Managing Temporomandibular Joint Osteoarthritis by Dental Stem Cell Secretome.

The potential therapeutic role of the Dental Pulp Stem Cells Secretome (SECR) in a rat model of experimentally induced Temporomandibular Joint (TMJ) Osteoarthritis (OA) was evaluated. Proteomic profiling of the human SECR under specific oxygen tension (5% O2) and stimulation with Tumor Necrosis Factor-alpha (TNF-α) was performed. SECR and respective cell lysates (CL) samples were collected and subjected to SDS-PAGE, followed by LC-MS/MS analysis. The identified proteins were analyzed with Bioinformatic tools. The anti-inflammatory properties of SECR were assessed via an in vitro murine macrophages model, and were further validated in vivo, in a rat model of chemically-induced TMJ-OA by weekly recording of the head withdrawal threshold, the food intake, and the weight change, and radiographically and histologically at 4- and 8-weeks post-treatment. SECR analysis revealed the presence of 50 proteins that were enriched and/or statistically significantly upregulated compared to CL, while many of those proteins were involved in pathways related to "extracellular matrix organization" and "immune system". SECR application in vitro led to a significant downregulation on the expression of pro-inflammatory genes (MMP-13, MMP-9, MMP-3 and MCP-1), while maintaining an increased expression of IL-10 and IL-6. SECR application in vivo had a significant positive effect on all the clinical parameters, resulting in improved food intake, weight, and pain suppression. Radiographically, SECR application had a significant positive effect on trabecular bone thickness and bone density compared to the saline-treated group. Histological analysis indicated that SECR administration reduced inflammation, enhanced ECM and subchondral bone repair and regeneration, thus alleviating TMJ degeneration.

Screwmentable implant-supported prostheses: A systematic review.

STATEMENT OF PROBLEM: Screwmentable prostheses were developed to combine the benefits of screw retention and cement retention. However, data are limited on the clinical performance of this type of prosthesis. PURPOSE: The purpose of this systematic review was to collect scientific evidence on screwmentable prostheses and evaluate their long-term clinical behavior. MATERIAL AND METHODS: An electronic search was conducted by 2 independent reviewers for articles published in scientific dental journals in English from 2004 to April 2020. The search strategy followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Inclusion criteria were scientific studies concerning the screwmentable type of prosthesis. RESULTS: The search provided 494 records. Of these, 24 studies fulfilled the inclusion criteria and were included in the review. The included articles presented significant heterogeneity concerning the manufacturing process and the materials used. One randomized clinical trial, 2 prospective clinical studies, 14 in vitro studies, 3 protocol descriptions, 1 case series, and 3 case reports were included. CONCLUSIONS: Based on the systematic search of the literature, it is concluded that the screwmentable prosthesis combines advantages of both cement-retained and screw-retained restorations, including passive fit, retrievability, excess cement control, tissue-friendly emergence profile, and improved esthetics. Nevertheless, data from well-designed clinical trials are limited, and further research is required to provide evidence on their long-term clinical behavior.

Effectiveness of conservative therapeutic modalities for temporomandibular disorders-related pain: a systematic review.

PURPOSE: The aim of this systematic review was to evaluate the effectiveness of conservative different therapeutic modalities for temporomandibular disorders (TMD) pain. MATERIALS AND METHODS: An electronic systematic search was conducted in the MEDLINE (PubMed) database to identify the randomized clinical trials (RCTs) published between 2001 and 2021. The following, simple or multiple conjunctions, search keywords were selected: TMD pain, TMD management or conservative treatment or treatment strategies and TMD pain, therapeutic modalities or interventions and TMD. Studies included must have patients older than 18 years, with painful TMD, which diagnosis was performed by Research Diagnostic Criteria for TMD or Diagnostic Criteria for TMD. Outcome variables were pain relief and post treatment pain intensity reduction. Data were analysed with non-parametric tests and the level of significance was set at p<.05. RESULTS: Out of 1599 articles obtained, 28 RCTs fulfilled all selection criteria and were included. The results of this study show that there was a significant decrease in short-term post-treatment TMD pain with the use of occlusal splint alone or in combination with other therapeutic modalities when compared with the control group. Statistically significant differences were also detected between laser and photobiomodulation group and the control, in short-term treatment TMD-related pain. CONCLUSIONS: The primary findings of the present systematic review showed that the occlusal splint alone or combined with other therapeutic intervention presented positive effect on short-term TMD pain reduction. Secondary outcome suggests that laser and photobiomodulation therapy had, also, a significant role in short term pain relief.

