JPWO2015194668A1 - Neutrophil removal column - Google Patents

Abstract

The number of platelets in the circulating blood can be controlled within the range that maintains the hemostatic ability, and the viable cell ratio of neutrophils in the circulating blood (the proportion of viable neutrophils in the total neutrophils) is increased. A neutrophil removal column is provided. A container having a capacity of 125 mL made of a polyethylene terephthalate fibrous carrier having a hydrophilic polymer (HM-3) obtained by solution radical polymerization of 2-hydroxyethyl methacrylate (HEMA) and dimethylaminoethyl methacrylate (DM) on the surface The total surface area of the fibrous carrier is 10.5 m 2 or more, and the fibrous carrier is composed of a carrier having an average fiber diameter of 1.4 μm or more. A neutrophil removal column for augmentation is provided.

Description

  The present invention relates to a neutrophil removal column.

  Neutrophils play a central role in the first line of infection defense and have a powerful intracellular bactericidal action. Neutrophil's intracellular bactericidal action, chemotaxis ability to chemotaxis to invading foreign substances, adherence of opsonized foreign substances, adherence to vascular endothelial cells, take in foreign substances and bacteria attached to the cells The phagocytic ability and the bactericidal ability to destroy and digest the eaten foreign substances are the typical functions.

  Neutrophils play an important role in biological defense and wound healing. On the other hand, neutrophils are inferior in their ability to discriminate between self and foreign bodies compared to other immune cells such as lymphocytes and macrophages. There is also a risk of causing damage to the organs due to abnormal accumulation in vital organs. Among them, surgical operation of the intestinal tract and parenchymal organs causes ischemia / reperfusion of the organs, and neutrophils can cause organ damage.

In the peripheral blood of an adult, neutrophils in the order of 10 to the 10th power are considered to exist, and there are approximately 8 billion to 30 billion neutrophils when the body weight is 50 kg. The lifespan of neutrophils in the blood is within one day, approximately 10 hours, and is several days in the tissue. In addition, neutrophils in the blood vessel wall, tissue, spleen, liver, etc. are present in the peripheral pool as a marginal pool. Furthermore, the bone marrow has a pool 10 to 30 times as much as that in peripheral blood, and there are hundreds of billions of neutrophils in the whole body in the order of 10 11.
Therefore, since there is a large pool, even if neutrophils are removed from the peripheral blood, the neutrophils in the pool are mobilized and the neutrophil count in the peripheral blood increases rapidly.

  On the other hand, the lifespan of platelets takes 8 to 10 days from production in the bone marrow to processing in organs such as the liver and spleen, and 1/8 to 1/10 is changed every day. About 1/3 of the peripheral platelets are pooled in the spleen. Normal blood contains about 100,000 to 400,000 platelets / μL, and when the platelet count is less than 100,000 / μL, bleeding time is prolonged. Compared to neutrophils, the pool is small, and removing platelets from peripheral blood takes time to recover the platelet count.

In digestive surgery, there are abdominal incisions and intestinal anastomoses, and appropriate wound healing needs to occur in these areas. Wound healing consists of a series of processes such as a blood coagulation phase, an inflammation phase, a proliferation phase, and a tissue remodeling phase.
In non-infectious advanced surgical invasion, the function of neutrophils in peripheral blood, that is, migratory ability, phagocytic ability, active oxygen production ability, and bactericidal ability is suppressed, and the suppression is considered to be easily infectious after invasion. It has been reported to be involved.
For example, Patent Document 1 discloses that patients with ulcerative colitis (UC) frequently experience postoperative infectious complications such as surgical site infection (SSI).
In UC patients, cytokine production in the perioperative period has accelerated, and this cytokine is mainly produced in neutrophils. It is known that high neutrophil elastase has a high risk of developing SSI after surgery. (Non-Patent Document 1).
It has been reported that leukocytapheresis (LCAP), which is recognized as an effective medical treatment in the active phase of UC, can be introduced immediately after surgery to suppress the onset of SSI (Non-patent Documents 2 and 3). ).

Patent No. 5048225

Dig Surg. 2006; 23 (3): 179-85 Japanese Journal of Gastroenterological Surgery 2007; 40 (7): 1007 Surg Today 2008; 38: 609-617

The existing leukocyte removal column removes almost 100% of granulocytes and monocytes, while removing 50% to 90% of platelets. Therefore, there is a possibility of promoting bleeding when used immediately after the operation.
Therefore, the problem to be solved by the present invention is that the number of platelets in the blood circulating in the body can be controlled within a range in which the hemostatic ability is maintained, and the viable cell ratio of neutrophils in the blood circulating in the body (total It is an object of the present invention to provide a neutrophil removal column capable of increasing the viable neutrophil ratio in neutrophils).

As a result of intensive studies to solve the above problems, the present inventors analyzed peripheral blood granulocytes of patients with a high risk of infection immediately after the operation with a flow cytometer, and caused neutrophils that caused early apoptosis or late apoptosis. It was found that there are a large number of spheres, and that the number of viable neutrophils increases when neutrophils that have undergone early or late apoptosis, which are present in large numbers on the neutrophil removal column, are removed.
When treated with a neutrophil removal column packed with a specific fibrous carrier, the number of platelets in the blood circulating in the body is controlled within a range that maintains hemostasis, and the living cells of neutrophils in the blood circulating in the body It was found that the rate (ratio of viable neutrophils in all neutrophils) can be increased, and the present invention was completed.

That is, the present invention is as follows.
(1)
Filled with a fibrous carrier having a hydrophilic polymer on its surface,
The total surface area of the fibrous carrier is 10.5 m ² or more;
A neutrophil removal column for increasing the viable cell ratio of neutrophils in blood circulating in the body, wherein the fibrous carrier is composed of a carrier having an average fiber diameter of 1.4 μm or more.
(2)
The neutrophil removal column according to (1), wherein the granulocyte removal rate is 80% or more and the platelet passage rate is 75% or more.
(3)
The neutrophil removal column according to (2), wherein the lymphocyte removal rate is 70% or less.
(4)
The neutrophil removal column according to any one of (1) to (3), wherein the column is sterilized by radiation in a state where the oxygen concentration in the surrounding atmosphere is 1% or less.
(5)
The neutrophil removal column according to any one of (1) to (4), wherein the hydrophilic polymer has a basic functional group and a nonionic hydrophilic group.
(6)
The neutrophil removal column according to (5), wherein the basic functional group is derived from dimethylaminoethyl methacrylate, and the nonionic hydrophilic group is derived from 2-hydroxyethyl methacrylate.
(7)
The neutrophil removal column according to any one of (1) to (6), wherein the fibrous carrier includes a carrier having an average fiber diameter of 1.4 μm or more and 2.6 μm or less.
(8)
The neutrophil removal column according to (7), wherein the fibrous carrier further includes a carrier having an average fiber diameter of 8 μm to 14 μm.
(9)
The neutrophil removal column according to any one of (1) to (8), wherein the fibrous carrier is a nonwoven fabric.
(10)
The neutrophil removal column according to any one of (1) to (9), wherein a critical wet surface tension of the fibrous carrier is not less than a surface tension of a physiological solution.
(11)
(1) For patients with ulcerative colitis, colon perforation, esophageal cancer, rectal cancer laparotomy, patients with wound class 2 or higher, patients with high surgical invasiveness, or patients with infection -The neutrophil removal column in any one of (10).

