The Treatment Efficiency and Microbiota Analysis of Sapindus mukorossi Seed Oil on the Ligature-Induced Periodontitis Rat Model.

Periodontitis is a common oral disease mainly caused by bacterial infection and inflammation of the gingiva. In the prevention or treatment of periodontitis, anti-bacterial agents are used to inhibit pathogen growth, despite increasing levels of bacterial resistance. Sapindus mukorossi Gaertn (SM) seed oil has proven anti-bacterial and anti-inflammation properties. However, the possibility of using this plant to prevent or treat periodontitis has not been reported previously. The aim of this study was to evaluate the effects of SM oil on experimental periodontitis in rats by using micro-CT and microbiota analysis. The distance between cementoenamel junction (CEJ) and alveolar bone crest (ABC) on the sagittal micro-CT slide showed that total bone loss (TBL) was significantly lower in CEJ-ABC distances between SM oil and SM oil-free groups on Day 14. Histology data also showed less alveolar bone resorption, a result consistent result with micro-CT imaging. The microbiota analyzed at phylum and class levels were compared between the SM oil and SM oil-free groups on Day 7 and Day 14. At the phylum level, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were the dominant bacterium. Firmicutes in box plot analysis was significantly less in the SM oil group than in the SM oil-free group on Day 7. At the class level, Bacteroidia, Gammaproteobacteria, Bacilli, Clostridia, and Erysipelotrichia were the dominant bacteria. The bacteria composition proportion of Bacilli, Clostridiay, and Erysipelotrichia could be seen in the SM oil group significantly less than in t SM oil-free group on Day 7. Overall, the present results show that topical application of SM oil can reduce bone resorption and change bacteria composition in the ligature-induced periodontitis model. According to these results, it is reasonable to suggest SM oil as a potential material for preventing oral disease.

The investigation of thermal behaviour and physical properties of several types of contemporary gutta-percha points.

AIM: To analyse the contents and thermal behaviour of several brands of contemporary gutta-percha points due to the variable nature of the components of gutta-percha, and the impact they can have on the physical properties of this unique material during canal filling. METHODOLOGY: Six brands of gutta-percha were investigated: Conform Fit TM Gutta-Percha Points for ProTaper Gold® (PTG) (Dentsply Sirona), ProTaper® Universal Gutta-Percha Points (PTU) (Dentsply Sirona), Autofit TM Feathered Tip Gutta Percha (Kerr), Mtwo® Gutta-Percha (VDW), Gutta Percha Root Canal Points (GC, GC Corporation) and Gutta-Percha Points ISO Color-Coded (ISO; Dentsply Sirona). The organic and inorganic fractions of gutta-percha points were separated by quantitative chemical analysis. Thermal conductivity was detected using a laser flash method. In addition, the thermal behaviour of gutta-percha in response to temperature variations was analysed using differential scanning calorimetry (DSC). Kruskal-Wallis and Dunn tests were applied for comparisons amongst groups for chemical compositions and temperature obtained from DSC. The associations between compositions and thermal conductivity were determined using simple linear regression. A p value <.05 was considered to be statistically significant. RESULTS: There were significant difference in the inorganic fractions of the gutta-percha points in percentage by weight amongst the groups (p < .05). PTG had the lowest thermal conductivity (0.42 W/m K). A positive correlation was observed between the percentage of inorganic fraction and thermal conductivity (r = 0.95). The initial phase changing temperature and peak temperature measured by DSC were significantly different when the ß-form transformed to α-form (p < .05), whereas no significant difference was found during the α-form to the amorphous-phase transition (p > .05). CONCLUSIONS: Chemical compositions and initial phase changing temperature by DSC varied according to the various brands of gutta-percha points. Conform Fit TM gutta-percha had the lowest percentage of inorganic fraction and thermal conductivity amongst these six brands of gutta-percha. Thermal conductivity had the strongest positive correlation with the percentage of inorganic components and zinc, whilst there was a negative correlation to the amount (ratio) of gutta-percha.

Combined Treatment of Heteronemin and Tetrac Induces Antiproliferation in Oral Cancer Cells.

