CN110787647A - Platelet leukocyte-removing filter membrane and preparation method thereof - Google Patents
Platelet leukocyte-removing filter membrane and preparation method thereof Download PDFInfo
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/12—Composite membranes; Ultra-thin membranes
- B01D69/125—In situ manufacturing by polymerisation, polycondensation, cross-linking or chemical reaction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
- B01D71/06—Organic material
- B01D71/76—Macromolecular material not specifically provided for in a single one of groups B01D71/08 - B01D71/74
- B01D71/78—Graft polymers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01D2323/00—Details relating to membrane preparation
- B01D2323/30—Cross-linking
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2323/00—Details relating to membrane preparation
- B01D2323/38—Graft polymerization
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2325/00—Details relating to properties of membranes
- B01D2325/24—Mechanical properties, e.g. strength
Abstract
The invention discloses a platelet leukocyte-removing filter membrane and a preparation method thereof, wherein the filter membrane comprises a PBT (polybutylene terephthalate) grafted non-woven fabric and a polymer coated on the PBT grafted non-woven fabric, the PBT grafted non-woven fabric is a PBT base material non-woven fabric, and the polymer is obtained by polymerizing vinyl acetate and vinyl pyrrolidone in an acetone solvent. The polymer has low washout property and excellent biocompatibility, can effectively remove leukocytes in platelet preparations, has a removal rate of over 99.9 percent, has a platelet recovery rate of over 85 percent, and does not influence the functions of the filtered platelets. Meanwhile, the filtering membrane has better flexibility, is not only suitable for a hard shell filter, but also meets the manufacturing requirement of a soft filter, and has low manufacturing cost.
Description
Technical Field
The invention relates to a filtering membrane and a preparation method thereof, in particular to a platelet leukocyte-removing filtering membrane and a preparation method thereof.
Background
Bleeding caused by thrombocytopenia is a common clinical symptom, and death of patients is caused by untimely treatment or poor curative effect. Currently, an effective therapeutic means for preventing and treating bleeding caused by a decrease in platelet count or a functional disorder is platelet infusion. Platelet products are classified into machine-harvested platelets and manually concentrated platelets. The collection and preparation process of the platelet product inevitably introduces a large amount of white blood cells, and particularly, the manual concentration of the platelets comes from the separation and mixing of a plurality of platelets in a plurality of parts, wherein the mixing amount of the white blood cells is larger. The transfusion of platelet products with a large amount of white blood cells is easy to cause transfusion adverse reactions such as nonhemolytic transfusion fever reaction, HLA alloimmunization, graft-versus-host disease after transfusion, ineffective platelet transfusion and the like. Therefore, the removal of leukocytes from platelets plays an important role in the clinical efficacy of platelet products.
With the rapid growth in clinical use of platelet products, there is an increasing demand for leukocyte-reduction treatments for platelet preparations. However, since platelets have high adhesion capability and are easily activated in vitro, the existing filter materials for leukocyte-removing treatment of whole blood or other blood components are hardly suitable for leukocyte-removing treatment of platelet preparations; and the white removing treatment of the platelet product needs to meet the recovery rate of the platelets and ensure the related functions of the filtered platelets.
Although the existing platelet leukocyte-removing filter on the market in China at present has high price and relatively low leukocyte removal rate, and the leukocyte removal rate is generally difficult to reach more than 99.9 percent. Therefore, there is a need for an improved platelet-leukocyte-removing filtration membrane and a method for producing the same, which are capable of obtaining a platelet filtration membrane with improved performance.
Disclosure of Invention
The technical problem to be solved by the invention is to provide the platelet leukocyte-removing filter membrane and the preparation method thereof, which have better leukocyte-removing effect, better flexibility and certain deformability, are suitable for a hard shell filter, meet the manufacturing requirement of a soft filter and have low manufacturing cost.