  The neutrophil removal column of the present invention is capable of controlling the number of platelets in the blood circulating in the body within a range in which the hemostatic ability is maintained, and the viable cell ratio of neutrophils in the blood circulating in the body (total neutrophils) The ratio of viable neutrophils to the total can be increased.

The relationship between the total surface area of a fibrous support | carrier and the removal rate of a granulocyte is shown. The horizontal axis represents the total surface area of the fibrous carrier, and the vertical axis represents the granulocyte removal rate. ◆ represents each example, and ■ represents each comparative example. The relationship between the total surface area of a fibrous carrier and the passage rate of platelets is shown. The horizontal axis represents the total surface area of the fibrous carrier, and the vertical axis represents the platelet passage rate. ◆ represents each example, and ■ represents each comparative example. The relationship between the total surface area of a fibrous carrier and the removal rate of lymphocytes is shown. The horizontal axis represents the total surface area of the fibrous carrier, and the vertical axis represents the lymphocyte removal rate. ◆ represents each example, and ■ represents each comparative example.

  Hereinafter, embodiments for carrying out the present invention (hereinafter referred to as “embodiments”) will be described in detail. Note that the embodiments described below do not limit the present invention.

The neutrophil removal column of this embodiment is filled with a fibrous carrier having a hydrophilic polymer on its surface, the total surface area of the fibrous carrier is 10.5 m ² or more, and the fibrous carrier has an average fiber diameter. It is composed of a carrier of 1.4 μm or more, and is used for increasing the viable cell rate of neutrophils in blood circulating in the body.
The neutrophil removal column of this embodiment is a neutrophil removal column that has a high affinity for neutrophils and a low affinity for platelets by filling a specific fibrous carrier. be able to.
And the neutrophil removal column of this embodiment can be used as a column for blood circulation.
In this embodiment, the blood circulation column is a column through which whole blood can pass, and the blood that circulates inside the body can be taken out of the body, passed through the column, and returned to the body again. Say.
Specifically, the neutrophil removal column of the present embodiment is composed of a container having an inlet and an outlet for blood to be passed, and a fibrous carrier filled in the container.

In the present embodiment, the fibrous carrier means a woven fabric, non-woven fabric, cotton-like, thread-like, bundle-like or the like fibrous structure that is water-insoluble and is made of a fibrous base material. Of these, woven fabrics and non-woven fabrics are preferable from the viewpoint of blood cell adsorbability and handling properties as a separating material, and non-woven fabrics are more preferable because they can be contacted with blood cells in many ways.
In the present embodiment, the fibrous carrier has a fibrous base material and a hydrophilic polymer fixed to the surface of the fibrous base material.
The fibrous base material is not particularly limited as long as it is capable of filtering blood and hardly damages blood cells, but the organic polymer material is excellent in workability such as cutting. preferable.
Examples of the organic polymer material include polyester, polyolefin, polyacrylonitrile, polyamide, polystyrene, polymethyl methacrylate, polyvinyl fluoride, polyurethane, polyvinyl alcohol, polyvinyl acetal, polysulfone, polyvinylidene fluoride, and polytrifluorochlorovinyl. , Vinylidene fluoride-tetrafluoroethylene copolymer, polyethersulfone, polyacrylate, butadiene-acrylonitrile copolymer, polyether-polyamide block copolymer, ethylene-vinyl alcohol copolymer, cellulose, cellulose acetate, etc. It is done.
The organic polymer material is preferably a polyester or polyolefin, more preferably a polyester, since it can easily be a fibrous base material.

The fibrous carrier has a hydrophilic polymer on its surface.
In this embodiment, the fibrous carrier has a hydrophilic polymer on its surface, the fibrous carrier has a total surface area of 10.5 m ² or more, and the fibrous carrier has an average fiber diameter of 1.4 μm or more. The removal rate of granulocytes is 80% or more, and the passage rate of platelets is 75% or more.
If the platelet passage rate is 75% or more, the number of platelets in the circulating blood can be controlled within the range that maintains hemostasis even after column operation, so the risk of promoting bleeding is low, and the patient with a high risk of infection (infection Neutrophil removal using a neutrophil removal column can also be safely performed.

The adhering amount of the hydrophilic polymer is preferably in the range of 1 mg to 90 mg per 1 g of the fibrous carrier, and more preferably 4 mg to 50 mg per 1 g of the fibrous carrier.
The hydrophilic polymer in the present embodiment is not particularly limited as long as it is a hydrophilic polymer that can be fixed to the surface of the fibrous base material, but has a nonionic hydrophilic group and a basic functional group. It is preferable that it is a compound.

The nonionic hydrophilic group means a functional group that is very difficult to ionize and has a highly hydrophilic structure.
Examples of the nonionic hydrophilic group include a hydroxyl group, an ethylene oxide chain, an amide group, etc. Among them, a hydroxyl group is preferable. C3b produced by activation of complement binds to the hydroxyl group, and adsorbs neutrophils having C3b receptor CR1 (CD35). When the nonionic hydrophilic group is present on the surface of the fibrous base material, the fibrous carrier is easily wetted when it comes into contact with blood, so that there is an effect of preventing a single flow of blood. As a result, the blood flow in the fibrous carrier The area increases, which can contribute to the improvement of neutrophil removal ability.

The basic functional group means a functional group having a positive charge.
Examples of basic functional groups include amino groups such as primary amino groups, secondary amino groups, tertiary amino groups, and quaternary ammonium groups, and nitrogen-containing aromatic groups such as pyridyl groups and imidazole groups. Among them, an amino group is preferable.
Since the basic functional group has a positive charge, it has an effect of adsorbing negatively charged neutrophils under physiological conditions by electrostatic interaction.

