Int Poster J Dent Oral Med 2003, Vol 5 No 03, Poster 191
Immunohistomorphometry of VEGF-Levels in Periodontitis and Systemic Sclerosis (Ssc)
Dr. med. dent. Georg Gassmann, Prof. Dr. med. dent. habil. Wolf-Dieter Grimm
ZMK Klinik, Abteilung für Parodontologie Universität Witten/Herdecke;
PhD Sarah Geva
School of Dentistry, University of North Carolina at Chapel Hill, USA;
Prof. Dr. Statis Philippou
Institut für Pathologie, Augustakrankenanstalt, Ruhr-Universität Bochum;
PD Dr. med. dent. Jochen Jackowski
ZMK Klinik, Abteilung für Zahnärztliche Chirurgie, Universität Witten/Herdecke.
International Congress of Periodontal Research, Molecular Determinants of Risk
University at Chapel Hill, North Carolina, USA
Vascular endothelial growth factor (VEGF) as an angiogeneticic cytokine is a potent growth factor for endothelial cell proliferation (Kikuchi et al. 1998). Its level has been proven to be elevated in systemic sclerosis (SSc) as well as in periodontitis (Booth et al. 1998;Chapple et al.2000; Harada et al. 1998).
It was the aim of this study to demonstrate whether VEGF-levels in immunostained gingival biopsies quantified by immunohistomorphometry may be regarded as a specific risk indicator for the progression of SSc with periodontitis in comparison to periodontitis alone.
Material and Methods
Within indicated surgical procedures (Fig. 1) biopsies were taken from 13 patients with SSc (Fig.2) leading to 29 preparations and from 8 patients with chronic periodontitis without SSc leading to 13 preparations.
Indicated surgical prodedure within a patient with advanced periodontitis.
Patient with systemical sclerosis showing microstomia.
Patient with systemical sclerosis showing perioral pleating.
Immunohistochemical testing was performed applying
the monoclonal antibody anti-human VEGF Clone G 153-694 (Pharmingen, San Diego, CA92121,USA)(Fig. 3 and 4).
|Fig.3: Positive Immunohisto-chemical staining in a patient with periodontitis. The arrows are pointig to VEGF expression within the wall of a bloodvessel and around fibroblasts.
||Fig.4: The slide preparation of a patient with SSc and periodontitis performs positively stained areas within blood-vessel walls and around fibroblasts too.
Transverse gingival sections from each patient sample were analyzed for VEGF expression evaluation.
The area of positive immunostaining in the gingival subepithelial connective tissue was measured using a microscope connected to a computerized video digital system (JAVA video analysis software, Jandel Scientific, Corte Madera, Ca) (Fig.5a and b) at a magnification of x100.
As an example this figure demonstrates that the black areas in A show the positively stained areas from Fig.3 in a patient with periodontitis and Fig. 5b figuring the areas stained in the slide preparation of a patient with SSc and periodontitis from Fig.4.
For each sample five different areas were analyzed and the data were pooled to represent a mean value. The results were expressed in percentage of the positively immunostained area per total connective tissue area measuring 0.3615mm2. The data gained were combined in each group of patients to create a group mean and pooled estimate of standard error. The significance was evaluated applying the Mann-Whitney-U-test.
The mean percentage of the positively immunostained area for VEGF in SSc was 7.6%±2.2 while measuring 5.3%±1.2 for chronic periodontitis. The mean rank for SSc was 26.6 and 10.12 for periodontitis. The rank sum for SSc was 771.5 and 131.5 for periodontitis. Thus the difference was highly significant (p<0.001).
|Fig.6: Periodontitis constantly provokes less staining.
|Fig.7: SSc and Periodontitis lead to a significantly higher expression of VEGF than periodontitis alone.
Using quantitative immuno-histomorphometry reveals VEGF expression in gingival biopsies to be elevated more in SSc than in chronic periodontitis. Our results suggest to use VEGF as a mole-cular marker to distinguish between SSc associated with periodontitis and periodontal diseases alone. Thus further studies have been initiated with larger groups of patients to corroborate or confute that VEGF is a sufficient parameter for risk assessement in SSc progression in comparison to progression in periodontal diseases (Johnson et al 1999) .
Kikuchi K, Kubo M, Kadono T, Yazawa N, IHN H, Tamaki K.: Serum concentrations of vascular endothelial growth factor in collagen diseases. Br J Dermatol 1998 Dec;139(6):1049-51
Harada M, Mitsuyama K, Yoshida H, Sakisaka S, Taniguchi E, Kawaguchi T, Ariyoshi M, Saiki T, Sakamoto M, Nagata K, Sata M, Matsuo K, Tanikawa K.: Vascular endothelial growth factor in patients with rheumatoid arthritis. Scand J Rheumatol 1998;27(5):377-80
Chapple CC, Kumar RK, Hunter N.: Vascular remodelling in chronic inflammatory periodontal disease. J Oral Pathol Med 2000 Nov;29(10):500-6
Johnson RB, Serio FG, Dai X.: Vascular endothelial growth factors and progression of periodontal diseases. J Periodontol 1999 Aug;70(8):848-52
Booth V, Young S, Cruchley A, Taichman NS, Paleolog E.: Vascular endothelial growth factor in human periodontal disease. J Periodontal Res 1998 Nov;33(8):491-9
SSc = Systemic Sclerosis
VEGF = Vascular Endothelial Growth Factor
This Poster was submitted by Prof. Dr. med. dent. habil. Wolf-Dieter Grimm.
Dr. med. dent. Georg Gassmann
ZMK Klinik, Abteilung für Parodontologie