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Apatite/Amelogenin Coating on Titanium Promotes Osteogenic Gene Expression

¹ Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, 2250 Alcazar Street, CSA 103, Los Angeles, CA 90033; and
² Tissue Engineering and Bone Biology Lab, Dows Institute for Dental Research, University of Iowa College of Dentistry, Iowa City, IA, USA;

View larger version (34K): [in this window] [in a new window]   Figure 1. Characterization of amelogenin incorporation into the coating and its release kinetics into solutions. (A) Fluorescent light micrographs of the FITC labeling on the apatite coatings prepared in the absence (left) and the presence of rM179 (25 µg/mL, right). (B) Analytical HPLC profiles of the elution solutions in distilled water at different time points (2–90 hrs) and the dissolved coating solution after 90 hrs. The scale used for the elution solutions was enlarged 10 times. The samples were prepared at 50 µg/mL rM179 by superficial adsorption (left) or co-precipitation (right) processes. (C) The cumulative release profile of a sample prepared by co-precipitation process.

View larger version (109K): [in this window] [in a new window]   Figure 2. Morphology of HEPM pre-osteoblast cells. Fluorescent light micrographs of cell attachment and spreading on glass coverslips (A), blank apatite coating (B), and rM179-containing coating prepared at a protein concentration of 100 µg/mL (C). The cells were immunofluorescently labeled for actin and the focal adhesion protein vinculin. Original magnification, 430X. SEM micrographs of cells cultured on cpTi (D), blank apatite coating (E), and rM179-containing coating prepared at a protein concentration of 50 µg/mL (F). Square indicates a nodule formation. Hollow arrows indicate the filopodia of the cells.

View larger version (22K): [in this window] [in a new window]   Figure 3. Normalized real-time RT-PCR analysis of gene expression levels by HEPM cells. The cells were cultured in triplicate, and RNA extracts for each culture were analyzed in triplicate. The data are presented as mean ± SD. (A) The cells were cultured on tissue culture plastic (TCP), commercially pure titanium (cpTi), and biomimetic apatite coating on titanium, without (HAP) and with different quantities of amelogenin incorporated into the coating. The x-axis numbers are concentrations of rM179 in µg/mL in the calcifying solution during the coating preparation. (B) The cells were cultured on tissue culture plastic in the absence (TCP) and the presence of 50 and 1000 ng/mL recombinant amelogenin (rM179) or bovine serum albumin (BSA) in the culture media.

Journal of Dental Research, Vol. 84, No. 11, 1070-1074 (2005)
DOI: 10.1177/154405910508401120


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