The technical scheme adopted by the invention for solving the technical problems is to provide a platelet leukocyte-removing filter membrane, which comprises PBT (polybutylene terephthalate) grafted non-woven fabric and a polymer coated on the PBT grafted non-woven fabric, wherein the polymer is obtained by polymerizing vinyl acetate and vinyl pyrrolidone in an acetone solvent.
Further, the PBT grafted non-woven fabric is a PBT base material non-woven fabric which is modified by ethyl acrylate and has a grafting rate of 10-20%.
Further, the weight ratio of the vinyl pyrrolidone to the vinyl acetate is 1: 6-1: 9.
furthermore, the average pore diameter of the PBT base material non-woven fabric is 12-21 mu m.
Furthermore, the mass content of the polymer in the filtering membrane is 15% -20%, and the coverage rate of the polymer on the surface of the PBT grafted non-woven fabric is 80% -100%.
Furthermore, the number of layers of the PBT base material non-woven fabric is multiple, the pore diameter of the multiple layers of PBT base material non-woven fabrics is sequentially reduced from top to bottom, the polymer content is sequentially increased from top to bottom, and the surface coverage rate of the polymer on the lowest layer of PBT base material non-woven fabric is 100%.
Further, the number of layers of the PBT base material non-woven fabric is 10, the pore diameters of the PBT base material non-woven fabric are respectively 21 micrometers, 17 micrometers and 12 micrometers, the layer number ratio of the PBT base material non-woven fabric with three different pore diameters is 2:3:5, wherein the 2 layers of the inlet face are the PBT base material non-woven fabric with the pore diameter of 21 micrometers, the polymer content is 18.11%, and the surface coverage rate is 95%; the middle 3 layers are non-woven fabrics with 17 mu m aperture, the polymer content is 19.23 percent, and the surface coverage rate is 100 percent; the outlet face 5 layer is 12 μm aperture non-woven fabric, polymer content is 20%, and surface coverage is 100%.
The invention also provides a preparation method of the platelet leukocyte-removing filter membrane for solving the technical problems, which comprises the following steps: s1) soaking and cleaning the PBT base material non-woven fabric; s2) adding vinyl acetate and vinyl pyrrolidone into an acetone solvent, stirring and reacting in a constant-temperature water bath under the protection of nitrogen to obtain viscous liquid, extracting with n-hexane, dissolving the bottom liquid in acetone, pouring the solution into water to precipitate a polymer, dissolving with acetone for multiple times, precipitating with water to remove unreacted monomers, drying and drying to obtain the polymer; s3) dissolving the polymer in an acetone solution, immersing the grafted non-woven fabric in the acetone solution, airing, drying and drying to obtain the filter membrane.
Further, the weight ratio of the vinyl pyrrolidone to the vinyl acetate in the step S2 is 1: 6-1: 9, the step S2 is stirred and reacted for 72 hours under the protection of nitrogen at the constant temperature water bath of 54 ℃ to obtain viscous liquid.
Further, the percentage concentration of the polymer in the acetone solution of the step S3 is 4 wt%, the step S3 is to soak the grafted non-woven fabric in the acetone solution containing the polymer for 1 hour, dry and dry at 80 ℃, and the content of the azobisisobutyronitrile in the acetone solution of the step S3 is 2.95 g/L.
Compared with the prior art, the invention has the following beneficial effects: the polymer has low elution and excellent biocompatibility, can effectively remove leukocytes in platelet preparations, has a removal rate of over 99.9 percent, has a platelet recovery rate of over 85 percent, and does not influence the functions of filtered platelets. Meanwhile, the filtering membrane has good flexibility and certain deformability, is suitable for a hard shell filter, meets the manufacturing requirement of a soft filter, and is low in manufacturing cost.
Drawings
FIG. 1 is a schematic flow chart illustrating the preparation of a platelet leukocyte-removing filtration membrane according to an embodiment of the present invention;
FIG. 2 is a schematic representation of the morphology of the filter membrane in an embodiment of the invention.
