Preparation And Biological Evaluation Of Fibrous Carrier Coated With Type ? Collagen In Hepatocyte Culture

Chapter ? Preparation and characterization of fibrous carrier coated with type ? collagenObjective:Preparing a suitable carrier,provides a favorable environment for hepatocyte growing and maintains high growth density and function of hepatocyte,is the key to study bioartificial liver.The purpose of this chapter is to prepare a type of fibrous carrier coated with type I collagen for hepatocyte growth.Methods:1.The surface of PP fibers were hydroxylated using "Piranha" solution.Process parameters including temperature,time and concentration were used for orthogonal test.The best process parameters were determined after samples charactering.2.The surface of hydroxylated PP fibers were aminated using amino silane solution.Process parameters including temperature,time,concentration and type of amino silane solution were used for orthogonal test.The best process parameters were determined after samples charactering.3.The surface of aminated PP fibers were cross-linked using glutaraldehyde solution.Process parameters including temperature,time and concentration were used for orthogonal test.The best process parameters were determined after samples charactering.4.The surface of cross-linked PP fibers were cross-linked using type ? collagen.Process parameters including temperature,time and concentration were used for orthogonal test.The best process parameters were determined after samples charactering.Results:1.The hydroxyl group was successfully introduced on the surface of PP fibers using "Piranha" solution.The best process parameters are temperature:90?,time:60min,"Piranha":H2SO4/30%H2O2=3/1.2.The amino group was successfully introduced on the surface of hydroxylated PP fibers using amino silane solution.The best process parameters are APTES,temperature:60?,time:60min,concentration:10%.3.The aldehyde group was successfully introduced on the surface of aminated PP fibers using glutaraldehyde solution.The best process parameters are temperature:20?,time:120min,concentration:5%.4.The type I collagen was successfully introduced on the surface of cross-linked PP fibers using type I collagen.The best process parameters are temperature:30?,time:24h,concentration:2.5%.Conclusion:An optimum process of coating type I collagen on PP fibers was established.The PP fibrous scaffold firmly coated with I collagen was successfully prepared.Chapter ? Biological evaluation of fibrous carrier coated with type ? collagenObjective:The biological evaluation of the fibrous carrier coated with type ?collagen(PP-OKGC)provides a basis of safety and functionality for further applications.Methods:1.Extracts of PP-OKGC fiber material were extracted and subjected to glutaraldehyde residuals,cytotoxicity,hemolytic reaction,skin irritation reaction,intracutaneous irritant reaction,acute and subacute systemic toxicity test respectively.Samples were tested using HPLC,MTT,spectrophotometer and histological sections respectively to evaluate the biocompatibility of PP-OKGC.2.HepG2 cells were planted on PP-OKGC and untreated PP fibers(PP-N).The fibrous carriers and culture fluid were extracted on the 1st,3rd,5th and 7th days.The adhesion,proliferation,cell viability,cell morphology and cell function of HepG2 cultured on PP-OKC and PP-N fibrous carriers were observed to evaluate the biological activity of PP-OKGC.Results:1.PP-OKGC materials has good biocompatibility.2.The HepG2 cells cultured on PP-OKGC fibrous carriers maintain normal cell morphology.The cell adhesion,proliferation,cell activity and cell function of HepG2 in PP-OKGC group are better than PP-N group.Conclusion:The fibrous carrier coated with type I collagen has good biocompatibility and biological activity.