Tumor neoangiogenesis detection by confocal laser endomicroscopy and anti-CD105 antibody:Pilot study

AIM: 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 preexisting parent tissue vessels and newly formed vessels.CD105(endoglin) represents a proliferation-associated endothelial cell adhesion molecule.In contrast to panendothelial 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/mm3 in only four patients).The average diameter of antiCD105 stained vessels was 10.97 ± 0.6 μm in tumor tissue,and the vessel density was 2787.40 ± 134.8 vessels/mm3.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/mm3,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/mm3.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.CONCLUSION: Specific imaging and quantification of tumor microvessels are feasible in human rectal cancer using CLE examination and CD105 immunostaining of fresh tissue samples.

Effect of growth hormone on colonic anastomosis after intraperitoneal administration of 5-fluorouracil, bleomycin and cisplatin: An experimental study

BACKGROUND Growth hormone(GH)plays a crucial role in wound healing and tissue repair in postoperative patients.In particular,colonic anastomosis healing following colorectal surgery is impaired by numerous chemotherapy agents.AIM To investigate whether GH can improve the healing of a colonic anastomosis following the adverse effects of intraperitoneal administration of 5-fluorouracil(5-FU),bleomycin and cisplatin.METHODS Eighty Wistar rats underwent laparotomy and a 1 cm-resection of the transverse colon,followed by an end-to-end anastomosis under general anesthesia.The rats were blindly allocated into four equal groups and administered a different daily intraperitoneal therapeutic regimen for 6 days.The control group(A)received normal saline.Group B received chemotherapy with 5-FU(20 mg/kg),bleomycin(4 mg/kg)and cisplatin(0.7 mg/kg).Group C received GH(2 mg/kg),and group D received the aforementioned combination chemotherapy and GH,as described.The rats were sacrificed on the 7th postoperative day and the anastomoses were macroscopically and microscop-ically examined.Body weight,bursting pressure,hydroxyproline levels and inflammation markers were measured.RESULTS All rats survived until the day of sacrifice,with no infections or other complications.A decrease in the body weight of group D rats was observed,not statistically significant compared to group A(P=1),but significantly different to groups C(P=0.001)and B(P<0.01).Anastomotic dehiscence rate was not statistically different between the groups.Bursting pressure was not significantly different between groups A and D(P=1.0),whereas group B had a significantly lower bursting pressure compared to group D(P<0.001).All groups had significantly more adhesions than group A.Hydroxyproline,as a measurement of collagen deposition,was significantly higher in group D compared to group B(P<0.05),and higher,but not statistically significant,compared to group A.Significant changes in group D were recorded,compared to group A regarding inflammation(3.450 vs 2.900,P=0.016)and fibroblast activity(2.75 vs 3.25,P=0.021).Neoangiogenesis and collagen deposition were not signifi-cantly different between groups A and D.Collagen deposition was significantly increased in group D compared to group B(P<0.001).CONCLUSION Intraperitoneal administration of chemotherapy has an adverse effect on the healing process of colonic anastomosis.However,GH can inhibit the deleterious effect of administered chemotherapy agents and induce colonic healing in rats.

Prefabricated flaps and neoangiogenesis initiated via venous grafts in arteriovenous Ioops

New developments in regenerative medicine are bound to revolutionize the way we approach loss of function and form in human organisms. Especially in the field of reconstructive plastic surgery new biotechnologies find their way from bench to bed. Biofabrication is an evolving field that aims to combine natural biologic processes with bioartificial constructs with the scope of reconstituting tissue without having to rely on autotransplantation. In this brief review we present the concepts of intrinsic vs. extrinsic neovascularization and we discuss the use of neovascularization in three dimensional matrices. In a clinical context matrix flaps for application in reconstructive surgery can be fabricated this way.

Morphological substantiation of application of cellular technologies for correction of striae

The possibility of the application of autologic pluripotent cells from peripheral blood (APCPB) for correction of striae after pregnancy was studied. Visually first signs of improvement of atrophic scars can be noticed in 6-8 weeks after injection of APCPB and gradually progress during 6 months. The atrophy of epithelium, hyperkeratosis, sclerosis of all dermis layers with smoothing of papillae, reduction of number of blood vessels and skin appendages were found before treatment by method of light mi-croscopy. In all observations after injection of APCPB in a connective tissue, which replaces of dermis, an increase of number of vessels at the expense of processes of neoangiogenesis, leukocytic infiltration reduction, thinning and ordering of arrangement of collagen and elastin fibers were detected. These results give a chance for more successful application of cosmetology procedures for correction of these defects of skin.