Detailed Description
The invention is further described below with reference to the figures and examples.
Platelet adhesion is obviously influenced by factors such as plasma proteins Fg, Fn and VWF, and surface chemical groups and surface charges of the filter material and hydrophilicity of the surface of the material have important influence on the plasma protein adsorption of the material. Therefore, the PBT (polybutylene terephthalate) non-woven fabric is selected as the filtering material, and the embodiment of the invention adopts the commercially available PBT base material non-woven fabric for related research, so that the surface modification of the material with poor adsorption on related proteins is searched while the leukocyte removing effect is ensured.
The platelet leukocyte-removing filter membrane provided by the invention comprises PBT grafted non-woven fabric and a polymer coated on the PBT grafted non-woven fabric, wherein the polymer is obtained by polymerizing vinyl acetate and vinyl pyrrolidone in an acetone solvent. The vinyl pyrrolidone is an important fine chemical product, is widely used for crosslinking and polymerization reaction, and is an excellent modified grafting agent. The invention utilizes the use of the substance on the surface of the base material to increase the hydrophilic property of the membrane material, and the hydrophilic property plays an important role in the filtering effect of the blood platelet and the function after filtering. However, since the elution is easy because of the simple use of vinylpyrrolidone grafting, the present invention performs grafting using a polymer formed from vinylpyrrolidone and vinyl acetate, and the preparation process is shown in FIG. 1.
The polymer of the invention is prepared by the following steps:
preparing AIBN (azodiisobutyronitrile) into an acetone solution with the AIBN content of 2.95g/L to obtain the acetone solvent; adding vinyl acetate and vinyl pyrrolidone in a certain weight ratio into an acetone solvent, and stirring and reacting for 72 hours in a constant-temperature water bath at 54 ℃ under the protection of nitrogen to obtain viscous liquid; extracting the viscous liquid with n-hexane, dissolving the bottom liquid in acetone, and pouring the acetone-dissolved liquid into water to precipitate the polymer; and dissolving the precipitate by acetone for many times, precipitating by water to remove unreacted monomers, and drying at 80 ℃ to obtain the polymer.
Polymer example 1
In accordance with the above procedure, vinylpyrrolidone and vinyl acetate were polymerized in an acetone solvent at a weight ratio of 1:4 in a final yield of 21.42% and a polymer weight-average molecular weight of 18520 g/mol.
Polymer example 2
Vinyl pyrrolidone and vinyl acetate were polymerized in a 1:7 weight ratio in acetone solvent as described above, with a final yield of 85.71% and a polymer weight average molecular weight of 40430 g/mol.
Polymer example 3
Vinyl pyrrolidone and vinyl acetate were polymerized in a 1:12 weight ratio in acetone solvent as described above, with a final yield of 90.66% and a polymer weight average molecular weight of 43810 g/mol.
The weight average molecular weight of the polymer is closely related to the service performance and the processing performance of the material. The material has low mechanical strength and toughness and no application value; too high, increases melt viscosity and makes processing difficult. According to repeated experiments, the weight average molecular weight of the polymer is preferably 20000 to 100000g/mol, so that 1:4 is not preferable. Preparing 4 wt% acetone solution with the polymer, infiltrating the grafted non-woven fabric, manufacturing a final filter membrane, and mixing the filter material with water for injection 1: 20 at 37 ℃ for 2h, and performing chemical property detection (including ultraviolet absorbance, pH value, reducing substances, metal ions, evaporation residues and the like) on the leachate, wherein the results show that the ratio of 1:12 fail to yield evaporation residue. The combined consideration is given as 1:7 and adjacent weight ratios, i.e. the weight ratio of vinylpyrrolidone and vinyl acetate of the present invention is preferably 1: 6-1: 9.
the filter membrane is prepared by the following steps:
preparing the polymer into an acetone solution with a certain weight ratio; and (3) immersing the grafted non-woven fabric into the acetone solution, taking out the grafted non-woven fabric after 1 hour, airing the grafted non-woven fabric, and drying the grafted non-woven fabric at the temperature of 80 ℃ to constant weight to obtain the filter membrane.