Examples of the method for fixing the hydrophilic polymer to the surface of the fibrous base material include grafting methods such as radiation grafting and plasma grafting, and coating methods using polymers.
The coating method is preferable because the operation is simple and the productivity is excellent.
The polymer that can be used in the coating method can be synthesized from a monomer having a polymerizable functional group such as a vinyl group by ordinary radical polymerization, anionic polymerization, or the like. Moreover, you may synthesize | combine by carrying out random copolymerization and block copolymerization of 2 or more types of different types of monomers.
Examples of the monomer capable of synthesizing the coating polymer include 2-hydroxyethyl (meth) acrylate having a nonionic hydrophilic group, hydroxymethyl (meth) acrylate, methoxypolyethylene glycol (meth) acrylate, and (meth) acrylamide. Among them, 2-hydroxyethyl methacrylate is preferable.
Examples of the monomer capable of synthesizing the coating polymer include (meth) acrylic acid derivatives such as dialkylaminoethyl (meth) acrylate having a basic functional group, among which dimethylaminoethyl methacrylate is preferable. .
The hydrophilic polymer may be a copolymer of a monomer having a nonionic hydrophilic group and a polymerizable functional group and a monomer having a basic functional group and a polymerizable functional group. The copolymer of the (meth) acrylic acid derivative which has a hydrophilic group, and the (meth) acrylic acid derivative which has a basic functional group is mentioned.

The fibrous carrier is filled in the neutrophil remover so that the total surface area of the fibrous carrier is 10.5 m ² or more.
When the total surface area of the fibrous carrier is within this range, the removal rate of granulocytes is 80% or more. 90% to 95% of granulocytes are neutrophils. If the removal rate of granulocytes is 80% or more, neutrophils that have undergone early apoptosis, necrosis, and late apoptosis present in the circulating blood (collectively, these are collectively referred to as non-viable neutrophils) are removed. Then, neutrophils are mobilized from the internal pool to the blood circulating in the body, and the viable cell ratio of neutrophils in the blood circulating in the body increases. Increasing the viable cell rate of neutrophils in the systemic circulation increases resistance to bacterial infection in patients at high risk of infection and decreases risk of infection. In patients who have already been infected, the number of non-viable neutrophils in the inward circulation blood is large. However, by performing neutrophil removal using a neutrophil removal column, non-viable neutrophils can be obtained. The number of neutrophils decreases, the viable cell rate increases, and the elimination efficiency of infecting bacteria increases significantly. Non-viable neutrophils are Propidium
Since Iodide (PI) or Annexin V is bound, it can be measured by analyzing neutrophils double-stained with PI and fluorescent or biotin-labeled Annexin V using a flow cytometer.
From the viewpoint of setting the platelet passage rate to 75% or more, the total surface area of the fibrous carrier is more preferably 21 m ² or less.
In the present embodiment, the total surface area of the fibrous carrier can be measured by the method described in the examples.

The fibrous carrier filled in the neutrophil removal column can be of any shape, but removes a sufficient amount of granulocytes without causing clogging of the fibrous carrier during the extracorporeal circulation of blood. For this purpose, the fibrous carrier is composed of a carrier having an average fiber diameter of 1.4 μm or more as a filter for selectively removing granulocytes.
In the present embodiment, the phrase “consisting of a carrier having an average fiber diameter of 1.4 μm or more” means that a carrier having an average fiber diameter of less than 1.4 μm is not included.
In this embodiment, the fibrous carrier is composed of a fibrous base material, but may be composed of one type of fibrous base material, or may be composed of two or more types of fibrous base materials. Each carrier constituting the fibrous carrier may have a different average fiber diameter between the carriers.
An average fiber diameter of 1.4 μm or more means that, in each carrier constituting the fibrous carrier, the average fiber diameter is 1.4 μm or more, and the average fiber diameter for each carrier unit constituting the fibrous carrier. Is 1.4 μm or more.
Therefore, in the case of comprising two or more types of carriers, the average fiber diameter of the constituent carriers may be different, but even if the average fiber diameter is different, the average fiber diameter is 1.4 μm or more in each carrier.
In this embodiment, after separating the fibrous carrier into individual carriers, the average fiber diameter of each carrier is measured, and when the minimum average fiber diameter is 1.4 μm or more, the fibrous carrier is It can be said that it consists of a carrier having an average fiber diameter of 1.4 μm or more.
In the fibrous carrier, when the minimum average fiber diameter is 1.4 μm or more, an increase in pressure loss accompanying the passage of blood can be suppressed and blood can continue to flow.
In order to suppress the lymphocyte removal performance of the neutrophil removal column and more selectively remove granulocytes, it is preferable to comprise a carrier having an average fiber diameter of 2.0 μm or more.
As the lymphocyte removal performance, if the lymphocyte removal rate is 70% or less, it is more preferable from the viewpoint of suppressing the onset of infectious diseases because lymphocytes responsible for immune memory are not seriously damaged.
The fibrous carrier preferably contains a fibrous carrier having an average fiber diameter of 2.6 μm or less in order to enhance the removal performance of granulocytes. The average fiber diameter of the fibrous carrier is preferably 1.4 μm or more and 2.6 μm or less, and more preferably 2.0 μm or more and 2.6 μm or less.
In addition to the filter for removing neutrophils, the neutrophil removal column further includes a relatively coarse carrier having an average fiber diameter of 8 μm to 14 μm as a prefilter for removing aggregates. preferable. The neutrophil removal column is a filter for removing neutrophils, and has an average fiber diameter of preferably 1.4 μm or more and 2.6 μm or less, more preferably 2.0 μm or more and 2.6 μm or less, and an aggregate. A prefilter for removal may contain a carrier having an average fiber diameter of 8 μm or more and 14 μm or less in combination.
In the present embodiment, the average fiber diameter can be measured by the method described in the examples.

  In this embodiment, when the neutrophil removal column includes a prefilter, the prefilter is disposed on the blood inlet side of the container, and the filter for removing neutrophils is disposed on the blood outlet side of the container. Is preferred.