BACKGROUND: Heteronemin, a marine sesterterpenoid-type natural product, possesses an antiproliferative effect in cancer cells. In addition, heteronemin has been shown to inhibit p53 expression. Our laboratory has demonstrated that the thyroid hormone deaminated analogue, tetrac, activates p53 and induces antiproliferation in colorectal cancer. However, such drug mechanisms are still to be studied in oral cancer cells. METHODS: We investigated the antiproliferative effects by Cell Counting Kit-8 and flow cytometry. The signal transduction pathway was measured by Western blotting analyses. Quantitative PCR was used to evaluate gene expression regulated by heteronemin, 3,3',5,5'-tetraiodothyroacetic acid (tetrac), or their combined treatment in oral cancer cells. RESULTS: Heteronemin inhibited not only expression of proliferative genes and Homo Sapiens Thrombospondin 1 (THBS-1) but also cell proliferation in both OEC-M1 and SCC-25 cells. Remarkably, heteronemin increased TGF-ß1 expression in SCC-25 cells. Tetrac suppressed expression of THBS-1 but not p53 expression in both cancer cell lines. Furthermore, the synergistic effect of tetrac and heteronemin inhibited ERK1/2 activation and heteronemin also blocked STAT3 signaling. Combined treatment increased p53 protein and p53 activation accumulation although heteronemin inhibited p53 expression in both cancer cell lines. The combined treatment induced antiproliferation synergistically more than a single agent. CONCLUSIONS: Both heteronemin and tetrac inhibited ERK1/2 activation and increased p53 phosphorylation. They also inhibited THBS-1 expression. Moreover, tetrac suppressed TGF-ß expression combined with heteronemin to further enhance antiproliferation and anti-metastasis in oral cancer cells.

Modified surgical drilling protocols influence osseointegration performance and predict value of implant stability parameters during implant healing process.

OBJECTIVE: The aim of this study was to evaluate the effects of three different surgical drilling protocols on changes of implant stability parameters and osseointegration performance during the healing period in rabbit femoral condyles. MATERIAL AND METHODS: Thirty New Zealand white rabbits were used in this study. Three experimental groups according to different surgical drilling protocols (undersized, standard and oversized preparation) were designed. Measurements of implant stability parameters were performed immediately after implant insertion and then at 1, 2, 4, and 8 weeks after the operation. After the animals were sacrificed, the bone blocks with implant were prepared for histological evaluation and histomorphometric analysis. RESULTS: The results demonstrated that the ISQ values of each group increased gradually through the whole healing period, while the damping factor showed the opposite tendency. The histomorphometric analysis revealed that BIC (bone-implant contact) values gradually increased with time until 8 weeks of healing at each group. In addition, the undersized group has the highest initial BIC (25.16% ± 7.25%) and the lowest values were found in oversized group (9.13% ± 5.89%). Moreover, a higher correlation (R2 = 0. 9817) between ISQ and BIC values in oversized group and moderate correlations between DF and BIC values in undersized group (R2 = 0.823) were demonstrated. CONCLUSIONS: The undersized drilling protocol group presented the highest implant stability and BIC values in the whole healing period, while the similar tendency of results was found between standard and oversized drilling protocol groups. CLINICAL RELEVANCE: These results suggested that undersized drilling protocol is mechanically and biologically beneficial in low-density bone. The modifications of surgical drilling protocols would influence the predictive value of implant stability parameters for osseointegration performance.

Damping ratio analysis of tooth stability under various simulated degrees of vertical alveolar bone loss and different root types.

BACKGROUND: The purpose of this study was to evaluate the feasibility of using damping ratio (DR) analysis combined with resonance frequency (RF) and periotest (PTV) analyses to provide additional information about natural tooth stability under various simulated degrees of alveolar vertical bone loss and various root types. METHODS: Three experimental tooth models, including upper central incisor, upper first premolar, and upper first molar were fabricated using Ti6Al4V alloy. In the tooth models, the periodontal ligament and alveolar bone were simulated using a soft lining material and gypsum, respectively. Various degrees of vertical bone loss were simulated by decreasing the surrounding bone level apically from the cementoenamel junction in 2-mm steps incrementally downward for 10 mm. A commercially available RF analyzer was used to measure the RF and DR of impulse-forced vibrations on the tooth models. RESULTS: The results showed that DRs increased as alveolar vertical bone height decreased and had high coefficients of determination in the linear regression analysis. The damping ratio of the central incisor model without a simulated periodontal ligament were 11.95 ± 1.92 and 27.50 ± 0.67% respectively when their bone levels were set at 2 and 10 mm apically from the cementoenamel junction. These values significantly changed to 28.85 ± 2.54% (p = 0.000) and 51.25 ± 4.78% (p = 0.003) when the tooth model was covered with simulated periodontal ligament. Moreover, teeth with different root types showed different DR and RF patterns. Teeth with multiple roots had lower DRs than teeth with single roots. CONCLUSION: Damping ratio analysis combined with PTV and RF analysis provides more useful information on the assessment of changes in vertical alveolar bone loss than PTV or RF analysis alone.