Effect of different zirconia surface pretreatments on the flexural strength of veneered Y-TZP ceramic before and after in vitro aging.

PURPOSE: The investigation of zirconia core surface pretreatments on the flexural strength of bilayered zirconia ceramics before and after artificial accelerating aging. METHODS: Ninety bar-shaped specimens were manufactured from Yttria Stabilized Tetragonal Zirconia Polycrystal (Y-TZP) and divided in three groups depending on zirconia surface pretreatment before veneering: layering with liner, pretreatment with silane-containing gas flame (SGF) with the Silano-Pen device and alumina air-abrasion. Half of the veneered specimens in each group (n=15) underwent artificial accelerating aging. A 4-point bending test was performed to determine flexural strength. Three specimens from each group were further analyzed using Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR) before veneering and after fracture (aged and non-aged subgroups). RESULTS: Alumina air-abrasion was correlated to increased phase transformation from tetragonal to monoclinic zirconia phase. Qualitative analysis revealed that with the majority of the specimens pretreated with the silane-containing gas flame, areas of the veneering material remained firmly attached to the zirconia core after flexural strength testing. There was no statistically significant difference on the flexural strength among the groups before or after aging. Artificial accelerating aging resulted in statistically significant higher flexural strength of the specimens after aging. CONCLUSION: SGF pretreatment can be an acceptable and feasible alternative method before the veneering of Y-TZP zirconia as it presented slightly higher bond strength compared with alumina air-abrasion which was associated with higher tetragonal to monoclinic (t→m) phase transformation. Accelerating aging leads to an increase of the mechanical properties under in vitro conditions.

Effect of in vitro aging and acidic storage on color, translucency, and contrast ratio of monolithic zirconia and lithium disilicate ceramics.

STATEMENT OF PROBLEM: How the translucency and color of ceramic restorations are affected by surface changes from the corrosive environment in the oral cavity and aging of materials is unclear. PURPOSE: The purpose of this in vitro study was to compare the impact of acidic exposure and aging on the color and translucency of monolithic zirconia and lithium disilicate ceramics. MATERIAL AND METHODS: Twenty computer-aided design and computer-aided manufacturing (CAD-CAM) monolithic zirconia specimens and 20 pressed lithium disilicate specimens were fabricated. Half of the specimens of each group were aged in an autoclave (7.5 hours, 134 °C, 0.2 MPa), and the rest were immersed in hydrochloric acid to simulate the acidic conditions in the oral cavity from gastric reflux. The color coordinates L∗, a∗, and b∗ were measured with an ultraviolet spectrophotometer before and after aging or acidic storage. The translucency parameters and contrast ratios were calculated, and the CIEDE2000 color difference formula was used to determine color differences before and after each treatment. ANOVA and ANCOVA test models were used for data analysis (α=.05), while differences of color parameters in respect to acceptability and perceptibility thresholds were evaluated with the 1-sample t test (α=.05). RESULTS: Lithium disilicate presented a significantly higher translucency parameter and lower contrast ratio at baseline compared with monolithic zirconia specimens (P<.001). Acidic storage significantly impacted all parameters compared with aging, especially for the lithium disilicate group. Color differences were above the acceptability ΔΕ00 threshold for lithium disilicate after acidic storage (P=.001) and below for monolithic zirconia after acidic storage (P=.003). CONCLUSIONS: The performance of lithium disilicate was slightly inferior compared with that of monolithic zirconia specimens, as they presented significant and clinically observable differences for the translucency parameter and ΔΕ00 after acidic storage and aging.

Effect of Water Storage on Hardness and Interfacial Strength of Resin Composite Luting Agents Bonded to Surface-Treated Monolithic Zirconia.