The critical wet surface tension (CWST) of the fibrous carrier is preferably equal to or greater than the surface tension of the physiological solution used when priming the neutrophil removal column. The surface tension of a physiological solution is a so-called Wilhelmy method in which a platinum plate whose surface is sufficiently red-hot and cleaned with an alcohol lamp or the like is prepared, and the resistance when the platinum plate is immersed in a physiological solution is measured. Use to measure. As the surface tension meter, for example, FACE SURFACE TENSIOMETER CBVP-A3 (Kyowa Interface Science Co., Ltd.) can be used.
When the surface tension of a physiological solution is measured by this method, for example, physiological saline, titramine (Fuso Pharmaceutical Co., Ltd.), ACD-A solution (Terumo Co., Ltd.), Fusan (registered trademark: Torii Pharmaceutical Co., Ltd.), etc. The surface tension of the physiological saline containing the anticoagulant is 72 dyn / cm.
In the present embodiment, the CWST of the fibrous carrier is preferably 72 dyn / cm or more, more preferably 72 dyn / cm or more and 115 dyn / cm or less.

In the present embodiment, CWST of the fibrous carrier refers to a value obtained by the following method.
Aqueous solutions having different concentrations of sodium hydroxide, calcium chloride, sodium nitrate, acetic acid and ethanol are prepared so that the surface tension changes by 2 to 4 dyn / cm. The surface tension (dyn / cm) of each aqueous solution is 94 to 115 with an aqueous sodium hydroxide solution, 90 to 94 with an aqueous calcium chloride solution, 75 to 87 with an aqueous sodium nitrate solution, 72.4 with pure water, and 38 to 38 with an aqueous acetic acid solution. 69, 22 to 35 ethanol aqueous solution can be obtained ("Chemical Handbook Basic Edition II" revised 2nd edition, The Chemical Society of Japan, Maruzen, 164 (1975)). Moreover, you may prepare the aqueous solution whose surface tension is 74-100 dyn / cm with sodium hydroxide. The aqueous solutions having different surface tensions of 2 to 4 dyn / cm thus obtained are placed on the filter material in order of decreasing surface tension and left for 10 minutes. After standing for 10 minutes, it is defined as a wet state when 9 or more of 10 drops are absorbed by the filter material, and is defined as a non-wet state when the absorption is less than 9 of 10 drops. In this manner, when the liquid is sequentially measured from the liquid having a small surface tension on the filter material, a wet state and a non-wet state appear. At this time, the average value of the surface tension value of the liquid observed in the wet state and the surface tension value of the liquid observed in the non-wet state is defined as the CWST value of the filter material.
For example, when wetted with a liquid having a surface tension of 64 dyn / cm and non-wet with a liquid having a surface tension of 66 dyn / cm, the CWST value of the filter material is 65 dyn / cm.
In the present embodiment, the fibrous carrier may be made of the same filter material or different filter materials.

  In the present embodiment, the internal volume of the neutrophil removal column is not particularly limited as long as it is within a range that can be used as a blood purifier for treating blood extracorporeally, but is usually 50 mL to 500 mL, preferably 70 mL to 400 mL. And more preferably 90 mL to 300 mL.

The neutrophil removal column of this embodiment is preferably sterilized by any one of radiation sterilization, ethylene oxide gas sterilization, and autoclave sterilization. It is more preferable that radiation sterilization is performed from the viewpoint of safety, environmental load, and influence of heat on the column.
When the neutrophil removal column is sterilized by radiation, it is preferably sterilized by radiation in a state where the fibrous carrier is in an atmosphere having an oxygen concentration of 1% or less.
Examples of radiation used for sterilization include γ-rays, electron beams, and X-rays, and the line type is not particularly limited.
From the viewpoint of neutrophil removal performance and platelet permeability of the neutrophil removal column, it is preferable that the fibrous carrier is maintained in an atmosphere having an oxygen concentration of 1% or less after sterilization.

The neutrophil removal column of this embodiment is used for increasing the viable cell ratio of neutrophils in the blood circulating in the body in blood circulation applications.
When blood is extracorporeally treated using the neutrophil removal column of this embodiment, an anticoagulant may be used, or an anticoagulant is used if there is no risk of blood coagulating in the extracorporeal circuit. It is not necessary to use it.
Examples of blood anticoagulants include those that exhibit antithrombin activity such as heparin, those that exhibit anticoagulant activity by forming a complex with divalent metal ions such as citric acid and its salts, and nafamostat mesylate (Nafamostat Mesilate). Any of proteolytic enzyme inhibitors such as) can be used, but preferably an anticoagulant that forms a complex with a divalent metal ion, more preferably an anticoagulant containing citric acid and its salt.
Examples of the anticoagulant mainly composed of citric acid and its salts include titramine (Fuso Pharmaceutical Co., Ltd.), ACD-A solution (Terumo Corporation) and the like.

When the neutrophil removal column of this embodiment is used for patients with high risk of infection such as UC, colon perforation, esophageal cancer, and post-rectal cancer laparotomy patients or patients with infection, The neutrophil survival rate is increased, and the neutrophil infection defense function is normalized, so that the risk of infection is reduced or the efficiency of eliminating infectious bacteria is significantly increased.
Patients with high risk of infection such as UC, colon perforation, esophageal cancer, post-rectal cancer laparotomy patients, UC, colon perforation, esophageal cancer, patients after rectal cancer laparotomy, patients with wound classification class 2 or higher, nutritional status and Examples include patients with poor general condition and patients with high surgical invasion.
Here, patients with wound classification class 2 or higher refer to patients with semi-clean wounds (class 2), contaminated wounds (class 3), and infected / unclean wounds (class 4) other than the first stage of clean wounds.
Further, when the neutrophil removal column of the present embodiment is applied to a patient having an infection and the ratio of apoptotic cells in peripheral blood is higher than the normal value, the ratio of viable neutrophils increases, Since the ratio of apoptotic neutrophils is decreased and the ratio of viable neutrophils / apoptotic neutrophils is remarkably increased, the function of neutrophils is normalized and the elimination efficiency of infecting bacteria is remarkably increased.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not restrict | limited at all by these Examples. The measuring method in this example is as follows.

(1) Blood cell performance and pressure loss Porcine blood added with ACD-A solution (Terumo Corporation) is adjusted to 7 mass%, and before passing through the column with a blood analyzer (Sysmex Corporation TX-1800i) Each concentration of granulocytes, lymphocytes and platelets was measured.
Porcine blood was warmed to 37 ± 1 ° C., 2 L of warm porcine blood was flowed through the column at a flow rate of 50 mL / min with a blood pump, and all blood flowing out from the column outlet was collected. Concentrations of granulocytes, lymphocytes and platelets were measured for the collected blood.
The removal rate or passage rate of each component before and after passing through the column was determined.
The column inlet pressure and the column outlet pressure at the time when 2 L of blood was processed were measured, and the value obtained by subtracting the outlet pressure from the inlet pressure was defined as the pressure loss.

