Froghi Saied, Hall Andrew, Hanafi Bin Jalal Arif, Andrade Matheus Oliveira de, Mohammad Hadi Layla, Rashidi Hassan, Gélat Pierre, Saffari Nader, Davidson Brian, Quaglia Alberto
Department of HPB & Liver Transplantation Surgery, Royal Free London NHS Foundation Trust, Pond Street, Hampstead, London NW3 2QG, UK.
Centre for Surgical Innovation, Organ Regeneration and Transplantation, UCL Division of Surgery & Interventional Sciences, Royal Free Hospital Campus, Pond Street, Hampstead, London NW3 2QG, UK.
Bioengineering (Basel). 2023 Feb 21;10(3):278. doi: 10.3390/bioengineering10030278.
Non-invasive therapeutic-focused ultrasound (US) can be used for the mechanical dissociation of tissue and is described as histotripsy. We have performed US histotripsy in viable perfused ex vivo porcine livers as a step in the development of a novel approach to hepatocyte cell transplantation. The histotripsy nidus was created with a 2 MHz single-element focused US transducer, producing 50 pulses of 10 ms duration, with peak positive and negative pressure values of P+ = 77.7 MPa and P- = -13.7 MPaat focus, respectively, and a duty cycle of 1%. Here, we present the histological analysis, including 3D reconstruction of histotripsy sites. Five whole porcine livers were retrieved fresh from the abattoir using human transplant retrieval and cold static preservation techniques and were then perfused using an organ preservation circuit. Whilst under perfusion, histotripsy was performed to randomly selected sites on the live. Fifteen lesional sites were formalin-fixed and paraffin-embedded. Sections were stained with Haematoxylin and Eosin and picro-Sirius red, and they were also stained for reticulin. Additionally, two lesion sites were used for 3D reconstruction. The core of the typical lesion consisted of eosinophilic material associated with reticulin loss, collagen damage including loss of birefringence to fibrous septa, and perilesional portal tracts, including large portal vein branches, but intact peri-lesional hepatic plates. The 3D reconstruction of two histotripsy sites was successful and confirmed the feasibility of this approach to investigate the effects of histotripsy on tissue in detail.
非侵入性治疗聚焦超声（US）可用于组织的机械解离，被称为组织粉碎术。我们在有活力的灌注离体猪肝中进行了超声组织粉碎术，作为开发一种新型肝细胞移植方法的一个步骤。使用一个2兆赫的单元素聚焦超声换能器创建组织粉碎术病灶，产生50个持续时间为10毫秒的脉冲，焦点处的正峰值压力和负峰值压力分别为P+ = 77.7兆帕和P- = -13.7兆帕，占空比为1%。在此，我们展示了组织学分析，包括组织粉碎术部位的三维重建。使用人体移植获取和冷静态保存技术从屠宰场新鲜获取五个完整的猪肝，然后使用器官保存回路进行灌注。在灌注过程中，对肝脏上随机选择的部位进行组织粉碎术。15个病灶部位用福尔马林固定并石蜡包埋。切片用苏木精和伊红以及苦味酸天狼星红染色，还进行了网状纤维染色。此外，两个病灶部位用于三维重建。典型病灶的核心由与网状纤维丢失相关的嗜酸性物质、包括纤维间隔双折射丧失的胶原损伤以及病灶周围的门静脉区域组成，包括大型门静脉分支，但病灶周围的肝板完整。两个组织粉碎术部位的三维重建成功，证实了这种方法详细研究组织粉碎术对组织影响的可行性。