BACKGROUND: Durable bonding between resin composite luting agents (CLA) and zirconia is still a matter of controversy. The purpose of this study was to evaluate the effect of water storage on hardness and interfacial strength of three CLA, a non-adhesive (Multilink Automix/ML), an adhesive (Panavia F 2.0/PF) and a self-adhesive (PermaCem 2.0/PC), bonded to polished (CL) and grit-blasted (AL: 50 µm alumina, SJ: Sil-Jet + Monobond Plus silane) monolithic zirconia surfaces. METHODS: CLA specimens (n = 5/cement, condition) were prepared, stored under dry conditions or immersed in water, and Vickers hardness (VH) measurements were obtained at 1 h, 24 h, 1 week and 3 weeks intervals. Optical profilometry was used to determine the roughness parameters (Sa, Sz, Sdr, Sci) of zirconia surfaces (n = 5/treatment). A shear strength test (SBS, n = 10 × 2/cement) was performed to assess the strength and fractography of the cements bonded to zirconia after isothermal water storage and thermal-cycling (TC). RESULTS: PF demonstrated significantly lower VHN after water storage at all time intervals, PC at 1 w, 3 w and ML at 3 w. SJ and AL showed significantly higher values from CL in all roughness parameters. Weibull analysis revealed the following significance in σο ranking within the same material: AL, SJ, ALTC > SJTC, CL > CLTC (PF); SJ, SJTC, AL, ALTC > CL, CLTC (PC) and SJ, SJTC > AL > ALTC > CL, CLTC (ML). Within the same surface treatment subgroups, the significance in σo ranking was PC, ML > PF (before/after TC) for SJ; PC > PF > ML (before TC), PC, PF > ML (after TC) for AL, and PC > PF > ML (before/after TC) for CL. For the m ranking, the only significant difference within each material group was found in PC (AL > ALTC) and for the same surface treatment in AL (PC > ML). CONCLUSION: There are significant differences in the water plasticization susceptibility of the CLA tested; the materials with adhesive monomers were the most affected. Tribo-chemical silica coating combined with a silane coupling agent was the most efficient bonding treatment for the non-adhesive and the self-adhesive materials. The adhesive CLA performed better on alumina-blasted than on tribo-chemically coated surfaces.

The Use of Lasers in Dental Materials: A Review.

Lasers have been well integrated in clinical dentistry for the last two decades, providing clinical alternatives in the management of both soft and hard tissues with an expanding use in the field of dental materials. One of their main advantages is that they can deliver very low to very high concentrated power at an exact point on any substrate by all possible means. The aim of this review is to thoroughly analyze the use of lasers in the processing of dental materials and to enlighten the new trends in laser technology focused on dental material management. New approaches for the elaboration of dental materials that require high energy levels and delicate processing, such as metals, ceramics, and resins are provided, while time consuming laboratory procedures, such as cutting restorative materials, welding, and sintering are facilitated. In addition, surface characteristics of titanium alloys and high strength ceramics can be altered. Finally, the potential of lasers to increase the adhesion of zirconia ceramics to different substrates has been tested for all laser devices, including a new ultrafast generation of lasers.

Biocompatibility assessment of resin-based cements on vascularized dentin/pulp tissue-engineered analogues.

