Song C-L, Li Q, Zhang J-C, Wang J-P, Xue X, Wang G, Shi Y-F, Diao H-Y, Liu B
Department of Cardiology, the Second Hospital of Jilin University, Nanguan District, Changchun, China.
Eur Rev Med Pharmacol Sci. 2016;20(2):311-6.
To access the cytotoxicity and the effect on the endothelial progenitor cell (EPC) differentiation of stainless steel sheets simultaneously coated with VEGF and anti-CD34 antibody.
316L stainless steel sheets (diameter 6 mm, thickness 1 mm) were divided into the D-H (Bare metal), D-(H-V)10 (VEGF-coated metal) and D-(H-V)10-A (VEGF and anti-CD34 antibody co-coated metal) groups. The cytotoxicity effect of the three groups was measured using MTT assay. Percentage of EPC positive for CD34, CD133 and KDR were detected by flow cytometric assay. Endothelial cells positive for CD31 and VE-Cadherin were also detected by flow cytometric assay.
The percentages of isolated cells positive for CD133, CD34 and KDR were 89.9%, 91.3%, and 90.4%, respectively, suggesting that the EPCs were successfully isolated. MTT results showed that the stainless steel sheets coated with VEGF and anti-CD34 antibody have less toxicity on seeded EPCs than single VEGF coating or bare metal. We further found that with VEGF and anti-CD34 antibody co-coating could significantly promote the differentiation of EPCs in vitro when compared with that of single VEGF coating and bare metal.
Our study provided a preliminary evaluation of metallic steel sheet coated with VEGF and anti-CD34 antibody in vitro. Our findings suggest that simultaneously coating the stents with VEGF and anti-CD34 antibody might be a novel research direction for facilitating re-endothelialization in order to reduce ISR after stent implantation.
评估同时包被血管内皮生长因子（VEGF）和抗CD34抗体的不锈钢片的细胞毒性及其对内皮祖细胞（EPC）分化的影响。
将316L不锈钢片（直径6mm，厚度1mm）分为D-H（裸金属）、D-(H-V)10（包被VEGF的金属）和D-(H-V)10-A（包被VEGF和抗CD34抗体的金属）组。采用MTT法检测三组的细胞毒性作用。通过流式细胞术检测CD34、CD133和激酶插入区受体（KDR）阳性的EPC百分比。也通过流式细胞术检测CD31和血管内皮钙黏蛋白（VE-钙黏蛋白）阳性的内皮细胞。
分离得到的CD133、CD34和KDR阳性细胞百分比分别为89.9%、91.3%和90.4%，表明EPC成功分离。MTT结果显示，与单一VEGF包被或裸金属相比，包被VEGF和抗CD34抗体的不锈钢片对接种的EPC毒性更小。我们进一步发现，与单一VEGF包被和裸金属相比，VEGF和抗CD34抗体共同包被能显著促进EPC在体外的分化。
我们的研究对体外包被VEGF和抗CD34抗体的金属钢片进行了初步评估。我们的研究结果表明，同时用VEGF和抗CD34抗体包被支架可能是促进再内皮化以减少支架植入后支架内再狭窄（ISR）的一个新的研究方向。
