This study was performed to evaluate the effect of bone graft materials including demineralized freeze-dried bone, freeze-dried bone, deproteinized bovine bone on space-making capacity and bone formation in guided bone regeneration with titanium reinforced ePTFE membrane(TR-ePTFE). Adult male rabbits(mean BW 2kg) were used in this study. Intramarrow penetration defects were surgically created with round bur on calvaria of rabbits. TR-ePTFE membrane was adapted to calvarial defect and bone graft materials were placed. Animals were sacrificed at 2, 8, 12 weeks after surgery. Non-decalcified specimens were processed for histologic analysis and prepared with Villaneuva bone stain. The results of this study were as follows: 1. TR-ePTFE membrane was biocompatible and capable of maintaining the space-making. 2. Tissue integration was not good at TR-ePTFE membrane. Fixation was not enough. so, wound stabilization was not good. 3. In animals using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was little. 4. In animals using freeze-dried bone, bone formation was better. Within the above results, bone formation may be inhibited when wound stabilizafion was not good.
Adult ; Male ; Female ; Humans ; Rabbits ; Animals

Runx2 and Osterix, the transcription factors for osteoblast differentiation, are known as fundamental factors to regulate the development of calcified tissues. However, the biological functions of these factors in the development of the periodontal tissues remain unclear. In this study, we investigated the distribution of Runx2 and Osterix during periodontal tissue development of the mice. Mandibles from 14-day-old mice were prepared for paraffin section. Serial sections of the mandible containing 1st molar tooth germs were obtained as a thickness of 7 microm. Some sections were stained with hematoxylin and eosin. Others were used for immunohistochemistry for PCNA, Runx2, and Osterix. Epithelial cells in growing end of Hertwig's epithelial root sheath (HERS) and mesenchymal cells adjacent to the growing end of HERS expressed PCNA. Undifferentiated mesenchymal cells and hard tissue forming cells like cementoblasts and osteoblasts in early stage of differentiation expressed Runx2. Fully differentiated cementoblasts and osteoblasts secreting matrix proteins expressed Osterix. However, the cells terminated the matrix formation did not express Osterix. Periodontal ligament cells expressed Runx2 and Osterix. Pulp cells expressed Runx2 only.These results suggest that Runx2 and Osterix might regulate the differentiation of cementoblasts in the same manner as osteoblasts. Runx2 might participate in the process of cementoblast differentiation in early stage, whether Osterix might regulate the maturation and matrix synthesis of the cells.
Animals ; Dental Cementum ; Eosine Yellowish-(YS) ; Epithelial Cells ; Hematoxylin ; Immunohistochemistry ; Mandible ; Mice ; Molar ; Osteoblasts ; Paraffin ; Periodontal Ligament ; Proliferating Cell Nuclear Antigen ; Proteins ; Tooth Germ ; Transcription Factors

Our previous principal component analysis conducting on reference points, lines and angles, and a vectordeveloped polar coordinate system has elucidated that the components of eigenvectors had positive relationships in the curvature of anterior teeth segment, between the protrusion of canines and degree of arch roundness, and in the length-to-width ratio of 62 maxillary dentitions, which were preliminarily classified with reference to the conventional Thompson's morphological descriptions for dental arch forms. In the present study on morphological characters of the maxillary dentitions, we conducted a Fourier analysis on the previously obtained data. We observed that the amplitude of 2nd, 3rd and 4th Fourier harmonics were closely correlated with the length-to-width ratio, curvature of the anterior teeth segment, and the curvilinear contour of maxillary dental arches. In addition, the relationships between previously estimated data and the constant value and the amplitude of the Fourier series were examined by analysis of correlation coefficients (p<0.01). The results of the present study suggest that the morphology of maxillary dentitions consists of three essentials-the length-to-width ratio, the curvature of anterior teeth and the curvilinear contour of dental arches.
Dental Arch ; Dentition ; Fourier Analysis ; Principal Component Analysis ; Tooth