Static magnetic field attenuates lipopolysaccharide-induced inflammation in pulp cells by affecting cell membrane stability.

One of the causes of dental pulpitis is lipopolysaccharide- (LPS-) induced inflammatory response. Following pulp tissue inflammation, odontoblasts, dental pulp cells (DPCs), and dental pulp stem cells (DPSCs) will activate and repair damaged tissue to maintain homeostasis. However, when LPS infection is too serious, dental repair is impossible and disease may progress to irreversible pulpitis. Therefore, the aim of this study was to examine whether static magnetic field (SMF) can attenuate inflammatory response of dental pulp cells challenged with LPS. In methodology, dental pulp cells were isolated from extracted teeth. The population of DPSCs in the cultured DPCs was identified by phenotypes and multilineage differentiation. The effects of 0.4 T SMF on DPCs were observed through MTT assay and fluorescent anisotropy assay. Our results showed that the SMF exposure had no effect on surface markers or multilineage differentiation capability. However, SMF exposure increases cell viability by 15%. In addition, SMF increased cell membrane rigidity which is directly related to higher fluorescent anisotropy. In the LPS-challenged condition, DPCs treated with SMF demonstrated a higher tolerance to LPS-induced inflammatory response when compared to untreated controls. According to these results, we suggest that 0.4 T SMF attenuates LPS-induced inflammatory response to DPCs by changing cell membrane stability.

Static magnetic field increases survival rate of dental pulp stem cells during DMSO-free cryopreservation.

Successful and efficient cryopreservation of living cells and organs is a key clinical application of regenerative medicine. Recently, magnetic cryopreservation has been reported for intact tooth banking and cryopreservation of dental tissue. The aim of this study was to assess the cryoprotective effects of static magnetic fields (SMFs) on human dental pulp stem cells (DPSCs) during cryopreservation. Human DPSCs isolated from extracted teeth were frozen with a 0.4-T or 0.8-T SMF and then stored at -196 °C for 24 h. During freezing, the cells were suspended in freezing media containing with 0, 3 or 10% DMSO. After thawing, the changes in survival rate of the DPSCs were determined by flow cytometry. To understand the possible cryoprotective mechanisms of the SMF, the membrane fluidity of SMF-exposed DPSCs was tested. The results showed that when the freezing medium was DMSO-free, the survival rates of the thawed DPSCs increased 2- or 2.5-fold when the cells were exposed to 0.4-T or 0.8-T SMFs, respectively (p < 0.01). In addition, after exposure to the 0.4-T SMF, the fluorescence anisotropy of the DPSCs increased significantly (p < 0.01) in the hydrophilic region. These results show that SMF exposure improved DMSO-free cryopreservation. This phenomenon may be due to the improvement of membrane stability for resisting damage caused by ice crystals during the freezing procedure.

A real-time impedance-sensing chip for the detection of emulsion phase separation.

