Abstract
Cytapheresis is an extracorporeal separation technique widely used in medicine for elimination of specific classes of blood cells from circulating blood. It has been shown recently to have clinical efficacy in various disease states, such as leukaemia, autoimmune disorders, rheumatoid arthritis, renal allograft rejection and sickle–cell anaemia. The current study was undertaken to produce an affinity-binding column, based upon a macroporous monolithic cryogel with a structure of interconnected pores, with pore size and low flow resistance potentially suitable for use in cytapheresis. The affinity column was produced from poly (2-hydroxyethyl methacrylate) PHEMA cryogels synthesized by free radical polymerization at -12°C. This study involved assessing haemolytic potential, and functionalisation of polymer matrix with biological ligands. Haemolytic potential of poly (2-hydroxyethyl methacrylate) cryogel was established by measuring free haemoglobin after blood filtration through the column. The anti-human albumin (antibody) was chemically coupled to the epoxy derivatised monolithic cryogels and the binding efficiency of anti-human albumin (antibody) to the cryogel was determined. Our results show that approximately 100% of Red blood cells passed through the column with no evidence of haemolysis found in blood eluted. It was found that ~82% of human serum albumin was retained on the monolithic IgG anti-human albumin cryogel matrix. The obtained results suggest that poly (2-hydroxyethyl methacrylate) monolithic cryogel is a non-haemolytic material (haemocompatible matrix) capable of functionalisation with antibody and thus can be an appropriate matrix for use in extracorporeal apheresis system.
Highlights
Cytapheresis is aimed at separation of certain types of cells from blood by either centrifugation or adsorption
Considering the fact that the development of improved haemocompatible biomaterials is one of the most important challenges in material science, this paper demonstrated for the first time that PHEMA monolithic cryogel is a non-haemolytic material by passing whole blood through the column by mean of evaluating the free haemoglobin, and amount of red blood cells eluted with the use of Sysmex cell counter and Blakney and Dinewoodie (1975) method
The in-vitro studies on the haemocompatibility of the material produced in this paper reported that ~100% (4.77 x 106 cells/μL) of Red blood cells was eluted after passing whole blood through the monolithic cryogel column and that no haemolysis was observed
Summary
Cytapheresis is aimed at separation of certain types of cells from blood by either centrifugation or adsorption. Various physicians have reported the efficacy of cytapheresis in inflammatory bowel disease such as ulcerative colitis and Crohn’s disease [3,4,5,6,7,8]. There are mainly three methods of extracorporeal leukocyte removal therapy in use in the clinical field. These are the centrifugal method, filtration and the adsorptive method using fibre or beads. Leukocytapheresis using the leukocyte filter Cellsorba [11] and granulocytapheresis using the Adacolumn [12] have been proved to have reduced leukocyte load in patients with rheumatoid arthritis and inflammatory bowel disease, but still has major limitations of specificity and selectivity [13]
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