Perrod Guillaume, Pidial Laetitia, Camilleri Sophie, Bellucci Alexandre, Casanova Amaury, Viel Thomas, Tavitian Bertrand, Cellier Chirstophe, Clément Olivier, Rahmi Gabriel
Assistance Publique-Hôpitaux de Paris, Université Paris Descartes Sorbonne Paris Cité; Department of Gastroenterology, Hôpital Européen Georges Pompidou; UMR-S970, Université Paris Descartes Sorbonne Paris Cité.
UMR-S970, Université Paris Descartes Sorbonne Paris Cité.
J Vis Exp. 2017 Feb 10(120):55018. doi: 10.3791/55018.
In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The development of diversified technologies combining adipose tissue-derived stromal cells (ADSCs) with various biomaterials has led to the construction of numerous types of tissue-engineered substitutes, such as bone, cartilage, and adipose tissues from rodent, porcine, or human ADSCs. Extended esophageal endoscopic submucosal dissection (ESD) is responsible for esophageal stricture formation. Stricture prevention remains challenging, with no efficient treatments available. Previous studies reported the effectiveness of mucosal cell-sheet transplantation in a canine model and in humans. ADSCs are attributed anti-inflammatory properties, local immune modulating effects, neovascularization induction, and differentiation abilities into mesenchymal and non-mesenchymal lineages. This original study describes the endoscopic transplantation of an ADSC tissue-engineered construct to prevent esophageal stricture in a swine model. The ADSC construct was composed of two allogenic ADSC sheets layered upon each other on a paper support membrane. The ADSCs were labeled with the PKH67 fluorophore to allow probe-based confocal laser endomicroscopy (pCLE) monitoring. On the day of transplantation, a 5-cm and hemi-circumferential ESD known to induce esophageal stricture was performed. Animals were immediately endoscopically transplanted with 4 ADSC constructs. The complete adhesion of the ADSC constructs was obtained after 10 min of gentle application. Animals were sacrificed on day 28. All animals were successfully transplanted. Transplantation was confirmed on day 3 with a positive pCLE evaluation. Compared to transplanted animals, control animals developed severe strictures, with major fibrotic tissue development, more frequent alimentary trouble, and reduced weight gain. In our model, the transplantation of allogenic ADSCs, organized in double cell sheets, after extended ESD was successful and strongly associated with a lower esophageal stricture rate.
在过去几年中，细胞片构建体在再生医学领域引发了广泛关注，尤其是在重建外科手术中。将脂肪组织来源的基质细胞（ADSCs）与各种生物材料相结合的多样化技术发展，促使人们构建了多种类型的组织工程替代品，例如来自啮齿动物、猪或人类ADSCs的骨、软骨和脂肪组织。扩大的食管内镜黏膜下剥离术（ESD）会导致食管狭窄形成。预防狭窄仍然具有挑战性，目前尚无有效的治疗方法。先前的研究报道了黏膜细胞片移植在犬模型和人类中的有效性。ADSCs具有抗炎特性、局部免疫调节作用、诱导新血管形成以及向间充质和非间充质谱系分化的能力。这项原创研究描述了在猪模型中内镜下移植ADSC组织工程构建体以预防食管狭窄的情况。ADSC构建体由两张同种异体ADSC片在纸质支撑膜上相互层叠组成。ADSCs用PKH67荧光团标记，以便进行基于探针的共聚焦激光内镜显微镜检查（pCLE）监测。在移植当天，进行了一次已知会导致食管狭窄的5厘米半周径ESD。动物随即接受内镜下4个ADSC构建体的移植。轻轻放置10分钟后，ADSC构建体实现了完全黏附。在第28天对动物实施安乐死。所有动物均成功移植。在第3天通过pCLE阳性评估确认了移植情况。与移植动物相比，对照动物出现了严重狭窄，伴有大量纤维化组织形成、更频繁的消化问题以及体重增加减少。在我们的模型中，扩大ESD后移植双层组织的同种异体ADSCs是成功的，并且与较低的食管狭窄发生率密切相关。
