BACKGROUND/AIMS: Tumor necrosis factor-alpha (TNF-alpha) exerts its anti-tumor effect through direct cytotoxicity on tumor cells and damage to the tumor vasculature. However, its role in tumor angiogenesis is controversial. We evaluated the angiogenic effect of TNF-alpha on BALB/c mouse colon carcinoma homograft model. METHODS: Ten BALB/c mice were inoculated intraperitoneally with CT-26 mouse colon carcinoma cells. After a week, recombinant mouse TNF-alpha (2microgram/mL) were given four times on every other day to five animals and the same volume of phosphate buffered saline was given at the same interval to five animals as control. Harvested tumor tissues were stained by immunohistochemistry with CD31 and VEGF antibodies. Number of microvessels and VEGF expression were counted by light microscope. RESULTS: The mean microvessel counts per 200x field of TNF-alpha treated animals were 70.2+/-7.8 and those of nontreated animals were 83.8+/-8.3 (p<0.05). The VEGF score of both groups were 3. CONCLUSIONS: TNF-alpha treated animals showed decreased microvessel counts in tumor tissue but VEGF expression in both groups showed no difference. Therefore, TNF-alpha showed antiangiogenic effects on colon carcinoma homograft model.
Animals ; Antigens, CD31/analysis ; Carcinoma/chemistry/*physiopathology ; Cell Line, Tumor ; Colonic Neoplasms/chemistry/*physiopathology ; English Abstract ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Neovascularization, Pathologic/*physiopathology ; Tumor Necrosis Factor-alpha/*pharmacology ; Vascular Endothelial Growth Factor A/analysis

PURPOSE: Distant metastasis and recurrence are major prognostic factors associated with breast cancer. Both lymphovascular invasion (LVI) and blood vessel invasion (BVI) are important routes for metastasis to regional lymph nodes and for systemic metastasis. Despite the importance of vascular invasion as a prognostic factor, application of vascular invasion as a histopathological criterion is controversial. The aim of this study was to distinguish LVI from BVI in prognosis and recurrence of breast cancer using an endothelial subtype specific immunohistochemical stain (podoplanin, D2-40, and CD31). METHODS: Sections from 80 paraffin-embedded archival specimens of invasive breast cancer were stained for podoplanin, D2-40, or CD31 expression. Immunohistochemical staining results were correlated with clinicopathological features, such as tumor size, status of lymph node metastases, estrogen receptor status, progesterone receptor status, human epidermal growth factor receptor-2 expression, and recurrence. Patients with ductal carcinoma in situ and stage IV breast cancer were excluded. RESULTS: A significant correlation was found between D2-40 LVI positivity and lymph node metastasis (p=0.022). We found a significant correlation between D2-40 LVI positivity and recurrence of breast cancer (p=0.014). However, no significant correlation was found between BVI and recurrence. A poorer disease free survival was shown for D2-40 positive LVI (p=0.003). In a multivariate analysis, the presence of D2-40 LVI positivity revealed a significant association with decreased disease-free survival. CONCLUSION: D2-40 LVI positivity was a more prognostic predictor of breast cancer than BVI.
Antibodies, Monoclonal, Murine-Derived ; Antigens, CD31 ; Blood Vessels ; Breast ; Breast Neoplasms ; Carcinoma, Ductal ; Carcinoma, Intraductal, Noninfiltrating ; Disease-Free Survival ; Epidermal Growth Factor ; Estrogens ; Glycosaminoglycans ; Humans ; Lymph Nodes ; Multivariate Analysis ; Neoplasm Metastasis ; Prognosis ; Receptors, Progesterone ; Recurrence

