Effect of tension and compression on dynamic alveolar histomorphometry.

Here, we tested the hypothesis that tensile and compressive stresses generated in the alveolar bone proper regulate site-specific cellular and functional changes in osteoclasts and osteoblasts. Thirty-two 13-week-old male mice were randomly divided into four groups: two experimental groups with vertical loading obliquely from the palatal side to the buccal side of the maxillary molar (0.9 N) 30 min per day for 8 or 15 days and unloaded controls (n = 8). Calcein and alizarin were administered 8 and 2 days before euthanization, respectively, to detect the time of bone formation. Undecalcified sections parallel to the occlusal plane were prepared on the palatal root and the surrounding alveolar bone in the middle of the root length. The alveolar perimeter was divided into 12 equal regions for site analysis, and the bone histomorphometric parameters were obtained for each region. Data from in vivo microfocus computed tomography were used to construct animal-specific finite element models. 2D stress distribution images were overlain on histology images obtained from the same location. Significant differences in the total perimeter between groups and between loading and unloading in each region were statistically analyzed (α = 0.05). Osteoclast counts and the alizarin label ratio were significantly higher in the loaded group than in the unloaded group in regions where the maximum von Mises and principal tensile stresses were the highest along the perimeter. The label ratio of calcein was significantly lower in the 8-day loaded group than in the unloaded group, indicating that the calcein-labeled surface was resorbed by osteoclasts that appeared during the loading period. The effect of loading was mitigated by an increase in the maximum principal compressive stress. We conclude that bone resorption and bone formation are functions of site-specific tension and compression in the alveolar bone proper, confirming our hypothesis. This finding is critical for the advancement of diagnosis and treatment planning in clinical dentistry.

Effect of load-induced local mechanical strain on peri-implant bone cell activity related to bone resorption and formation in mice: An analysis of histology and strain distributions.

The purpose of this study was to investigate the effect of load-induced local mechanical strain on bone cell activity of peri-implant bone in mice. Titanium implants were placed in the maxillae of 13-week-old male C57BL/6J mice and subjected to intermittent 0.15 N, 0.3 N, or 0.6 N loads for 30 min/day for 6 days. The animals were sacrificed 2 days after the final loading. Unloaded mice were used as controls. An animal-specific three-dimensional finite element model was constructed based on morphological data retrieved from in vivo microfocus computed tomography for each mouse to calculate the mechanical strain distribution. Strain distribution images were overlaid on corresponding histological images of the same site in the same animal. The buccal cervical region of the peri-implant bone was predetermined as the region of interest (ROI). Each ROI was divided by four strain intensity levels: 0-20 µÎµ, 20-60 µÎµ, 60-100 µÎµ, and ≥100 µÎµ, and the bone histomorphometric parameters were analyzed by the total area of each strain range for all loaded samples. The distance between the calcified front and calcein labeling as a parameter representing the mineral apposition rate was significantly greater in the areas with strain intensity ≥100 µÎµ than in the area with strain intensity <100 µÎµ, suggesting that the bone formation activity of osteoblasts was locally enhanced by a higher mechanical strain. However, the shrunken osteocytes and the empty osteocyte lacunae were significantly lower in the highest strain area, suggesting that osteoclastogenesis was more retarded in higher strain areas than in lower strain areas. The histomorphometric parameters were not affected geometrically in the unloaded animals, suggesting that the load-induced mechanical strain caused differences in the histomorphometric parameters. Our findings support the hypothesis that bone cell activity related to bone resorption and formation is local strain-dependent on implant loading.

Amino acid residues modulating the activities of staphylococcal glutamyl endopeptidases.

The glutamyl endopeptidase family of enzymes from staphylococci has been shown to be important virulence determinants of pathogenic family members, such as Staphylococcus aureus. Previous studies have identified the N-terminus and residues from positions 185-195 as potentially important regions that determine the activity of three members of the family. Cloning and sequencing of the new family members from Staphylococcus caprae (GluScpr) and Staphylococcus cohnii (GluScoh) revealed that the N-terminal Val residue is maintained in all family members. Mutants of the GluV8 enzyme from S. aureus with altered N-terminal residues, including amino acids with similar properties, were inactive, indicating that the Val residue is specifically required at the N-terminus of this enzyme family in order for them to function correctly. Recombinant GluScpr was found to have peptidase activity intermediate between GluV8 and GluSE from Staphylococcus epidermis and to be somewhat less specific in its substrate requirements than other family members. The 185-195 region was found to contribute to the activity of GluScpr, although other regions of the enzyme must also play a role in defining the activity. Our results strongly indicate the importance of the N-terminal and the 185-195 region in the activity of the glutamyl endopeptidases of staphylococci.