No abstract

The aim of this study was to examine morphologically the participation of extraradicular biofilm in refractory periapical periodontitis. Six teeth and five extruded root filling gutta-percha points associated with refractory periapical periodontitis were investigated by scanning electron microscope. In nine of 11 samples examined, bacterial biofilms were seen at the extraradicular area. The gutta-percha surface was covered with glycocalyx-like structures, and filaments, long rods, and spirochete-shaped bacteria were predominant in the extraradicular sites. Planktonic cells, which were filaments and spirochete-shaped bacteria, emigrated from the glycocalyx structures in some spots. In the extracted teeth, biofilm consisting of both bacteria and glycocalyx-like structures were observed on the periapical root surfaces. Next to the residual periodontal ligament, a few filaments, rods, and fusiforms were attached on the healthy cementum surface. The present findings suggested that bacterial biofilms formed in the extraradicular areas were related to refractory periapical periodontitis.

No abstract

No abstract

No abstract

Hydroxyapatite/soluble calcium phosphate composites (HAp/SCaP) are novel HAp-based materials with enhanced solubility that have been developed by annealing HAp in a vacuum. This study compared the effects of HAp and HAp/SCaP on osteoblast proliferation, differentiation, and mineralization using an MC3T3-E1 cell culture system. MC3T3-E1 cells were cultured on HAp or HAp/SCaP, and the number of attached cells and their morphology were examined. The influence of the extract from HAp/SCaP on osteoblast differentiation was determined by the measurement of alkaline phosphatase activity and reverse transcriptase-polymerase chain reaction analysis of the expression of osteoblastic markers. In addition, mineralization was evaluated by the staining of calcium deposits with Alizarin red. Attachment of a greater number of cells exhibiting no degeneration in their morphology was observed on HAp/SCaP compared with HAp after incubation for 7 days or more. Culturing cells with the extract from HAp/SCaP resulted in promotion of alkaline phosphatase activity, the expression of type I collagen, and bone-like tissue formation. The results of the present study indicate that HAp/SCaP shows greater ability in osteogenesis than HAp by increasing collagen synthesis and calcification of the extracellular matrix.

The participation of bacterial biofilms in the over-filled gutta-percha points associated with refractory periapical periodontitis has recently been reported. This study investigated the initial biofilm-forming ability of root canal isolates (Enterococcus faecalis, Streptococcus sanguis, Strep. intermedius, Strep. pyogenes, Staphylococcus aureus, Fusobacterium nucleatum, Propionibacterium acnes, Porphyromonas gingivalis and Prevotella intermedia) on gutta-percha points in vitro. Each bacterial strain was suspended in 100% cell culture medium or in culture medium containing 4.5, 45 or 90% (vol/vol) serum. The bacterial suspensions were then co-incubated anaerobically with gutta-percha points for 7 d. The gutta-percha points were processed for scanning electron microscopic observation and examined for biofilm presence and thickness. E. faecalis, Strep. sanguis, Strep. intermedius, Strep. pyogenes and Staph. aureus biofilms were generated on the surfaces of the specimens incubated in culture medium supplemented with 45 or 90% (vol/vol) serum. The E. faecalis and Strep. sanguis biofilms were significantly thicker than those of Strep. intermedius, Strep. pyogenes and Staph. aureus. No biofilms were detected on the specimens incubated with F. nucleatum, Prop. acnes, Porph. gingivalis and Prev. intermedia. These findings suggest that Gram-positive facultative anaerobes have the ability to colonize and form extracellular matrices on gutta-percha points, while serum plays a crucial role in biofilm formation.

HAp and the related calcium phosphates have been of great interest as biological materials for regenerating hard tissues, but their solubility should be improved to fit the remodeling cycle of hard tissues in vivo before their practical use for bone grafts. Control of the grain size of hydroxyapatite (HAp) matrix and change in constituent phase of the surface layer were examined to improve the dissolution rate in HAp ceramics. Surface and boundary layers composed of calcium phosphate phases with high solubility were formed by annealing at 1350 • C in a vacuum, and the apparent mass transfer coefficient in the initial stage of the solubility experiment remarkably increased in an acetate buffer solution at pH of 4.0. The soluble surface and boundary layers are composed of tetracalcium diphosphate monoxide (TTCP), α-tricalcium phosphates (α-TCP) and amorphous calcium oxide (CaO) with high solubility, and their formation mechanism was discussed. Refinement of the grain size of HAp matrix also contributed to a slight increase in the apparent solubility.