双层可降解雷帕霉素洗脱支架在钬激光碎石术后输尿管狭窄中的体内外评价。

In vitro and in vivo assessment of a bilayered degradable rapamycin-eluting stent for ureteral stricture caused by holmium: YAG laser lithotripsy.

机构信息

Department of Urology, the First Affiliated Hospital of China Medical University, Shenyang, 110002, China; Department of Urology, General Hospital of Northern Theater Command, Shenyang, 110840, China.

Department of Urology, General Hospital of Northern Theater Command, Shenyang, 110840, China.

出版信息

Acta Biomater. 2023 Dec;172:321-329. doi: 10.1016/j.actbio.2023.10.009. Epub 2023 Oct 10.

Abstract

Ureteral stricture caused by holmium: YAG laser lithotripsy is one of the most challenging issues for urologists. Currently, evidence for rapamycin application in reducing ureterostenosis is not sufficient. This study aimed to assess the inhibition of ureteral stricture of rapamycin-eluting stents in vitro and in vivo. A bilayered drug-eluting ureteral stent consisted of drug blending with poly (lactic-co-glycolic acid) (PU/drug stent), which was over-layered by polycaprolactone (PCL) by ultrasonic atomizing spraying. Stent morphology was observed by scanning electron microscope. A kidney-ureter-bladder model was established to simulate the stents-releasing condition, and high-performance liquid chromatography was used to measure the drug release rate. The inhibitory proliferation was detected by CCK-8. The bladder of rats was injured through electro tome, and stents were implanted for 7, 14, and 28 days. The effects of drug-eluting stents was investigated by hematoxylin-eosin staining, immunofluorescence staining, real-time quantitative polymerase chain reaction and western blot. The bilayered stents could block the burst loss of the drug and maintained a sustained delivery period because of the 5.3 μm thickness of the PCL layer. The relative growth rates of cells plotted inhibitory effect on the proliferation of human urethral scar fibroblast cells. For in vivo results of 28 days, the bilayered stent maintained structural integrity and induced less deposition of crystals, thinner and less lamina propria connective tissues were formed, and α-SMA and TGF-β1 were downregulated. Bilayered rapamycin-eluting stent is significantly effective in alleviating fibrosis in in vitro and in vivo models. STATEMENT OF SIGNIFICANCE: The occurrence of ureteral stricture resulting from holmium: YAG laser lithotripsy presents a significant challenge for urologists. Traditional double J stents have not been proven to offer a shorter indwelling time or improved inhibition of tissue blocking. While drug-eluting stents containing rapamycin, paclitaxel, and other substances have been extensively used in treating artery stenosis, there is insufficient evidence supporting their application in reducing ureterostenosis. Consequently, a biodegradable polymer ureteric scaffold incorporating rapamycin was fabricated in this study, employing ultrasonic atomization spraying technology to optimize the bilayers composed of 75/25 poly (lactic-co-glycolic acid) (PLGA) and polycaprolactone (PCL). The efficacy of the scaffold was subsequently confirmed through in vitro and in vivo experiments.

摘要

钬激光碎石术后导致的输尿管狭窄是泌尿科医生面临的最具挑战性问题之一。目前,尚无充分的证据表明雷帕霉素在减少输尿管狭窄中的应用。本研究旨在评估雷帕霉素洗脱支架在体外和体内对输尿管狭窄的抑制作用。双层药物洗脱输尿管支架由与聚乳酸-羟基乙酸共聚物(PU/药物支架)混合的药物组成,通过超声雾化喷涂在聚己内酯(PCL)上层。通过扫描电子显微镜观察支架形态。建立肾-输尿管-膀胱模型模拟支架释放条件,采用高效液相色谱法测定药物释放率。CCK-8 检测抑制增殖。通过电切损伤大鼠膀胱,植入支架 7、14 和 28 天。通过苏木精-伊红染色、免疫荧光染色、实时定量聚合酶链反应和 Western blot 研究药物洗脱支架的作用。双层支架可阻止药物的爆发性损失,并保持持续释放期,因为 PCL 层的厚度为 5.3μm。细胞相对增长率图绘制了对人尿道瘢痕成纤维细胞增殖的抑制作用。28 天的体内结果显示,双层支架保持结构完整性并诱导晶体沉积减少,形成更薄、更少的固有层结缔组织,α-SMA 和 TGF-β1 下调。双层雷帕霉素洗脱支架在体外和体内模型中均能显著有效缓解纤维化。 意义声明:钬激光碎石术后发生的输尿管狭窄对泌尿科医生来说是一个重大挑战。传统的双 J 支架尚未被证明具有较短的留置时间或改善组织阻塞的抑制作用。虽然含有雷帕霉素、紫杉醇等物质的药物洗脱支架已广泛应用于治疗动脉狭窄,但尚无充分证据支持其在减少输尿管狭窄中的应用。因此,本研究采用超声雾化喷涂技术制备了一种载雷帕霉素可生物降解聚合物输尿管支架,优化了由 75/25 聚乳酸-羟基乙酸共聚物(PLGA)和聚己内酯(PCL)组成的双层结构。随后通过体外和体内实验证实了支架的疗效。

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