Cyclic pressure-induced cytokines from gingival fibroblasts stimulate osteoclast activity: Clinical implications for alveolar bone loss in denture wearers.

Purpose The involvement of oral mucosa cells in mechanical stress-induced bone resorption is unclear. The aim of this study was to investigate the effects of cyclic pressure-induced cytokines from oral mucosal cells (human gingival fibroblasts: hGFs) on osteoclast activity in vitro.Methods Cyclic pressure at 50 kPa, which represents high physiologic occlusal force of dentures on the molar area, was applied to hGFs. NFAT-reporter stable RAW264.7 preosteoclasts (NFAT/Luc-RAW cells) were cultured in conditioned medium collected from hGF cultures under cyclic pressure or static conditions. NFAT activity and osteoclast formation were determined by luciferase reporter assay and TRAP staining, respectively. Cyclic pressure-induced cytokines in hGF culture were detected by ELISA, real-time RT-PCR, and cytokine array analyses.Results Conditioned media from hGFs treated with 48 hours of cyclic pressure significantly induced NFAT activity and increased multinucleated osteoclast formation. Furthermore, the cyclic pressure significantly increased the bone resorption activity of RAW264.7 cells. Cyclic pressure significantly increased the expression of major inflammatory cytokines including IL-1ß/IL-1ß, IL-6/IL-6, IL-8/IL-8 and MCP-1/CCL2 in hGFs compared to hGFs cultured under static conditions, and it suppressed osteoprotegerin (OPG/OPG) expression. A cytokine array detected 12 cyclic pressure-induced candidates. Among them, IL-8, decorin, MCP-1 and ferritin increased, whereas IL-28A and PDGF-BB decreased, NFAT activation of NFAT/Luc-RAW cells.Conclusions These results suggest that cyclic pressure-induced cytokines from hGFs promote osteoclastogenesis, possibly including up-regulation of IL-1ß, IL-6, IL-8 and MCP-1, and down-regulation of OPG. These findings introduce the possible involvement of GFs in mechanical stress-induced alveolar ridge resorption, such as in denture wearers.

Cross-sectional and longitudinal assessment of subchondral cysts in temporomandibular joints: Clinical and MRI study with a mean follow-up of 66 months.

PURPOSE: This observational study aimed to elucidate the pathophysiology of subchondral cysts (SC) in the temporomandibular joint (TMJ) and examine the results of conservative therapy administered to patients with SCs in the TMJ. METHODS: The study included 41 patients with SCs, extracted from 684 consecutive patients who underwent magnetic resonance imaging (MRI). The anatomical features of SCs and positional abnormalities of the articular disc were initially evaluated using MRI. A second MRI examination was performed for 28/41 patients at 40-107 months (mean, 66 months) after the first MRI. The joint space, anteroposterior width of the condylar head (WiC), articular eminence angle (AEA), and visual analog scale of jaw pain (VAS) were assessed alongside the MRI examinations. RESULTS: Most SCs were present in the anterosuperior and central condyle. Disc displacement was observed in 100% of 42 TMJs with SCs. Of the 29 joints in 28 patients, SCs in 19 joints resolved with time, whereas SCs in 10 joints persisted. A significant increase in the WiC and a significant decrease in AEA and VAS scores were observed on the second MRI scan. CONCLUSIONS: SCs tended to form in the anterosuperior and central parts of the condyle, where mechanical loading was likely to be applied. SCs are strongly associated with articular disc displacement. Two-thirds of SCs resolved over time, accompanied by resorption and osteophytic deformation of the condyle. SC might not be an indicator for the start of surgical treatment, and nonsurgical treatment could improve the clinical symptoms of patients with SCs.

Multivariate analysis of causal factors influencing accuracy of guided implant surgery for partial edentulism: a retrospective clinical study.