Filtration Membrane example 1
The method comprises the following steps of 1: the polymer obtained by the polymerization was formulated into a 1 wt% acetone solution, and a PBT base nonwoven fabric having an average pore size of 12 μm was treated to finally obtain a filtration membrane having a polymer content of about 6.67%.
Filtration Membrane example 2
The method comprises the following steps of 1: the polymer obtained by polymerization was formulated into a 2 wt% acetone solution, and a PBT base nonwoven fabric having an average pore size of 12 μm was treated to finally obtain a filtration membrane having a polymer content of about 11.81%.
Filtration Membrane example 3
The method comprises the following steps of 1: the polymer obtained by polymerization was formulated into a 4 wt% acetone solution, and a PBT base nonwoven fabric having an average pore size of 12 μm was treated to finally obtain a filtration membrane having a polymer content of about 23.81%.
As the polymer concentration increases, the areal density of the membrane also gradually increases, with more polymer being coated onto the fibers of the membrane. Platelets are more readily adsorbed on hydrophobic surfaces, and the higher the degree of coverage of the membrane fibers by the polymer, the less likely the platelets will be adsorbed by the membrane and the more readily the polymer will be eluted. The shape of the filtering membrane is shown in FIG. 2, wherein a in FIG. 2 is a blank membrane; b: a filtering membrane after being immersed into 1% acetone solution to treat the PBT base material non-woven fabric; c: a filtering membrane after being immersed into 2% acetone solution to treat the PBT base material non-woven fabric; d: and (3) a filtering membrane after the PBT base material non-woven fabric is immersed in 4% acetone solution for treatment.
Mixing the filter material with water for injection 1: 20 at 37 ℃ for 2h, and detecting the soaking solution of the membrane soaked by 1%, 2% and 4% acetone solution according to the GB/14233.1-1998 method. The 1% and 2% soaked membranes had associated test data inferior to that of the 4% soaked membrane, but were still acceptable.
From the above results, 1) when the ratio of vinylpyrrolidone to vinyl acetate is 1: and when 7, the yield and the weight average molecular weight of the obtained polymer are higher, and various chemical detections such as reduced matter detection, absorbance and pH value, evaporation residues and metal ions meet the requirements. 2) From the density change before and after coating and the film appearance before and after coating, the fiber of the film becomes gradually thicker and the area density of the film also increases gradually as the concentration of the polymer increases, and more polymer is coated on the fiber of the film.
Therefore, the mixture ratio of the vinyl pyrrolidone to the vinyl acetate is 1:7, further processing by covering the membrane material with 4 wt% acetone solution.
The finally obtained filtering membrane is a 10-layer 12-micron filtering membrane, a round hard shell filter with the filtering diameter of 40mm is manufactured, and after high-temperature and high-pressure sterilization, physical and chemical properties are detected, and ultraviolet and hemolysis are unqualified; the platelet-rich plasma was prepared by filtering 1 unit of whole blood, and the experimental results showed a platelet recovery of 61.36% and a leukocyte residue of 3.52 x 104And the platelet aggregation rate is unqualified after the storage for one day.
