Phytocystatin CsinCPI-2 Reduces Osteoclastogenesis and Alveolar Bone Loss.

Periodontal disease (PD) is a polymicrobial chronic inflammatory condition of the supporting tissues around the teeth, leading to the destruction of surrounding connective tissue. During the progression of PD, osteoclasts play a crucial role in the resorption of alveolar bone that eventually leads to the loss of teeth if the PD is left untreated. Therefore, the development of antiresorptive therapies targeting bone-resorbing cells will significantly benefit the treatment of PD. Here, we demonstrate the inhibitory effect of CsinCPI-2, a novel cysteine peptidase inhibitor from the orange tree, on periodontitis-induced inflammation, alveolar bone loss, and osteoclast differentiation. Using the ligature-induced periodontitis model in mice, we show that treatment with CsinCPI-2 (0.8 µg/g of body weight) significantly reduced inflammatory cell infiltrate in the connective tissue and prevented the loss of alveolar bone mass (BV/TV) caused by PD, effects associated with diminished numbers of TRAP-positive multinucleated cells. Furthermore, CsinCPI-2 significantly downregulated the numbers of inflammatory cells expressing CD3, CD45, MAC387, and IL-1ß. In vitro, CsinCPI-2 inhibited RANKL-induced TRAP+ multinucleated osteoclast formation in mouse bone marrow macrophage cultures in a concentration-dependent manner. This effect was not due to cytotoxicity, as demonstrated by the MTT assay. CsinCPI-2 inhibited RANKL-induced mRNA expression of Acp5, Calcr, and Ctsk, as well as the RANKL-induced upregulation of Nfatc1, a crucial transcription factor for osteoclast differentiation. Based on our findings, CsinCPI-2 prevents bone loss induced by PD by controlling the inflammatory process and acting directly on osteoclastogenesis, suggesting an interesting potential for CsinCPI-2 in the strategy for PD treatment.

A randomized clinical trial evaluating maxillary sinus augmentation with different particle sizes of demineralized bovine bone mineral: histological and immunohistochemical analysis.

This study was performed to investigate sinus floor augmentation with two different particle sizes of demineralized bovine bone mineral (DBBM) by means of histological and immunohistochemical (IHC) analysis. A randomized clinical trial was conducted involving 10 individuals requiring two-stage bilateral maxillary sinus augmentation for implant installation. The patients were randomly divided into two groups following a split-mouth design: the maxillary sinus on one side was filled with small-sized particles (0.25-1mm) and on the contralateral side with large-sized particles (1-2mm). After a healing period of 8 months, 25 implants were placed. During implant site preparation, bone biopsies were obtained from each sinus, perpendicular to the long axis of the implant (buccal-palatal direction), for descriptive and histomorphometric analyses. IHC staining for protein expression of osteocalcin (OCN), vascular endothelial growth factor (VEGF), and tartrate-resistant acid phosphatase (TRAP) was also performed. Histomorphometric analysis revealed no statistically significant difference in the percentage of biomaterial (32.4±8.56% and 38.0±6.92%), newly formed bone (36.1±9.60% and 36.7±5.79%), or connective tissue (30.4±8.63% and 23.8±6.16%) between the small- and large-sized particle groups, respectively. IHC analysis did not reveal differences in the expression of OCN, VEGF, or TRAP. These findings suggest that both particle sizes of DBBM are effective for bone augmentation in the maxillary sinus.

Implant stability after sinus floor augmentation with deproteinized bovine bone mineral particles of different sizes: a prospective, randomized and controlled split-mouth clinical trial.

The aim of this study was to compare implant stability after maxillary sinus floor augmentation using small- or large-sized particles of Bio-Oss. Ten partially edentulous patients requiring bilateral maxillary sinus floor augmentation were enrolled. The subjects were assigned randomly to one of two experimental groups: maxillary sinus was filled with 0.25-1mm particle size (small particles) and the contralateral side was filled with 1-2mm particle size (large particles). After 8 months, a total of 25 implants were placed in the two maxillary sinuses. Primary implant stability was measured immediately after implant placement (T0) using a torque controller and resonance frequency analysis (RFA). Six months after implant placement (T1), the implant stability was measured again. There were no postoperative complications in either particle size group, and the success rate for implant survival was 100%. All implants showed good primary stability as evidenced by high torque for the implant insertion in both groups. RFA revealed high ISQ values for all implants installed in both groups at T0 and T1. These results indicate that the size of the Bio-Oss particles (small and large) did not influence implant stability in the maxillary sinus. Indeed, small and large particles of Bio-Oss presented optimal properties, supporting their possible use as osteoconductive grafts.

Comparison of changes in dental and bone radiographic densities in the presence of different soft-tissue simulators using pixel intensity and digital subtraction analyses.

OBJECTIVES: To evaluate the influence of soft-tissue simulation materials on dental and bone tissue radiographic densities using pixel intensity (PI) and digital subtraction radiography (DSR) analyses. METHODS: 15 dry human mandibles were divided into halves. Each half was radiographed using a charge-coupled device sensor without a soft-tissue simulation material (Wm) and with 5 types of materials: acrylic (Ac), wax (Wx), water (Wt), wood (Wd) and frozen bovine tissue (Bt). Three thicknesses were tested for each material: 10 mm, 15 mm and 20 mm. The material was positioned in front of the mandible and the sensor parallel to the molar region. The radiation beam was perpendicular to the sensor at 30 cm focal spot-to-object distance. The digital images of the bone and dental tissue were captured for PI analyses. The subtracted images were marked with 14 landmark magnifications, and 2 areas of analyses were defined, forming the regions of interest. Shapiro-Wilk and Kruskal-Wallis tests followed by Dunn's post-test were used (p < 0.05). RESULTS: DSR showed that both the material type and the thickness tested influenced the gain of density in bone tissue (p < 0.05). PI analyses of the bone region did not show these differences, except for the lower density observed in the image without soft-tissue simulation material. In the dental region, both DSR and PI showed that soft-tissue simulators did not influence the density in these regions. CONCLUSIONS: This study showed that the materials evaluated and their thicknesses significantly influenced the density-level gain in alveolar bone. In dental tissues, there was no density-level gain with any soft-tissue material tested.