In this study, author examined the effect of the concentration of the inorganic polyphosphate on the process of the bone regeneration by using the 6 weeks old rabbit with the weight of 2.0kg in average. we performed the experiment by using TR-ePTFE membrane filled with collagen immersed with 1%, 2%, and 4% of inorganic polyphosphate, respectively, after removing the proper sized cortical bones from the calvaria of rabbit. The experimental results were compared with the one of the following four groups: The control group for membrane only, experimental group I for membrane filled with collagen immersed with 1% of inorganic polyphosphate, experimental group II for membrane filled with collagen immerse with 2% of inorganic polyphosphate, experimental group III for membrane filled with collagen immersed with 4% of inorganic polyphosphate. The fragments of the tissue with membrane were obtained from each group of the sacrificed rabbits for 4 or 8 weeks sustained after surgery, were then prestained and coated. New bone formation was assessed by histomorphometric and statistical analysis. We may draw the conclusions from these experiments as following: 1. Collagen was an excellent carrier with a minimal inflammatory reaction and sustaining the form. 2. The sample of the 8th week group has shown the best bone regeneration compared with the cases of all groups including the control group. 3. The samples of collagen immersed with 2% and 4% of inorganic polyphosphate have shown more bone regeneration relative to the sample of the 1% inorganic polyphosphate. 4. The new bone regeneration was shown actively in the group for membrane filled with collagen immersed with 4% of inorganic polyphosphate. With above results, it is strongly suggested the use of inorganic polyphosphate with vehicle under TR-ePTFE membrane.
Bone Regeneration* ; Collagen ; Membranes ; Osteogenesis ; Rabbits ; Skull

Bone Matrix ; Cell Line ; Dental Cementum ; Dental Enamel ; Durapatite ; Fibroblasts ; Humans ; Immunoassay ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteocalcin ; Osteopontin ; Periodontal Ligament ; Periodontium ; Proteins ; Regeneration

Porphyromonas gingivalis has been implicated in periodontal diseases. Accumulating evidence suggests that cardiovascular disease is the most prevalent medical problem in patients with periodontal diseases. In order to check the possibility that P. gingivalis is involved in coronary heart disease, the present study was performed to observe P. gingivalis adherence and invasion of human coronary artery endothelial cells (HCAEC) and production of cytokines and growth factors by HCAEC upon P. gingivalis infection. 3H-labeled P. gingivalis 381 was incubated with HCAEC for 90 min. The radioactivity of the washed HCAEC was a measure of the absorbed (adhering and invading) P. gingivalis. The absorption radioactivity of the HCAEC infected by P. gingivalis was determined to be 59.58% of the input bacterial cells. In contrast, the absorption radioactivity of the cells infected by S. gordonii Challis which was employed as a control was negligible (0.59%). DPG3, a P. gingivalis mutant defective of fimbriae, appeared to be impaired to some extent in capability of adherence/invasion as compared to that of the parental strain 381, showing 43.04% of the absorption radioactivity. The absorption radioactivity of the HCAEC infected by P. gingivalis 381 in the presence of excessive fimbriae at the concentrations of 50 mug and 100 mug/ml was 57.27 and 45.44%, respectively. Invasion of HCAEC by P. gingivalis 381 was observed by an antibiotic (metronidazole) protection assay and transmission electron microscopy (TEM). In the antibiotic protection assay, invasion by the bacterium was measured to be 0.73, 1.09, and 1.51% of the input bacterial cells after incubation for 30, 60, and 90 min, respectively. Invasion by DPG3 was shown to be 0.16% after 90-min incubation. In comparison of invasion efficiency at 90 min of the incubation, the invasion efficiency of DPG3 was 0.37% while that of its parental strain 381 was 2.54%. The immunoblot analysis revealed fimbriae of P. gingivalis did not interact with the surface of HCAEC. These results suggest that fimbriae are not the major contribution to the adherence of P. gingivalis to HCAEC but may be important in the invasion of HCAEC by the bacterium. The presence of cytochalasin D (1 mug/ml) and staurosporine (1 muM) reduced the invasion of HCAEC by P. gingivalis 381 by 78.86 and 53.76%, respectively, indicating that cytoskeletal rearrangement and protein kinase of HCAEC are essential for the invasion. Infection of P. gingivalis induced HCAEC to increase the production of TNF-alpha by 60.6%. At 90 min of the incubation, the HCAEC infected with P. gingivalis cells was apparently atypical in the shape, showing loss of the nuclear membrane and subcellular organelles. The overall results suggest that P. gingivalis may cause coronary heart disease by adhering to and invading endothelial cells, and subsequently damaging the cells.
Absorption ; Cardiovascular Diseases ; Coronary Disease* ; Coronary Vessels ; Cytochalasin D ; Cytokines ; Endothelial Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; Microscopy, Electron, Transmission ; Nuclear Envelope ; Organelles ; Parents ; Periodontal Diseases ; Porphyromonas gingivalis* ; Porphyromonas* ; Protein Kinases ; Radioactivity ; Staurosporine ; Tumor Necrosis Factor-alpha