Synthetic ePTFE Grafts Coated with an Anti-CD133 Antibody-Functionalized Heparin/Collagen Multilayer with Rapid in vivo Endothelialization Properties

Abstract

An anti-CD133 antibody multilayer functionalized by heparin/collagen on an expanded polytetrafluoroethylene (ePTFE) graft was developed to accelerate early endothelialization. The surface modification of ePTFE grafts demonstrated that the multilayer is stable in static incubation and shaking conditions and that the anti-CD133 antibodies were successfully cross-linked onto the surface. Blood compatibility tests revealed that the coimmobilized heparin/collagen films in the presence or absence of anti-CD133 antibodies prolonged the blood coagulation time and that there was less platelet activation and aggregation, whereas the hemolysis rate was comparable with the bare ePTFE grafts. Cellular proliferation was not inhibited, as the heparin/collagen synthetic vascular grafts coated with CD133 antibody showed little cytotoxicity. The endothelial cells adhered well to the modified ePTFE grafts during a cell adhesion assay. A porcine carotid artery transplantation model was used to evaluate the modified ePTFE grafts in vivo. The results of histopathological staining and scanning electron microscopy indicated that the anti-CD133 antibody was able to accelerate the attachment of vascular endothelial cells onto the ePTFE grafts, resulting in early rapid endothelialization. The success of the anti-CD133 antibody-functionalized heparin/collagen multilayer will provide an effective selection system for the surface modification of synthetic vascular grafts and improve their use in clinical applications.

Vascular Grafting Strategies in Coronary Intervention

“…Consequently, research has also focused on improving endothelial cell growth, retention, and confluency on the luminal surface of synthetic grafts. Due to electrostatic and hydrophobic interactions with ePTFE vascular grafts, incomplete endothelialization is typically encountered (Lu et al, 2013). Williams et al have recently studied the effect of conjugated mouse laminin type 1 on endothelialization and neovascularization of ePTFE grafts as interpositional aortic grafts in rats.…”

“…In addition to the in situ endothelialization, the authors reported significantly decreased platelet adhesion in the heparin/collagen coated ePTFE grafts. Despite the promise of the reported work, future studies will need to address two issues -appropriately sized ePTFE grafts (<6 mm) and longer in vivo studies to ensure no late stage thrombosis occurs due to a loss of functional endothelium (Lu et al, 2013).…”

Desenvolvimento e fabricação de filmes ultra-­finos, obtidos pela técnica layer-by-layer, para aplicações na entrega direcionada de fármacos e na captura seletiva de bio-­marcadores

“…In the future, other approaches as applying external stimuli, such as ultrasound, magnetic field, and light irradiation, can be investigated in films containing vesicles, in order to trigger controlled drug delivery (VOLODKIN et al, 2009 DEROCHER et al, 2012). Such coatings have been developed for substrates ranging from standard solid surfaces to highly porous membranes (HARRIS, STAIR, BRUENING, 2000;MANNARINO et al, 2013), degradable scaffolds (LU et al, 2008;NEAL et al, 2013), microneedles (SAURER et al, 2010;DEMUTH et al, 2012), carbon nanotubes (CARRILLO et al, 2003;ARTYUKHIN et al, 2004;KANG et al, 2014), and microfluidic devices (WANG, XU, CHEN, 2006;KUMLANGDUDSANA et al, 2012;LEE, AHN, 2013 (LIU et al, 2007;BRUENING et al, 2008), and especially conjugation with antibodies (KOMATSU, 2012). Such biofunctional coatings have already seen applications in promoting endothelialization and cell growth (LIN et al, 2010;LU et al, 2013), the capture and detection of circulating tumor cells (WEN et al, 2014), and to protein--based nanotubes for the selective capture of nanoparticles, proteins and viruses (KOMATSU, 2012).…”

“…Such coatings have been developed for substrates ranging from standard solid surfaces to highly porous membranes (HARRIS, STAIR, BRUENING, 2000;MANNARINO et al, 2013), degradable scaffolds (LU et al, 2008;NEAL et al, 2013), microneedles (SAURER et al, 2010;DEMUTH et al, 2012), carbon nanotubes (CARRILLO et al, 2003;ARTYUKHIN et al, 2004;KANG et al, 2014), and microfluidic devices (WANG, XU, CHEN, 2006;KUMLANGDUDSANA et al, 2012;LEE, AHN, 2013 (LIU et al, 2007;BRUENING et al, 2008), and especially conjugation with antibodies (KOMATSU, 2012). Such biofunctional coatings have already seen applications in promoting endothelialization and cell growth (LIN et al, 2010;LU et al, 2013), the capture and detection of circulating tumor cells (WEN et al, 2014), and to protein--based nanotubes for the selective capture of nanoparticles, proteins and viruses (KOMATSU, 2012). The detection of dilute protein biomarkers can facilitate early disease diagnosis and consequently treatment, thereby improving patient survival (JUSTINO, ROCHA--SANTOS, DUARTE, 2013).…”

“…PSA is a blood biomarker secreted by normal and tumor cells (AMIRRASOULI et al, 2010;GREENE et al, 2013) with its concentration of ~ 4 ng/mL in serum (GREENE et al, 2013) having been attributed to the presence of prostate cancer. However, the detection of PSA alone is not decisive for cancer diagnose (BOTCHORISHVILI, MATIKAINEN, LILJA, 2009), nor can it be correlated with tumor size (LUTZ et al, 2008 which usually uses an EDC--NHS amidification to anchor specific antibodies to materials surfaces (SARDESAI et al, 2013;POON, CHAN, LI, 2014). Such devices have been recently investigated in the capture of important biomarkers like interleukin--6 and prostate specific antigen (MALHOTRA et al, 2012;SARDESAI et al, 2013).…”