(2) Total surface area of fibrous carrier The total surface area of the fibrous carrier packed in the neutrophil removal column is the specific surface area of the fibrous carrier calculated from the average fiber diameter of the fibrous carrier, and the fibrous shape to be filled Calculated as the sum of the product with the weight of the carrier. When a fibrous carrier is used by using a plurality of types of filter materials, the surface area is calculated for each filter material, and the sum is taken as the total surface area of the fibrous carrier.
The specific surface area (m ² / g) of the fibrous carrier was calculated by 4 / (ρ × D) where D is a mean fiber diameter and ρ is a substrate density of the fibrous carrier. The substrate density of the fibrous carrier is measured according to JIS Z 8807: 2012.

(3) Coating amount of hydrophilic polymer A certain amount of the fibrous carrier coated with the hydrophilic polymer was sampled for each filter material, and the weight (W0) was measured.
The fibrous carrier after weight measurement was immersed in a solvent in which the hydrophilic polymer was dissolved and the substrate was not dissolved, and the mixture was shaken and stirred for 5 minutes. After stirring, the solvent was changed, and the mixture was again stirred with shaking for 5 minutes. After exchanging the solvent and shaking and stirring a total of three times, the solvent was discarded and the fibrous carrier was vacuum-dried at 50 ° C. for 24 hours. The weight (W1) after drying was measured.
The coating amount was calculated by (W1-W0) / W0.

(4) Average fiber diameter For each filter material, photographs of the fibrous carrier were taken using a scanning electron microscope at a magnification of 2500 times and changing the field of view until the total number exceeded 100. The width of the fiber taken perpendicular to the fiber axis of each fiber was measured as the diameter of the fiber. A value obtained by dividing the total diameter of the measured fibers by the total number of fibers whose diameters were measured was calculated as an average fiber diameter.

(5) CWST
The fibrous carrier was cut into 100 mm squares for each filter material. When the surface tension was in the range of 86 dyn / cm to 100 dyn / cm, 10 aqueous sodium hydroxide solutions differing by 2 dyn / cm were placed on the filter material in descending order of surface tension and left for 10 minutes. After standing for 10 minutes, when 9 or more drops out of 10 drops are absorbed by the filter material, it is assumed to be in a wet state, and when absorption is less than 9 drops out of 10 drops, it is assumed to be in a non-wet state. Among the observed values, the maximum surface tension value of the aqueous sodium hydroxide solution and the minimum surface tension value of sodium hydroxide among the observed non-wet conditions were averaged and measured as the CWST value of the fibrous carrier. .

(6) Method for measuring neutrophil fraction A blood cell separation solution was put in a centrifuge tube, and the same amount of collected blood was overlaid. Centrifugation was performed to collect a suspension of polynuclear granulocytes. Phosphate buffered saline
(saline, PBS) was added and mixed, centrifuged again, the supernatant was removed, and the remaining multinucleated granulocytes were suspended in PBS to prepare samples.
Next, Propidium iodide (PI) and FITC-labeled Annexin-V were added to the specimen for reaction and staining. The proportion of cells that were negative for both PI and Annexin-V (cells that were not stained with either PI or Annexin-V) was measured with a flow cytometer, and this was defined as the viability of the cells.
In addition, PI-negative and Annexin-V-positive cells were designated as early apoptotic neutrophils, PI-positive and Annexin-V-negative cells were designated as necrotic neutrophils, and PI and Annexin-V-positive cells were designated as late apoptotic neutrophils. .

(7) Use of neutrophil removal column for patients with high risk of infection or patients with infections Neutrophil removal column has high risk of infection such as UC, colon perforation, esophageal cancer, patients after rectal cancer laparotomy An open-label study was conducted to evaluate the function of neutrophils in patients and patients with infectious diseases.
A patient with a high risk of infection or a patient with an infection is a patient who meets 1) any of the selection criteria and 2) does not meet any of the exclusion criteria.
1) Selection criteria ・ UC, colon perforation, esophageal cancer, patients after rectal cancer laparotomy, wound classification class 2 or more ・ UC, colon perforation, esophageal cancer, patients after rectal cancer laparotomy, nutritional status and Patients with poor general condition, UC, colon perforation, esophageal cancer, patients after rectal cancer laparotomy, patients with high surgical invasion, patients with infectious disease 2) exclusion criteria, heart disease (after onset Patients with acute myocardial infarction within 6 months, ischemic heart disease, arrhythmia requiring treatment), patients with cerebral infarction, cerebrovascular disorder such as cerebral hemorrhage, etc., patients with hypotension or systolic blood pressure of 80 mmHg or less Pregnant, breastfeeding women, women who may be pregnant, patients with dementia, patients with a history of shock during extracorporeal circulation treatment, taking angiotensin converting enzyme (ACE) inhibitor, Difficult to discontinue taking before extracorporeal circulation Patients and here wound classification class 2 or more patients, semi-clean wounds other than clean wounds one level (class 2), contaminated wounds (class 3), refers to a patient infected, unclean wounds (Class 4).
The choice of implementation will be explained before the operation for patients scheduled to undergo surgery, and during the treatment of infection in cases where infection has occurred.If consent is obtained to use a neutrophil removal column, When consent was not obtained but consent was obtained for neutrophil function evaluation, it was set as a non-implementation group (control group).
The time of administration was immediately after the operation, and the number of times of execution was one. As an anticoagulant, titramine (Fuso Pharmaceutical Co., Ltd.) was used and added at 6% of the blood flow. The enforcement conditions were a blood flow rate of 30 to 50 mL / min, an extracorporeal circulation time of about 60 minutes, and a target blood throughput of 3,000 mL.

<Synthesis of hydrophilic polymer>
2-hydroxyethyl methacrylate (HEMA) and dimethylaminoethyl methacrylate (DM) are mixed at a molar ratio of 97: 3, and the total monomer concentration in ethanol is 1.0 mol / L, 1/200 mol / L. A hydrophilic polymer (hereinafter abbreviated as HM-3) was synthesized by solution radical polymerization at 60 ° C. for 8 hours in the presence of a polymerization initiator of L azobisisobutyronitrile.

