在挤压伤绵羊模型中使用荧光寿命成像技术定义急性周围神经损伤区域
Defining the Zone of Acute Peripheral Nerve Injury Using Fluorescence Lifetime Imaging in a Crush Injury Sheep Model.
作者信息
Dave Dattesh R, Alfonso Garcia Alba, Kraft Lisanne, Marcu Laura, Pereira Clifford T
机构信息
Hand and Upper Extremity Division of Plastic and Reconstructive Surgery, University of California Davis, Sacramento, CA.
Biomedical Engineering Department, University of California Davis, Davis, CA.
出版信息
J Hand Surg Am. 2025 Jan 4. doi: 10.1016/j.jhsa.2024.11.020.
PURPOSE
Current technologies to define the zone of acute peripheral nerve injury intraoperatively are limited by surgical experience, time, cumbersome electrodiagnostic equipment, and interpreter reliability. In this pilot study, we evaluated a real-time, label-free optical technique for intraoperative nerve injury imaging. We hypothesize that fluorescence lifetime imaging (FLIm) will detect a difference between the time-resolved fluorescence signatures for acute crush injuries versus uninjured segments of peripheral nerves in sheep.
METHODS
Label-free FLIm uses ultraviolet laser pulses to excite endogenous tissue fluorophores and detect their fluorescent decay over time, generating real-time tissue-specific signatures. A crush injury was produced in eight peripheral nerves of two sheep. A hand-held FLIm instrument captured the time-resolved fluorescence signatures of injured and uninjured nerve segments across three spectral emission channels (390/40 nm, 470/28 nm, and 540/50 nm). The average FLIm parameters (ie, lifetime and intensity ratios) for injured and uninjured nerve segments were compared. We used linear discriminant analysis to differentiate between crushed and uninjured nerve segments.
RESULTS
A total of 23,692 point measurements were collected from eight crushed peripheral nerves of two sheep. Histology confirmed the zone of injury. Average lifetime at 470 nm and 540 nm were significantly different between crushed and uninjured sheep nerve segments. The linear discriminant analysis differentiated between crushed and uninjured areas of eight nerve segments with 92% sensitivity, 85% specificity, and 88% accuracy.
CONCLUSIONS
In this pilot study, FLIm detected differing average lifetime values for crushed versus uninjured sheep peripheral nerves with high sensitivity, specificity, and accuracy.
CLINICAL RELEVANCE
With further investigation, FLIm may guide the peripheral nerve surgeon to the precise zone of injury for reconstruction.
目的
目前术中确定急性周围神经损伤区域的技术受到手术经验、时间、笨重的电诊断设备以及解释器可靠性的限制。在这项初步研究中，我们评估了一种用于术中神经损伤成像的实时、无标记光学技术。我们假设荧光寿命成像（FLIm）将检测到绵羊急性挤压伤与周围神经未损伤节段的时间分辨荧光特征之间的差异。
方法
无标记FLIm使用紫外激光脉冲激发内源性组织荧光团，并检测其随时间的荧光衰减，生成实时的组织特异性特征。对两只绵羊的八条周围神经造成挤压伤。一台手持式FLIm仪器在三个光谱发射通道（390/40nm、470/28nm和540/50nm）上捕获损伤和未损伤神经节段的时间分辨荧光特征。比较损伤和未损伤神经节段的平均FLIm参数（即寿命和强度比）。我们使用线性判别分析来区分挤压和未损伤的神经节段。
结果
从两只绵羊的八条挤压周围神经中总共收集了23692个点测量值。组织学证实了损伤区域。挤压和未损伤的绵羊神经节段在470nm和540nm处的平均寿命有显著差异。线性判别分析区分了八个神经节段的挤压和未损伤区域，灵敏度为92%，特异性为85%，准确率为88%。
结论
在这项初步研究中，FLIm以高灵敏度、特异性和准确率检测到挤压与未损伤的绵羊周围神经的平均寿命值不同。
临床意义
通过进一步研究，FLIm可能会引导周围神经外科医生找到精确的损伤区域进行重建。
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