新型微纳秒脉冲电场消融仪器的研发及动物实验研究

doi:10.3877/cma.j.issn.1672-6448.2024.12.005

目的

探讨微纳秒脉冲电场（μs-nsPEFs）消融新技术的安全性和有效性，主要包含消融范围、肌肉收缩程度及对重要脉管结构的保护作用等。

方法

本研究利用3D 电场仿真软件建立三维模型，再进行超声引导下开腹消融手术。选用4 头雌性白猪，将白猪随机分为μs-nsPEFs 和纳秒脉冲电场（nsPEFs）两组（每组各2 头），比较两组术前仿真模拟电场分布、术中量化肌肉收缩程度、术后造影面积和组织病理学表现等，探索μs-nsPEFs 消融猪肝脏组织的安全性和有效性。计量资料组间比较采用Student’s t 检验。

结果

使用COMSOL 软件绘制的3D 仿真电场分布图显示，μs-nsPEFs的最大消融面积为（6.06±0.02）cm²；nsPEFs 的最大消融面积为（5.00±0.03）cm²（P＜0.01）。术后10 min 的超声造影（CEUS）结果显示，μs-nsPEFs 组消融面积为（4.70±1.62）cm²，nsPEFs 组为（4.33±1.55）cm²（P＞0.05）。术后第7 天超声探查μs-nsPEFs 组存在低回声区域，而nsPEFs 组未发现明显的低回声区域。术后第7 天的HE 染色切片评估提示μs-nsPEFs 消融区边界明确，面积为（0.15±0.08）cm²，而nsPEFs 组未提示消融灶。胆囊壁HE 染色切片提示μs-nsPEFs 术后第7 天消融病灶周围血流通畅，胆囊整体结构保持完整，肝肾功能指标未出现异常。

结论

新型μs-nsPEFs 技术不仅能够有效控制术中肌肉收缩和保护脉管结构，还能使肝脏消融效果持久有效，具有良好的临床应用前景。

关键词: 肝脏消融, 不可逆电穿孔, 微纳秒脉冲电场, 超声引导, 动物实验

Objective

To investigate the safety and efficacy of the new microsecond-nanosecond pulsed electric field (μs-nsPEF) ablation technology, focusing on ablation range, degree of muscle contraction,and protection of vital vascular structures.

Methods

A three-dimensional (3D) model was established using 3D electric field simulation software, followed by an ultrasound-guided open-abdomen ablation procedure.Four pigs were randomly assigned to μs-nsPEF (n=2) and nanosecond pulsed electric field (nsPEF) groups(n=2).Safety and efficacy of μs-nsPEFs in porcine liver tissue were evaluated by comparing preoperative simulated electric field distributions, intraoperative muscle contraction measurements, postoperative ablation area, and histopathological findings between the two groups.Student's t-test was used for comparisons between groups.

Results

3D electric field distribution maps created with COMSOL software showed a maximum ablation area of 6.06±0.02 cm² for the μs-nsPEF group and 5.00±0.03 cm² for the nsPEF group (P＜0.01).Contrast-enhanced ultrasound (CEUS) imaging 10 minutes post-ablation indicated an ablation area of 4.70±1.62 cm² for the μs-nsPEF group and 4.33±1.55 cm² for the nsPEF group (P＞0.05).Ultrasound examination on postoperative day 7 revealed a hypoechoic area in the μs-nsPEF group, whereas no such area was observed in the nsPEF group.H&E staining on postoperative day 7 showed a well-defined ablation boundary in the μs-nsPEF group with an area of 0.15±0.08 cm², while no ablation lesion was observed in the nsPEF group.H&E staining of the gallbladder wall indicated that on postoperative day 7, blood flow around the μs-nsPEF ablation site remained unobstructed, the gallbladder structure was intact, and liver and kidney function indicators showed no abnormalities.

Conclusion

The novel μs-nsPEF technique demonstrates sustained and effective liver ablation while ensuring muscle contraction and vascular protection during surgery, indicating promising clinical applications.

