RU2610361C2 - Method for experimental modelling of autotransplantation of splenic tissue into liver - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to experimental surgery and deals with modelling of autotransplantation of splenic tissue into liver. For this purpose after laparoscopic splenectomy and fragmentation of decapsulated splenic tissue homogenate is prepared by addition of physiological solution to crushed splenic tissue in ratio 2:1. After that, obtained homogenate is introduced subcapsularly into anterior-outer surface of right liver lobe in volume from 1/14 to 1/20 of initial splenic tissue mass.

EFFECT: method makes it possible to study viability of autolientransplant under liver capsule to prevent postoperative hyposplenism with possibility to use such transplantation technique in laparoscopic surgery.

2 ex, 4 tbl, 8 dwg

Description

The invention relates to medicine and will find application in experimental surgery for modeling autologous transplantation of spleen tissue into the liver, studying the pathogenesis and the possibility of determining the viability of an autoglant transplant under a liver capsule.

The only way to prevent postoperative hyposplenism syndrome is autologous splenic tissue transplantation. Most of the current methods for autolient transplantation are available with wide laparotomy. While the active development of laparoscopy makes us look for new surgical solutions that are available for minimally invasive surgery. The controversial question remains about the optimal mass of the autolient transplant, which allows to stop the development of postoperative hyposplenism syndrome.

A known method of preventing post-splenectomy syndrome in an experiment (RF patent No. 2247567, published March 10, 2005), in which the spleen is collected from an embryo, which is washed in nutrient medium No. 199, placed in fresh medium No. 199, and a homogenate are obtained in a Teflon homogenizer, which centrifuged; the upper, middle and lower layers are distinguished; sucking off the middle layer and the upper part of the lower layer; the resulting mixture of cells is diluted in nutrient medium No. 199, which is then injected into the mesentery of the small intestine or rectus abdominis muscle. The known method is time-consuming, not applicable in emergency surgery.

There is a method of autologous transplantation of spleen tissue into the liver (RF patent No. 2305502, published September 10, 2007), in which autotransplantation of a perforated fragment of the splenic tissue into the wound of the liver is carried out, followed by fixation of this fragment with U-shaped sutures through the capsule of the splenic transplant stored in the peripheral part. However, this method is only available with simultaneous damage to the spleen and liver and wide laparotomic access.

A known method of modeling foci of disseminated post-splenectomy splenosis (RF patent No. 2481645, published 05/10/2013), in which open splenectomy and contamination of the abdominal organs with blood from a removed spleen is performed. At the same time, conditions are created that are similar to the outflow of blood of the spleen into the abdominal cavity when it is damaged, and the conditions for activating the growth mechanisms of “non-spleen” tissue, which are closest to those occurring in human pathology. The disadvantage of this method is uncontrolled splenolysis, which can lead to the development of hypersplenism, adhesive intestinal obstruction.

The objective of the invention is the modeling of the method of autotransplantation of splenic tissue into the liver, applicable in laparoscopic surgery.

The task is achieved in that after laparoscopic splenectomy and fragmentation of the decapsulated splenic tissue up to 0.2 cm, homogenate is prepared by adding physiological solution to the crushed splenic tissue in a ratio of 2: 1. Then, the obtained homogenate is introduced subcapsularly into the anteroposterior surface of the right lobe of the liver in a volume of 1/14 to 1/20 of the initial mass of the spleen tissue, which allows preventing the development of postoperative hyposplenism.

The invention is illustrated by photographs.

Photo 1 shows the stage of immobilization of the spleen.

Photo 2 shows the splenectomy stage.

Photo 3 shows the stage of autolient transplantation under the liver capsule.

On a photo 4 - a view of a liver with a zone of an autolient transplant.

In photo 5, a histological preparation of the rabbit liver with an autolient transplant 30 days after surgery, an increase of 1 × 100.

Photo 6 shows a histological preparation of the rabbit liver with an autolient transplant site 60 days after surgery, an increase of 1 × 40.

Photo 7 shows a histological preparation of a rabbit liver with an autolient transplant 30 days after surgery, an increase of 1 × 100.

