Medical macromolecular microsphere adsorbent for a blood perfusion apparatus and a preparation method thereof

Abstract

A medical macromolecular microsphere adsorbent for a blood perfusion apparatus and a preparation method thereof are provided. The polystyrene-divinylbenzene microspheres are graded by different pore sizes and specific surface areas, medically purified, and grafted by a bioactivity-controlled grafting technology. In the microsphere adsorbent, the volume ratio of microspheres with pore sizes of 1-10 m, 10-100 m and 100 m is 1:(1-10):(1-20), the content of residual monomers in the microsphere has O.D.sub.190-400 nm0.03. The microsphere adsorbent not only can adsorb harmful micromolecules in blood but also can effectively adsorb harmful medium-molecules and macromolecules in blood, thereby meeting clinical application demands.

Claims

1. A medical macromolecular microsphere adsorbent for a blood perfusion apparatus, wherein the microsphere adsorbent consists of three polystyrene-divinylbenzene microspheres having diameters being 1-10 m, 10-100 m and 100-500 m respectively in a volume ratio of 1:(1-10):(1-20); the surface of the microsphere adsorbent is grafted with vinyl pyrrolidone and/or 2-methylacryloxy ethyl phosphorylcholline; the content of residual monomers in the microsphere adsorbent has O.D190-400 nm0.03.

2. The medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 1, wherein the surface area of the microsphere adsorbent is 400 m.sup.2/g-1800 m.sup.2/g.

3. The medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 1, wherein a raw material of the polystyrene-divinylbenzene macromolecular microsphere meets regulations of FDA, the mass content of divinylbenzene is larger than 79%, and the polystyrene-divinylbenzene macromolecular microsphere can bear a heavy load of 300-450 g and has a suspension double-bond content of 2.2-3.1 mmol/g.

4. A method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 1, characterized by comprising the following steps: 1) grading of a pore size and a specific surface area of a microsphere adsorbent: separating polystyrene-divinylbenzene macromolecular microspheres according to diameters being 1-10 m, 10-100 m and 100-500 m respectively; and mixing the microspheres within three size ranges in a volume ratio of 1:(1-10):(1-20); 2) medical purification of the microsphere adsorbent: washing the microsphere adsorbent obtained in step 1) utilizing hydrochloric acid, ammonium hydroxide and ethanol water in turn, and then carrying out rinsing combined treatment and gradient elution on the microsphere adsorbent; 3) grafting a bioactive aglucon on the surface of the microsphere adsorbent utilizing a bioactivity-controlled grafting technology: immersing the microsphere adsorbent purified in step 2) into a bioactive aglucon solution for 1-24 h, the bioactive aglucon being polyvinylpyrrolidone and/or 2-methylacryloxy ethyl phosphorylcholine; and then grafting the microsphere under ultraviolet light after being taken out, wherein, the intensity of light is 50 uw/cm2-150 uw/cm2, and grafting time is 1-4 h; and 4) posttreatment of the microsphere adsorbent: immersing the grafted microsphere for 1-3 h utilizing buffer solution after being taken out, and washing with deionized water; carrying out freeze drying to obtain the medical macromolecular microsphere adsorbent for the blood perfusion apparatus.

5. The method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 4, wherein the mass concentration of hydrochloric acid is 1%-5%; the mass concentration of ammonium hydroxide is 5%-10%; the mass concentration of ethanol water is 10-20%.

6. The method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 4, wherein the mass concentration of the bioactive aglucon solution is 1%-10%.

7. The method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 4, wherein the ultraviolet light is distanced from the to-be-treated solution by 1 m.

8. The method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 4, wherein the freeze drying is carried out in a freeze drying machine, and the freeze drying time is 12-24 h.

9. The method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 4, wherein the buffer solution is phosphate buffer saline (PBS).

10. The method for preparing for a medical macromolecular microsphere adsorbent for a blood perfusion apparatus according to claim 4, wherein the number of times of washing with deionized water is 3-10 times.

Description

DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is an infrared test diagram before and after the microsphere is grafted with NVP and MPC according to embodiment 1.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

(2) To better understanding the disclosure, the disclosure will be further described in combination with drawings and embodiments below, but, the embodiments of the disclosure are not limited thereto.

Embodiment 1

(3) A polystyrene-divinylbenzene macromolecular microsphere meeting regulations of FDA and containing divinylbenzene whose mass content is larger than 79% is selected as a raw material, and the microsphere can bear a heavy load of 300-450 g and has a suspension double-bond content of 2.2-3.1 mmol/g.

(4) Step 1: Grading of a Pore Size and a Specific Surface Area of the Microsphere Adsorbent

(5) (1) Microspheres within three size ranges of 1-10 m, 10-100 m and 100-500 m are mixed in a volume ratio of 1:8:10, and after mixing, the specific surface area is about 951.8 m2/g.