This paper describes a novel real-time impedance chip for the detection of squalene-water emulsion phase separation. Each impedance chip contains eight pairs of indium tin oxide microelectrode arrays for detecting eight samples, and six chips can be connected with the switch relay to measure 48 samples in the system simultaneously. The proposed impedance chip has the advantages of needing only a small sample volume (0.5 mL), and provides parallel, continuous, and real-time detection. The effects of the surfactant concentration on the stability of a squalene/water emulsion were studied by means of a visual inspection, a conductance probe, and by impedance chip. Three different concentrations of Tween 20 surfactant (9, 17, and 29 wt%) were employed for the examinations. The results indicated that the phase separation rate was faster in the lower surfactant concentration. However, the emulsion of 29 wt% Tween 20 was fairly stable for more than 2 days since there were no signal changes according to the three detection methods. The reaction time (TR) for completing the measured phase separation process differed for each of the three methods (measuring aqueous phase height, conductance, and impedance, respectively). For the 9 wt% Tween 20, the reaction times were 24 h, 20 min, and 5 min in the tests using visual inspection, conductance probe, and impedance chip, respectively. For the 17 wt% Tween 20, the TR was also shorter when using the impedance chip method compared to the other two methods. Therefore the proposed impedance chip has a quick reaction response and provides an alternative and effective method to detect emulsion stability.

Functionalization of zirconia ceramic with fibronectin proteins enhanced bioactivity and osteogenic response of osteoblast-like cells.

Introduction: To overcome the genuine bioinert properties of zirconia ceramic, functionalization of the surface with the bioactive protein fibronectin was conducted. Methods: Glow discharge plasma (GDP)-Argon was first used to clean the zirconia surface. Then allylamine was treated at three different powers of 50 W, 75 W, and 85 W and immersed into 2 different fibronectin concentrations (5 µg/ml and 10 µg/ml). Results and Discussion: After surface treatment, irregularly folded protein-like substances were attached on the fibronectin coated disks, and a granular pattern was observed for allylamine grafted samples. Infrared spectroscopy detected C-O, N-O, N-H, C-H, and O-H functional groups for fibronectin treated samples. Surface roughness rose and hydrophilicity improved after the surface modification, with MTT assay showing the highest level of cell viability for the A50F10 group. Cell differentiation markers also showed that fibronectin grafted disks with A50F10 and A85F10 were the most active, which in turn encouraged late-stage mineralization activity on 21d. Up-regulation of osteogenic related mRNA expression from 1d to 10d can be observed in RT-qPCR data for ALP, OC, DLX5, SP7, OPG and RANK biomarkers. These physical and biological properties clearly indicate that an allylamine and fibronectin composite grafted surface significantly stimulated the bioactivity of osteoblast-like cells, and can be utilized for future dental implant applications.

A Novel Endodontic Approach in Removing Smear Layer Using Nano and Submicron Diamonds with Intracanal Oscillation Irrigation.

Sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA) are commonly recommended for effectively removing organic and inorganic components in the smear layer. This layer is found on root canal walls after root canal instrumentation. However, high-concentration EDTA reduces the strength of dentin and the dissolution efficacy of organic substances in NaOCl solution. The objective of this study was to investigate the efficacy of applying nano and submicron diamonds in irrigation solutions with sonic and ultrasonic oscillation for removing the smear layer during endodontic treatment. Extracted single-rooted human teeth were instrumented with ProTaper® Gold (Dentsply Sirona) nickel-titanium rotary instruments. Subsequently, each canal was irrigated with 3% NaOCl, 17% EDTA, distilled water, and 10-1000 nm-sized nano and submicron diamond irrigation solutions, respectively. Sonic and ultrasonic instruments were compared for oscillating the irrigation solutions. The teeth were processed for scanning electron microscopy to observe the efficiency of smear layer removal on the canal walls. Our results indicated that diamond sizes of 50 nm and above irrigation solutions showed significant effectiveness in removing the smear layer following the oscillation of sonic instruments for 10 s. Ultrasonic assisted 500 nm and 1000 nm diamond solutions significantly differed from the other diamond-sized solution in their ability to remove the smear layer. These results suggest that sonic and ultrasonic oscillation with specific sizes of nano and submicron diamond irrigation solution can be used as an alternative approach to removing the smear layer during endodontic treatment. The potential clinical application of root canal treatments can be expected.

Fabrication of Low-Molecular-Weight Hyaluronic Acid-Carboxymethyl Cellulose Hybrid to Promote Bone Growth in Guided Bone Regeneration Surgery: An Animal Study.