BACKGROUND: In dental implant treatment, the placement position of the implant body is important. The hypothesis is that there are factors that have a greater impact than the factors that have been studied so far. MATERIAL AND METHODS: The deviation between planned and actually placed implants was measured three-dimensionally by modified treatment evaluation method in 110 patients who underwent implant placement with guided surgery for partial edentulism. Ten factors that seemed to affect errors in placement were selected: the type of tooth, type of edentulism, distance from the remaining teeth, the type of implant, implant length, number of implants, method of guidance, the number of teeth supporting the surgical guide, number of anchor pins, and presence or absence of a reinforcement structure. The effect of each factor that corrected each confounding was calculated using multivariate analysis. RESULTS: In this study, 188 implant bodies were set to target, and the errors measurement data of the implant position were as follows: average Angle, 2.5 ± 1.6° (95% CI 2.25-2.69); Base, 0.67 ± 0.37 mm (95% CI 0.62-0.72); and Apex, 0.92 ± 0.47 mm (95% CI 0.86-0.98). As the result of multivariate analysis, larger errors were present in the partially guided group than the fully guided group. The number of teeth supporting the surgical guide significantly influenced the error in placement position. The error caused by the number of anchor pins was significantly different for the Angle. Similarly, the presence of the reinforcement structure influenced the error significantly for the Angle. CONCLUSIONS: It was suggested that the smaller errors could be present by performing guided surgery with full guidance and devising the design of the guide such as the number of teeth supporting the surgical guide, the setting of the anchor pin, and the reinforcement structure.

Evaluation of the effect of keratinized mucosa on peri-implant tissue health using a multivariate analysis.

PURPOSE: This study investigated the relationship between peri-implant tissue health and the presence of keratinized mucosa (≥ 2 mm) using multivariate analysis. METHODS: A total of 334 dental implants placed in 111 partially edentulous patients (34 males, 77 females) and restored with fixed prostheses were included in this study. The patients were recalled 12-146 months after completion of the prosthodontic treatment. Clinical parameters included modified plaque index (mPI), modified bleeding index (mBI), probing pocket depth (PPD), and radiographic bone loss (BL). The effects of the following potential explanatory variables on these parameters were analyzed: the presence of keratinized mucosa, age, sex, oral hygiene status, history of periodontitis, cigarette smoking, implant site, and time elapsed since prosthesis delivery. Statistical analysis included multivariate ordinal logistic regression and generalized estimating equations. Significance wa s established when two-sided p-values were less than 0.05. RESULTS: The mPI, mBI, and PPD in the presence or absence of keratinized mucosa did not show statistically significant differences. However, the presence of keratinized mucosa was significantly related to BL (odds ratio 4.33, p < 0.01). CONCLUSIONS: The results of our study suggest that the presence of keratinized mucosa is useful for reducing bone resorption and can help to maintain peri-implant tissue health.

Marginal Bone Loss Around Dental Implants Inserted with Static Computer Assistance in Healed Sites: A Systematic Review and Meta-analysis.

PURPOSE: The radiologic outcomes of implants placed using static computer-guided surgery have not yet been systematically investigated. The purpose of this study was to evaluate the marginal bone loss (MBL) around dental implants inserted with static computer assistance in healed sites. MATERIALS AND METHODS: An electronic search of publications in English from three databases (from 2000 to March 2015), including PubMed, Web of Science, and Cochrane Oral Health Group Trials Register, and a hand search of peerreviewed journals for relevant articles were performed. Only clinical human studies, either randomized or nonrandomized, with at least 10 cases and a minimum follow-up time of 12 months, reporting on MBL were included. RESULTS: The search strategy resulted in 18 publications, with 2,675 implants inserted with static computer assistance in healed sites. The pooled mean MBL at 1-year follow-up was 1.06 mm (95% CI: 0.83 to 1.30 mm; heterogeneity: random-effects model, I² = 99.38%; P < .01). Moreover, when considering studies with a 3-year follow-up only (n = 5; 748 implants), the pooled MBL was 1.48 mm (95% CI: 0.81 to 2.15 mm; heterogeneity: random-effects model, I² = 99%; P < .01). CONCLUSION: Within the limitations of this review, the MBL around dental implants placed in healed sites with computer-guided surgery seems to be a well-functioning one-stage alternative to extended two-stage conventional procedures if patients are appropriately selected and an appropriate width of bone is available for implant placement. However, current evidence is limited by the quality of available studies and the lack of comparative long-term clinical trials.

