Study On The Biocompatibility Of A Novel Bioartificial Liver Reactor Material (Propylene-acidamide Grafted Polypropylene Membrane)

Background and Objective:The premise and background of biomaterials or wares to be used for implantation in vivo or contacting with blood of human body was to detect its reliability, which was the biocompatibility of the biomaterials. A novel bioartifical liver reactor material was manufactured by high polymer institution of Zhejiang Univercity, which was heterogeneous polymer on the interface of polypropylene (propylene-acidamide grafted polypropylene membrane, PP-g-AAm), and was evaluated its biocombatibility by a series experiment according the evaluation criterion and requirement of medical apparatus and instruments which was made by the International Standard Organization (ISO 10993-1:1992), so as to acquire the operative evaluation of toxicology, acute total body toxicity, blood compatibility and immucompatibility of this material and establish basement for the clinical use of it.Methods:1. Establish the PP-g-AAm and its extractsThe polypropylene(PP) membrane was sheared into a pieces of 2cm × 5cm, and was dipped into a solution including 30% hydrogen dioxide. Then was oxidized under ultraviolet for 4 hours(the wave-length was 254nm, the intensity was 5000 μ W/cm and the distance was 20cm). Later, it was dipped into a solutions including 5% propylene-acidamide 10 ml and was filled with nitrogen and excluded with oxygen. Then 1 ml errous sulphate amines were put in to cause the grafted reaction of propylene-acidamide on the PP membrance. The grafted reaction was continued for 30 min with 30℃, and the PP-g-AAm was established.And the extracts of the materials was made by this condition, media(culture medium of RPIM-1640 or saline):surface of sample(cm~2)=10:6, and the condition was 37℃ , 72h±2h.2. Test of toxicologyL929 cells on the logarithm growth period were produced into cell suspension with ten kinds of different concentration, and the standard curve of growth and metabolism of theL929 cells of our lab was established. The number of 6000 cells/well L929 cell was defined as the inoculation number according the growth curve and was inoculated into 96 pore plate. And the extracts of materials, negative culture media and positive culture media were added in. After 24, 48 and 72 hours, 20 μl MTT (5mg/ml) was added in the 96 pores plate. Then after cultured for 4 hours, the solution in the 96 pores plate was give up and added in 150 ul DMSO. The absorption value of every pore was measured at 570 nm wave-length by used a enzyme unite immune meter. Then the cell proliferation rate and the grade of toxicity were calculated.3. Acute test of total body toxicity30 healthy mouse about 20 g and half were male were selected and divided into three groups randomly, their initials body weight was recorded. Mouse of the test group were injected the extracts of the two materials through vein, the dosage was 50 g/kg. And the control group mice were injected with saline. After instant, 4 hours, 24 hours, 48 hours, 72 hours, the general behavior, toxicity manifestation and death number was observed. The change of body weight was observed after 24 hours, 48 hours and 72 hours. The toxicity and death was observed in the following 3 weeks.4. Hemolytic test10 ml of extracts of the two materials, sterilized distilled water and saline were acquired and were in water bath at 37 ℃ for 30 min, then 0.2 ml anti-coagulated diluted fresh blood of rabbit was added in each group. After 60 min in water bath, they were centrifugated for 5 minutes (l000prm). The super suspension was acquired and was measured at 545 nm by use a spectrophotomete.5. Establish PRP and PT, APTT testVenous blood of some healthy male volunteer was drawn and anti-coagulated by using 2 % EDTA-Na. Which was centrifugated for 8 minutes at 800 prms instantly, light yellow plasm at superior layer was sucked, which was platelet rich plasm (PRP). 3 ml PRP from one volunteer was divided into 3 silicon test tubes. 1 cm~2 membrane of the two materials was put into the two tubes respectively and the three tubes were as control group without any material. Incubated on h...