Key words: Liver ablation, Irreversible electroporation, Microsecond-nanosecond pulsed electric fields, Ultrasound-guided, Animal experiment

图1 电场消融系统图。图a 为Curaway 集成式消融能量平台；图b 为用于消融的双针不锈钢正负电极；图c 为双针电极的针尖裸露端；图d 为超声引导下肝脏消融实物图；图e 为纳秒脉冲电场（nsPEFs）电脉冲参数示意图；图f 为微纳秒脉冲电场（μs-nsPEFs）电脉冲参数示意图

图2 3D 仿真脉冲电场分布示意图。图a 为仿真电场框架示意图；图b 为微纳秒脉冲电场（μs-nsPEFs）仿真电场场强分布示意图；图c 为纳秒脉冲电场（nsPEFs）仿真电场场强分布示意图

图3 猪肝脏消融术前及术后超声检查图像。图a ~ c 为纳秒脉冲电场（nsPEFs）术前超声引导、术后10 min 超声造影与术后第7 天超声探查图像；图d ~ f 为微纳秒脉冲电场（μs-nsPEFs）术前超声引导、术后10 min 超声造影与术后第7 天超声探查图像

图4 μs-nsPEFs 和nsPEFs 消融术后第7 天HE 染色病理图像。图a、d、g 为μs-nsPEFs 术后第7 天肝脏HE 染色图像；图b、e、h 为nsPEFs 术后第7 天肝脏HE 染色图像；图c、f、i 为μs-nsPEFs 术后第7 天胆囊壁HE 染色图像（物镜放大倍数分别为×1、×4、×20，目镜为×10；黑色虚线为消融边界）注：μs-nsPEFs 为微纳秒脉冲电场；nsPEFs 为纳秒脉冲电场

表1 μs-nsPEFs 消融前后P01 号白猪血清电解质及肝肾生化指标分析

电解质及生化指标	术前2h	术后2h	术后7d
天门冬氨酸氨基转移酶（U/L）	36.00	88.00	29.20
丙氨酸氨基转移酶（U/L）	77.60	99.30	68.20
白蛋白（g/L）	39.16	37.97	35.28
总蛋白（g/L）	60.11	67.63	64.97
球蛋白（g/L）	29.82	29.66	29.69
碱性磷酸酶（U/L）	77.70	78.20	58.90
胆固醇（mmol/L）	1.69	1.42	1.77
肌酐（μmol/L）	106.10	110.50	106.40
谷氨酰转肽酶（U/L）	35.10	41.00	36.70
葡萄糖（mmol/L）	5.99	4.77	5.13
总胆红素（μmol/L）	1.22	1.30	1.20
甘油三酯（mmol/L）	0.25	0.60	0.22
尿素（mmol/L）	3.37	8.23	3.46
钙（mmol/L）	2.33	2.37	2.29
无机磷（μmol/L）	1.99	1.98	1.96
钾（mmol/L）	4.53	4.33	4.49
钠（mmol/L）	144.50	141.70	141.20
氯（mmol/L）	98.20	97.80	98.90

表1 μs-nsPEFs 消融前后P01 号白猪血清电解质及肝肾生化指标分析

电解质及生化指标	术前2h	术后2h	术后7d
天门冬氨酸氨基转移酶（U/L）	36.00	88.00	29.20
丙氨酸氨基转移酶（U/L）	77.60	99.30	68.20
白蛋白（g/L）	39.16	37.97	35.28
总蛋白（g/L）	60.11	67.63	64.97
球蛋白（g/L）	29.82	29.66	29.69
碱性磷酸酶（U/L）	77.70	78.20	58.90
胆固醇（mmol/L）	1.69	1.42	1.77
肌酐（μmol/L）	106.10	110.50	106.40
谷氨酰转肽酶（U/L）	35.10	41.00	36.70
葡萄糖（mmol/L）	5.99	4.77	5.13
总胆红素（μmol/L）	1.22	1.30	1.20
甘油三酯（mmol/L）	0.25	0.60	0.22
尿素（mmol/L）	3.37	8.23	3.46
钙（mmol/L）	2.33	2.37	2.29
无机磷（μmol/L）	1.99	1.98	1.96
钾（mmol/L）	4.53	4.33	4.49
钠（mmol/L）	144.50	141.70	141.20
氯（mmol/L）	98.20	97.80	98.90

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Development of novel microsecond-nanosecond pulsed electric field ablation equipment and its animal experimental study