OBJECTIVES: A three-dimensional (3D) dentin/pulp tissue analogue, resembling the human natural tissue has been engineered in an in vitro setup, aiming to assess the cytocompatibility of resin-based dental restorative cements. METHODS: Stem Cells from Apical Papilla (SCAP) and Human Umbilical Vein Endothelial Cells (HUVEC) were embedded in Collagen-I/Fibrin hydrogels at 1:3 ratio within 24-well plates. Hanging culture inserts were placed over the hydrogels, housing an odontoblast-like cell layer and a human treated-dentin barrier. Shear modulus of the hydrogels at 3.5 and 5 mg/ml was evaluated by dynamic mechanical analysis. Eluates of two resin-based cements, a dual-cure- (Breeze™, Pentron: Cement-1/C1), and a self-adhesive cement (SpeedCEMplus™, Ivoclar-Vivadent: Cement-2/C2) were applied into the dentin/pulp tissue analogue after pre-stimulation with LPS. Cytocompatibility was assessed by MTT assay, live/dead staining and real-time PCR analysis. RESULTS: Both hydrogel concentrations showed similar shear moduli to the natural pulp until day (D) 7, while the 5 mg/ml-hydrogel substantially increased stiffness by D14. Both cements caused no significant toxicity to the dentin/pulp tissue analogue. C1 induced stimulation (p < 0.01) of cell viability (158 ± 3%, 72 h), while pre-stimulation with LPS attenuated this effect. C2 (±LPS) caused minor reduction of viability (15-20%, 24 h) that recovered at 72 h for the LPS+ group. Both cements caused upregulation of VEGF, ANGP-1, and downregulation of the respective receptors VEGFR-2 and Tie-1. SIGNIFICANCE: Both resin-based cements showed good cytocompatibility and triggered angiogenic response within the dentin/pulp tissue analogue, indicating initiation of pulp repair responses to the released xenobiotics.

Fracture strength of endodontically treated premolars restored with different post systems and metal-ceramic or monolithic zirconia crowns.

The purpose of this study was to investigate the fracture strength of endodontically treated human maxillary premolars (ETP) restored with posts and metal ceramic (MC) or monolithic zirconia (MZ) crowns. Sixty ETP were randomly divided into 3 groups. Teeth in control group (C) received a resin filling. ETP in the MC group were restored with prefabricated metal posts, composite cores and MC crowns while in the MZ group with glass-fiber posts, composite cores and MZ crowns. Half of the specimens were loaded at a 135° angle and half under axial loading until fracture. The fracture modes were divided in repairable and irreparable using optical microscopy. Mean fracture strength was significantly higher for MC than for MZ crowns and control group only under axial loading. The distribution of repairable and irreparable failures presented no significant differences. Crown placement significantly improved the fracture strength of ETP irrespectively of post and crown type.

Hybrid chitosan/gelatin/nanohydroxyapatite scaffolds promote odontogenic differentiation of dental pulp stem cells and in vitro biomineralization.

OBJECTIVE: Hybrid chitosan/gelatin/nanohydroxyapatite (CS/Gel/nHA) scaffolds have attracted considerable interest in tissue engineering (TE) of mineralized tissues. The present study aimed to investigate the potential of CS/Gel/nHA scaffolds loaded with dental pulp stem cells (DPSCs) to induce odontogenic differentiation and in vitro biomineralization. METHODS: CS/Gel/nHA scaffolds were synthesized by freeze-drying, seeded with DPSCs, and characterized with flow cytometry. Scanning Electron Microscopy (SEM), live/dead staining, and MTT assays were used to evaluate cell morphology and viability; real-time PCR for odontogenesis-related gene expression analysis; SEM-EDS (Energy Dispersive X-ray spectroscopy), and X-ray Diffraction analysis (XRD) for structural and chemical characterization of the mineralized constructs, respectively. RESULTS: CS/Gel/nHA scaffolds supported viability and proliferation of DPSCs over 14 days in culture. Gene expression patterns indicated pronounced odontogenic shift of DPSCs, evidenced by upregulation of DSPP, BMP-2, ALP, and the transcription factors RunX2 and Osterix. SEM-EDS showed the production of a nanocrystalline mineralized matrix inside the cell-based and - to a lesser extent - the cell-free constructs, with a time-dependent production of net-like nanocrystals (appr. 25-30nm in diameter). XRD analysis gave the crystallite size (D=50nm) but could not distinguish between the initially incorporated and the biologically produced nHA. SIGNIFICANCE: This is the first study validating the potential of CS/Gel/nHA scaffolds to support viability and proliferation of DPSCs, and to provide a biomimetic microenvironment favoring odontogenic differentiation and in vitro biomineralization without the addition of any inductive factors, including dexamethasone and/or growth/morphogenetic factors. These results reveal a promising strategy towards TE of mineralized dental tissues.

Advanced in Vitro Experimental Models for Tissue Engineering-based Reconstruction of a 3D Dentin/pulp Complex: a Literature Review.

