纤维蛋白胶释放血管内皮生长因子加速损伤动脉血管内皮化

背景血管去内皮化,血栓形成和内膜增生极大的限制了血管治疗方法的临床效果.目的利用含血管内皮细胞生长因子的纤维蛋白胶作用于球囊损伤血管内膜来评估其对血管内皮化、细胞增殖和内膜增生的影响.设计随机对照、重复测量设计.单位解放军第四军医大学西京医院血管外科及神经外科研究所.材料实验于2002-10/2004-06在解放军第四军医大学西京医院神经外科研究所完成.15只健康杂种犬(雌雄不限,体质量12.5～18.9 kg)由西京医院动物实验外科提供.方法损伤犬双侧颈动脉,一侧为纤维蛋白胶/血管内皮细胞生长因子/肝素处理组,另一侧为生理盐水对照组.损伤后10,30和90d观察对比内膜损伤和处理的结果.切片应用Bioquant BQ OS/2计算机形态测量仪测量血管内膜和中层厚度.5-溴脱氧尿苷掺入免疫组化切片用于计算细胞增殖率,40倍光镜下计算5-溴脱氧尿苷阳性细胞数.电镜检查血管腔内表面内皮细胞覆盖率.主要观察指标内皮细胞覆盖情况、新生内膜和中层厚度及细胞增殖率.结果所有犬均存活至标本收集时,无脱失值.①内皮细胞覆盖率在10 d和30 d时,处理侧颈动脉的内皮细胞覆盖率明显高于对照侧[(66.73±30.78)%,(40.8±27.74)%,P=0.04;(96.67±10.29)%,(82.07±22.82)%,P=0.048].②增殖率10 d时,血管各层细胞增殖达到高峰,90d恢复正常.和对照组相比,处理组在30 d时新生内膜和血管中层内侧半中层及血管中层的细胞增殖率明显增强[(7.41±6.75)%,(3.56±2.72)%;(2.81±2.65)%,(0.83±0.59)%;(2.06±1.81)%,(0.62±0.31)%,P＜0.05].③新生内膜厚度和对照组相比,处理组在30 d和90 d时双侧近端和中段的内膜厚度/中层厚度比值明显增加(0.18±0.22,0.10±0.06;0.21±0.23,0.14±0.14;0.12±0.08,0.09±0.08;0.29±0.40,0.12±0.12,P＜0.05,P＜0.01),但远端没有变化.结论纤维蛋白胶能将细胞因子释放入动脉血管壁并保持生物学活性.纤维蛋白胶释放血管内皮细胞生长因子加肝素能有效加速损伤血管内皮化但伴有平滑肌细胞增殖和内膜增生.

Vascular endothelial growth factor delivered by fibrin glue accelerating arterial endothelialization

BACKGROUND: Clinical efficacy of vascular therapeutic intervention is limited by the resultant de-endothelialization, thrombogenecity and intimal hyperplasia.OBJECTIVE: To evaluate the effect of fibrin glue (FG) containing vascular endothelial cell growth factor (VEGF) on re-endothelialization, cellular proliferation and intimal hyperplasia by using canine model of balloon angioplasty.DESIGN: A randomized controlled repeated measurement design.SETTING: Department of Vascular Surgery; Institute of Neurosurgery,Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: The study was carried out in the Institute of Neurosurgery,Xijing Hospital Affiliated to Fourth Military Medical University of the Chinese PLA, between October 2002 and June 2004. Fifteen healthy adult mongrel dogs of either gender, with body mass of 12.5 to 18.9 kg, were provided by the Surgery Laboratory for Experimental Animals, Xijing Hospital.METHODS: In the bilateral carotid artery, FG/VEGF/heparin of one side was set as treatment group and the other side was set as control group. The intimal injury and the treatment results were observed at three time points 10, 30 and 90 days after injury. Thickness of vascular intima and medial layer was measured with Bioquant BQ OS/2 computer morphology measuring instrument. Cell proliferation rate was quantitated by 5-bromodeoxyuridine (BrdU) incorporation by immunohistochemistry. BrdU positive cells were counted using 40 × magnification. Scanning electron microscopy (SEM) was used to evaluate the percentage of endothelial cell coverage on the luminal surface.MAIN OUTCOME MEASURES: Coverage of endothelial cells, neointimal and medial thickness, and cellular proliferation.RESULTS: All the dogs survived till the collection of samples with no erage rate at the treated side at days 10 and 30 was significantly higher than that at the control side [(66.73±30.78)%, (40.8±27.74)%, P=0.04;cular layer: It reached the peak at day 10 and recovered to normal at day 90. Compared with that of control group, cellular proliferation rate of neointima and the 1/2 of inside of media as well as media was significantly increased [(7.41±6.75)%, (3.56±2.72)%; (2.81±2.65)%, (0.83±0.59)%;Compared with that of control group, the thickness of intima/thickness of medial layer in both the proximal and the middle segments was significantly increased (0.18 ±0.22, 0.10 ±0.06; 0.21 ±0.23, 0.14 ±0.14;0.12±0.08, 0.09±0.08; 0.29±0.40, 0.12±0.12, P ＜ 0.05, P ＜ 0.01), but there was no change in the distal segments.CONCLUSION: FG can distribute cytokines into the wall of injured arteries and retain the biological function of cytokines. VEGF plus heparin delivered by FG accelerates re-endothelialization concomitant with the proliferation of smooth muscle cell and intimal hyperplasia.