Abstract
IntroductionNumerous anti-angiogenic agents are currently developed to limit tumor growth and metastasis. While these drugs offer hope for cancer patients, their transient effect on tumor vasculature is difficult to assess in clinical settings. Confocal laser endomicroscopy (CLE) is a novel endoscopic imaging technology that enables histological examination of the gastrointestinal mucosa. The aim of the present study was to evaluate the feasibility of using CLE to image the vascular network in fresh biopsies of human colorectal tissue. For this purpose we have imaged normal and malignant biopsy tissue samples and compared the vascular network parameters obtained with CLE with established histopathology techniques.Materials and MethodsFresh non-fixed biopsy samples of both normal and malignant colorectal mucosa were stained with fluorescently labeled anti-CD31 antibodies and imaged by CLE using a dedicated endomicroscopy system. Corresponding biopsy samples underwent immunohistochemical staining for CD31, assessing the microvessel density (MVD) and vascular areas for comparison with CLE data, which were measured offline using specific software.ResultsThe vessels were imaged by CLE in both normal and tumor samples. The average diameter of normal vessels was 8.5±0.9 µm whereas in tumor samples it was 13.5±0.7 µm (p = 0.0049). Vascular density was 188.7±24.9 vessels/mm2 in the normal tissue vs. 242.4±16.1 vessels/mm2 in the colorectal cancer samples (p = 0.1201). In the immunohistochemistry samples, the MVD was 211.2±42.9/mm2 and 351.3±39.6/mm2 for normal and malignant mucosa, respectively. The vascular area was 2.9±0.5% of total tissue area for the normal mucosa and 8.5±2.1% for primary colorectal cancer tissue.ConclusionSelective imaging of blood vessels with CLE is feasible in normal and tumor colorectal tissue by using fluorescently labeled antibodies targeted against an endothelial marker. The method could be translated into the clinical setting for monitoring of anti-angiogenic therapy.
Highlights
Numerous anti-angiogenic agents are currently developed to limit tumor growth and metastasis
Materials and Methods: Fresh non-fixed biopsy samples of both normal and malignant colorectal mucosa were stained with fluorescently labeled anti-CD31 antibodies and imaged by Confocal laser endomicroscopy (CLE) using a dedicated endomicroscopy system
Selective imaging of blood vessels with CLE is feasible in normal and tumor colorectal tissue by using fluorescently labeled antibodies targeted against an endothelial marker
Summary
Numerous anti-angiogenic agents are currently developed to limit tumor growth and metastasis. The aim of the present study was to evaluate the feasibility of using CLE to image the vascular network in fresh biopsies of human colorectal tissue. The advent of new anti-angiogenic agents into the oncological clinical practice has generated the need for enhanced imaging methods for evaluation of the microvascular network during treatment. A variety of clinical applications of the technique have already been described in lesions of both the upper and lower gastrointestinal tract, with particular interest on neoplasia, where CLE generates real-time histological diagnosis and targeted biopsies of relevant areas for a higher diagnostic yield than random biopsies [7,8,9,10,11,12]. CLE has shown high accuracy in detecting intraepithelial neoplasia based on the pattern of the vascular network and crypt architecture [7]
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