OBJECTIVE: Experimental procedures have been used to monitor cellular responses at the dentin/pulp interface. Aiming to divert from in vivo studies and oversimplified two-dimensional assays, three-dimensional (3D) models have been developed. This review provides an overview of existing literature, regarding 3D in vitro dentin/pulp reconstruction. MATERIAL & METHODS: PubMed, Scopus, Cochrane Library and Web of Science- were systematically searched for attributes between 1998 and 2020. The search focused on articles on the development of three-dimensional tools for the reconstruction of a dentin/pulp complex under in vitro conditions, which were then screened and qualitatively assessed. Article grouping according to mode of implementation, resulted in five categories: the customised cell perfusion chamber (CPC) (n = 8), the tooth bud model (TBM) (n = 3), the 3D dentin/pulp complex manufactured by tissue engineering (DPC) (n = 6), the entire tooth culture (ETC) (n = 4) and the tooth slice culture model (TSC) (n = 5). RESULTS: A total of 26 publications, applying nine and eight substances for pulp and dentin representation respectively, were included. Natural materials and dentin components were the most widely utilized. The most diverse category was the DPC, while the CPC group was the test with the highest longevity. The most consistent categories were the ETC and TSC models, while the TBM presented as the most complete de novo approach. CONCLUSIONS: All studies presented with experimental protocols with potential upgrades. Solving the limitations of each category will provide a complete in vitro testing and monitoring tool of dental responses to exogenous inputs. CLINICAL RELEVANCE: The 3D dentin/pulp complexes are valid supplementary tools for in vivo studies and clinical testing. Graphical Abstract.

The effect of different aging protocols on the flexural strength and phase transformations of two monolithic zirconia ceramics.

INTRODUCTION: The aim of the present study was to investigate how different aging protocols can affect the flexural strength and phase transformations of yttrium-stabilized zirconia ceramics (Y-TZP) for monolithic restorations. MATERIALS AND METHODS: Bar-shaped specimens from two zirconia ceramics bars were divided into three groups: a. no treatment (c), b. aging in an autoclave (a), and c. thermal cycling (t). The flexural strength was determined by the 3-point bending test and statistical analysis was performed to determine significant differences (p< 0.05). Weibull statistics was used to analyze the dispersion of strength values while surface microstructural analysis was performed through X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). RESULTS: Aging did not significantly affect the flexural strength but differences were recorded between the two groups, with group A presenting higher strength values and m-phase percentages. CONCLUSIONS: The observed differences between the two ceramics could be attributed to variations in composition and processing.

The Efficacy of Stem Cells Secretome Application in Osteoarthritis: A Systematic Review of In Vivo Studies.

PURPOSE: Mesenchymal stem cells (MSCs) have appeared as a promising regenerative cell-based therapeutic, for degenerative conditions, such as OA, while the beneficial results from the application of MSCs have been attributed to the MSCs-derived secretome, which is the sum of cytoprotective factors produced by the MSCs. Aim of this study was to systematically review the literature in order to assess whether stem cell secretome (conditioned medium-CM, exosome-Exos or microvesicles-MV)(CM/Exos/MVs) treatment reduces inflammation and enhances cartilage regeneration in preclinical studies of experimental arthritis. MATERIALS AND METHODS: An extensive electronic search was conducted by 2 independent reviewers by using the PubMed, Cochrane Library, Web of Science, and Scopus database, as well as Google Scholar, in order to identify the studies that met our inclusion criteria until August 2019. Included studies were assessed for quality and Risk of Bias (RoB) using the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines and a modification of Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) RoB tool for animal studies, respectively. RESULTS: The initial search provided 525 records, with 28 fulfilling the inclusion criteria. The included studies presented great heterogeneity regarding the stem cells used, the preparation of therapeutic agent as well as the animal models used for testing. In addition, most studies presented with an unclear or high risk bias. CONCLUSION: In summary, the positive results of CM/Exos/MVs application in preclinical models of experimentally induced OA in terms of resolution of inflammation and cartilage regeneration are highlighted in this review, presenting a promising therapeutic solution for OA.

