title: Tumor neoangiogenesis detection by confocal laser endomicroscopy and anti-CD105 antibody: Pilot study.
abstract: OBJECTIVE
  To evaluate neoangiogenesis in patients with colon cancer by two fluorescently labeled antibodies on fresh biopsy samples imaged with confocal laser endomicroscopy (CLE).
  
  
  METHODS
  CLE is an imaging technique for gastrointestinal endoscopy providing in vivo microscopy at subcellular resolution. An important question in validating tumor angiogenesis is what proportion of the tumor vascular network is represented by pre-existing parent tissue vessels and newly formed vessels. CD105 (endoglin) represents a proliferation-associated endothelial cell adhesion molecule. In contrast to pan-endothelial markers, such as CD31, CD105 is preferentially expressed in activated endothelial cells that participate in neovascularization. Thus, we evaluated CD105 and CD31 expression from samples of ten patients with primary rectal adenocarcinoma, using a dedicated endomicroscopy system. A imaging software was used to obtain the Z projection of the confocal serial images from each biopsy sample previously combined into stacks. Vascular density and vessel diameters were measured within two 50 μm x 475 μm rectangular regions of interest centered in the middle of each image in the horizontal and vertical direction. The results were averaged over all the patients and were expressed as the mean ± SE.
  
  
  RESULTS
  The use of an anti-CD105 antibody was found to be suitable for the detection of blood vessels in colon cancer. Whereas anti-CD31 antibodies stained blood vessels in both normal and pathologic colon equally, CD105 expression was observed primarily in malignant lesions, with little or no expression in the vessels of the normal mucosa (244.21 ± 130.7 vessels/mm(3) in only four patients). The average diameter of anti-CD105 stained vessels was 10.97 ± 0.6 μm in tumor tissue, and the vessel density was 2787.40 ± 134.8 vessels/mm(3). When using the anti-CD31 antibody, the average diameter of vessels in the normal colon tissue was 7.67 ± 0.5 μm and the vessel density was 3191.60 ± 387.8 vessels/mm(3), while in the tumors we obtained an average diameter of 10.88 ± 0.8 μm and a vessel density of 4707.30 ± 448.85 vessels/mm(3). Thus, there were more vessels stained with CD31 than CD105 (P < 0.05). The average vessel diameter was similar for both CD31 and CD105 staining. A qualitative comparison between CLE vs immunohistochemistry lead to similar results.
  
  
  CONCLUSIONS
  Specific imaging and quantification of tumor microvessels are feasible in human rectal cancer using CLE examination and CD105 immunostaining of fresh tissue samples.
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Adriana
authors.initials: A
authors.lastname: Ciocâlteu
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Adrian
authors.initials: A
authors.lastname: Săftoiu
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Daniel
authors.initials: D
authors.lastname: Pirici
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Claudia-Valentina
authors.initials: CV
authors.lastname: Georgescu
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Cârţână
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Dan Ionuț
authors.initials: DI
authors.lastname: Gheonea
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Lucian Gheorghe
authors.initials: LG
authors.lastname: Gruionu
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Cosmin Gabriel
authors.initials: CG
authors.lastname: Cristea
authors.affiliation: Adriana Ciocâlteu, Adrian Săftoiu, Tatiana Cârţână, Dan Ionuț Gheonea, Lucian Gheorghe Gruionu, Cosmin Gabriel Cristea, Gabriel Gruionu, Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, 200349 Craiova, Romania.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Gruionu
keywords: Anti-CD105 antibody; Confocal laser endomicroscopy; Neoangiogenesis; Panendothelial markers; Rectal cancer