RU2349350C2 - Method of pre-operation autoerythrocyte mass reservation - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention concerns medicine, particularly hematology and blood transfusion, and can be applied in high-quality autoerythrocyte containing hemocomponent collection. Invention claims method of pre-operation autoerythrocyte mass reservation by patient's blood exfusion not later than 4 days before surgical intervention, blood component fractioning by centrifugation with further plasma separation, labelling and erythrocyte mass storage at +2°C to +6°C. Patient's blood is exfused to first hemocontainer with CPDA-1 conservation agent, stirred and passed under gravity through built-in leukofilter Leukotrap Al (Pall) to the second of coupled hemocontainers. Obtained leukocyte-lean blood is centrifuged at +20°C, 900 g for 15 minutes, plasma is separated into single polymer container, autoerythrocyte mass remaining in hemocontainer is stored with conservation agent till surgical intervention but not more than 10-14 days.

EFFECT: preserved quality of autoerythrocyte mass at collection and long-term storage.

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Description

The invention relates to medicine, in particular hematology and blood transfusion, and can be used for the procurement of high-quality autoerythrocyte-containing hemocomponents.

A known method of harvesting autoerythrocyte mass without the use of leukocyte filters (Vilyaninov V.N. Preoperative reservation of autologous blood components in patients with endoprosthetics of large joints / V.N.Vilyaninov, A.V. Chechetkin, R.M. Tikhilov // Surgery. - 2005. - No. 2. - S.54-57).

The disadvantage of this method is that the long-term storage of autologous red blood cells at a positive temperature negatively affects the morphofunctional properties of red blood cells. This fact significantly limits the implementation of the method of autologous donation in surgical hospitals to a greater extent where there is no technical ability to separate the blood into components and it is impossible to organize autologous blood procurement in an outpatient setting.

The purpose of the invention is to ensure the preservation of the quality of autoerythrocyte mass during its procurement and long-term storage.

The goal is achieved by conducting blood exfusion of the patient no later than 4 days before surgery, fractionating the blood components by centrifugation followed by separation of the plasma, marking and storing the erythrocyte mass at a temperature of + 2 ° C to + 6 ° C, and the patient is exfused blood into the first blood container with CPDA-1 preservative, mixed and transferred by gravity through the built-in Leukotap Al (Pall) leukofilter to the second of the dual blood containers, the obtained leukocyte-depleted blood is centrifuged at + 20 ° С, 900 g within 15 min, the plasma is separated into a single polymer container, the autoerythrocyte mass remaining with the preservative in the blood container is stored for no more than 10-14 days until the surgical intervention.

The method is implemented as follows.

The patient’s blood is exfused no later than 4 days before the surgical intervention, blood components are fractionated by centrifugation followed by plasma separation, labeling and storage of the erythrocyte mass at a temperature of + 2 ° C to + 6 ° C, and blood is exfused into the patient’s first blood container with CPDA-1 preservative, mixed and transferred by gravity through the built-in Leukotap Al (Pall) leukofilter to the second of the dual blood containers, the obtained leukocyte-depleted blood is centrifuged at + 20 ° С, 900g for 15 min, separated by Lazma in a single polymer container, the remaining red blood cell mass with a preservative is stored in the blood container until the time of surgery for no more than 10-14 days.

For filtration, Leukotrap Al (Pall) leukofilters are used, which are dual plastic blood containers with an integrated leukofilter. In patients, 450 ml of autologous blood was exfused into the first blood container with CPDA-1. The mixed blood was transferred by gravity through the built-in leukofilter to the second of the dual blood containers. Leukocyte depleted canned autologous blood was centrifuged using a PC-6 refrigerator centrifuge at a temperature of + 20 ° С, g = 900 for 15 min. A single polymer container of the Compoplast-300 type was connected to the outlet of the container with autoblood in compliance with aseptic rules, into which the plasma was transferred using a manual plasma extractor. The leukofiltered autoerythrocyte mass (AEM) remaining in the blood container, preserved on CPDA-1, was labeled according to the established procedure, placed in a medical refrigerator and stored at a temperature of + 2 ° С to + 6 ° С until the time of surgery no more than 35 days.

Autoblood canned;

Autologous blood is passed through a leukofilter;

Centrifuged;

The erythrocyte mass is passed through a leukofilter;

The mass obtained after filtration is stored at a temperature of + 2 ° C to + 6 ° C until use until 35 days.

