HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text Full Text (PDF) Services Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (4) Citing Articles via Google Scholar Google Scholar Articles by Tran-Hung, L. Articles by About, I. Search for Related Content PubMed PubMed Citation Articles by Tran-Hung, L. Articles by About, I.

Role of Human Pulp Fibroblasts in Angiogenesis

¹ Laboratoire IMEB-ERT 30, Faculté d’Odontologie, Université de la Méditerranée, 27 Boulevard Jean Moulin, 13355 Marseille Cedex 05, France; and
² INSERM U559, Faculté de Médecine, Marseille, France

View larger version (119K): [in a new window]   Figure 1. Co-culture of human umbilical vein endothelial cells and human pulp fibroblasts. Human pulp fibroblasts and L929 fibroblastic cell line were fluorescence-labeled by transduction with the Enhanced Green Fluorescent Protein; endothelial cells were labeled with the Discosoma Red Fluorescent Protein-2. In co-cultures, pulp fibroblasts and endothelial cells remained separated after 3 hrs (A). Endothelial cells started to elongate and form closed structures at 24 hours (B,C). At 48 hrs, most endothelial cells were organized (D), and at 6 days, they formed a complete tubular network (E,F). Endothelial cells co-cultured with L929 cells remained separated from each other after 3 hrs (G), and no organization was found even after day 6 (H,I). Green and red fluorescence was simultaneously examined (A,B,D,E,G,H). The other photographs (C,F,I) were visualized by the red fluorescence only. Scale bar = 200 µm.

View larger version (51K): [in a new window]   Figure 2. Effect of indirect contact between pulp fibroblasts and endothelial cells. Morphological features of endothelial cells incubated after 24 hrs in control medium without contact with pulp fibroblasts (A), and after contact with conditioned media from intact pulp cells (B) and injured pulp fibroblasts (C). The capillary network of capillary-like structure formation on the Matrigel was photographed in fresh medium under a phase-contrast microscope. Scale bar = 200 µm.

View larger version (18K): [in a new window]   Figure 3. Effects of soluble factors secreted by pulp fibroblasts on the tubular perimeters of capillary-like structures formed by endothelial cells. The quantitative evaluation was done by measurment of the tubular perimeters in 30 fields of view per group in 3 independent experiments. Results are expressed as mean ± SD. Endothelial cells were seeded on the 24-well culture plate on Matrigel and cultured in conditioned media obtained from intact or injured pulp fibroblasts. Control medium (not in contact with pulp fibroblasts) was also used. After endothelial cells were incubated with these media, the formation of the capillary network was observed by phase-contrast microscopy. (A) The perimeters of tubular structures formed by endothelial cells were measured and expressed as a percentage of the perimeters obtained after the incubation of endothelial cells with the control medium. A significant increase in tube perimeter was observed with the conditioned media from intact and injured pulp fibroblasts. There was a significant increase in the tubular perimeter with the conditioned medium obtained from injured as compared with that obtained from intact fibroblasts (*p < 0.05). (B) The addition of FGF-2 or VEGF neutralizing antibodies to the conditioned media obtained from injured pulp fibroblasts resulted in a reduction in the tubular perimeters as compared with those observed without neutralizing antibodies. This difference was statistically significant when both neutralizing antibodies were used. The tubular perimeters were measured and expressed as a percentage of those obtained from injured pulp cells without neutralizing antibodies (*p < 0.05).

View larger version (76K): [in a new window]   Figure 4. Expression of FGF-2 and VEGF in human pulp cells. Pulp fibroblasts were cultured in four-well culture chambers. Injuries were made with scalpels on confluent cultures. After a five-hour culture period, immunohistochemistry showed the expression of FGF-2 (A) and VEGF (B) at a low level in pulp fibroblasts. Stronger expression of FGF-2 (D) and VEGF (E) could be observed after injury. Controls with no primary antibodies were negative in intact (C) and injured cells (F). Arrows indicate injury sites. Scale bars = 100 µm.