Photo 8 shows a histological preparation of the rabbit liver with an autolient transplant site 60 days after surgery, an increase of 1 × 40.

The method of experimental modeling of autologous transplantation of splenic tissue into the liver is as follows.

Crushed after laparoscopic splenectomy (photo 1, photo 2), splenic tissue up to 0.2 cm in volume from 1/14 to 1/20 of the initial mass of splenic tissue is placed in the cylinder of the system for puncture biopsy of the liver with an inner diameter of the needle lumen of 1.8 mm (a disposable sterile injection syringe and a needle with an internal lumen diameter of 2 mm and a needle cut angle of 45 ° can be used). Fragments of the spleen tissue in the cylinder (syringe) are mixed with saline in a ratio of 2: 1, mixed with gentle swaying until a homogeneous mass. Then, puncture introduction of the obtained splenic tissue homogenate into the anteroposterior part of the right lobe of the liver is performed subcapsularly as follows. After introducing the needle to a depth of 0.1 cm-0.2 cm over 3.0 cm-5.0 cm (photo 3) under the capsule of the liver, the needle begins to be removed with the simultaneous introduction of homogenate. Thus, a transplant of splenic tissue is obtained subcapsularly (3-5) cm long, (1.0-1.5) cm wide, (photo 4). In a similar manner, the remainder of the splenic tissue homogenate is introduced.

Case study No. 1.

Rabbit, 6 months old, weight 3500 g.

After drug sedation with veterinary medicine and appropriate treatment of the surgical field, a median laparotomy was performed with a length of 10.0 cm, the spleen vessels were clamped and crossed. The splenic tissue is crushed and placed in the cylinder of a device for puncture biopsy of the liver. Then, according to the above method, puncture introduction of homogenate into the anteroposterior surface of the liver was performed. Control of hemostasis. Laparotomic wound sutured tightly.

Histological examination of the autotransplantation zone:

after 1 month, a weakly defined border of the autograft expressed by fibroblasts, proliferation of lymphoid tissue, single sinuses is determined (photo 5).

After 2 months, newly formed sinuses with a small clearance and moderate blood filling are determined (photo 6).

Case study No. 2.

Rabbit, 6 months old, 3750 gr.

After appropriate treatment of the surgical field through a 1.0 cm incision at the midpoint of the anterior abdominal wall and the application of carboperitoneum, additional accesses were applied at points typical of splenectomy. After splenectomy, placement in the reservoir and removal of the spleen from the abdominal cavity, it was crushed and the homogenate prepared by adding 0.9% saline in a 2: 1 ratio. Under video control in the projection of the anterior external surface of the liver, a puncture of the anterior abdominal wall was performed by the system for puncture biopsy of the liver, the homogenate was introduced subcapsularly over 3.0 cm, the puncture system was removed. Control of hemostasis. The trocars are removed from the abdominal cavity. Stitches to the skin.

Histological examination of the autotransplantation zone:

after 1 month, a weakly defined border of the autograft, expressed by fibroblasts, proliferation of lymphoid tissue, single sinuses is determined (photo 7).

After 2 months, newly formed sinuses with a small clearance and moderate blood filling are determined (photo 8).

Thus, this method has significant advantages over existing autolient transplantation methods, consisting in the possibility of performing laparoscopically. So, after splenectomy by the laparoscopic method and preparation of a homogenate from splenectomized tissue under video control, the anterior abdominal wall is punctured in the projection of the anterior external surface of the liver and the homogenate is injected in the manner described above, which minimizes the invasiveness of surgical intervention, reduces the risk of purulent-septic complications, and facilitates the postoperative period.

Claims (1)

A method for experimental modeling of autologous transplantation of spleen tissue into the liver by laparoscopic splenectomy, decapsulation and fragmentation of spleen tissue, characterized in that the mixture obtained by adding physiological solution to the crushed splenic tissue in a 2: 1 ratio is introduced subcapsularly into the anterior surface of the right liver lobe in a volume of 1 / 14 to 1/20 of the original mass of splenic tissue.

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