Guided bone regeneration surgery is an important dental operation used to regenerate enough bone to successfully heal dental implants. When this technique is performed on maxilla sinuses, hyaluronic acid (HLA) can be used as an auxiliary material to improve the graft material handling properties. Recent studies have indicated that low-molecular hyaluronic acid (L-HLA) provides a better regeneration ability than high-molecular-weight (H-HLA) analogues. The aim of this study was to fabricate an L-HLA-carboxymethyl cellulose (CMC) hybrid to promote bone regeneration while maintaining viscosity. The proliferation effect of fabricated L-HLA was tested using dental pulp stem cells (DPSCs). The mitogen-activated protein kinase (MAPK) pathway was examined using cells cultured with L-HLA combined with extracellular-signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 inhibitors. The bone growth promotion of fabricated L-HLA/CMC hybrids was tested using an animal model. Micro-computer tomography (Micro-CT) and histological images were evaluated quantitatively to compare the differences in the osteogenesis between the H-HLA and L-HLA. Our results show that the fabricated L-HLA can bind to CD44 on the DPSC cell membranes and affect MAPK pathways, resulting in a prompt proliferation rate increase. Micro CT images show that new bone formation in rabbit calvaria defects treated with L-HLA/CMC was almost two times higher than in defects filled with H-HLA/CMC (p < 0.05) at 4 weeks, a trend that remained at 8 weeks and was confirmed by HE-stained images. According to these findings, it is reasonable to conclude that L-HLA provides better bone healing than H-HLA, and that the L-HLA/CMC fabricated in this study is a potential candidate for improving bone healing efficiency when a guided bone regeneration surgery was performed.

Comparison of 2,3,5,4'-tetrahydroxystilbene-2-O-b-D-glucoside-induced proliferation and differentiation of dental pulp stem cells in 2D and 3D culture systems-gene analysis.

BACKGROUND/PURPOSE: Culture environments play a critical role in stem cell expansion. This study aimed to evaluate the effects of 2,3,5,4'-tetrahydroxystilbene-2-O-b-D-glucoside (THSG) on the proliferation and differentiation of human dental pulp stem cells (DPSCs) in 2-dimensional (2D) and 3-dimensional (3D) culture systems. MATERIALS AND METHODS: Human DPSCs were seeded in T25 flasks for 2D cultivation. For the 3D culture system, DPSCs were mixed with microcarriers and cultured in spinner flasks. Cells in both culture systems were treated with THSG, and cell proliferation was determined using a cell counter and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. In THSG-treated DPSCs, the genes associated with proliferation, adipogenesis, neurogenesis, osteogenesis, pluripotency, oncogenesis, and apoptosis were analyzed using real-time polymerase chain reactions. RESULTS: The spinner flask time-dependently improved cell numbers, cell viability, and expansion rates in THSG-treated DPSCs. In both the T25 and spinner flasks, the messenger RNA (mRNA) levels of proliferation, osteogenesis, and pluripotent-related genes had a significant maximum expression with 10 µM THSG treatment. However, 0.1 µM of THSG may be the most suitable condition for triggering neurogenesis and adipogenesis gene expression when DPSCs were cultured in spinner flasks. Furthermore, the number of oncogenes and apoptotic genes decreased considerably in the presence of THSG in both the T25 and spinner flasks. CONCLUSION: The spinner flask bioreactor combined with THSG may upregulate proliferation and lineage-specific differentiation in DPSCs. Thus, the combination can be used to mass-produce and cultivate human DPSCs for regenerative dentistry.

Melatonin enhances osteogenic differentiation of dental pulp mesenchymal stem cells by regulating MAPK pathways and promotes the efficiency of bone regeneration in calvarial bone defects.