Scaffold-Free Fabrication of Osteoinductive Cellular Constructs Using Mouse Gingiva-Derived Induced Pluripotent Stem Cells.

Three-dimensional (3D) cell constructs are expected to provide osteoinductive materials to develop cell-based therapies for bone regeneration. The proliferation and spontaneous aggregation capability of induced pluripotent stem cells (iPSCs) thus prompted us to fabricate a scaffold-free iPSC construct as a transplantation vehicle. Embryoid bodies of mouse gingival fibroblast-derived iPSCs (GF-iPSCs) were seeded in a cell chamber with a round-bottom well made of a thermoresponsive hydrogel. Collected ball-like cell constructs were cultured in osteogenic induction medium for 30 days with gentle shaking, resulting in significant upregulation of osteogenic marker genes. The constructs consisted of an inner region of unstructured cell mass and an outer osseous tissue region that was surrounded by osteoblast progenitor-like cells. The outer osseous tissue was robustly calcified with elemental calcium and phosphorous as well as hydroxyapatite. Subcutaneous transplantation of the GF-iPSC constructs into immunodeficient mice contributed to extensive ectopic bone formation surrounded by teratoma tissue. These results suggest that mouse GF-iPSCs could facilitate the fabrication of osteoinductive scaffold-free 3D cell constructs, in which the calcified regions and surrounding osteoblasts may function as scaffolds and drivers of osteoinduction, respectively. With incorporation of technologies to inhibit teratoma formation, this system could provide a promising strategy for bone regenerative therapies.

Bonding effectiveness of self-adhesive and conventional-type adhesive resin cements to CAD/CAM resin blocks. Part 1: Effects of sandblasting and silanization.

The present study assessed the effect of sandblasting and silanization on resin cement bond strengths to CAD/CAM resin blocks. Twenty four blocks (KATANA AVENCIA BLOCK) were divided into two resin cement groups (PANAVIA V5 [PV5] and PANAVIA SA CEMENT HANDMIX [PSA]), and further divided into four subgroups representing different surface treatment methods: no treatment (Ctl), silanization (Si), sandblasting (Sb), and Sb+Si. After resin application, microtensile bond strengths (µTBSs) were measured immediately, 1, 3 and 6 months after water storage. In addition, surfaces resulting from each of the treatment methods were analyzed by scanning electron microscopy (SEM). Three-way analysis of variance revealed a statistically significant effect for the parameters 'surface treatment' (p<0.001, F=370), 'resin cement' (p<0.001, F=103, PSA

Bonding effectiveness of self-adhesive and conventional-type adhesive resin cements to CAD/CAM resin blocks. Part 2: Effect of ultrasonic and acid cleaning.

The present study assessed the effect of ultrasonic and acid cleaning on resin cement bonding to CAD/CAM resin blocks. One of two resin cements, PANAVIA V5 (PV5) or PANAVIA SA CEMENT HANDMIX (PSA), were bonded to one of 24 CAD/CAM blocks (KATANA AVENCIA BLOCK). Each cement group was divided into four subgroups: no cleaning (Ctl), ultrasonic cleaning (Uc), acid cleaning (Ac) and Uc+Ac. Micro-tensile bond strengths (µTBSs) were measured immediately and 1, 3, and 6 months after water storage. Block surfaces after each treatment were analyzed by scanning electron microscopy. Analysis of variance revealed a statistically significant effect for the parameters 'surface treatment' (p<0.001, F=40), 'resin cement' (p<0.001, F=696) and 'water aging' (p<0.001, F=71). The PV5 group exhibited higher µTBS values than the PSA group. Although cleaning after sandblasting was effective in removing residual alumina particles, it did not affect the long-term bonding durability with non-contaminated CAD/CAM resin blocks.

Controlled Osteogenic Differentiation of Mouse Mesenchymal Stem Cells by Tetracycline-Controlled Transcriptional Activation of Amelogenin.

