A novel approach of optical biopsy using probe-based confocal laser endomicroscopy for peritoneal metastasis

作者：Hara Hisashi; Takahashi Tsuyoshi; Nakatsuka Rie; Higashi Shigeyoshi; Naka Tetsuji; Sumiyama Kazuki; Miyazaki Yasuhiro; Makino Tomoki; Kurokawa Yukinori; Yamasaki Makoto; Takiguchi Shuji; Mori Masaki; Doki Yuichiro; Nakajima Kiyokazu^*

来源：Surgical Endoscopy and Other Interventional Techniques, 2016, 30(8): 3437-3446.

DOI：10.1007/s00464-015-4626-7

• Confocal laser endomicroscopy
• pCLE
• Fluorescein
• 5-Aminolevulinic acid
• Peritoneal metastasis
• Optical biopsy

In digestive cancers, it is mandatory to diagnose peritoneal metastasis prior to selecting therapy. Therefore, exploratory laparoscopy has gained wider clinical acceptance. In laparoscopy, the peritoneal metastasis is pathologically confirmed by excisional biopsy; however, there remain technical difficulties in performing precise diagnosis and adequate biopsy on small peritoneal lesions without damaging organs. We have focused on "optical biopsy" using probe-based confocal laser endomicroscopy (pCLE). The aims of this study were (1) to optimize current CLE system for real-time observation of peritoneal metastases and (2) to assess its potential usefulness as diagnostic modality in preclinical settings.
To optimize condition and evaluate feasibility, we prepared peritoneal metastasis mice model with gastric cancer cell line (MKN-45). On Day 10 after seeding, the mice were laparotomized and performed pCLE observations with CellvizioLAB (LSU-F 400/488 nm, Mauna Kea Technologies, Paris, France). We evaluated two different CLE probes, three different dyes, and optimal interval time. The detected sites were excised and pathologically evaluated on its morphology. Next, the feasibility and safety were validated in porcine model for clinical usage. After injection of fluorescein, pCLE was applied for the observation of intra-abdominal organs.
A miniature probe-type pCLE system with 60 mu m focal depth (UltraMini O) and 1 % fluorescein dye was chosen for good visualization in mice model. The irregular microarchitecture images suspected to malignancy were obtained from the metastases. In the porcine model, observation of abdominal organs was feasible without any organ injury in the laparoscopic procedures. The dosage of 1 % fluorescein (3 ml/body) was appropriate in observing intra-abdominal organs, and each intra-abdominal organ was clearly observed with the same imaging quality we obtained in mice model.
The pCLE was feasible and safe and potentially useful for the diagnosis of the peritoneal metastasis in in vivo animal models.

• 出版日期2016-8