Prognostic Significance of Pigment Epithelium-Derived Factor Expression in Patients with Non-Small-Cell-Lung Cancer

The aims of this study were to examine prognostic significance of pigment epithelium-derived factor (PEDF) in patients with stage IA-IIIB non-small cell lung cancer (NSCLC). Using immunohistochemistry and multivariate analysis, we set out to investigate whether PEDF expression could provide prognostic information in NSCLC in a cohort of 69 patients who had undergone radical resection for NSCLC. The correlation between PEDF and the clinical pathological features of stage I-III NSCLC after radical surgery were analyzed as well as influence on long term survival. No correlation between PEDF intensity, PEDF area or PEDF area index and clinic opathologic parameters was seen. PEDF values showed a slight correlation to the tumor stage. There was a significant negative correlation (T = -0.288, p = 0.002) between pathologic T-stage and median PEDF area and vice versa a positive correlation (T = 0.227, p = 0.016) with median PEDF intensity. We could not detect any correlation between PEDF and long term survival. For PEDF analysis, there was only a slight correlation between expression and T-stage of the tumor.

Prognostic Significance of CD31 Expression in Patients with Non-Small-Cell-Lung Cancer

Non-small cell lung cancer (NSCLC) is the primary cause of cancer related death worldwide. After resection of early stage NSCLC, the benefit of adjuvant chemotherapy for patient survival still remains unclear and investigations for further risk stratification are needed for an improved treatment decision. Microvessel density (MVD) influences both the nutrition of the cancer and the access to the bloodstream for the development of distant metastasis. The aim of this study was to examine the prognostic significance of microvessel density by CD31 staining in patients with resected stage IA-IIIB NSCLC. We used immunohistochemistry (IHC) of CD31 to examine the microvessel density in a cohort of 69 patients who had undergone radical resection for NSCLC. Correlation of IHC values and standard clinicopathologic parameters was analyzed as well as influence on long term survival. Survival analysis revealed a significant better overall survival for patients with higher median microvessel density (log rank p = 0.031) independent of clinicopathologic parameters. Regarding primary cancer related death, the survival was again significantly longer in patients with high CD31 count (log rank p = 0.036). A higher microvessel density was a strong predictor for a longer tumor related survival and could be used for therapeutic decisions of adjuvant chemotherapy after resection of early stage NSCLC.

Interplay between Platelet Endothelial Cell Adhesion Molecule-1 and Pigment Epithelium-Derived Factor in Non-Small-Cell-Lung Cancer

Pigment epithelium-derived factor (PEDF), a potent antiangiogenesis agent, is a multifunctional protein with important roles in regulation of inflammation and angiogenesis. It has recently attracted attention for targeting tumor cells in several types of tumors. PECAM-1 is an integral membrane protein, a cell adhesion molecule with proangiogenic activity and plays an important role in the process of angiogenesis. The correlation between proangiogenic activity PECAM-1 and antiangiogenic activity PEDF in Non-Small-Cell-Lung Cancer has not been reported. The present study was designed to evaluate using immunohistochemical techniques and multivariate analysis the interplay between PECAM-1 and PEDF in NSCLC, especially in adenocarcinoma and in squamous cell carcinoma stage IA-IIIB. Analyzing the mixed study collectively (n = 69), there was no significant correlation (p = 0.553) between PECAM-1 signal and PEDF area. Only including patients with adenocarcinoma (Figure 2), we found a positive correlation between PECAM-1 signal and PEDF area (p = 0.025). In patients with squamous cell carcinoma, we did not find a significant correlation between PECAM-1 signal and PEDF area (p = 0.530). In patients with squamous cell carcinoma, PECAM-1 and PEDF show a significant different expression pattern, measured via staining intensity (p = 0.013). These results might support the hypothesis that squamous cell carcinomas heavily rely on angiogenic processes.