Regenerative dental therapies for bone tissues rely on efficient targeting of endogenous and transplanted mesenchymal stem cells (MSCs) to guide bone formation. Amelogenin is the primary component of Emdogain, which is used to regenerate periodontal defects; however, the mechanisms underlying the therapeutic effects on alveolar bone remain unclear. The tetracycline (Tet)-dependent transcriptional regulatory system is a good candidate to investigate distinct roles of genes of interest during stem cell differentiation. Here, we investigated amelogenin-dependent regulation of osteogenesis in MSCs by establishing a Tet-controlled transcriptional activation system. Clonal mouse bone marrow-derived MSCs were lentivirally transduced with the Tet repressor (TetR) expression vector followed by drug selection to obtain MSCs constitutively expressing TetR (MSCs-TetR). Expression vectors that contained the Tet operator and amelogenin-coding (Amelx) cDNA fragments were constructed using the Gateway system and lentivirally introduced into MSCs-TetR to generate a Tet regulation system in MSCs (MSCs-TetR/Amelx). MSCs-TetR/Amelx significantly overexpressed the Amelx gene and protein in the presence of the tetracycline derivative doxycycline. Concomitant expression of osterix, bone sialoprotein (BSP), osteopontin, and osteocalcin was modulated by addition or removal of doxycycline under osteogenic guidance. During osteogenic induction, MSCs-TetR/Amelx treated with doxycycline showed significantly increased gene expression of osterix, type I collagen, BSP, and osteocalcin in addition to increased alkaline phosphatase activity and mineralized nodule formation. Enhanced extracellular matrix calcification was observed when forced Amelx expression commenced at the early stage but not at the intermediate or late stages of osteogenesis. These results suggest that a Tet-controlled Amelx gene regulation system for mouse MSCs was successfully established, in which transcriptional activation of Amelx was associated with enhanced osteogenic differentiation, especially in the early stage of biomineralization.

Clinical application of removable partial dentures using thermoplastic resin. Part II: Material properties and clinical features of non-metal clasp dentures.

This position paper reviews physical and mechanical properties of thermoplastic resin used for non-metal clasp dentures, and describes feature of each thermoplastic resin in clinical application of non-metal clasp dentures and complications based on clinical experience of expert panels. Since products of thermoplastic resin have great variability in physical and mechanical properties, clinicians should utilize them with careful consideration of the specific properties of each product. In general, thermoplastic resin has lower color-stability and higher risk for fracture than polymethyl methacrylate. Additionally, the surface of thermoplastic resin becomes roughened more easily than polymethyl methacrylate. Studies related to material properties of thermoplastic resin, treatment efficacy and follow-up are insufficient to provide definitive conclusions at this time. Therefore, this position paper should be revised based on future studies and a clinical guideline should be provided.

Comparative analysis of mouse-induced pluripotent stem cells and mesenchymal stem cells during osteogenic differentiation in vitro.

Induced pluripotent stem cells (iPSCs) can differentiate into mineralizing cells and are, therefore, expected to be useful for bone regenerative medicine; however, the characteristics of iPSC-derived osteogenic cells remain unclear. Here, we provide a direct in vitro comparison of the osteogenic differentiation process in mesenchymal stem cells (MSCs) and iPSCs from adult C57BL/6J mice. After 30 days of culture in osteogenic medium, both MSCs and iPSCs produced robustly mineralized bone nodules that contained abundant calcium phosphate with hydroxyapatite crystal formation. Mineral deposition was significantly higher in iPSC cultures than in MSC cultures. Scanning electron microscopy revealed budding matrix vesicles in early osteogenic iPSCs; subsequently, the vesicles propagated to exhibit robust mineralization without rich fibrous structures. Early osteogenic MSCs showed deposition of many matrix vesicles in abundant collagen fibrils that became solid mineralized structures. Both cell types demonstrated increased expression of osteogenic marker genes, such as runx2, osterix, dlx5, bone sialoprotein (BSP), and osteocalcin, during osteogenesis; however, real-time reverse transcription-polymerase chain reaction array analysis revealed that osteogenesis-related genes encoding mineralization-associated molecules, bone morphogenetic proteins, and extracellular matrix collagens were differentially expressed between iPSCs and MSCs. These data suggest that iPSCs are capable of differentiation into mature osteoblasts whose associated hydroxyapatite has a crystal structure similar to that of MSC-associated hydroxyapatite; however, the transcriptional differences between iPSCs and MSCs could result in differences in the mineral and matrix environments of the bone nodules. Determining the biological mechanisms underlying cell-specific differences in mineralization during in vitro iPSC osteogenesis may facilitate the development of clinically effective engineered bone.

