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Effects of Echistatin and an RGD Peptide on Orthodontic Tooth Movement

Department of Orthodontics, College of Dentistry, University of Florida, Box 100444, JHMHC, Gainesville, FL 32610-0444;

View larger version (52K): [in a new window]   Figure 1. RGD peptide and echistatin released from ELVAX disrupt osteoclasts on coverslips or bone slices. Mouse marrow osteoclasts were grown to maturity, loaded on slices of sperm whale dentin, and cultured for 2 days to allow the osteoclasts to adhere and become active. The osteoclasts were then left untreated (A), treated with ELVAX alone (B), or with ELVAX loaded with echistatin (C) or RGD peptide (D). After 90 min, the cells were fixed with 2% formaldehyde in PBS, permeabilized with 0.5% Triton X-100 in PBS, and stained with fluorescein-tagged phalloidin. Representative cells from the various treatment conditions are shown. Arrows point to the edges of the respective cells. The cell in each pane is approximately the same size, and all were photographed at the same magnification. The dramatic difference in appearance is the result of disruption of actin rings resulting from RGD-peptide and echistatin treatment. The bar is 5 µm. "I" shows quantitative analysis of the number of osteoclasts with actin rings on dentin slices after 90 min. The data represent mean ± SD of 4 experiments.

View larger version (88K): [in a new window]   Figure 2. Localization of the biotinylated RGD peptide in alveolar tissues. ELVAX was loaded with synthetic biotinylated RGD peptide (A) or alone (B) and implanted next to the maxillary first molars in rats. The rats were killed at day 6, and the diffusion of this biotinylated peptide was assessed. Tissue sections were incubated with avidin conjugated to horseradish peroxidase (HRP) and then visualized by treatment with 3,3'-diaminobenzidine (DAB). PDL, periodontal ligament. Note the increased staining in (A), indicating the presence of the biotinylated RGD peptide.

View larger version (17K): [in a new window]   Figure 3. Tooth movement kinetics in the control, echistatin-, or RGD-peptide-treated rats. Each point represents the mean ± SEM of 5–7 animals. *p < 0.01.

View larger version (15K): [in a new window]   Figure 4. Tooth drift kinetics in the control or RGD-peptide-treated rats. Each point represents the mean ± SEM of 5–7 animals. *p < 0.05.