Distinctive effects of CD34- and CD133-specific antibody-coated stents on re-endothelialization and in-stent restenosis at the early phase of vascular injury

Abstract

It is not clear what effects of CD34- and CD133-specific antibody-coated stents have on re-endothelialization and in-stent restenosis (ISR) at the early phase of vascular injury. This study aims at determining the capabilities of different coatings on stents (e.g. gelatin, anti-CD133 and anti-CD34 antibodies) to promote adhesion and proliferation of endothelial progenitor cells (EPCs). The in vitro study revealed that the adhesion force enabled the EPCs coated on glass slides to withstand flow-induced shear stress, so that allowing for the growth of the cells on the slides for 48 h. The in vivo experiment using a rabbit model in which the coated stents with different substrates were implanted showed that anti-CD34 and anti-CD133 antibody-coated stents markedly reduced the intima area and restenosis than bare mental stents (BMS) and gelatin-coated stents. Compared with the anti-CD34 antibody-coated stents, the time of cells adhesion was longer and earlier present in the anti-CD133 antibody-coated stents and anti-CD133 antibody-coated stents have superiority in re-endothelialization and inhibition of ISR. In conclusion, this study demonstrated that anti-CD133 antibody as a stent coating for capturing EPCs is better than anti-CD34 antibody in promoting endothelialization and reducing ISR.

Keywords: CD133; CD34; endothelial progenitor cells; re-endothelialization; stent.

Fig. 1.

Characterization of EPCs. (A) EPCs were isolated from bone marrow for 4 days. (B) Secondary colony formation of EPCs after seeding 7 days. Scale bars = 500 m

Fig. 2.

Identification of EPCs by immunocytochemistry and immunofluorescence. (A) Immunocytochemistry, scale bars = 500 µm. (B) Immunofluorescence, scale bars = 200 µm

Fig. 3.

The in vitro tube formation assay of EPCs. (A) Without coating, scale bars = 500 µm; (a): without coating, scale bars = 200 µm. (B) Anti-CD34 antibody coating, scale bars = 500 µm; (b): anti-CD34 antibody coating, scale bars = 200 µm. (C) Anti-CD133 antibody coating, scale bars = 500 µm; (c): anti-CD133 antibody, scale bars = 200 µm (*P < 0.05; **P < 0.01)

Fig. 4.

Preparation of coating stents. (A) SEM observed the surface of stents before expanded. (B) SEM observed the surface of stents after expanded. BMS, bare metal stent; glutin, gelatin-coated stent; CD34, anti-CD34 antibody-coated stent; CD133, anti-CD133 antibody-coated stent

Fig. 5.

The capability to capture EPCs in vitro. (A) Phase-contrast microscopy observed the surface of stents after static culturing, scale bars = 500 µm. (B) SEM observed the surface of stents after culturing with shear force. (C) The extracorporeal circulation system. (D) Quantification of number of captured cells in the static condition. (E) Quantification of number of captured cells with shearforce (shear rate 2.5 dynes/cm²). The arrow showed cells adhesion on the surface of stents

Fig. 6.

Angiograms obtained immediately after stents implantation. (A) All deployment procedures were successful. (B) All vessels implanted stents were competent. The arrows show the position of stents implantation

Fig. 7.

SEM of stents demonstrated the rate of endothelial coverage after 1 week. (A, a): bare metal stent; (B, b): gelatin-coated stent; (C, c): anti-CD34 antibody-coated stent and (D, d): anti-CD133 antibody-coated stent

Fig. 8.

SEM of stents demonstrated the rate of endothelial coverage after 4 weeks. (A, a): bare metal stent; (B, b): gelatin-coated stent; (C, c): anti-CD34-coated stent and (D, d): anti-CD133-coated stent

Fig. 9.

SEM of stents demonstrated the rate of endothelial coverage after 12 weeks. (A, a): bare metal stent; (B, b): gelatin-coated stent; (C, c): anti-CD34-coated stent and (D, d): anti-CD133-coated stent

Fig. 10.

Results of the tissue slice. Representative sections of arteries at the sites of stent struts demonstrating neointimal growth at 12 weeks and histomorphometric measurements. (A) Bare metal stent; (B) gelatin-coated stent; (C) anti-CD34-coated stent; (D) anti-CD133-coated stent; (E) statistical result of intima area for four kinds of stents; (F) statistical result of lumen area for four kinds of stents; (G) statistical result of intima thickness for four kinds of stents and (H) statistical result of stenosis for four kinds of stents