Example 1A Filter Membrane for platelet preparations
Adopting PBT non-woven fabric and 1: the filtration membrane described in example 1 was prepared from a 4 wt% acetone solution prepared from the polymer obtained by polymerization, and the polymer content in the filtration membrane was 15.52%, 17.10% and 17.21% respectively from large to small according to the pore size of the nonwoven fabric, and the surface coverage was 81%, 85% and 85% respectively. The finally obtained filtering membrane is prepared according to the proportion of the number of the pore diameter layers: a round hard shell filter with a filtration diameter of 40mm was manufactured with a diameter of 21 μm, 17 μm, 12 μm, 2:3:5, i.e. the inlet face 2 layer was a 21 μm pore size nonwoven fabric, the polymer content was 15.52%, and the surface coverage was 81%; the middle 3 layers are non-woven fabrics with 17 mu m aperture, the polymer content is 17.10 percent, and the surface coverage rate is 85 percent; the outlet face 5 layer was a 12 μm pore size nonwoven with a polymer content of 17.21% and a surface coverage of 85%. After high-temperature and high-pressure sterilization, the related performance tests are all qualified; the platelet-rich plasma was prepared by filtering 1 unit of whole blood, and the experimental results showed a platelet recovery of 86.07% and a leukocyte residue of 1.62 x 104The leukocyte removal rate reaches more than 99.9 percent, and the related functions of the filtered platelets meet the requirements.
Example 2A Filter Membrane for platelet preparation
Adopting PBT non-woven fabric and 1:7 wt% of an acetone solution prepared from the polymer obtained by polymerization, the filtration membrane described in example 1 was obtained, and the polymer content in the filtration membrane was 18.11%, 19.23% and 20% respectively from the large to the small pore size of the nonwoven fabric, and the surface coverage was 95%, 100% and 100% respectively. The finally obtained filtering membrane is prepared according to the proportion of the number of the pore diameter layers: a round hard shell filter with a filtration diameter of 40mm is manufactured with a filtration diameter of 21 μm, 17 μm, 12 μm, 2:3:5, namely, a non-woven fabric with a pore diameter of 21 μm is arranged on the inlet face 2 layer, the polymer content is 18.11 percent, and the surface coverage rate is 95 percent; the middle 3 layers are non-woven fabrics with 17 mu m aperture, the polymer content is 19.23 percent, and the surface coverage rate is 100 percent; the outlet face 5 layer is a 12 μm pore size nonwoven fabric with a polymer content of 20% and a surface coverage of 100%. After high-temperature and high-pressure sterilization, the related performance tests are all qualified; the platelet-rich plasma is prepared by filtering 1 unit of whole blood, and the experimental result shows that the recovery rate of the platelets is 89.41 percentThe leukocyte remains 2.33 x 104The leukocyte removal rate reaches more than 99.9 percent, and the related functions of the filtered platelets meet the requirements.
Example 3A Filter Membrane for platelet preparation
Adopting PBT non-woven fabric and 1: 9 wt% of an acetone solution prepared from the polymer obtained by polymerization, the filtration membrane described in example 1 was obtained, and the polymer content in the filtration membrane was 18.78%, 19.07% and 19.31% respectively from the large to the small of the pore size of the nonwoven fabric, and the surface coverage was 97%, 100% and 100% respectively. The finally obtained filtering membrane is prepared according to the proportion of the number of the pore diameter layers: a round hard shell filter with a filtration diameter of 40mm is manufactured with a filtration diameter of 21 μm, 17 μm, 12 μm, 2:3:5, namely, a non-woven fabric with a pore diameter of 21 μm is arranged on the inlet face 2 layer, the polymer content is 18.78 percent, and the surface coverage rate is 97 percent; the middle 3 layers are non-woven fabrics with 17 mu m aperture, the polymer content is 19.07 percent, and the surface coverage rate is 100 percent; the outlet face 5 layer was a 12 μm pore size nonwoven with a polymer content of 19.31% and a surface coverage of 100%. After high-temperature and high-pressure sterilization, the related performance tests are all qualified; the platelet-rich plasma was prepared by filtering 1 unit of whole blood, and the experimental results showed a platelet recovery of 87.32% and a leukocyte residue of 3.59 x 104The leukocyte removal rate reaches more than 99.9 percent, and the related functions of the filtered platelets meet the requirements.