BACKGROUND: Mesenchymal stem cell (MSC)-based tissue engineering plays a major role in regenerative medicine. However, the efficiency of MSC transplantation and survival of engrafted stem cells remain challenging. Melatonin can regulate MSC biology. However, its function in the osteogenic differentiation of dental pulp-derived MSCs (DPSCs) remains unclear. We investigated the effects and mechanisms of melatonin on the osteogenic differentiation and bone regeneration capacities of DPSCs. METHODS: The biological effects and signaling mechanisms of melatonin with different concentrations on DPSCs were evaluated using a proliferation assay, the quantitative alkaline phosphatase (ALP) activity, Alizarin red staining, a real-time polymerase chain reaction, and a western blot in vitro cell culture model. The in vivo bone regeneration capacities were assessed among empty control, MBCP, MBCP + DPSCs, and MBCP + DPSCs + melatonin preconditioning in four-created calvarial bone defects by using micro-computed tomographic, histological, histomorphometric, and immunohistochemical analyses after 4 and 8 weeks of healing. RESULTS: In vitro experiments revealed that melatonin (1, 10, and 100 µM) significantly and concentration-dependently promoted proliferation, surface marker expression (CD 146), ALP activity and extracellular calcium deposition, and osteogenic gene expression of DPSCs (p < 0.05). Melatonin activated the protein expression of ALP, OCN, and RUNX-2 and inhibited COX-2/NF-κB expression. Furthermore, the phosphorylation of mitogen-activated protein kinase (MAPK) p38/ERK signaling was significantly increased in DPSCs treated with 100 µM melatonin, and their inhibitors significantly decreased osteogenic differentiation. In vivo experiments demonstrated that bone defects implanted with MBCP bone-grafting materials and melatonin-preconditioned DPSCs exhibited significantly greater bone volume fraction, trabecular bone structural modeling, new bone formation, and osteogenesis-related protein expression than the other three groups at 4 and 8 weeks postoperatively (p < 0.05). CONCLUSIONS: These results suggest that melatonin promotes the proliferation and osteogenic differentiation of DPSCs by regulating COX-2/NF-κB and p38/ERK MAPK signaling pathways. Preconditioning DPSCs with melatonin before transplantation can efficiently enhance MSCs function and regenerative capacities.

Orthodontic bonding with self-etching primer and self-adhesive systems.

The purpose of this study was to compare the tensile bond strengths of orthodontic brackets bonded to enamel using conventional multi-step adhesive, self-etching primer (SEP), which combines etching and priming into a single step, and self-adhesive systems, which combine etchant, primer, and adhesive. Metal brackets were bonded to 90 extracted human premolars according to three experimental protocols: group 1, conventional multi-step adhesive system; group 2, SEP; and group 3, self-adhesive system. All specimens were debonded using an Instron universal machine and failures between the tooth surface and bracket base were observed by scanning electron microscopy (SEM). The bracket bases were then analysed by mapping of energy-dispersive X-ray (EDX) spectrometry to calculate the distributive percentages of enamel or resin. The bond strength, percentage distribution, and calcium on the debonded interface were determined and analysed by one-way analysis of variance, and means were ranked by a Tukey interval, calculated at the 95 per cent confidence level. Group 1 produced the greatest bond strength, followed by groups 2 and 3. Group 3 showed the highest debonded interface between resin and enamel or within the resin itself, followed by groups 2 and 1. Groups 1 and 2 displayed significantly more debond failures at the interface between the bracket and the resin than group 3. More calcium particles were observed on the bracket base after debonding in group 3 than in groups 2 and 1. The simplified bonding procedures caused an undesirable decrease in tensile bond strength.

The Review of Bioeffects of Static Magnetic Fields on the Oral Tissue-Derived Cells and Its Application in Regenerative Medicine.

Magnets have been widely used in dentistry for orthodontic tooth movement and denture retention. Nevertheless, criticisms have arisen regarding the biosafety of static magnetic field (SMF) effects on surrounding tissues. Various controversial pieces of evidence have been discussed regarding SMFs on cellular biophysics, but little consensus has been reached, especially in the field of dentistry. Thus, the present paper will first review the safe use of SMFs in the oral cavity and as an additive therapy to orthodontic tooth movement and periodontium regeneration. Then, studies regarding SMF-incorporated implants are reviewed to investigate the advantageous effects of SMFs on osseointegration and the underlying mechanisms. Finally, a review of current developments in dentistry surrounding the combination of magnetic nanoparticles (MNPs) and SMFs is made to clarify potential future clinical applications.

2,3,5,4'-Tetrahydroxystilbene-2-O-ß-d-glucoside promotes the effects of dental pulp stem cells on rebuilding periodontal tissues in experimental periodontal defects.