PIH1D1 interacts with mTOR complex 1 and enhances ribosome RNA transcription.

PIH1D1 is the defining component of the R2TP complex. Recently, R2TP has been reported to stabilize mTOR (mammalian target of rapamycin), an important regulator of cell growth and protein synthesis. Two complexes of mTOR, mTORC1 and mTORC2, have been identified. We demonstrate that immunoprecipitation (IP) of PIH1D1 results in the co-IP of Raptor (mTORC1 specific), but not Rictor (mTORC2 specific), and that knockdown of PIH1D1 decreases mTORC1 assembly, S6 kinase phosphorylation (indicator of mTORC1 activity), and rRNA transcription without affecting mTORC2 in human breast cancer MCF-7 cells. In addition, we provide evidence that PIH1D1 is overexpressed in various breast cancer cell lines. These findings collectively suggest that PIH1D1 may have an important role in mTORC1 regulation in breast cancers.

Exosome-bound WD repeat protein Monad inhibits breast cancer cell invasion by degrading amphiregulin mRNA.

Increased stabilization of mRNA coding for key cancer genes can contribute to invasiveness. This is achieved by down-regulation of exosome cofactors, which bind to 3'-UTR in cancer-related genes. Here, we identified amphiregulin, an EGFR ligand, as a target of WD repeat protein Monad, a component of R2TP/prefoldin-like complex, in MDA-MB-231 breast cancer cells. Monad specifically interacted with both the 3'-UTR of amphiregulin mRNA and the RNA degrading exosome, and enhanced decay of amphiregulin transcripts. Knockdown of Monad increased invasion and this effect was abolished with anti-amphiregulin neutralizing antibody. These results suggest that Monad could prevent amphiregulin-mediated invasion by degrading amphiregulin mRNA.

Application of cyclic strain for accelerated skeletal myogenic differentiation of mouse bone marrow-derived mesenchymal stromal cells with cell alignment.

The fabrication of biomimetic skeletal myocyte constructs continues to present a challenge to functional tissue engineering. The skeletal myogenesis of bone marrow-derived mesenchymal stromal cells (BMSCs) to mimic the native tissue architecture offers great therapeutic promise, but remains particularly difficult. The aim of this study was to examine the possibility of accelerating the skeletal myogenic differentiation of BMSCs with an aligned structure by applying cyclic strain. Mouse BMSCs (mBMSCs) were plated on silicone sheets that were coated with fibronectin and subjected to cyclic 10% uniaxial strain when they reached 80%-90% cell confluency. Cells cultured in a growth medium that were subjected to cyclic strain at a frequency of 0.17 Hz (10 times/min) demonstrated a shift of alignment within 48 h from a completely random orientation to a well-aligned morphology with well-organized actin stress fibers that were parallel to the strain vector. The cyclic strain restricted the motility and proliferation of the aligned mBMSCs in the growth medium, which resulted in tight cellular contact in the cell population. When mBMSCs were subjected to cyclic strain in a myogenic medium, reverse transcription-polymerase chain reaction analysis demonstrated the upregulation of skeletal myogenic marker genes (myogenic factor 5 [Myf5], myogenin, and myogenic regulatory factor 4 [MRF4]), but not smooth muscle marker genes (myocardin and α-smooth muscle actin). In addition, immunocytochemistry showed that the mBMSCs fused to form multinucleated myosin- and myogenin-positive myotubes in the direction of the applied tension within 5 days. These results demonstrate that our simple method of applying of cyclic strain to cells cultured in a myogenic medium greatly accelerates the skeletal myogenic differentiation of mBMSCs with an aligned structure, and they highlight the importance of cellular alignment for creating physiologically relevant environments to study the myogenesis of BMSCs and engineer skeletal muscle.

In vitro reproduction of endochondral ossification using a 3D mesenchymal stem cell construct.