(6) Step 2: Medical Purification of the Microsphere Adsorbent: The Following Washings are Carried Out in Turn

(7) (1) The microsphere adsorbent is washed for 2 min utilizing hydrochloric acid having a mass concentration of 2%;

(8) (2) The microsphere adsorbent is washed for 9 min utilizing ammonium hydroxide having a mass concentration of 5%;

(9) (3) The microsphere adsorbent is washed for 2 min utilizing ethanol having a mass concentration of 20%;

(10) (4) The microsphere adsorbent is subjected to rinsing combined treatment and gradient elution.

(11) By detection, the washed microsphere has O.D190-400 nm=0.03

(12) Step 3: Grafting a Bioactive Aglucon on the Surface of the Microsphere Adsorbent Utilizing a Bioactivity-Controlled Grafting Technology.

(13) (1) The microspheres are immersed into a bioactive aglucon solution containing vinyl pyrrolidone (NVP) having a mass concentration of 6% and 2-methylacryloxy ethyl phosphorylcholine (MPC) having a mass concentration of 7% for 16 h;

(14) The structural formula of NVP is:

(15) ##STR00001##

(16) The structural formula of MPC is:

(17) ##STR00002##

(18) (2) The microspheres are grafted under ultraviolet light after being taken out, wherein, the intensity of the light is 80 uw/cm2 (a position distanced from 1 m), and grafting time is 3 h.

(19) Step 4: Posttreatment of the Microsphere Adsorbent

(20) (1) The grafted microspheres are soaked for 3 h utilizing PBS buffer solution after being taken out, and washed 10 times with deionized water;

(21) (2) The microspheres are placed in a freeze drying machine to carry out freeze drying for 18 h, and finally, a novel microsphere adsorbent for a blood perfusion apparatus is obtained.

(22) Test result: an infrared test result before and after the microspheres are grafted is as shown in FIG. 1; specifically, FIG. 1 is a test result diagram obtained by carrying out infrared spectroscopy analysis (a sample is prepared using a tabletting method) on the microsphere sample before and after modification, wherein, PS-DVB represents the polystyrene-divinylbenzene macromolecule microsphere; NVP represents vinyl pyrrolidone; MPC represents 2-methylacryloxy ethyl phosphorylcholine.

(23) In order to illustrate the removal efficiency of the microspheres on toxins in blood, and through this embodiment, removal rates of the modified microspheres on micromolecular toxins creatinine and pentobarbital sodium and medium-molecular toxins B12 and 2 microglobulin are as shown Table 1.

(24) TABLE-US-00001 TABLE 1 Removal Rates of Microspheres on Toxins Removal rate % Microsphere Pentobarbital Vitamin 2 proportion Creatinine sodium B12 microglobulin 1:8:10 32.15 85.18 95.23 86.31

(25) In Table 1, an initial concentration of creatinine is 35 mg/L, an initial concentration of pentobarbital sodium is 80 mg/L, an initial concentration of vitamin B12 is 25 mg/L, and an initial concentration of 2 microglobulin is 1 mg/mL. Table 1 illustrates the adsorbent prepared in this embodiment not only can adsorb harmful micromolecules in blood but also can effectively adsorb harmful medium-molecules and macromolecules in blood so as to meet clinical application demands. At present, application of polystyrene-divinylbenzene microspheres existing on the market to blood perfusion needs to be combined with hematodialysis so as to achieve effective removal of micromolecules and medium-molecules (Sun Tao, comparison of different blood purification manners on toxin removal and dialysis sufficiency [D]; Jilin University; 2009). The microsphere prepared by the disclosure can be applied to blood perfusion, which can be separately used to effectively removing both of micromolecules and medium molecules.

(26) A hemolysis rate represents a destruction degree of a material on red cells in blood when a material applied to biomedicine contacts with blood, and is expressed by measuring degrees of red cell dissolution and hemoglobin dissociation. Test results of hemolysis rates before and after the microspheres are modified are as shown in Table 2. Table 2 lists results of a test carried out according to regulations of International standard GB/T16886.4-2003, and the hemolysis rate is required to be less than 5%.

(27) TABLE-US-00002 TABLE 2 Results of Microsphere Hemolysis Rate Samples Average Hemolysis Sample 1 Sample 2 Sample 3 value rate (%) Embodiment 0.024 0.022 0.023 0.023 0.82 0.10 1 Negative 0.016 0.015 0.017 0.016 control Positive 0.882 0.842 0.875 0.866 control

(28) Hemolysis rate (HR) is one of extremely important indexes for blood compatibility of a material, and is an important in-vitro coarse screening experiment. A national hemolytic experiment standard of a biomedical material contacting with blood is that a material has a standard hemolysis rate (HR) of <5%. The test illustrates the result of the hemolysis rate of the material prepared by the disclosure is basically similar to that of the product on the market, and achieves the national standard.

