Akira Hasuike

In vivo bone regenerative effect of low-intensity pulsed ultrasound in rat calvarial defects.

OBJECTIVES Low-intensity pulsed ultrasound (LIPUS) is a biophysical intervention in a bone repair process. However, neither the healing events of the flat bones of the skull using LIPUS nor the optimal stimulation settings are fully understood. The aim of this study was to evaluate the bone regenerative effect of LIPUS in rat calvarial flat bone defects by using in vivo microfocus computerized tomography (micro-CT). STUDY DESIGN The calvarium was exposed, and noncritical-sized 2.7-mm defects were prepared. LIPUS (1.6 MHz, repeating 1.0-kHz pulsation, and 30-mW/cm(2) intensity) was applied for 20 minutes daily. Bone regeneration was evaluated by image analysis using micro-CT and histologic examination. RESULTS Within 2 weeks, LIPUS-treated rats demonstrated 7.0% reossification of the original surgical defect, whereas control rats demonstrated 3.6%. At 3 and 4 weeks, a significant difference in the reossification ratio was observed (12.0% vs. 5.8% and 18.1% vs. 9.8%, respectively; P < .05). CONCLUSIONS LIPUS accelerated bone regeneration of noncritical rat calvarial defects as confirmed by micro-CT.

Micro‐CT observation of angiogenesis in bone regeneration

OBJECTIVES Restoration of an adequate blood supply is essential for the bone healing process and is key to the success of bone augmentation procedures. In this study, we evaluated angiogenesis in rat calvarial flat bone defects using in vivo microfocus computed tomography (micro-CT). MATERIALS AND METHODS Twenty rats were used. The calvarium was exposed and calvarial bone defects of critical (5-mm diameter) and non-critical (2.7-mm diameter) sizes were prepared. Bone regeneration and angiogenesis were evaluated by image analysis using micro-CT and histological examination. RESULTS Critical- and non-critical-sized calvarial bone defects showed bone regeneration and angiogenesis around the midsagittal suture. Critical-sized calvarial bone defects showed approximately 1.2% reossification of the original surgical defect, whereas the non-critical-sized defects showed approximately 43.3% reossification at day 28. Furthermore, angiogenesis was observed later in the critical-sized calvarial bone defects (about 38.2%), whereas angiogenesis was observed early in the non-critical-sized calvarial bone defects (about 75.5%) at day 28. New blood vessel networks were observed around defects of both sizes. CONCLUSIONS Angiogenesis preceded bone regeneration around critical- and non-critical-sized calvarial bone defects. Angiogenesis led to full bone formation in non-critical-sized defects.

Effects of intermittent administration of parathyroid hormone on bone augmentation in rat calvarium.

Objectives:This study examined the effects of parathyroid hormone (PTH) on bone augmentation beyond the skeletal envelope within a plastic cap in rat calvaria. Materials and Methods:The calvaria of 30 rats were exposed, and 2 plastic caps were placed on each. Each of the 10 rats was treated with 35 or 105 &mgr;g/kg (PTH-35, PTH-105) PTH 3 times per week. The control group was injected with sterile saline 3 times per week. Micro-computed tomography (CT) imaging was performed every 2 weeks for 12 weeks. Micro-CT and histological sections were used to determine the amount of bone augmentation within the plastic caps. Bone volume (BV) was calculated using BV-measuring software. Results:The histomorphometric and histological analyses showed that the amount of bone augmentation was increased significantly in the PTH groups compared with the controls at 12 weeks. The PTH-105 group showed significantly more bone augmentation and osteoblasts compared with the PTH-35 group. Conclusions:These results indicate that the higher the dose of intermittent PTH administered, the greater the amount of bone formation beyond the skeletal envelop in the rat calvarium.

