the humane treatment of animals.Surgical operations in two series of experiments were performed under operating conditions under general anesthesia on 10 white sexually mature linear rats – Wistar males, 12-18 months old, weighing about 250 grams. All morphological material (the first and second series of experiments) was marked and fixed in a 10% solution of neutral formalin. Subsequently, paraffin sections were prepared according to the generally accepted histotechnical method and stained with hematoxylin - eosin; according to Van Gieson and Mallory, an immunohistochemical study was used to study microenvironment factors.
 Results and Discussion. Immunohistochemical examination of the intestine was carried out in several stages, at the first stage we took intestinal tissue during the surgery to form the intestinal anastomosis, before the introduction of allogeneic cell transplants. Also carried out immunohistochemical studies of the emerging mucous membrane at various times after the formation of insolvency of intestinal anastomoses in both studied groups. When using a cell allograft, the mechanisms of repair of the intestinal mucosa include the appearance of cells that carry the expression of markers of mesenchymal and epithelial differentiation (Vim, CD34, CKW, Ecad), and are usually localized in the walls of blood vessels of granulation tissue, as well as in areas of the repairing epithelium. In addition, in the main group, only 7 days after the reaction to cytokeratins (CKW) and Ecad, areas of the repairing epithelium were detected, while the visualization of the epithelium in the control group did not begin even after 10 days. The analysis of immunophenotypes of repairing cells allows us to suggest the participation of mesenchymal-epithelial transformation mechanisms in this process.
 Conclusions.Thus, studies have shown that the use of cell transplantation of culture of allogeneic cells of the intestine leads to the reconstruction of the intestinal mucosa in a fairly short time. Conducted histological and immunohistochemical studies showed that in the vast majority of animals (77.2%) 10-14 days after the operation, an almost unchanged mucous membrane forms with the preservation of its elasticity.