<Creation of fibrous carrier>
HM-3 was dissolved in a 50% aqueous ethanol solution so that the HM-3 concentration was 0.4 mass%. A polyethylene terephthalate nonwoven fabric having an average fiber diameter of 12 μm, a basis weight of 100 g / m ² , and a thickness of 0.47 mm is immersed in the obtained solution to remove excess liquid, and then dried at 50 ° C. for 20 minutes to obtain a filter material ( A) was obtained. The density of the polyethylene terephthalate was 1.38 g / cm ³ .
The CWST value of the filter material (A) was 95 dyn / cm. When the coating amount of the hydrophilic polymer of the filter material (A) was measured using a 50% aqueous ethanol solution as a solvent, it was 17 mg per 1 g of the fibrous carrier.

HM-3 was dissolved in a 50% aqueous ethanol solution so that the HM-3 concentration was 0.4 mass%. A polyethylene terephthalate nonwoven fabric having an average fiber diameter of 12 μm, a basis weight of 30 g / m ² , and a thickness of 0.20 mm is immersed in the obtained solution, and after removing excess liquid, it is dried at 50 ° C. for 20 minutes to obtain a filter material (B )
The CWST value of the filter material (B) was 95 dyn / cm. When the coating amount of the hydrophilic polymer of the filter material (B) was measured using a 50% ethanol aqueous solution as a solvent, it was 8 mg per 1 g of the fibrous carrier.

HM-3 was dissolved in a 50% aqueous ethanol solution so that the HM-3 concentration was 0.4 mass%. A polyethylene terephthalate nonwoven fabric having an average fiber diameter of 2.3 μm, a basis weight of 60 g / m ² , and a thickness of 0.30 mm is immersed in the obtained solution, and the excess liquid is removed, followed by drying at 50 ° C. for 20 minutes. (C) was obtained.
The CWST value of the filter material (C) was 95 dyn / cm. When the coating amount of the hydrophilic polymer of the filter material (C) was measured using a 50% ethanol aqueous solution as a solvent, it was 32 mg per 1 g of the fibrous carrier.

HM-3 was dissolved in a 50% aqueous ethanol solution so that the HM-3 concentration was 0.4 mass%. A non-woven polyethylene terephthalate nonwoven fabric having an average fiber diameter of 1.4 μm, a basis weight of 66 g / m ² and a thickness of 0.40 mm is immersed in the obtained solution, and the excess liquid is removed, followed by drying at 50 ° C. for 20 minutes. (D) was obtained.
The CWST value of the filter material (D) was 91 dyn / cm. When the coating amount of the hydrophilic polymer of the filter material (D) was measured using a 50% ethanol aqueous solution as a solvent, it was 27 mg per 1 g of the fibrous carrier.

HM-3 was dissolved in a 50% aqueous ethanol solution so that the HM-3 concentration was 0.4 mass%. A polyethylene terephthalate nonwoven fabric having an average fiber diameter of 1.1 μm, a basis weight of 40 g / m ² , and a thickness of 0.24 mm is immersed in the obtained solution, and after removing excess liquid, it is dried at 50 ° C. for 20 minutes. (E) was obtained.
The CWST value of the filter material (E) was 87 dyn / cm. When the coating amount of the hydrophilic polymer of the filter material (E) was measured using a 50% ethanol aqueous solution as a solvent, it was 17 mg per 1 g of the fibrous carrier.

[Example 1]
Eighteen filter materials (C) were laminated, and nine filter materials (A) and four filter materials (B) were stacked thereon to form a sheet-like filter. The sheet-like filter is cut into a 97 mm square, and the filter material (A) is moved to the second inlet / outlet side into a 125 mL square flat container having a first inlet / outlet and a second inlet / outlet at the opposite apex portions. A flat column was prepared by performing ultrasonic welding. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Example 2]
Eighteen filter materials (D) were stacked, and nine filter materials (A) and four filter materials (B) were stacked thereon to form a sheet-like filter. The sheet-like filter is cut into a 97 mm square, and the filter material (A) is moved to the second inlet / outlet side into a 125 mL square flat container having a first inlet / outlet and a second inlet / outlet at the opposite apex portions. A flat column was prepared by performing ultrasonic welding. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Example 3]
Thirteen filter materials (C) were stacked, and nine filter materials (A) and four filter materials (B) were stacked thereon to form a sheet-like filter. The sheet filter is cut into a 97 mm square, and the filter material (C) is moved to the second inlet / outlet side into a 125 mL square flat container having a first inlet / outlet and a second inlet / outlet at opposite apex portions. A flat column was prepared by performing ultrasonic welding. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Example 4]
Eighteen filter materials (C) were laminated to form a sheet filter. By cutting the sheet filter into a 97 mm square, filling it into a 125 mL square flat container having a first inlet and a second inlet and a second inlet and outlet at opposite corners, respectively, and performing ultrasonic welding. A flat column was produced. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Example 5]
27 filter materials (C) were laminated to prepare a sheet-like filter. By cutting the sheet filter into a 97 mm square, filling it into a 125 mL square flat container having a first inlet and a second inlet and a second inlet and outlet at opposite corners, respectively, and performing ultrasonic welding. A flat column was produced. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Example 6]
Eighteen filter materials (C) were laminated, and nine filter materials (A) and four filter materials (B) were stacked thereon to form a sheet-like filter. The sheet-like filter is cut into a 97 mm square, and the filter material (A) is moved to the second inlet / outlet side into a 125 mL square flat container having a first inlet / outlet and a second inlet / outlet at the opposite apex portions. A flat column was prepared by performing ultrasonic welding. The column was put into a packaging bag made of a laminated film of nylon, aluminum and polyethylene, and the packaging bag was sealed by heat sealing. Sterilization was performed by irradiation with 25 kGy of γ-rays to obtain a neutrophil removal column.

