Indicative effect of indocyanine green-near-infrared fluorescence imaging in the thickness of necrotic dermal tissue in porcine burn wounds
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摘要:
目的 探讨吲哚菁绿近红外荧光显影（ICG-NIFI）对猪烧伤创面真皮层坏死组织厚度的指示效果。 方法 该研究为医工交叉应用基础研究。选择1头3个月龄雄性巴马香猪,将烫伤仪（致伤温度75 ℃）与猪背部皮肤分别接触3、7、11 s造成直径2 cm的圆形烧伤创面,每个致伤时长2个创面。采用ICG-NIFI技术检测推注吲哚菁绿后（下称给药后）700 s内创面和创周正常皮肤感兴趣区的荧光强度,观察归一化处理的荧光强度的变化趋势并确定创面和创周正常皮肤感兴趣区的荧光强度达到峰值的时间（下称达峰时间）。另外选择2头3个月龄雄性巴马香猪,将烫伤仪（致伤温度75 ℃）与两侧胸壁皮肤分别接触5、7、9、11、13、15、17、19 s造成直径2 cm的圆形烧伤创面,每个致伤时长共4个创面。采用激光散斑对比成像技术检测创面感兴趣区的血流强度；采用ICG-NIFI技术检测并计算创面和创周正常皮肤感兴趣区的达峰时间时创面感兴趣区的相对荧光强度。取创面全层皮肤组织,行苏木精-伊红染色,测量真皮层坏死组织厚度。分析创面感兴趣区的血流强度及创面和创周皮肤达峰时间时创面感兴趣区的相对荧光强度与创面真皮层坏死组织厚度的相关性。 结果 创周正常皮肤感兴趣区的荧光强度呈先快速升高、后缓慢下降的趋势,达峰时间约为给药后60 s；创面感兴趣区的荧光强度呈先缓慢上升、后持续平稳的趋势,达峰时间约为给药后600 s。致伤时长5、7、9、11、13、15、17、19 s造成的创面真皮层坏死组织厚度分别为（101±8）、（130±6）、（201±19）、（197±30）、（204±21）、（280±39）、（302±35）和（366±27）μm,创面感兴趣区的血流强度与创面真皮层坏死组织厚度的相关性不显著（P>0.05）。给药后60 s,创面感兴趣区的相对荧光强度与创面真皮层坏死组织厚度具有显著的负相关性（R²=0.97,P<0.05）；给药后600 s,创面感兴趣区的相对荧光强度与创面真皮层坏死组织厚度具有显著的正相关性（R²=0.96,P<0.05）。 结论 ICG-NIFI检测的猪烧伤创面感兴趣区在给药后60和600 s时的相对荧光强度均与创面真皮层坏死组织厚度具有显著的相关性,说明ICG-NIFI对猪烧伤创面真皮层坏死组织厚度具有显著灵敏的指示作用。 Abstract:Objective To explore the indicative effects of indocyanine green-near-infrared fluorescence imaging (ICG-NIFI) in the thickness of necrotic dermal tissue in porcine burn wounds. Methods This study was a medical-engineering interdisciplinary basic research. One 3-month-old male Bama miniature pig was selected, and circular burn wounds with a diameter of 2 cm were created by applying a scalding instrument with inflicting temperature of 75 ℃ to the skin on the dorsal side of the pig for 3, 7, and 11 seconds, respectively, with two wounds for each duration of injury. The fluorescence intensity of the regions of interest in the wounds and the surrounding normal skin was detected by ICG-NIFI technology within 700 seconds after injecting indocyanine green (hereinafter referred to as after administration). The trend of the normalized fluorescence intensity was observed, and the time when the fluorescence intensity of the regions of interest in the wounds and the surrounding normal skin reached the peak (hereinafter referred to as the peak time) was determined. Additional two 3-month-old male Bama miniature pigs were selected, and circular burn wounds with a diameter of 2 cm were created on the skin of bilateral thoracic walls by applying a scalding instrument with inflicting temperature of 75 ℃ for 5, 7, 9, 11, 13, 15, 17, and 19 seconds, respectively, with 4 wounds for each duration of injury. The blood flow intensity of the regions of interest in the wounds was detected by laser speckle contrast imaging technology; the relative fluorescence intensity of the regions of interest in the wounds was detected by ICG-NIFI technology at the peak time of the wounds and the surrounding normal skin. The full-thickness skin tissue of the wounds was taken for hematoxylin-eosin