RU2801037C1 - Method of stimulating the regeneration of intestinal anastomoses using autogenous growth factors - Google Patents

Abstract

FIELD: medicine; abdominal surgery.

SUBSTANCE: before applying intestinal sutures to the wound edges of the distal and proximal parts of the intestine of the formed anastomosis, 3 injections of 0.5 ml of platelet-enriched autoplasma containing autogenous growth factors are performed. Injections are performed into the muscle layer to a depth of 0.5 cm at 3, 9 and 12 o’clock position of the conventional dial, taking the mesenteric part of the wound edge of the intestine as 6 o’clock position.

EFFECT: method provides stimulation of regeneration of the intestinal anastomosis by accelerating neoangiogenesis, increasing the rate of local blood flow and migration of cellular elements, thereby reducing the risk of postoperative complications, including anastomosis failure.

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Description

SUBSTANCE: invention relates to medicine, namely to surgery, and can be used as a method for stimulating regeneration and preventing intestinal anastomoses from leaking using autogenous growth factors.

There is a method aimed at improving the conditions for the healing of the intestinal anastomosis and sealing the intestinal sutures using the medical adhesive "Sulfacrylate". However, this method has the following disadvantages: the absence of biologically active substances in the adhesive determines the absence of the effect of stimulating the regeneration of the intestinal anastomosis; poor adhesive adhesion to the tissues of the intestinal anastomosis zone; during the polymerization of the adhesive in the area of the intestinal sutures, it is possible to form a zone of aseptic coagulation necrosis, which can prevent the regeneration of the intestinal suture and increase the risk of its failure [Registration certificate No. FS 01031444/2587-06 dated February 07, 2006, Marchenko V.T. Clinical and morphological aspects of the use of the new adhesive "Sulfacrylate" in surgery of the thoracic and abdominal organs. Dis. doc. honey. Sciences. - Novosibirsk. - 2004. - 212 p.].

A known method of intraoperative prevention of failure of anastomoses of the hollow organs of the gastrointestinal tract and improving the conditions for their regeneration by intraoperative injection into the walls of the hollow organs subserously, 1.0 cm away from the suture line, a 25% solution of 5-hydroxymethyluracil in an amount of 0.5 ml per 1 cm ² [US Pat. 2304972 Russian Federation, IPC A61K 31/7072. The method of intraoperative prevention of insolvency of anastomoses of hollow organs in surgery of the gastrointestinal tract / V.V. Plechev [and others]; applicant and patent holder GOU VPO BSMU ROSZDRAVA - No. 2005141843/14; dec. 27.12. 2005; publ. 27.08.2007, Bull. No. 24]. This method provides an increase in the rate of restoration of regional blood flow in the area of the anastomosis, allows you to form an interintestinal anastomosis in conditions of violation of the microcirculation of the intestinal wall. However, when injected 1 cm away from the suture line, additional subserous damage to the intestinal wall occurs, and the use of 5-hydroxymethyluracil does not provide sufficient stimulation of the regeneration of the intestinal anastomosis with its low availability and cost.

Closest to the claimed method is the method of immersing the edges of the enterotomy wound in platelet-rich plasma (PRP-Platelet Rich Plasma) and keeping them for 5 minutes until the formation of a gelatinous clot, followed by the imposition of intestinal anastomosis [Fresno L, Fondevila D, Bambo O, Chacaltana A, García F, Andaluz A. Effects of platelet-rich plasma on intestinal wound healing in pigs. Vet J. 2010 Sep;185(3):322-7. doi: 10.1016/j.tvjl.2009.06.009. Epub 2009 Jul 17. PMID: 19616459; Giusto G, Vercelli C, Iussich S, Tursi M, Perona G, Gandini M. Comparison of the effects of platelet-rich or growth factor-rich plasma on intestinal anastomosis healing in pigs. BMC Vet Res. 2017 Jun 19;13(1):188. doi: 10.1186/s12917-017-1102-8. PMID: 28629420; PMCID: PMC5477282]. However, this method has a number of disadvantages associated with the application of the gel on the surface of the anastomosis: in the area of the anastomosis, there is an increase in the adhesion formation process; there are great technical difficulties in the imposition of the anastomosis associated with the prevention of removal from the surface of the edges of the anastomosis formed gelatinous clot from platelet-rich plasma; immersion of bleeding and contaminated intestinal contents into the edges of the intestine in platelet-rich plasma adversely affects the effectiveness and concentration of its initial composition.

Objective: to develop a new method for stimulating regeneration and preventing intestinal anastomosis failure using intraoperative injection of autologous growth factors as part of platelet-rich plasma.

This goal is achieved by the fact that before the imposition of a digestive anastomosis into the muscle layer of its distal and proximal edges, autogenous growth factors are injected by injection, contained in the autoplasma, previously prepared from the patient's own blood, enriched with platelets.

The technical result when using the invention is the stimulation of regeneration and the prevention of insolvency of intestinal anastomoses by accelerating neoangiogenesis, increasing the rate of local blood flow and migration of cellular elements.

The method is carried out as follows.

