The Preparation Of A Novel Matrix Modified Filter Membrane And The Measurement Of Its Anticoagulant Properties

Objective:In this study, polyethersulfone (PES) and argatroban (Arg) are used as raw materials, we use the method of matrix modifying and immersion precipitation phase transformation to construct a novel type of matrix modified filter membrane with anticoagulant properties.Methods:(1)Firstly, covalent grafting the argatroban modify the polyethersulfone material successively by acetylation, oxidation and amidation reaction, and then alloying modify the polyethersulfone material and modified polyethersulfone according to different scheme, thirdly, preparing the polyethersulfone membrane (PES), matrix modified filter membrane without argatroban (PES/CPES) and novel type of matrix modified filter membrane with argatroban (Arg-PES/CPES) by immersion precipitation phase transformation method.(2) Synchronous representation the modifying process of novel type of matrix modified filter membrane by Fourier infrared spectra analysis, nuclear magnetic resonance spectroscopy and scanning electron microscope. (3) In this study, healthy normal plasma is the negative control group (Control group), and plasma containing Arg is the positive control group (Arg group). We use automatic coagulation detector to detect the activated partial thromboplastin time, prothrombin time and thrombin time of the novel type of matrix modified membrane, Observe and evaluate their coagulant properties.Results:1. Track and detect the preparation process of novel type of matrix modified membrane materials:1) Fourier infrared spectra (FTIR) analysis:①After acetylation reaction, from the FTIR of PES-COCH3, wave number at1678.45cm-1was attributed to the stretching vibration peak of C=O of acetyl group (-COCH3). It suggests-COCH3has been successfully introducted to PES.②After oxidation reaction, from the FTIR of PES-COOH, wave number at3377.45cm-1was attributed to the stretching vibration peak of O-H of carboxy group (-COOH). It suggests-COOH has been successfully grafted.③After amidation reaction, from the FTIR of Arg-PES, wave number at1653.92cm-1was attributed to the stretching vibration peak of C=O of-CONH-. It suggests the carboxyl group of PES-COOH and amino group of Arg has reacted, and Arg has successfully grafted to PES.2) nuclear magnetic resonance (NMR) spectroscopy:①After acetylation reaction, from the1HNMR spectra of PES-COCH3, chemical shift at2.39was attributed to the vibration peak of H of-COCH3. It suggests-COCH3has been successfully introducted to PES, the grafted-ratio is13.13%.②After oxidation reaction, from the’HNMR spectra of PES-COOH, chemical shift at10.69was attributed to the vibration peak of H of-COOH. After adding10μl D2O, the1HNMR spectrum signal disappears. It suggests that-COOH has been successfully grafted, the grafted-ratio is7.19%.③After amidation reaction, from the’HNMR spectra of Arg-PES, chemical shift between0.4and3.5was attributed to the vibration peak of H of-NH2or-NH. It suggests that Arg has successfully grafted to PES.2. Using scanning electron microscope (SEM) to observe the surface and cross section shape of membranes:The polyethersulfone membrane and the matrix modified filter membrane both have the asymmetry morphology characteristics which make up thinner cortex and looser supporting layer. There are a lot of finger holes in the membranes, and the finger pore structure of novel type of matrix modified filter membrane is more stability, widely distribute different sizes of aperture between fingers. The membrane has met the basic form requirements of filter membrane.3. Anticoagulant properties:1) polyethersulfone membrane (PES membrane)Compared with the control group, the PT and TT value of PES membrane were shorter (P<0.05), suggesting the PES membrane can cause clotting.2) Matrix modified filter membrane (PES/CPES membrane (2:1)):①Compared with the group of control group and PES group, PES/CPES membrane (2:1) group’s APTT and PT values are extended (P<0.05), suggesting matrix modified membrane can inhibit the activation of intrinsic and exogenous coagulation pathway.②Compared with the group of control group, PES/CPES membrane (2:1) group’s TT values have no statistical significance (P>0.05), suggesting matrix modified membrane don’t has antithrombotic effect.③Compared with the group of PES group, PES/CPES membrane (2:1) group’s APTT\PT and TT values are extended (P<0.05), suggesting matrix modification method can inhibit the procoagulant and prothrombotic effect of PES membrane.3) Novel matrix modified filter membrane (PES/Arg-PES membrane(2:1)、PES/Arg-PES membrane (1:1)):①Compared with the group of control group, APTT、PT and TT values of PES/Arg-PES membrane(2:1) group are extended (P<0.05), suggesting novel matrix modified membrane not only can inhibit the activation of intrinsic and exogenous coagulation pathway, but also can inhibit thrombosis.②The APTT, PT and TT values of PES/Arg(1:1) significantly longer than Control group、PES/CPE (2:1) membrane group and PES/Arg-PES (2:1) membrane group (P<0.05), suggesting that when the blending ratio of PES/Arg-PES is1:1, the anticoagulant effect of novel matrix modification membrane is better. Conclusion:(1) In this study, polyethersulfone (PES) and argatroban (Arg) are used as raw materials, we can successfully prepare novel matrix modified polyethersulfone membrane by matrix modification and immersion precipitation phase inversion methods.(2) Novel matrix modified polyethersulfone membrane has good anticoagulant properties, the best blending ratio of PES/Arg-PES is1:1.