Alteplase stability and bioactivity after low-power ultrasonic energy delivery with the OmniSonics Resolution System
Alteplase stability and bioactivity after low-power ultrasonic energy delivery with the OmniSonics Resolution System
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DOI：
10.1097/01.rvi.0000147066.97599.87
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发表时间：
2005-03-01
影响因子：
2.9
通讯作者：
Semba, CP
中科院分区：
文献类型：
--
作者：
Smikahl, J;Yeung, D;Semba, CP
目的：低功率超声波（US）能量能够溶解体内的凝块。 US能量加阿替普酶的组合可能会进一步加速血栓溶解；然而，空化的影响可能会导致裂解蛋白变性和失活。本研究的目的是在体外模型中使用新型血管内超声线确定阿替普酶在暴露于超声能量时的生物活性和稳定性。 材料和方法：该模型由一根一端封闭的 6.4 毫米直径硅胶管组成，并在水浴（37 摄氏度）中填充阿替普酶 (1 mg/mL)。将 95 厘米美国电线（直径 0.025 英寸，20 kHz）插入管中并连接到可变发电机。该线在其 20 厘米有效长度周围 360 度提供低功率声能，并通过连续注入纯净水进行灌溉。将新鲜的 6 mL 阿替普酶等分试样暴露于美国能量中并一式两份进行测试。将零（对照）、1 W 或 2 W 的能量传送至各个测试样本，持续零（对照）、0.5、3 或 6 分钟。使用紫外分光光度法测定阿替普酶样品的光学透明度和蛋白质浓度，使用高性能尺寸排阻色谱法测定蛋白质单体百分比，并测定体外凝块溶解活性。 结果：在对照样品中，所有样品的光学透明度都是透明或无色；蛋白质浓度为 1.02 mg/mL +/- 0；蛋白质单体为98%；血块溶解活性为 108%/mg +/- 1。在测试样品中，所有样品的光学透明度均为透明或无色； 1 W 时 0.5、3 和 6 分钟的蛋白质浓度分别为 0.98 mg/mL +/- 0.02、0.93 mg/mL +/- 0.01 和 0.86 mg/mL +/- 0.02，1.00 mg/mL +/- 0.03、0.94 mg/mL +/- 0.10 和 0.84 mg/mL +/- 2 W 时分别为 0.17。所有样品的蛋白质单体均为 98%。 1 W 时 0.5、3 和 6 分钟的凝块溶解活性水平分别为 111%/mg +/- 1、110%/mg +/- 1 和 115%/mg +/- 1，2 W 时分别为 110%/mg +/- 0、111%/mg +/- 1 和 116%/mg +/- 2。结论：根据蛋白质测定结果，阿替普酶溶液暴露于低功率美国能量长达 6 分钟，仍保持完全活性和稳定。有必要结合使用美国能源和阿替普酶进行进一步研究。
PURPOSE: Low-power ultrasonic (US) energy is capable of clot dissolution in vivo. The combination of US energy plus alteplase may further accelerate clot lysis; however, the effects of cavitation could potentially denature and inactivate the lytic protein. The purpose of this study was to determine the bioactivity and stability of alteplase when exposed to US energy with use of a novel intravascular US wire in an in vitro model.MATERIALS AND METHODS: The model consisted of a 6.4-mm-diameter silicone tube closed at one end and filled with alteplase (1 mg/mL) in a water bath (37 degrees C). A 95-cm US wire (0.025-inch diameter, 20 kHz) was inserted into the tube and connected to a variable power generator. The wire delivers low-power acoustic energy 360 degrees around its 20-cm active length and was irrigated by a continuous infusion of purified water. Fresh 6-mL alteplase aliquots were exposed to US energy and tested in duplicates. Zero (control), 1 W, or 2 W of energy was delivered to individual test samples for zero (control), 0.5,3, or 6 minutes. Alteplase samples were assayed for optical clarity and protein concentration with use of UV spectrophotometry, for percent protein monomer with use of high-performance size-exclusion chromatography, and for in vitro clot lysis activity.RESULTS: In the control samples, optical clarity was clear or colorless in all samples; protein concentration was 1.02 mg/mL +/- 0; protein monomer was 98%; and clot lysis activity was 108% per mg +/- 1. In the test samples, optical clarity was clear or colorless in all samples; protein concentrations at 0.5, 3, and 6 minutes were 0.98 mg/mL +/- 0.02, 0.93 mg/mL +/- 0.01, and 0.86 mg/mL +/- 0.02, respectively, at 1 W, and 1.00 mg/mL +/- 0.03, 0.94 mg/mL +/- 0.10, and 0.84 mg/mL +/- 0.17, respectively, at 2 W. Protein monomer was 98% for all samples. Clot lysis activity levels at 0.5, 3, and 6 minutes were 111% per mg +/- 1, 110% per mg +/- 1, and 115% per mg +/- 1, respectively, at 1 W, and 110% per mg +/- 0, 111% per mg +/- 1, and 116% per mg +/- 2. respectively, at 2 W.CONCLUSIONS: Alteplase solutions exposed to low-power US energy for as long as 6 minutes remained fully active and stable as determined by protein assays. Further investigation is warranted with use of combinations of US energy and alteplase.