BACKGROUND: This study aimed to investigate the regenerative effects of 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucoside (THSG)-treated human dental pulp stem cells (DPSC) on the healing of experimental periodontal defects in rats. METHODS: The maxillary first molars of 30 male Sprague-Dawley rats were extracted, and after healing, bilateral periodontal defects were surgically created mesially in second molars. The defects were treated with Matrigel (as control), DPSC, or DPSC + THSG. After 2 weeks, the healed defects were evaluated using microcomputed tomography and through histological and immunohistochemical analyses. RESULTS: In the microcomputed tomography analysis, more new bone formation in the DPSC and DPSC + THSG groups was observed compared with the control group. The periodontal bone supporting ratio in site with DPSC + THSG was significantly higher than that in DPSC. Histologically, an enhanced new bone formation and more significant periodontal attachment were observed in the DPSC + THSG group. The expression levels of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), and osteopontin (OPN) in the DPSC + THSG group were significantly greater than those in other groups. CONCLUSIONS: THSG-revolutionized DPSCs significantly shortened the regenerative period of periodontal defects by enhancing the cell recruitment and possibly the angiogenesis in rat models, which illustrate the critical implications for a clinical application and provide a novel tactic for periodontitis treatment.

2,3,5,4'-tetrahydroxystilbene-2-O-b-D-glucoside triggers the pluripotent-like possibility of dental pulp stem cells by activating the JAK2/STAT3 axis: Preliminary observations.

Abstract. BACKGROUND/PURPOSE: Although 2,3,5,4'-Tetrahydroxystilbene-2-O-beta-glucoside (THSG) reportedly has anti-inflammatory properties, its role in inducing the dedifferentiation of human dental pulp stem cells (DPSC) into pluripotent-like stem cells remains to be determined. The purpose of this study is to evaluate the effects of THSG on the pluripotent-like possibility and mechanism of DPSC. MATERIALS AND METHODS: DPSCs were treated with THSG, and cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTS) assay. Real-time polymerase chain reaction was used to analyze the mRNA expression levels of pluripotency-associated genes and oncogenes and to detect telomerase activity in the cells. Embryoid body formation assay was conducted, and pluripotency-related proteins were identified using Western blotting. Data were analyzed using one-way analysis of variance. RESULTS: Cell viability, telomerase activity, and embryoid body formation were enhanced in THSG-treated DPSCs. The mRNA expression levels of pluripotent-like genes (including Nanog homeobox [NANOG], SRY-box 2 [SOX2], and POU class 5 homeobox 1 [POU5F1/OCT4]) significantly increased after THSG treatment. The expression levels of pluripotency-related genes (Janus kinase-signal transducer 2 [JAK2] and signal transducer and activator of transcription 3 [STAT3]) increased, whereas those of oncogenes (Ras, SRC, HER2, and C-sis) decreased. Furthermore, the expression levels of the phosphorylated JAK2 and STAT3 proteins significantly increased after THSG treatment. CONCLUSION: THSG treatment may enhance the pluripotent-like possibility of DPSC through the JAK2/STAT3 axis. Hence, it may be used as an alternative cell-based therapeutic strategy in regenerative dentistry.

Surface Modified ß-Tricalcium phosphate enhanced stem cell osteogenic differentiation in vitro and bone regeneration in vivo.

A major number of studies have demonstrated Beta-tricalcium phosphate (ß-TCP) biocompatibility, bioactivity, and osteoconductivity characteristics in bone regeneration. The aim of this research was to enhance ß-TCP's biocompatibility, and evaluate its physicochemical properties by argon glow discharge plasma (GDP) plasma surface treatment without modifying its surface. Treated ß-TCP was analyzed by scanning electron microscopy (SEM), energy-dispersive spectrometry, X-ray photoelectron spectroscopy (XPS), X-ray diffraction analysis, and Fourier transform infrared spectroscopy characterization. To evaluate treated ß-TCP biocompatibility and osteoblastic differentiation, water-soluble tetrazolium salts-1 (WST-1), immunofluorescence, alkaline phosphatase (ALP) assay, and quantitative real-time polymerase chain reaction (QPCR) were done using human mesenchymal stem cells (hMSCs). The results indicated a slight enhancement of the ß-TCP by GDP sputtering, which resulted in a higher Ca/P ratio (2.05) than the control. Furthermore, when compared with control ß-TCP, we observed an improvement of WST-1 on all days (p < 0.05) as well as of ALP activity (day 7, p < 0.05), with up-regulation of ALP, osteocalcin, and Osteoprotegerin osteogenic genes in cells cultured with the treated ß-TCP. XPS and SEM results indicated that treated ß-TCP's surface was not modified. In vivo, micro-computed tomography and histomorphometric analysis indicated that the ß-TCP test managed to regenerate more new bone than the untreated ß-TCP and control defects at 8 weeks (p < 0.05). Argon GDP treatment is a viable method for removing macro and micro particles of < 7 µm in size from ß-TCP bigger particles surfaces and therefore improving its biocompatibility with slight surface roughness modification, enhancing hMSCs proliferation, osteoblastic differentiation, and stimulating more new bone formation.

