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Imaging hamster model of bile duct cancer in vivo using fluorescent L-glucose derivatives.

Yokoyama H, Sasaki A, Yoshizawa T, Kijima H, Hakamada K, Yamada K - Hum. Cell (2016)

Bottom Line: In addition, a combined use of 2-TRLG, a membrane-impermeable fLG, with 2-NBDLG visualized membrane integrity as well.Extrahepatic cholangiocarcinoma developed in mid common duct in ~20 % of the animals subjected to cholecystoduodenostomy with the ligation at the distal end of the common duct followed by injection of a carcinogen N-nitrosobis(2-oxopropyl)amine.After infusing bile duct with a solution containing 2-NBDLG and 2-TRLG, the lumen was surgically exposed and examined by pCLE.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterological Surgery, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori, 036-8562, Japan.

ABSTRACT
Extrahepatic bile duct cancer (cholangiocarcinoma) has a poor prognosis. Since surgical resection is the only way to prolong the patient's life, it is of critical importance to correctly determine the extent of lesions. However, conventional pre-operative assessments have insufficient spatial resolution for determining the surgical margin. A fluorescent contrast agent might provide a more precise measure to identify anomalies in biliary surface, when combined with probe-based confocal laser endomicroscopy (pCLE). We have previously shown that 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-L-glucose (2-NBDLG), a fluorescent derivative of L-glucose (fLG), is specifically taken up into spheroids consisting of cells showing heterogeneous nuclear-cytoplasm ratio, a feature of malignant cells in clinical settings. In addition, a combined use of 2-TRLG, a membrane-impermeable fLG, with 2-NBDLG visualized membrane integrity as well. We therefore explored in the present study the availability of the fLGs in vivo as a contrast agent for pCLE by using a hamster model of cholangiocarcinoma. Extrahepatic cholangiocarcinoma developed in mid common duct in ~20 % of the animals subjected to cholecystoduodenostomy with the ligation at the distal end of the common duct followed by injection of a carcinogen N-nitrosobis(2-oxopropyl)amine. After infusing bile duct with a solution containing 2-NBDLG and 2-TRLG, the lumen was surgically exposed and examined by pCLE. Fluorescence pattern characterized by bright spots and dark clumps was detected in the areas diagnosed with cholangiocarcinoma in later histopathology, whereas no such pattern was detected in control animals. These findings may form a basis for elucidating a potential availability of fLGs in imaging cholangiocarcinoma by pCLE.

No MeSH data available.


Related in: MedlinePlus

Autofluorescence images of the bile duct surface of a hamster in CDDB/BOP group taken in a condition with no fLG infusion, and the corresponding histological specimens. a A macro zoom microscopic image taken during autofluorescence imaging of the biliary surface. b A representative autofluorescence image taken with pCLE at the site shown in a. c H&E section processed from the area corresponding to the site imaged in a. The cytoarchitecture changes gradually from the right to the left side, non-neoplastic simple epithelium, hyperplasia with angular-shaped small papillary protrusion, to hyperplasia composed of thinner papillary structures. d A reduced view of c. Demarcated area was magnified. e–h Similar to a–d, but taken at a different site in the bile duct of the same animal. A heterogeneous but coarse fluorescence pattern was detected (f). Classified as dysplasia characterized by thin papillary protrusions composed of cells bearing heterogeneously enlarged nuclei with coarse granular chromatin (g). i–l Similar to e–h, but taken at a site much closer to duodenum side. Only dim homogeneous fluorescence is seen (j) for lesions diagnosed with hyperplasia composed of uniform angular-shaped papillary structures (k, l)
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Fig4: Autofluorescence images of the bile duct surface of a hamster in CDDB/BOP group taken in a condition with no fLG infusion, and the corresponding histological specimens. a A macro zoom microscopic image taken during autofluorescence imaging of the biliary surface. b A representative autofluorescence image taken with pCLE at the site shown in a. c H&E section processed from the area corresponding to the site imaged in a. The cytoarchitecture changes gradually from the right to the left side, non-neoplastic simple epithelium, hyperplasia with angular-shaped small papillary protrusion, to hyperplasia composed of thinner papillary structures. d A reduced view of c. Demarcated area was magnified. e–h Similar to a–d, but taken at a different site in the bile duct of the same animal. A heterogeneous but coarse fluorescence pattern was detected (f). Classified as dysplasia characterized by thin papillary protrusions composed of cells bearing heterogeneously enlarged nuclei with coarse granular chromatin (g). i–l Similar to e–h, but taken at a site much closer to duodenum side. Only dim homogeneous fluorescence is seen (j) for lesions diagnosed with hyperplasia composed of uniform angular-shaped papillary structures (k, l)

Mentions: To assess an involvement of autofluorescence in fLG imaging, the bile duct of three animals subjected to CDDB/BOP procedure was imaged in a condition with no fLG infusion in the preliminary experiment (Fig. 4). As depicted, ambiguous (Fig. 4a–d) or almost no (Fig. 4i–l) fluorescence was detected in the epithelia corresponding to hyperplastic lesions. Although somewhat irregular autofluorescence pattern was detected in dysplastic lesions of the same animal, the characteristic fluorescence pattern typically seen in biliary epithelia after the fLG infusion has not been detected (Fig. 4e–h).