[Comparative Example 1]
Twelve filter materials (C) were laminated, and nine filter materials (A) and four filter materials (B) were stacked thereon to form a sheet-like filter. The sheet filter is cut into a 97 mm square, and the filter material (C) is moved to the second inlet / outlet side into a 125 mL square flat container having a first inlet / outlet and a second inlet / outlet at opposite apex portions. A flat column was prepared by performing ultrasonic welding. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Comparative Example 2]
18 non-woven polyethylene terephthalate nonwoven fabrics having an average fiber diameter of 2.3 μm, a basis weight of 60 g / m ² and a thickness of 0.30 mm are laminated, and an average fiber diameter of 12 μm, a basis weight of 100 g / m ² and a thickness of 0.47 mm made of polyethylene terephthalate Nine nonwoven fabrics, 4 nonwoven fabrics made of polyethylene terephthalate having an average fiber diameter of 12 μm, a basis weight of 30 g / m ² , and a thickness of 0.20 mm were stacked to form a sheet-like filter. A sheet-like filter is cut into a 97 mm square, and a nonwoven fabric having an average fiber diameter of 2.3 μm is applied to a 125 mL square flat container having a first entrance and a second entrance of liquid at the opposite apex portions, respectively. The flat column was produced by packing so that it might become 2 entrance-and-exit sides, and performing ultrasonic welding. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

[Comparative Example 3]
Eighteen filter materials (E) were stacked, and nine filter materials (A) and four filter materials (B) were stacked thereon to form a sheet-like filter. The sheet-like filter is cut into a 97 mm square, and the filter material (A) is moved to the second inlet / outlet side into a 125 mL square flat container having a first inlet / outlet and a second inlet / outlet at the opposite apex portions. A flat column was prepared by performing ultrasonic welding. The column was put in a packaging bag made of a laminated film of nylon, aluminum, and polyethylene together with an oxygen scavenger (Mitsubishi Gas Chemical Co., Ltd., Ageless (registered trademark) SS200), and the packaging bag was sealed by heat sealing.
The oxygen concentration in the packaging bag was measured using an oxygen concentration meter (Iijima Electronics Co., Ltd., RO-102-SDP). After the oxygen concentration became 1% or less, sterilization was performed by irradiating 25 kGy of γ rays. A neutrophil removal column was obtained.

The granulocyte / lymphocyte removal rate, platelet passage rate, and pressure loss of the obtained neutrophil removal column were measured, and the total surface area of the fibrous carrier was calculated. The results are shown in Table 1.
From Table 1, if the neutrophil removal column is composed of a filter material having a hydrophilic polymer on the surface, a total surface area of 10.5 m ² or more and an average fiber diameter of 1.4 μm or more, It can be seen that the removal rate is 80% or more and the platelet passage rate is 75% or more.

Figure shows the relationship between the total surface area of the fibrous carrier and the removal rate of granulocytes, the relationship between the total surface area of the fibrous carrier and the passage rate of platelets, and the relationship between the total surface area of the fibrous carrier and the removal rate of lymphocytes. 1 to FIG.
From FIG. 1, it is preferable that the total surface area of the fibrous carrier is 10.5 m ² or more from the viewpoint of making the granulocyte removal rate 80% or more.
Leukocyte removal therapy (LCAP) also has a function of improving inflammation in the peripheral blood and a vicious cycle of immune function. In order to obtain this function, the removal rate of lymphocytes needs to be 30% or more, and the total surface area of the fibrous carrier is preferably 10.5 m ² or more also from the function of the removal rate of lymphocytes.
From the viewpoint of the removal rate of granulocytes and the removal rate of lymphocytes, in the neutrophil removal column composed of a fibrous carrier having a hydrophilic polymer on the surface and an average fiber diameter of 1.4 μm or more It can be seen that the total surface area of the fibrous carrier is 10.5 m ² or more.
From FIG. 2, it is preferable that the total surface area of the fibrous carrier is 21 m ² or less from the viewpoint of setting the platelet passage rate to 75% or more.

[Example 7]
A elective surgery was performed on a 40-year-old male, wound classification class 2, good nutritional state, ASA classification 1 clone patient.
Here, the ASA (American Society of. Anesthesiologists) classification is a general state classification in the American Academy of Anesthesiology. ASA category 1 has no organic, physiological, biochemical or mental abnormalities, and the disease targeted for surgery is localized and does not cause systemic (systematic) damage. is there. ASA classification 2 refers to those with mild to moderate systematic impairment. The cause is due to a disease subjected to surgical treatment or other pathophysiological causes. ASA classification 3 refers to those with severe systemic disease. In this case, the cause of systematic failure may be anything, and even if the degree of clear failure cannot be determined. ASA classification 4 has a high degree of systematic disease that is being threatened by life, and just because an operation is performed, the lesion cannot always be treated. ASA Category 5 refers to those who are less likely to be saved in moribund patients but who must be operated on. ASA classification 6 refers to patients with brain death.
The operation was an endoscopic operation, and the operation time was 439 minutes. The patient had a slightly higher surgical invasiveness and no blood transfusion. 68 minutes after completion of the operation, neutrophil removal was started using the neutrophil removal column prepared in Example 1. Titoramine (Fuso Yakuhin Kogyo Co., Ltd.) was used as an anticoagulant, the blood throughput was 3.0 L, and the operation time was 79 minutes.
Peripheral blood was collected before and after neutrophil removal, and the viable cell ratio of peripheral blood neutrophils, late apoptotic neutrophils, and platelet count were measured.

[Example 8]
A elective surgery was performed on a 47-year-old woman, wound classification class 2, nutritional condition, ASA classification 1 clone patient. The operation time was 277 minutes, and the patient was slightly invasive. There was no blood transfusion. 72 minutes after the completion of the operation, neutrophil removal was started using the neutrophil removal column prepared in Example 1. Titoramine (Fuso Pharmaceutical Co., Ltd.) was used as an anticoagulant, the blood throughput was 3.0 L, and the enforcement time was 60 minutes.
Peripheral blood was collected before and after neutrophil removal, and the viable cell ratio of peripheral blood neutrophils, late apoptotic neutrophils, and platelet count were measured.

[Example 9]
A 44-year-old male, wound classification class 3, slightly poor nutritional state, ASA classification 3 all colitis type UC patient was subjected to elective surgery. The operation time was 276 minutes, and the patient had a slightly higher surgical invasion. The transfusion was MAP4 units. 77 minutes after completion of the operation, neutrophil removal was started using the neutrophil removal column prepared in Example 1. Titoramine (Fuso Yakuhin Kogyo Co., Ltd.) was used as an anticoagulant, the blood throughput was 3.0 L, and the operation time was 80 minutes.
Peripheral blood was collected before and after neutrophil removal, and the viable cell ratio of peripheral blood neutrophils, late apoptotic neutrophils, and platelet count were measured.

[Example 10]
A 53-year-old male, wound classification class 3, nutritional status, ASA classification 2 intraperitoneal abscess patient was subjected to elective surgery. The operation time was 137 minutes, and the patient was slightly more invasive. There was no blood transfusion. 66 minutes after completion of the operation, neutrophil removal was started using the neutrophil removal column prepared in Example 1. Titoramine (Fuso Yakuhin Kogyo Co., Ltd.) was used as an anticoagulant, the blood throughput was 3.0 L, and the enforcement time was 75 minutes.
Peripheral blood was collected before and after neutrophil removal, and the viable cell ratio of peripheral blood neutrophils and late apoptotic neutrophils were measured.

