他克莫司洗脱缝线抑制新生内膜增生：大鼠体内实验研究

Tacrolimus-Eluting Suture Inhibits Neointimal Hyperplasia: An Experimental In Vivo Study in Rats.

作者信息

Ak K, Ak E, Dericioglu O, Canak T, Akbuga J, Ozkan N, Cetinel S, Isbir S, Arsan S, Cobanoglu A

机构信息

Marmara University Faculty of Medicine, Department of Cardiovascular Surgery, Marmara Universitesi Pendik Egitim ve Arastırma Hastanesi, Ustkaynarca Pendik, Istanbul, Turkey; Eastern Mediterranean University, Faculty of Medicine, Gazimagusa, Cyprus.

Faculty of Dentistry, Department of Basic Sciences, Histology and Embryology, Marmara Universitesi Pendik Egitim ve Arastırma Hastanesi, Ustkaynarca Pendik, Istanbul, Turkey.

出版信息

Eur J Vasc Endovasc Surg. 2017 Mar;53(3):431-437. doi: 10.1016/j.ejvs.2016.11.027. Epub 2017 Jan 4.

DOI:10.1016/j.ejvs.2016.11.027

PMID:28065442

Abstract

OBJECTIVE/BACKGROUND: Neointimal hyperplasia (NIH) remains one of the leading causes of graft failure after vascular anastomoses. Cytotoxic drugs, such as rapamycin and tacrolimus, have been shown to inhibit the development of NIH. In this study, the aim was to test the impact of a sustained releasing tacrolimus-chitosan-eluting suture on the development of NIH in a rat model.

METHODS

After tacrolimus-chitosan coating of a 7/0 polyvinylidene difluoride (PVDF) Trofilen suture, the tacrolimus concentration on the coated suture and in vitro release trials were performed spectrophotometrically. Twelve Wistar rats were included. After midline laparotomy, a 7-8 mm longitudinal aortotomy in the infrarenal aorta was made and then closed by a bare 7/0 PVDF (group C, n = 6) and a 7/0 tacrolimus-chitosan coated PVDF suture (0.65 μg/cm tacrolimus [0.9 wt%] + 1.82 μg/cm chitosan [2.28 wt%]) (group T, n = 6). After 1 month, rats were sacrificed and aortotomy sites were examined histologically by ratio of intimal area (including neointima) and immunohistochemically by α-smooth muscle actin (ASMA) and proliferating cell nuclear antigen (PCNA) immunostaining. The PCNA positive cells were indexed to total cell number and expressed as percentage.

RESULTS

In vitro tacrolimus release tests for a 7/0 tacrolimus-chitosan coated PVDF suture were confirmed for 1 month without an initial burst release. Endothelialisation over the aortotomy line occurred in both groups. The area of neointima was significantly reduced in group T compared with group C (ratio 0.22 ± 0.12 vs. 0.42 ± 0.11; p = .017) 1 month post-operatively. Likewise, the percentage of PCNA immunostaining significantly decreased in group C compared with group T (3.83 ± 2.85% vs. 11.17 ± 7.78%; p = .026). The cells constituting NIH were positive for ASMA immunostaining.

CONCLUSIONS

Tacrolimus-chitosan-eluting suture is shown to be an effective way to reduce NIH without interfering with normal endothelialisation.

摘要

目的/背景：血管吻合术后新生内膜增生（NIH）仍是移植物失败的主要原因之一。已证实细胞毒性药物，如雷帕霉素和他克莫司，可抑制NIH的发展。在本研究中，目的是在大鼠模型中测试持续释放他克莫司-壳聚糖洗脱缝线对NIH发展的影响。

方法

在7/0聚偏二氟乙烯（PVDF）Trofilen缝线上涂覆他克莫司-壳聚糖后，采用分光光度法对涂覆缝线上的他克莫司浓度和体外释放试验进行检测。纳入12只Wistar大鼠。经腹正中切口后，在肾下腹主动脉做一个7-8毫米的纵向主动脉切口，然后分别用裸7/0 PVDF缝线（C组，n = 6）和7/0他克莫司-壳聚糖涂覆的PVDF缝线（0.65μg/cm他克莫司[0.9 wt%]+1.82μg/cm壳聚糖[2.28 wt%]）（T组，n = 6）进行缝合。1个月后，处死大鼠，通过内膜面积（包括新生内膜）比值对主动脉切口部位进行组织学检查，并通过α-平滑肌肌动蛋白（ASMA）和增殖细胞核抗原（PCNA）免疫染色进行免疫组织化学检查。将PCNA阳性细胞数与总细胞数进行指数化，并以百分比表示。

结果

7/0他克莫司-壳聚糖涂覆的PVDF缝线的体外他克莫司释放试验证实持续1个月无初始突释。两组主动脉切口处均发生内皮化。术后1个月，T组新生内膜面积较C组显著减小（比值0.22±0.12 vs. 0.42±0.11；p = 0.017）。同样，C组PCNA免疫染色百分比与T组相比显著降低（3.83±2.85% vs. 11.17±7.78%；p = 0.026）。构成NIH的细胞ASMA免疫染色呈阳性。