Embodiment 2

(29) A polystyrene-divinylbenzene macromolecular microsphere meeting regulations of FDA and containing divinylbenzene whose mass content is larger than 79% is selected as a raw material, and the microsphere can bear a heavy load of 300-450 g and has a suspension double-bond content of 2.2-3.1 mmol/g.

(30) Step 1: Grading of a Pore Size and a Specific Surface Area of the Microsphere Adsorbent

(31) (1) Microspheres within three size ranges of 1-10 m, 10-100 m and 100-500 m are mixed in a volume ratio of 1:8:18, and after mixing, the specific surface area is about 578.5 m2/g.

(32) Step 2: Medical Purification of the Microsphere Adsorbent

(33) (1) The microsphere adsorbent is washed for 1 min utilizing hydrochloric acid having a mass concentration of 5%;

(34) (2) The microsphere adsorbent is washed for 8 min utilizing ammonium hydroxide having a mass concentration of 10%;

(35) (3) The microsphere adsorbent is washed for 19 min utilizing ethanol having a mass concentration of 10%;

(36) (4) The microsphere adsorbent is subjected to rinsing combined treatment and gradient elution.

(37) After washing, O.D190-400 nm=0.025.

(38) Step 3: A Bioactive Aglucon is Grafted on the Surface of the Microsphere Adsorbent Utilizing a Bioactivity-Controlled Grafting Technology.

(39) (1) The microspheres are immersed into a bioactive aglucon solution containing vinyl pyrrolidone (NVP) having a mass concentration of 7% and 2-methylacryloxy ethyl phosphorylcholine (MPC) having a mass concentration of 2% for 12 h;

(40) (2) The microspheres are grafted under ultraviolet light after being taken out, wherein, the intensity of the light is 100 uw/cm2 (a position distanced from 1 m), and grafting time is 1 h.

(41) Step 4: Posttreatment of the Microsphere Adsorbent

(42) (1) The grafted microspheres are soaked for 1 h utilizing PBS buffer solution by being taken out, and washed 8 times with deionized water;

(43) (2) The microspheres are placed in a freeze drying machine to carry out freeze drying for 20 h, and finally, a novel microsphere adsorbent for a blood perfusion apparatus is obtained.

Embodiment 3

(44) A polystyrene-divinylbenzene macromolecular microsphere meeting regulations of FDA and containing divinylbenzene whose mass content is larger than 79% is selected as a raw material, and the microsphere can bear a heavy load of 300-450 g and has a suspension double-bond content of 2.2-3.1 mmol/g.

(45) Step 1: Grading of a Pore Size and a Specific Surface Area of the Microsphere Adsorbent

(46) (1) Microspheres within three size ranges of 1-10 m, 10-100 m and 100-500 m are mixed in a volume ratio of 1:5:12, and after mixing, the specific surface area is about 732.5 m2/g.

(47) Step 2: Medical Purification of the Microsphere Adsorbent

(48) (1) The microsphere adsorbent is washed for 10 min utilizing hydrochloric acid having a mass concentration of 1%;

(49) (2) The microsphere adsorbent is washed for 10 min utilizing ammonium hydroxide having a mass concentration of 7%;

(50) (3) The microsphere adsorbent is washed for 20 min utilizing ethanol having a mass concentration of 12%;

(51) (4) The microsphere adsorbent is subjected to rinsing combined treatment and gradient elution.

(52) After washing, O.D.sub.190-400 nm=0.01

(53) Step 3: A Bioactive Aglucon is Grafted on the Surface of the Microsphere Adsorbent Utilizing a Bioactivity-Controlled Grafting Technology.

(54) (1) The microspheres are immersed into a bioactive aglucon solution containing vinyl pyrrolidone (NVP) having a mass concentration of 3% and 2-methylacryloxy ethyl phosphorylcholine (MPC) having a mass concentration of 2% for 24 h;

(55) (2) The microspheres are grafted under ultraviolet light after being taken out, wherein, the intensity of the light is 150 uw/cm2 (a position distanced from 1 m), and grafting time is 4 h.

(56) Step 4: Posttreatment of the Microsphere Adsorbent

(57) (1) The grafted microspheres are soaked for 3 h utilizing PBS buffer solution after being taken out, and washed 5 times with deionized water;

(58) (2) The microspheres are placed in a freeze drying machine to carry out freeze drying for 14 h, and finally, a novel microsphere adsorbent for a blood perfusion apparatus is obtained.

(59) The test results of removal effects of novel medical macromolecular microsphere adsorbents for the blood perfusion apparatus obtained in embodiment 2 and embodiment 3 on micromolecular toxins creatinine and pentobarbital sodium and medium-molecular toxins vitamin B12 and 2 microglobulin are as shown in Table 3.

(60) TABLE-US-00003 TABLE 3 Removal rate % Pentobarbital 2 Embodiments Creatinine sodium Vitamin B12 microglobulin Embodiment 2 91 52 25 34 Embodiment 3 13 20 29 62