staining, and the thickness of necrotic dermal tissue was measured. The correlation between the blood flow intensity of the regions of interest in the wounds and the relative fluorescence intensity at the peak time of the regions of interest in the wounds and the surrounding normal skin and the thickness of necrotic dermal tissue in the wounds was analyzed. Results The fluorescence intensity of the regions of interest in the surrounding normal skin showed a rapid increase followed by a slow decrease, with the peak time of approximately 60 seconds after administration. The fluorescence intensity of the regions of interest in the wounds showed a slow increase followed by a sustained stable trend, with the peak time of approximately 600 seconds after administration. The thickness of necrotic dermal tissue in the wounds caused by durations of injury of 5, 7, 9, 11, 13, 15, 17, and 19 seconds was (101±8), (130±6), (201±19), (197±30), (204±21), (280±39), (302±35), and (366±27) μm, respectively. The correlation between the blood flow intensity of the regions of interest in the wounds and the thickness of necrotic dermal tissue in the wounds was not significant (P>0.05). The relative fluorescence intensity of the regions of interest in the wounds at 60 seconds after administration was significantly negatively correlated with the thickness of necrotic dermal tissue in the wounds (R²=0.97, P<0.05), and the relative fluorescence intensity of the regions of interest in the wounds at 600 seconds after administration was significantly positively correlated with the thickness of necrotic dermal tissue in the wounds (R²=0.96, P<0.05). Conclusions The relative fluorescence intensity of the regions of interest in porcine burn wounds detected by ICG-NIFI at 60 and 600 seconds after administration was significantly correlated with the thickness of necrotic dermal tissue in the wounds, indicating that ICG-NIFI has a significantly sensitive indicative effects for the thickness of necrotic dermal tissue in porcine burn wounds. -
Key words:
- Burns /
- Dermis /
- Laser speckle contrast imaging /
- Laser-Doppler flowmetry /
- Fluorescence imaging /
- Indocyanine green
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参考文献
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图 1 不同致伤时长造成的猪烧伤创面及用吲哚菁绿近红外荧光显影技术检测的创面和创周正常皮肤感兴趣区。1A.左侧2个创面的致伤时长为11 s,中间2个创面的致伤时长为7 s,右侧2个创面的致伤时长为3 s；1B.共16个感兴趣区,其中1~4、5~8、9~12分别表示致伤时长为11、7、3 s造成的创面的感兴趣区,13~16表示创周正常皮肤感兴趣区
图 2 猪烧伤创面和创周正常皮肤感兴趣区的荧光强度-时间曲线
注：ROI1~4、5~8、9~12分别表示致伤时长为11、7、3 s造成的创面的感兴趣区,ROI13~16表示创周正常皮肤感兴趣区；采用吲哚菁绿近红外荧光显影技术测量荧光强度,所有荧光强度数据进行了归一化处理
图 3 不同致伤时长造成的猪烧伤创面真皮层坏死组织厚度 苏木精-伊红×20。3A、3B、3C、3D、3E、3F、3G、3H.分别为致伤时长5、7、9、11、13、15、17、19 s造成的创面
注：图中黑色线段和数据表示真皮层坏死组织的厚度
图 4 不同致伤时长造成的猪烧伤创面感兴趣区的血流强度与创面真皮层坏死组织厚度的相关性（总样本数为8×4）
注：烧伤创面致伤时长分别为5、7、9、11、13、15、17、19 s；采用激光散斑对比成像技术检测创面感兴趣区的血流强度；图中灰色为95%置信区间；创面感兴趣区的血流强度与创面真皮层坏死组织厚度无显著相关性（R2=0.29,P=0.170）
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