Systematic review and assessment of systematic reviews examining the effect of periodontal treatment on glycemic control in patients with diabetes

Objetives There have been several systematic reviews(SRs) on whether periodontal treatment for an individual with both periodontal disease and diabetes can improve diabetes outcomes. The purpose of this investigation was to conduct a systematic review (SR) of previous meta-analyses, and to assess the methodological quality of the SRs examining the effects of periodontal treatment and diabetes. (PROSPERO Registration # CRD 42015023470). Study Design We searched five electronic databases and identified previous meta-analyses of randomized controlled trials published through July 2015. In cases where the meta-analysis did not meet our criteria, the meta-analyses were recalculated. General characteristics of each included trial were abstracted, analyzed, and compared. The mean difference, 95% confidence intervals (CIs) and the I2 statistic were abstracted or recalculated. The Assessment of Multiple Systematic Reviews Instrument (AMSTAR) was used to assess methodological quality. Results Of the 475 citations screened, nine systematic reviews were included. In total, 13 meta-analyses included in nine SRs were examined. In comparability analyses, meta-analyses in four SRs did not meet our criteria, and were recalcuated. Of these 13 meta-analyses, 10 suggested significant effects of periodontal treatment on HbA1c improvement. Mean differences found in the 13 meta-analyses ranged from -0.93 to 0.13. AMSTAR assessment revealed six SRs with moderate and three with high overall quality. Conclusions We can conclude that there is a significant effect of periodontal treatment on improvement of HbA1c in diabetes patients, although the effect size is extremely small. In addition to the small effect size, not all SRs could be considered of high quality. Key words:Periodontal treatment, diabetes, HbA1c, systematic review, systematic review of systematic reviews, evidence-based medicine, AMSTAR.

Melatonin enhances vertical bone augmentation in rat calvaria secluded spaces

Background Melatonin has many roles, including bone remodeling and osseointegration of dental implants. The topical application of melatonin facilitated bone regeneration in bone defects. We evaluated the effects of topical application of melatonin on vertical bone augmentation in rat calvaria secluded spaces. Material and Methods In total, 12 male Fischer rats were used and two plastic caps were fixed in the calvarium. One plastic cap was filled with melatonin powder and the other was left empty. Results Newly generated bone at bone defects and within the plastic caps was evaluated using micro-CT and histological sections. New bone regeneration within the plastic cap was increased significantly in the melatonin versus the control group. Conclusions Melatonin promoted vertical bone regeneration in rat calvaria in the secluded space within the plastic cap. Key words:Melatonin, bone regeneration, bone defects, secluded space, rat calvarium.

Influence of estrogen deficiency on guided bone augmentation: investigation of rat calvarial model and osteoblast-like MC3T3-E1 cells

The effect of estrogen deficiency in bone augmentation, and the mechanisms by which estrogen deficiency impedes osteoblast differentiation and collagen matrix production, were examined. Twenty female Jcl:Wistar rats were divided into two groups: ovariectomized rats; and control rats. Guided bone augmentation was performed by positioning plastic caps in the calvarium of all animals at 8 wk after ovariectomy or sham surgery. Micro-computed tomography and histological sections were used to determine the amount of bone augmentation within the plastic caps. At 8 wk, there was statistically significantly less newly formed bone volume in ovariectomized rats. Immunohistological staining revealed the rare alignment of runt-related transcription factor 2-positive osteoblast-like cells and collagen I-positive bundle fibers in ovariectomized rats. In cell culture experiments, pre-osteoblast-like cells, MC3T3-E1, were treated with the estrogen receptor antagonist, fulvestrant. In treated cells, alkaline phosphatase activity remained high, whereas Alizarin Red staining was completely inhibited. Extracellular staining intensity of collagen I was decreased after fulvestrant treatment. Consistent with these observations, gene-expression analysis confirmed that fulvestrant treatment led to weaker expression of mRNA for osteogenic transcription factors and bone matrix protein-related genes. The results demonstrate that estrogen deficiency suppresses osteoblast differentiation and collagen matrix production in bone augmentation.