Although the present invention has been described with respect to the preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. The platelet leukocyte-removing filter membrane is characterized by comprising a PBT (polybutylene terephthalate) grafted non-woven fabric and a polymer coated on the PBT grafted non-woven fabric, wherein the polymer is obtained by polymerizing vinyl acetate and vinyl pyrrolidone in an acetone solvent.
2. The platelet leukocyte-removing filter membrane according to claim 1, wherein the PBT grafted nonwoven fabric is a PBT-based nonwoven fabric modified with ethyl acrylate in a grafting ratio of 10% to 20%.
3. The platelet leukocyte-removing filter membrane according to claim 1 wherein the weight ratio of vinylpyrrolidone to vinyl acetate is from 1: 6-1: 9.
4. the platelet leukocyte-removing filter membrane according to claim 2, wherein the PBT-base nonwoven fabric has an average pore size of 12 to 21 μm.
5. The platelet leukocyte-removing filter membrane according to claim 1, wherein the polymer has a mass content of 15% to 20% in the filter membrane, and the polymer has a coverage of 80% to 100% on the surface of the PBT-grafted nonwoven fabric.
6. The platelet leukocyte-removing filter membrane according to claim 1, wherein the number of layers of the PBT base nonwoven fabric is plural, the pore size of the plural layers of PBT base nonwoven fabrics becomes smaller from top to bottom, the polymer content increases from top to bottom, and the surface coverage of the polymer on the lowest layer of PBT base nonwoven fabric is 100%.
7. The platelet leukocyte-removing filtration membrane according to claim 6, wherein the number of layers of the PBT-base nonwoven fabric is 10, the pore sizes are 21 μm, 17 μm and 12 μm, respectively, and the ratio of the number of layers of the PBT-base nonwoven fabrics with different pore sizes is 2:3:5, wherein the inlet face 2 layers are 21 μm pore size nonwoven fabrics, the polymer content is 18.11%, and the surface coverage is 95%; the middle 3 layers are non-woven fabrics with 17 mu m aperture, the polymer content is 19.23 percent, and the surface coverage rate is 100 percent; the outlet face 5 layer is a 12 μm pore size nonwoven fabric with a polymer content of 20% and a surface coverage of 100%.
8. A method of preparing a platelet leukocyte-removing filter membrane according to any one of claims 1 to 7, comprising the steps of:
s1) soaking and cleaning the PBT base material non-woven fabric;
s2) adding vinyl acetate and vinyl pyrrolidone into an acetone solvent, stirring and reacting in a constant-temperature water bath under the protection of nitrogen to obtain viscous liquid, extracting with n-hexane, dissolving the bottom liquid in acetone, pouring the solution into water to precipitate a polymer, dissolving with acetone for multiple times, precipitating with water to remove unreacted monomers, drying and drying to obtain the polymer;
s3) dissolving the polymer in an acetone solution, immersing the grafted non-woven fabric in the acetone solution, airing, drying and drying to obtain the filter membrane.
9. The method for producing a platelet leukocyte-removing filter membrane according to claim 8, wherein a weight ratio of vinylpyrrolidone to vinyl acetate in step S2 is 1: 6-1: 9, the step S2 is stirred and reacted for 72 hours under the protection of nitrogen at the constant temperature water bath of 54 ℃ to obtain viscous liquid.
10. The method of preparing a platelet leukocyte-removing filter membrane according to claim 8, wherein the percentage concentration of the polymer in the acetone solution of the step S3 is 4 wt%, and the step S3 comprises immersing the grafted nonwoven fabric in the acetone solution containing the polymer for 1 hour, drying the soaked grafted nonwoven fabric, and drying the dried grafted nonwoven fabric at 80 ℃ to obtain a solution containing azobisisobutyronitrile of 2.95g/L in the acetone solution of the step S3.
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| CN115245709A (en) * | 2022-07-28 | 2022-10-28 | 中国医学科学院输血研究所 | Filter material for selectively removing leukocytes and simultaneously retaining platelets and preparation method thereof |
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