After performing a laparotomy, bowel resection or enterotomy, the surgical procedure is completed with the imposition of an intestinal anastomosis in a certain way (end-to-end, side-to-side, end-to-side) and rows (single-row, double-row). When forming the intestinal anastomosis, the classical technique of its implementation is used. However, before intestinal sutures are applied, platelet-enriched autoplasma containing autogenous growth factors is injected into the muscle layer of the wall of the distal and proximal edges of the anastomosis using a 32G injection needle and an insulin syringe (1 ml) (Fig.). In this case, the introduction of platelet-rich autoplasma containing autogenic growth factors is carried out from the side of the wound edge of the intestine, excluding trauma to the serous and mucous membranes, by inserting a needle to a depth of 5 mm. from the edge. In the wound edges of the distal and proximal sections of the intestine, forming the intestinal anastomosis, 3 injections are carried out with the introduction of 0.5 ml of autoplasma enriched with platelets, in the projection of 3, 9 and 12 hours of the conventional dial, taking the mesenteric part of the wound edge of the intestine for 6 hours. After injection of platelet-enriched autoplasma containing autogenous growth factors into the wound edges of the distal and proximal parts of the intestine, intestinal sutures are applied with the final formation of the intestinal anastomosis. Platelet-enriched autoplasma containing autogenous growth factors is prepared intraoperatively, immediately before its introduction into the wound edges of the distal and proximal parts of the intestine of the formed anastomosis. At the same time, the patient's venous blood was taken in a volume of 6-9 ml. After filling with blood a standard tube used to prepare platelet-enriched autoplasma, it is centrifuged at 3500 rpm for 5 minutes. From centrifuged venous blood in a test tube, an accurate sampling of the released plasma layer is carried out, into 6 syringes of 0.5 ml of plasma each (3 syringes per wound edge of the distal and proximal sections of the intestine of the formed anastomosis).

Brief description of the drawing.

The number "1" indicates the injection sites of platelet-rich autoplasma into the wound edges of the distal and proximal sections of the intestine, in the projection of 3, 9 and 12 hours of the conventional dial, taking the mesenteric part of the wound edge of the intestine as 6 hours.

Example 1. The experiment was carried out on 30 sexually mature laboratory male rabbits of the Soviet Chinchilla breed (weight 2.0 ± 0.5 kg). Enterotomy was performed under intravenous anesthesia with Zoletil 100 (0.5 mg/kg), followed by the imposition of an end-to-end small bowel anastomosis (control anastomosis) at a distance of 15 cm from the ileocecal angle, then retreating 30 cm from the imposed anastomosis, enterotomy followed by injection of platelet-rich autoplasma into the muscle layer (as described above) and intestinal anastomosis (all anastomoses were performed with a single-row interrupted suture with atraumatic VICRYL 4-0 material). Comparative assessment of the state of the area of intestinal anastomosis was carried out on the 3rd, 7th, 14th day. A morphological (macro- and microscopic) assessment of the state of the intestinal anastomosis area was performed, as well as an assessment of the state of microcirculation by laser Doppler flowmetry (LDF) using a two-channel laser analyzer. When performing LDF, the following were determined: the value of the average blood perfusion flow - "M" in the registration time interval in perfusion units (pf units), the average perfusion fluctuation relative to the value of M (σ - "Flux"), calculated by the formula for the standard deviation (pf units), coefficient of variation - Kv (Kv=σ/M·100%).

A comparative assessment of indicators of the state of local blood flow is presented in Table 1.

Table 1 - Indicators of the state of local blood flow.

Indicators Control anastomosis Anastomosis using platelet-rich autoplasma (experimental group) 3 days 7 days 14 day 3 days 7 days 14 day M, pf.un. 4.28±1.21* 14.2±1.52** 19.75±2.87** 10.06±3.17* 17.93±1.37** 21.75±2.84** Kv 7.82±1.61** 6.92±2.42*** 5.98±2.93 9.19±2.38** 8.82±2.64*** 6.08±2.32 σ, pf.un. 0.49±0.42** 1.24±0.6*** 1.18±0.42*** 1.01±0.81** 1.6±0.57*** 1.53±0.5***

Note: * - (p<0.001), ** - (p<0.01), *** - (p<0.05), significance was assessed using the Mann-Whitney U-test.

The performed macroscopic study showed no significant difference with the control anastomosis in assessing the level of adhesion formation, while in the experimental group there were fewer complications and significant macroscopic signs of anastomosis formation. Microscopic (histological) examination revealed in the experimental group, compared with the control group, an acceleration in the dynamics of reparation, an increase in angiogenesis, a decrease in inflammatory infiltration, and an acceleration of signs of completion of the connective tissue organization. When performing LDF in the experimental group, a significant increase in the average blood perfusion flow and local blood flow velocity was determined.

As follows from the experimental results, the use of a new method of stimulating regeneration using autogenous growth factors, compared with the control anastomosis, shows a functional improvement in microcirculation, a decrease in tissue ischemia, no signs of increased adhesion formation, acceleration of neoangiogenesis and the repair process in the area of the digestive anastomosis, demonstrating the potential for application this method to reduce the frequency of possible complications associated with its failure.

Claims (1)

A method for stimulating the regeneration of intestinal anastomoses using autogenous growth factors, characterized in that before applying intestinal sutures to the wound edges of the distal and proximal sections of the intestine of the formed anastomosis, 3 injections of 0.5 ml of platelet-rich autoplasma containing autogenous growth factors are performed into the muscle layer with sides of the wound edge of the intestine, excluding trauma to the serous and mucous membranes of the intestine, to a depth of 0.5 cm at 3, 9 and 12 hours of the conventional dial, taking 6 hours the mesenteric part of the wound edge of the intestine, followed by the application of intestinal sutures and the completion of the formation of the intestinal anastomosis.

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