Endochondral ossification is one of the essential bone development processes in vertebrates. Although researchers from a variety of fields, including cellular/molecular biology, chemistry, and materials science, have worked to gain a better understanding of the tissue development, integration of findings from these different fields remains a major challenge. An in vitro model system that reproduces endochondral ossification would be a valuable tool for overcoming this problem, because an in vitro standardized model system can be easily accessed by researchers from different fields. Here, we fabricated a large 3D mesenchymal stem cell (MSC) construct with a ball-like morphology, which is termed a cell ball, and cultured it under a hypoxia condition, since hypoxia causes chondrogenic differentiation of MSCs in primordial cartilage, which is crucial for endochondral ossification. Region-specific chondrogenic differentiation of MSCs and mineralization within the cartilage tissue were observed in the cell ball. The precipitated minerals were detected as hydroxyapatite. Consequently, a 3D construct consisting of mineralized tissue surrounded by cartilage tissue was obtained. Moreover, the angiogenic activity of this synthesized tissue changed depending on the chondrogenic phenotype remains in the tissue, which is similar to what happens in the ossification process. Thus, this MSC cell ball system clearly reproduced the initial stage of endochondral ossification in vitro. This system is a promising tool for use as an in vitro model for investigating bone tissue development.

Gingival fibroblasts as a promising source of induced pluripotent stem cells.

BACKGROUND: Induced pluripotent stem (iPS) cells efficiently generated from accessible tissues have the potential for clinical applications. Oral gingiva, which is often resected during general dental treatments and treated as biomedical waste, is an easily obtainable tissue, and cells can be isolated from patients with minimal discomfort. METHODOLOGY/PRINCIPAL FINDINGS: We herein demonstrate iPS cell generation from adult wild-type mouse gingival fibroblasts (GFs) via introduction of four factors (Oct3/4, Sox2, Klf4 and c-Myc; GF-iPS-4F cells) or three factors (the same as GF-iPS-4F cells, but without the c-Myc oncogene; GF-iPS-3F cells) without drug selection. iPS cells were also generated from primary human gingival fibroblasts via four-factor transduction. These cells exhibited the morphology and growth properties of embryonic stem (ES) cells and expressed ES cell marker genes, with a decreased CpG methylation ratio in promoter regions of Nanog and Oct3/4. Additionally, teratoma formation assays showed ES cell-like derivation of cells and tissues representative of all three germ layers. In comparison to mouse GF-iPS-4F cells, GF-iPS-3F cells showed consistently more ES cell-like characteristics in terms of DNA methylation status and gene expression, although the reprogramming process was substantially delayed and the overall efficiency was also reduced. When transplanted into blastocysts, GF-iPS-3F cells gave rise to chimeras and contributed to the development of the germline. Notably, the four-factor reprogramming efficiency of mouse GFs was more than 7-fold higher than that of fibroblasts from tail-tips, possibly because of their high proliferative capacity. CONCLUSIONS/SIGNIFICANCE: These results suggest that GFs from the easily obtainable gingival tissues can be readily reprogrammed into iPS cells, thus making them a promising cell source for investigating the basis of cellular reprogramming and pluripotency for future clinical applications. In addition, high-quality iPS cells were generated from mouse GFs without Myc transduction or a specific system for reprogrammed cell selection.

Zoledronic acid delays wound healing of the tooth extraction socket, inhibits oral epithelial cell migration, and promotes proliferation and adhesion to hydroxyapatite of oral bacteria, without causing osteonecrosis of the jaw, in mice.