[Example 11]
Standby surgery was performed on a 30-year-old woman, wound classification class 2, good nutritional condition, ASA classification 2 UC patient. The operation time was 525 minutes, and the patient was highly invasive. There was no blood transfusion. 73 minutes after completion of the operation, neutrophil removal was started using the neutrophil removal column prepared in Example 1. Titoramine (Fuso Yakuhin Kogyo Co., Ltd.) was used as an anticoagulant, the blood throughput was 3.0 L, and the enforcement time was 67 minutes.
Peripheral blood was collected before and after neutrophil removal, and the viable cell ratio of peripheral blood neutrophils and late apoptotic neutrophils were measured.

[Comparative Example 4]
A elective surgery was performed on a 49-year-old male, all-colitis UC patient of wound classification class 2, ASA classification 1. The operation was an endoscopic operation, and the operation time was 396 minutes. There was no blood transfusion.
Peripheral blood was collected 1 hour and 2 hours after the end of surgery, and the viable cell ratio of peripheral blood neutrophils, late apoptotic neutrophils, and platelet count were measured.

[Comparative Example 5]
A 61-year-old male, wound classification class 2, ASA classification 2 esophageal cancer patient was subjected to elective surgery. The operation was an endoscopic operation, and the operation time was 910 minutes. There was no blood transfusion.
Peripheral blood was collected 1 hour and 2 hours after the end of surgery, and the viable cell ratio of peripheral blood neutrophils, late apoptotic neutrophils, and platelet count were measured.

The above results are summarized in Table 2.
In 3 cases, 1 UC patient and 2 Crohn's disease patients, after neutrophil removal, the viability of neutrophils increased, late apoptotic neutrophil count decreased, platelet count Was maintained at 90% or more before surgery. By performing neutrophil removal using a neutrophil removal column, the platelet count in the patient's peripheral blood was controlled to a range that maintains the hemostatic function, and neutrophils It has been shown that the viable cell rate increases. In particular, in cases where the percentage of viable neutrophils before the operation was less than 90%, the increase in the neutrophil viable cell rate was remarkable. It was also confirmed that the number of late-stage apoptotic neutrophils in the peripheral blood of patients decreased as a result of neutrophil removal.

[Example 12]
A 74-year-old male, who had a well-nutritioned abdominal abscess and iliopsoas abscess (no surgery, had infection), was neutral using the neutrophil removal column prepared in Example 1 Sphere removal started. Titoramine (Fuso Yakuhin Kogyo Co., Ltd.) was used as an anticoagulant, the blood throughput was 2.0 L, and the operation time was 40 minutes.
Peripheral blood was collected before neutrophil removal and one day after neutrophil removal. Viable neutrophil count, early apoptotic neutrophil count, necrotic neutrophil count, late apoptotic neutrophil in peripheral blood Number was measured. The results are shown in Table 3.

  This case is a patient having an infection, and the ratio of early apoptotic neutrophils in peripheral blood before neutrophil removal has reached 85%, and the ratio of viable neutrophils is only 9%. It was 1%. This ratio changed dramatically one day after the removal of neutrophils, and the ratio of early apoptotic neutrophils was 0.8% and that of viable neutrophils was 95.2%. When neutrophil removal is performed on patients with a low percentage of viable neutrophils (a high percentage of apoptotic neutrophils), the number of viable neutrophils increases and the number of apoptotic neutrophils decreases after the first day, It was found that the ratio of viable neutrophils / apoptotic neutrophils was significantly increased.

  When the peripheral blood of a patient with a high risk of infection is treated by extracorporeal circulation using the neutrophil removal column of the present invention, the viable cell ratio of neutrophils in the peripheral blood increases and the risk of infection can be reduced. In addition, when the peripheral blood of a patient who has already been infected using the neutrophil removal column of the present invention is subjected to extracorporeal circulation treatment, the viable cell ratio of neutrophils in the peripheral blood increases, and the elimination efficiency of infectious bacteria is increased. Can be significantly increased. Furthermore, when the neutrophil removal column of the present invention is used, the platelet passage rate is high, and the number of platelets in the blood circulating in the body can be controlled within a range that maintains the hemostatic ability.

  This application is based on a Japanese patent application (Japanese Patent Application No. 2014-127710) filed on June 20, 2014, the contents of which are incorporated herein by reference.

  The neutrophil removal column of the present invention has industrial applicability as a blood circulation column.

Claims (11)

Filled with a fibrous carrier having a hydrophilic polymer on its surface,
The total surface area of the fibrous carrier is 10.5 m ² or more;
A neutrophil removal column for increasing the viable cell ratio of neutrophils in blood circulating in the body, wherein the fibrous carrier is composed of a carrier having an average fiber diameter of 1.4 μm or more.   The neutrophil removal column according to claim 1, wherein the granulocyte removal rate is 80% or more and the platelet passage rate is 75% or more.   The neutrophil removal column according to claim 2, wherein the lymphocyte removal rate is 70% or less.   The neutrophil removal column according to any one of claims 1 to 3, wherein the column is sterilized by radiation in a state where the oxygen concentration in the surrounding atmosphere is 1% or less.   The neutrophil removal column according to any one of claims 1 to 4, wherein the hydrophilic polymer has a basic functional group and a nonionic hydrophilic group.   The neutrophil removal column according to claim 5, wherein the basic functional group is derived from dimethylaminoethyl methacrylate, and the nonionic hydrophilic group is derived from 2-hydroxyethyl methacrylate.   The neutrophil removal column according to any one of claims 1 to 6, wherein the fibrous carrier includes a carrier having an average fiber diameter of 1.4 µm or more and 2.6 µm or less.   The neutrophil removal column according to claim 7, wherein the fibrous carrier further comprises a carrier having an average fiber diameter of 8 µm or more and 14 µm or less.   The neutrophil removal column according to any one of claims 1 to 8, wherein the fibrous carrier is a nonwoven fabric.   The neutrophil removal column according to any one of claims 1 to 9, wherein a critical wet surface tension of the fibrous carrier is not less than a surface tension of a physiological solution.   For patients with ulcerative colitis, colon perforation, esophageal cancer, rectal cancer laparotomy, patients with wound class 2 or higher, patients with high surgical invasiveness, or patients with infection The neutrophil removal column of any one of 10-10.

Images