Practical implementation of the method. Preoperative backup of autologous components was performed in 37 patients with an orthopedic profile using the method of “selective exfusion-autohemotransfusion” (Vilyaninov V.N. et al., 2005). An indication for the procurement of components of autologous blood was the predicted value of intraoperative blood loss above 10% of the volume of circulating blood. Based on the data of the clinical examination of the patient, as well as hematological and biochemical parameters, they decided on the possibility of procuring a certain type and volume of autologous hemocomponents from the patient. The safety of preoperative backups was ensured by a thorough assessment of contraindications to autodonation. Hemoexfusion was performed no later than 4 days before surgery. The total accumulation of autologous hemocomponents was determined depending on the predicted value of intraoperative blood loss, preoperative temporary reserve and physiological state of the patient’s body, which was evaluated by a set of clinical, instrumental and laboratory parameters.

A comparative study of the qualitative and quantitative characteristics of a leukofiltered AEM (group I) and an AEM obtained by the standard method (group II) was carried out at different storage periods. Media samples were examined immediately after harvesting, on days 5–7 and 10–14 days of storage of hemocomponents.

Table 1 Dynamics of indicators of the rheological properties of red blood cells in an autologous red blood cell mass Indicators Initial values 5-7 days of storage 10-14 days of storage I group II group I group II group I group II group KV _e , cu 1.74 ± 0.06 1.82 ± 0.03 1.86 ± 0.05 1.97 ± 0.02 1.93 ± 0.05 * 2.20 ± 0.03 ID _e , cu 1.47 ± 0.01 1.51 ± 0.02 1.58 ± 0.01 1.61 ± 0.01 2.25 ± 0.02 * 1.74 ± 0.02 Note: * significance of differences between groups (p <0.05).

KV _e - coefficient of viscosity of red blood cells;

ID _e - the index of deformability of red blood cells.

Note: * significance of differences between groups (p <0.05).

From the data of Table 1 it can be seen that the rheological properties of the erythrocytes of the studied groups in the initial state did not differ significantly. During storage, a significant decrease in the viscosity coefficient of erythrocytes (KV _e ) was noted at 10-14 days of storage in group I by 14% compared with the control group. When storing AEM on days 5–7, KV _e in group I increased by 6.9%, and in group II by 8.2%, on day 10–14, storage in group I KV _e increased by 11%, and in group II 20% compared with the original values. An increase in the erythrocyte deformability index (ID _e ) was recorded by 5–7% in group I by 7%, in group II by 6%, by 10-14 days in group I by 47%, and in group II by 15% compared with the original values. On 10-14 days of storage, the ID of _e- leukofiltered AEM was 22.6% higher compared to standard AEM.

When analyzing the spontaneous level of malondialdehyde (MDA) in the studied AEM samples, a significant increase of 10-14 days in the second (control) group by 46% was revealed (Table 2). In the leukofiltered AEM on days 10-14 in the I group, the MDA level decreased by 12%, while in the control group there was an increase compared to the initial values by 33%. When analyzing the stimulated MDA level in the initial values, there were no significant differences between the groups. At the same time, on days 5–7 in the leukofiltered AEM, the content of MDA decreased by 6%, and in the control group it increased by 33% compared to the initial values. On 10-14 days of storage, the content of MDA in group I increased by 7%, and in group II by 40% compared with 5-7 days of storage. On days 5–7, the stimulated level of MDA in the leukofiltered AEM was lower by 30%, by 10–14 - by 40% compared to the standard AEM. Therefore, leukofiltration positively influenced the quality of AEM, reducing the intensity of lipid peroxidation (LP) in the blood components.

table 2 The dynamics of the content of MDA (μmol / l) in an autologous red blood cell during storage Observation groups Initial values 5-7 days 10-14 days Spontaneous level I group 2.21 ± 0.18 2.40 ± 0.18 1.94 ± 0.08 * II group 2.17 ± 0.28 2.10 ± 0.13 2.90 ± 0.19 Stimulated level I group 0.94 ± 0.10 0.88 ± 0.07 * 0.97 ± 0.02 * II group 0.86 ± 0.05 1.15 ± 0.01 1,60 ± 0,18 Note: * significance of differences between groups (p <0.05).

Note: * significance of differences between groups (p <0.05).

The elimination of leukocytes from autologous red blood cells helps to reduce the intensity of hemolysis, lipid peroxidation, the accumulation of pro-inflammatory cytokines, increase the stability of erythrocyte membranes in storage at a temperature of + 4 ° C for 14 days.

Claims (1)

The method of preoperative reservation of autoerythrocyte mass by exfusion of the patient’s blood no later than 4 days before surgery, fractionation of blood components by centrifugation followed by plasma separation, labeling and storage of the erythrocyte mass at a temperature of +2 to + 6 ° C, characterized in that the patient blood is perfused into the first blood container with CPDA-1 preservative, mixed and transferred by gravity through the built-in Leukotrap Al (Pall) leukofilter to the second of the dual blood containers, obtained both the blood donated by leukocytes is centrifuged at + 20 ° C, 900 g for 15 minutes, the plasma is separated into a single polymer container, the autoerythrocyte mass remaining with the preservative in the blood container is stored for no more than 10-14 days.