Evaluation Of Specificity To Capture Hemopoietic Stem Cells By CD133 Antibody-coated Stent

Cardiovascular and cerebrovascular diseases are the biggest killers against human life and health now, and the most rapid and effective treatment is stent implantation, which has saved countless lives of patients with such diseases. However, the first generation stent bare-metal stent prones to cause rejection and leads the rate of restenosis to reach above 25%. Although the second generation stent drug-coating stent can inhibit rejection effectively, it causes long-term restenosis and other adverse effects, such as hypersensitive response and intimal hyperplasia. The third generation stent bio-engineering stent captures repair cells in peripheral blood onto the surface of stent in order to accelerate the natural repair of blood vessels.Stent coating and whether the antibody can capture haemopoietic stem cells(HSC) specially are key problems of the third generation biological engineering stent. Analytical method for the quantification of antibody loaded on CD133 antibody-coated stent was established and validated. Influential factors to the preparation of CD 133 antibody-coated stent with electrostatic self-assembled method were investigated in this thesis. With flourescence immunifaction and scanning electron microscope(SEM), the compatibility of chitosan(CH)/hyaluronic acid(HA) base coating, the specificity of capturing HSC by CD133 antibody-coated stent in peripheral blood and the differentiation results of stem cells captured were evaluated.Te results showed that the quantities of antibody loaded on per 316L stainless steel coronary artery stent and on per Ni-Ti alloy renal artery stent are 50±15ng and 150±20ng respectively; the biological activity of the antibody kept unchanged after CD133 antibody-coated stent was stored at 4℃for half a year; the surface of the base coating was still compact after incubating in the blood for 1 hour and no platelet or other types of cells were found. CD133 antibody-coating only captured bird nest like cells in peripheral blood, which could neither be dyed by Hoechst33342 nor expressed CD20, CD7 and CD61 antigen and they could differentiate into endothelial progenitor cells(EPCs) under the induction of low molecular weight HA; CD34 antibody-coating captured bird nest like cells, spindle like cells and tricholith like cells in peripheral blood, which could be dyed by Hoechst33342 and expressed CD20, CD7, CD61 antigen. VEGFR-2 antibody-coating captured bird nest like cells, spindle like cells, pebble like cells with pseudopodium, CD271 antibody-coating captured mulberry like cells, folding scabbard like cells and sternzellen, which could neither be dyed by Hoechst33342 nor expressed CD20, CD7 and CD61 antigen.The results indicated that the long-term stability of CD133 antibody-coating and the compatibility of CH/HA base coating were excellent; CD133 antibody-coating only captured HSC, which could differentiate into EPCs directionally under the induction of low molecular weight HA, repairing injured blood vessels. CD34 antibody-coating and VEGFR-2 antibody-coating could capture stem cells and haemopoietic progenitor cells as well. CD271 antibody-coating only captured mesenchyma stem cells rather than haemopoietic stem/progenitor cellsIn conclusion, stent with CD 133 antibody-coating loading on CH/HA base coating with electrostatic self-assembled method is a kind of stent which can capture HSC specially and is superior to commercially available CD34 antibody-coating stent, providing a new method for healing cardiovascular and cerebrovascular diseases more effectively.