Influences of mechanical barrier permeability on guided bone augmentation in the rat calvarium

We used radiological and histological analyses to evaluate the effects of mechanical barrier permeability in a rat model of calvarial guided bone augmentation (GBA). The calvaria of 20 rats were exposed, and one of four types of plastic caps (an occlusive cylindrical plastic cap; a plastic cap with no top; a plastic cap with three holes; and a plastic cap with four holes) was randomly placed on both sides. Newly generated bone in the plastic caps was evaluated with micro-computed tomography (micro-CT) and histological analysis. Micro-CT volumetric analysis and decalcified hematoxylin and eosin-stained sections showed that GBA barrier permeability was inversely associated with the quantity of augmented bone obtained. Massons trichrome staining showed that collagen in newly generated bony tissue was more mature in plastic caps with three holes than in those with more-permeable or more-occlusive barriers. Bone augmentation was inhibited in specimens exhibiting invasion of soft tissue through penetrating holes, and barrier permeability was associated with the quantity of augmented bone developed. In conclusion, moderate barrier permeability is optimal for development of mature augmented bone.

Enhancement of Bone Augmentation in Osteoporotic Conditions by the Intermittent Parathyroid Hormone: An Animal Study in the Calvarium of Ovariectomized Rat

PURPOSE Intermittent parathyroid hormone (PTH) is the commonly used therapeutic approach for patients with severe osteoporosis. The goal of this study was to elucidate the effect of the intermittent PTH treatment on guided bone augmentation (GBA) in the calvarium of ovariectomized (OVX) rats. MATERIALS AND METHODS Surgical ovariectomy on 14 rats and sham surgery on 7 rats were conducted on all rats as the first surgery. GBA surgery was conducted 8 weeks following the first surgery in the rat calvarium by placing 5-mm-diameter cylindrical plastic caps. Following surgery, rats were treated with 40 μg/kg PTH (OVX-PTH) or saline (Sham-Saline, OVX-Saline) via intraperitoneal injection three times per week during the all-observational period. Longitudinal microcomputed tomography (micro-CT) imaging was performed every 2 weeks following the GBA surgery without euthanasia, and the amount of newly generated bone volume (BV) was calculated. All rats were euthanized 12 weeks after GBA surgery, and histology was obtained. Sections stained with hematoxylin and eosin were used for the quantitative analysis of newly generated tissue, and immunohistology was used to visualize Runx2-positive cells and TRAP-positive cells. RESULTS Throughout the monitoring period, the BVs of OVX rats without PTH treatment (OVX-Saline) were significantly lower than that of the other two groups at weeks 8 and 12 in micro-CT analysis. During all experimental periods, the BV was highest in the OVX rats that were treated with PTH (OVX-PTH). Histologic analysis confirmed the result of micro-CT, and determined that the OVX-PTH presented a greater number of Runx2-positive cells. The number of TRAP-positive multinucleated osteoclasts was highest in OVX-PTH rats; there were no significant differences between the other two groups. CONCLUSION The results of this study suggest that treatment with intermittent PTH was associated with increased newly regenerated bone volume in ovariectomized rat calvarial bone augmentation, which may have important clinical implications.

CXCR4 signaling contributes to alveolar bone resorption in Porphyromonas gingivalis -induced periodontitis in mice

Periodontitis caused by bacterial infection gradually progresses accompanied by periodontal tissue destruction. As a result, teeth lose their supporting structures, and this leads to tooth exfoliation. CXC-chemokine receptor 4 (CXCR4) is known to be expressed in lymphocytes, fibroblasts and osteoclasts in periodontal tissues, suggesting that periodontal CXCR4 signaling contributes to alveolar bone resorption in the milieu of periodontitis. However, the role of CXCR4 signaling in the pathogenesis of periodontitis has remained unknown. We established a mouse model of periodontitis by inoculation of Porphyromonas gingivalis (P.g.) into a silk ligature placed around the maxillary molar. Although there was no significant difference in the mechanical sensitivity in the periodontal tissue between P.g. treatment and sham treatment during the experimental period, mechanical allodynia in the periodontal tissue was induced after gingival injection of complete Freunds adjuvant compared with that resulting from sham and P.g. treatment alone. Moreover, CXCR4 neutralization in the periodontal tissue following P.g. treatment enhanced periodontal inflammatory cell infiltration and depressed alveolar bone resorption. These findings suggest that periodontal CXCR4 signaling in several cell types in P.g.-induced periodontal inflammation depresses alveolar bone resorption in periodontitis. CXCR4 signaling might be a target for therapeutic intervention to prevent alveolar bone resorption in periodontitis.