Nitrogen-containing bisphosphonates such as zoledronic acid (ZOL) and pamidronate have been widely and successfully used for the treatment of cancer patients with bone metastases and/or hypercalcemia. Accumulating recent reports have shown that cancer patients who have received these bisphosphonates occasionally manifest bisphosphonate-related osteonecrosis of the jaw (BRONJ) following dental treatments, including tooth extraction. However, little is known about the pathogenesis of BRONJ to date. Here, to understand the underlying pathogenesis of BRONJ, we examined the effects of ZOL on wound healing of the tooth extraction socket using a mouse tooth extraction model. Histomorphometrical analysis revealed that the amount of new bone and the numbers of blood vessels in the socket were significantly decreased in ZOL-treated mice compared to control mice. Consistent with these results, ZOL significantly inhibited angiogenesis induced by vascular endothelial growth factor in vivo and the proliferation of endothelial cells in culture in a dose-dependent manner. In contrast, etidronate, a non-nitrogen-containing bisphosphonate, showed no effects on osteogenesis and angiogenesis in the socket. ZOL also suppressed the migration of oral epithelial cells, which is a crucial step for tooth socket closure. In addition, ZOL promoted the adherence of Streptococcus mutans to hydroxyapatite and the proliferation of oral bacteria obtained from healthy individuals, suggesting that ZOL may increase the bacterial infection. In conclusion, our data suggest that ZOL delays wound healing of the tooth extraction socket by inhibiting osteogenesis and angiogenesis. Our data also suggest that ZOL alters oral bacterial behaviors. These actions of ZOL may be relevant to the pathogenesis of BRONJ.

Predictors of gastric myoelectrical activity in type 2 diabetes mellitus.

BACKGROUND: Previous studies have clearly demonstrated the delayed gastric emptying of solid meals in diabetics, whereas their gastric myoelectrical activity, which primarily determines gastric motility, has not yet been fully confirmed. GOALS: This study aimed to clarify the characteristics and potential predictors of gastric myoelectrical activity in type 2 diabetics. STUDY: Twenty-eight diabetics and 18 healthy controls participated. Duodenal biopsy sample was used for reverse transcription-polymerase chain reaction to evaluate cholecystokinin and motilin mRNA contents. Electrogastrography was performed before and after the test meal, and was assessed in terms of dominant frequency; dominant frequency instability coefficient; and the percentage of bradygastria, normogastria, and tachygastria. RESULTS: Over the entire recording period, dominant frequency was significantly lower, and dominant frequency instability coefficient and the percentage of bradygastria were significantly higher in diabetics than in controls. In diabetics, the multiple regression analysis demonstrated that dominant frequency instability coefficient and the percentage of tachygastria in the fasting period were dependent on fasting plasma glucose level and HbA1c, respectively. Moreover, dominant frequency over the entire period and the postprandial percentage of bradygastria were significantly associated with body mass index; the fasting percentage of bradygastria and postprandial dominant frequency instability coefficient were associated with fasting serum leptin level; the postprandial percentage of bradygastria was also associated with cholecystokinin mRNA content. CONCLUSIONS: Gastric myoelectrical activity in type 2 diabetics is impaired on dominant frequency, dominant frequency instability coefficient, and the percentage of bradygastria and predicted by body mass index, fasting serum leptin level, and cholecystokinin mRNA content besides the glycemic status.

Enhanced bone regeneration via multimodal actions of synthetic peptide SVVYGLR on osteoprogenitors and osteoclasts.

Recently, the binding sequence Ser-Val-Val-Tyr-Gly-Leu-Arg (SVVYGLR) was found adjacent to the RGD sequence in osteopontin, suggesting involvement in osteo-immune cross-talk. The aim of this study was to investigate bioactive functions of a synthetic SVVYGLR peptide in osteoprogenitor cells and osteoclasts, and to examine potential applications in bone regeneration. The SVVYGLR peptide significantly enhanced the adhesion and proliferation of several human mesenchymal cells including bone marrow-derived mesenchymal stem cells, and alphavbeta3 integrin was involved in cell attachment to the peptide. Additionally, the peptide reduced the number of TRAP-positive multinucleated cells during osteoclastogenesis of RAW264.7 cells and normal murine pre-osteoclasts, and also suppressed NFAT activity and expression of osteoclastogenesis-related mRNAs. When standardized bone defects in rat calvariae were filled with a collagen sponge containing the peptide or PBS (control), the number of TRAP-positive osteoclasts in the grafted sites after 3 weeks was significantly lower in the peptide group. By the 5th week, significantly enhanced resorption of the grafted collagen sponge and new bone formation was observed within and surrounding the sponge in the peptide group. These data suggest that SVVYGLR is an effective bioactive peptide for bone tissue regeneration that promotes attachment and proliferation of osteogenic cells while also suppressing osteoclastogenesis.