Effects of Sapindus mukorossi Seed Oil on Proliferation, Osteogenetic/Odontogenetic Differentiation and Matrix Vesicle Secretion of Human Dental Pulp Mesenchymal Stem Cells.

Stem cells have attracted great interest in the development of tissue engineering. However, the self-regeneration and multi-differentiation capabilities of stem cells are easily impaired during cell transplantation. Recent studies have demonstrated that Sapindus mukorossi (S. mukorossi) seed oil has various positive biological effects. However, it is not yet clear whether S. mukorossi seed oil can increase the growth and differentiation of dental pulp mesenchymal stem cells (DPSCs). The aim of this study is to investigate the effects of S. mukorossi seed oil on the proliferation and differentiation of DPSCs. DPSCs with and without S. mukorossi seed oil, respectively, were evaluated and compared. The viabilities of the cells were assessed by MTT tests. The osteogenetic and odontogenetic capacities of the DPSCs were tested using Alizarin red S staining and alkaline phosphatase (ALP) activity assays. In addition, real-time PCR was performed to examine the gene expression of ALP, BMP-2 and DMP-1. Finally, extracellular matrix vesicle secretion was detected via scanning electron microscopy. No significant difference was observed in the viabilities of the DPSCs with and without S. mukorossi seed oil, respectively. However, under osteogenic and odontogenic induction, S. mukorossi seed oil increased the secretion of mineralized nodules and the ALP activity of the DPSCs (p < 0.05). The ALP gene expression of the differentiation-induced DPSCs was also enhanced. Finally, a greater secretion of extracellular matrix vesicles was detected in the DPSCs following odontogenic induction complemented with S. mukorossi seed oil. Overall, the present results show that S. mukorossi seed oil promotes the osteogenic/odontogenic differentiation and matrix vesicle secretion of DPSCs.

Er,Cr:YSGG Laser Performance Improves Biological Response on Titanium Surfaces.

Porphyromonas gingivalis infection is one of the causes of implant failures, which can lead to peri-implantitis. Implant surface roughness is reportedly related strongly to P. gingivalis adhesion, which can lead to peri-implantitis and, later, cell adhesion. Our aim was to evaluate the effects of Er,Cr:YSGG laser on titanium (Ti) disc surfaces and its interaction with bacterial adhesion and fibroblast viability. Ti discs underwent two treatments: autoclaving (control) and erbium, chromium-doped yttrium scandium gallium garnet (Er,Cr:YSGG) laser treatment (test). Ti disc surfaces were examined with scanning electronic microscope (SEM), Energy-dispersive spectrometry (EDX), X-ray photoelectron spectroscopy (XPS). The surface roughness same as wettability were also investigated. Fibroblast viability was assessed with the water-soluble tetrazolium 1 (WST-1) test, and osteoblast differentiation was assessed with the alkaline phosphatase (ALP) assay. Bacterial structure and colony formation were detected with scanning electron microscopy and Gram stain. In comparison to control discs, the test discs showed smoother surfaces, with 0.25-µm decrease in surface roughness (p < 0.05); lower P. gingivalis adhesion (p < 0.01); less P. gingivalis colonization (p < 0.05); and increased fibroblast viability and osteoblast differentiation (p < 0.05). Er,Cr:YSGG laser treatment improved disc surfaces by making them slightly smoother, which reduced P. gingivalis adhesion and increased fibroblast viability and osteoblast differentiation. Er,Cr:YSGG laser treatment can be considered a good option for managing peri-implantitis. Further investigations of laser-assisted therapy are necessary for better guidelines in the treatment of peri-implantitis.