Imaging hamster model of bile duct cancer in vivo using fluorescent L-glucose derivatives.

Yokoyama H, Sasaki A, Yoshizawa T, Kijima H, Hakamada K, Yamada K - Hum. Cell (2016)

Autofluorescence images of the bile duct surface of a hamster in CDDB/BOP group taken in a condition with no fLG infusion, and the corresponding histological specimens. a A macro zoom microscopic image taken during autofluorescence imaging of the biliary surface. b A representative autofluorescence image taken with pCLE at the site shown in a. c H&E section processed from the area corresponding to the site imaged in a. The cytoarchitecture changes gradually from the right to the left side, non-neoplastic simple epithelium, hyperplasia with angular-shaped small papillary protrusion, to hyperplasia composed of thinner papillary structures. d A reduced view of c. Demarcated area was magnified. e–h Similar to a–d, but taken at a different site in the bile duct of the same animal. A heterogeneous but coarse fluorescence pattern was detected (f). Classified as dysplasia characterized by thin papillary protrusions composed of cells bearing heterogeneously enlarged nuclei with coarse granular chromatin (g). i–l Similar to e–h, but taken at a site much closer to duodenum side. Only dim homogeneous fluorescence is seen (j) for lesions diagnosed with hyperplasia composed of uniform angular-shaped papillary structures (k, l)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4930486&req=5

Fig4: Autofluorescence images of the bile duct surface of a hamster in CDDB/BOP group taken in a condition with no fLG infusion, and the corresponding histological specimens. a A macro zoom microscopic image taken during autofluorescence imaging of the biliary surface. b A representative autofluorescence image taken with pCLE at the site shown in a. c H&E section processed from the area corresponding to the site imaged in a. The cytoarchitecture changes gradually from the right to the left side, non-neoplastic simple epithelium, hyperplasia with angular-shaped small papillary protrusion, to hyperplasia composed of thinner papillary structures. d A reduced view of c. Demarcated area was magnified. e–h Similar to a–d, but taken at a different site in the bile duct of the same animal. A heterogeneous but coarse fluorescence pattern was detected (f). Classified as dysplasia characterized by thin papillary protrusions composed of cells bearing heterogeneously enlarged nuclei with coarse granular chromatin (g). i–l Similar to e–h, but taken at a site much closer to duodenum side. Only dim homogeneous fluorescence is seen (j) for lesions diagnosed with hyperplasia composed of uniform angular-shaped papillary structures (k, l)
Mentions: To assess an involvement of autofluorescence in fLG imaging, the bile duct of three animals subjected to CDDB/BOP procedure was imaged in a condition with no fLG infusion in the preliminary experiment (Fig. 4). As depicted, ambiguous (Fig. 4a–d) or almost no (Fig. 4i–l) fluorescence was detected in the epithelia corresponding to hyperplastic lesions. Although somewhat irregular autofluorescence pattern was detected in dysplastic lesions of the same animal, the characteristic fluorescence pattern typically seen in biliary epithelia after the fLG infusion has not been detected (Fig. 4e–h).

Bottom Line: In addition, a combined use of 2-TRLG, a membrane-impermeable fLG, with 2-NBDLG visualized membrane integrity as well.Extrahepatic cholangiocarcinoma developed in mid common duct in ~20 % of the animals subjected to cholecystoduodenostomy with the ligation at the distal end of the common duct followed by injection of a carcinogen N-nitrosobis(2-oxopropyl)amine.After infusing bile duct with a solution containing 2-NBDLG and 2-TRLG, the lumen was surgically exposed and examined by pCLE.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterological Surgery, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori, 036-8562, Japan.

ABSTRACT
Extrahepatic bile duct cancer (cholangiocarcinoma) has a poor prognosis. Since surgical resection is the only way to prolong the patient's life, it is of critical importance to correctly determine the extent of lesions. However, conventional pre-operative assessments have insufficient spatial resolution for determining the surgical margin. A fluorescent contrast agent might provide a more precise measure to identify anomalies in biliary surface, when combined with probe-based confocal laser endomicroscopy (pCLE). We have previously shown that 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-L-glucose (2-NBDLG), a fluorescent derivative of L-glucose (fLG), is specifically taken up into spheroids consisting of cells showing heterogeneous nuclear-cytoplasm ratio, a feature of malignant cells in clinical settings. In addition, a combined use of 2-TRLG, a membrane-impermeable fLG, with 2-NBDLG visualized membrane integrity as well. We therefore explored in the present study the availability of the fLGs in vivo as a contrast agent for pCLE by using a hamster model of cholangiocarcinoma. Extrahepatic cholangiocarcinoma developed in mid common duct in ~20 % of the animals subjected to cholecystoduodenostomy with the ligation at the distal end of the common duct followed by injection of a carcinogen N-nitrosobis(2-oxopropyl)amine. After infusing bile duct with a solution containing 2-NBDLG and 2-TRLG, the lumen was surgically exposed and examined by pCLE. Fluorescence pattern characterized by bright spots and dark clumps was detected in the areas diagnosed with cholangiocarcinoma in later histopathology, whereas no such pattern was detected in control animals. These findings may form a basis for elucidating a potential availability of fLGs in imaging cholangiocarcinoma by pCLE.

No MeSH data available.


Related in: MedlinePlus