Stainless steel surface functionalization for immobilization of antibody fragments for cardiovascular applications

Abstract

Stainless steel 316 L material is commonly used for the production of coronary and peripheral vessel stents. Effective biofunctionalization is a key to improving the performance and safety of the stents after implantation. This paper reports the method for the immobilization of recombinant antibody fragments (scFv) on stainless steel 316 L to facilitate human endothelial progenitor cell (EPC) growth and thus improve cell viability of the implanted stents for cardiovascular applications. The modification of stent surface was conducted in three steps. First the stent surface was coated with titania based coating to increase the density of hydroxyl groups for successful silanization. Then silanization with 3 aminopropyltriethoxysilane (APTS) was performed to provide the surface with amine groups which presence was verified using FTIR, XPS, and fluorescence microscopy. The maximum density of amine groups (4.8*10(-5) mol/cm(2)) on the surface was reached after reaction taking place in ethanol for 1 h at 60 °C and 0.04M APTS. On such prepared surface the glycosylated scFv were subsequently successfully immobilized. The influence of oxidation of scFv glycan moieties and the temperature on scFv coating were investigated. The fluorescence and confocal microscopy study indicated that the densest and most uniformly coated surface with scFv was obtained at 37 °C after oxidation of glycan chain. The results demonstrate that the scFv cannot be efficiently immobilized without prior aminosilanization of the surface. The effect of the chemical modification on the cell viability of EPC line 55.1 (HucPEC-55.1) was performed indicating that the modifications to the 316 L stainless steel are non-toxic to EPCs.

A promising biodegradable magnesium alloy suitable for clinical vascular stent application

“…Moreover, polymer stents are also associated with an increased risk of late stent thrombosis (ST)5. The conventional metallic implants based on stainless steel678, Co-Cr alloy91011, Ni-Ti alloy121314 and Ta15 are the commonly used materials for coronary artery reconstruction because of their high mechanical properties and excellent corrosion resistance. However, the long-term host foreign body response (FBR) to permanent stent implantation often results in restenosis or lumen encroachment, brought about by the recruitment of inflammatory and smooth muscle cells to the intima with subsequent proliferation and extracellular matrix (ECM) deposition1617.…”

Surface treatments and functionalization of metal‐ceramic membranes for improved enzyme immobilization performance

“…However, efficient functionalization of stainless steel with APTES or PEI also often requires introduction of hydroxyl groups on its surface, e.g. by etching or by coating with a titania‐based material . Both support functionalization methods were investigated in the current work and compared to the simplest enzyme immobilization method, physical adsorption (Table ).…”

“…by etching or by coating with a titania-based material. 39,41,42 Both support functionalization methods were investigated in the current work and compared to the simplest enzyme immobilization method, physical adsorption ( Table 4). As established earlier, the initial water permeability of the C-Y0.25M membranes was 57% of the water permeability of the heat-treated membranes, indicating that the Y 2 O 3 -coating reduced the open porosity and pore-neck size resulting in a lower water flux (Table 4).…”

Functionalization with a VEGFR2‐binding antibody fragment leads to enhanced endothelialization of a cardiovascular stent in vitro and in vivo

“…Single‐chain Fragment variable (scFv) antibody fragment 32 (binding VEGFR2) and scFv 4M5.3 (binding fluorescein isothiocyanate [FITC]) were each cloned with C‐terminal glycosylation motifs into the expression vector pIG6 and cotransformed into Escherichia coli CLM37 with the pACYC pgl vector for protein glycosylation. Expression in the E. coli periplasm was carried out using autoinduction in ZYP‐5052 medium for 24–48 h at 25°C with appropriate antibiotics.…”