To increase the biocompatibility and durability of glutaraldehyde (GA)-fixed valves, a biological coating with viable endothelial cells (ECs) has been proposed. However, stable EC layers have not been formed successfully on GA-fixed valves due to their inability to repopulate. In this study, to improve cellular adhesion and proliferation, the GA-fixed prostheses were detoxified by treatment with citric acid to remove free aldehyde groups. Canine bone marrow mononuclear cells (MNCs) were differentiated into EC-like cells and myofibroblast-like cells in vitro. Detoxified prostheses were seeded and recellularized with differentiated bone marrow-derived cells (BMCs) for seven days. Untreated GA-fixed prostheses were used as controls. Cell attachment, proliferation, metabolic activity, and viability were investigated and cell-seeded leaflets were histologically analyzed. On detoxified GA-fixed prostheses, BMC seeding resulted in uninhibited cell proliferation after seven days. In contrast, on untreated GA-fixed prostheses, cell attachment was poor and no viable cells were observed. Positive staining for smooth muscle a-actin, CD31, and proliferating cell nuclear antigen was observed on the luminal side of the detoxified valve leaflets, indicating differentiation and proliferation of the seeded BMCs. These results demonstrate that the treatment of GA-fixed valves with citric acid established a surface more suitable for cellular attachment and proliferation. Engineering heart valves by seeding detoxified GA-fixed biological valve prostheses with BMCs may increase biocompatibility and durability of the prostheses. This method could be utilized as a new approach for the restoration of heart valve structure and function in the treatment of end-stage heart valve disease.
Tissue Fixation ; Tissue Engineering/*methods ; Swine ; Proliferating Cell Nuclear Antigen/analysis ; Muscle, Smooth/chemistry ; Microscopy, Electron, Scanning ; Immunohistochemistry ; Heart Valves/cytology/*physiology ; Heart Valve Prosthesis ; Glutaral/*chemistry ; Endothelial Cells/cytology/physiology ; Dogs ; Cell Survival/physiology ; Cell Proliferation ; Cell Differentiation/physiology ; Cell Culture Techniques/*methods ; Cell Adhesion/physiology ; Bone Marrow Cells/chemistry/*physiology/ultrastructure ; Antigens, CD31/analysis ; Animals ; Actins/analysis

Cartilage oligomeric matrix protein-angiopoietin-1 (COMP-Ang1) is an angiogenic factor for vascular angiogenesis. The aim was to investigate the effect of an intracavernosal injection of COMP-Ang1 on cavernosal angiogenesis in a diabetic rat model. Male Otsuka Long-Evans Tokushima Fatty (OLETF) rats made up the experimental group (1 yr old) and Long-Evans Tokushima Otsuka (LETO) rats made up the control group. The experimental group was divided into vehicle only, 10 microg COMP-Ang1, and 20 microg COMP-Ang1. COMP-Ang1 was injected into the corpus cavernosum of the penis. After 4 weeks, the penile tissues of the rats were obtained for immunohistochemistry and Western blot analysis. The immunoreactivity of PECAM-1 and VEGF was increased in the COMP-Ang1 group compared with the vehicle only group. Moreover, the expression of PECAM-1 and VEGF was notably augmented in the 20 microg Comp Ang-1 group. In the immunoblotting study, the expression of PECAM-1 and VEGF protein was significantly less in the OLEFT rats than in the control LETO rats. However, this expression was restored to control level after intracavernosal injection of COMP-Ang1. These results show that an intracavernosal injection of COMP-Ang1 enhances cavernous angiogenesis by structurally reinforcing the cavernosal endothelium.
Angiopoietin-1/genetics/*metabolism ; Animals ; Antigens, CD31/metabolism ; Blood Glucose/analysis ; Blotting, Western ; Body Weight ; Cartilage Oligomeric Matrix Protein/genetics/*metabolism ; Diabetes Mellitus, Experimental/*pathology ; Immunohistochemistry ; Male ; Neovascularization, Physiologic/*drug effects ; Penis/metabolism/pathology ; Rats ; Rats, Long-Evans ; Recombinant Fusion Proteins/biosynthesis/genetics/*pharmacology ; Vascular Endothelial Growth Factor A/metabolism