Variables affecting the fit of zirconia fixed partial dentures: A systematic review.

STATEMENT OF PROBLEM: Different parameters affect the marginal and internal fit of zirconia fixed partial dentures (FPDs) on natural teeth. Determining a way to optimize these restorations is essential. PURPOSE: The purpose of this systematic review was to determine the variables affecting the marginal fit and internal accuracy of zirconia FPDs on natural teeth. MATERIAL AND METHODS: An electronic search was conducted by 2 independent reviewers by using the MEDLINE, Cochrane, Web of Science, and Scopus databases, as well as Google Scholar, for studies published up to July 2018, and a manual search was conducted from the reference lists of related articles. Eligibility criteria included articles in English published in peer-reviewed journals that assessed the marginal and/or internal adaptation of zirconia FPDs on teeth with 3 or more units, with at least 1 of the experimental groups being frameworks or FPDs fabricated from zirconia. Risk of bias was assessed with the aid of the Cochrane Handbook for Systematic Reviews of Interventions. RESULTS: The search provided 418 records, with 41 fulfilling the inclusion criteria. The selected studies showed considerable heterogeneity regarding materials, state of sintering, manufacturer and computer-aided design and computer-aided manufacturing (CAD-CAM) system, experimental methodology, sample size, and span length. Of the included articles, 36 were in vitro studies and 5 were clinical studies, most of which exhibited high-performance and detection biases. CAD-CAM systems provided more precise marginal and internal fit than CAM systems. An increase of framework span length to 6 or more units decreased both marginal and internal fit. The reported marginal gap tended to increase after the veneering process. The introduction of a conventional impression into an otherwise digital workflow seems to have a negative effect on the marginal fit. CONCLUSIONS: The accuracy of zirconia FPDs or frameworks is considerably influenced by the processing procedure used and the choice of CAD-CAM system.

Color alterations of a PMMA resin for fixed interim prostheses reinforced with silica nanoparticles.

PURPOSE: The aim of the present study was to evaluate the color changes of an autopolymerizing PMMA resin used for interim fixed restorations, reinforced with SiO2 nanoparticles. MATERIALS AND METHODS: Silica nanoparticles were blended with the PMMA resin powder through high-energy ball milling. Four shades of PMMA resin were used (A3, B3, C3, D3) and total color differences were calculated through the equations ΔEab= [(ΔL*)2 + (Δa*)2 + (Δb*)2]½ and Δ E 00 = [ Δ L ' K L S L 2 + Δ C ' K C S C 2 + Δ H ' K H S H 2 + R T Δ C ' K C S C Δ H ' K H S H ] 1 / 2 . Statistically significant differences between ΔEab and the clinically acceptable values of 3.3 and 2.7 and those between ΔE00 and the clinically acceptable value of 1.8 were evaluated with one sample t-test (P<.05). Differences among the different shades were assessed through One-Way ANOVA and Bonferroni multiple comparison tests. RESULTS: Significantly lower values were detected for all groups concerning ΔEab compared to the intraorally clinical acceptable values of 3.3 and 2.7. Significantly lower mean values were detected for groups B3, C3, and D3, concerning ΔE00 compared to the intraorally clinical acceptant value of 1.8. Color pigments in red-brown (A3) and red-grey (D3) shades affect the total color change to a greater extent after the reinforcement with SiO2 nanoparticles compared to the red-yellow (B3) shade. CONCLUSION: Within the limitations of this in vitro study, it can be suggested that reinforcing PMMA with SiO2 nanoparticles at 0.25 wt% slightly affects the optical properties of the PMMA resin without being clinically perceivable.

In vitro evaluation of the shear bond strength and bioactivity of a bioceramic cement for bonding monolithic zirconia.

STATEMENT OF PROBLEM: Adhesive cementation is the most common bonding strategy for zirconia restorations. Although cementation with a bioactive luting agent has been proposed as an alternative, how the bond strength compares is unclear. PURPOSE: The purpose of this in vitro study was to evaluate shear bond strength after cementing a monolithic zirconia ceramic to human dentin with a bioceramic cement, compare it with a traditional cement, and evaluate its bioactive properties. MATERIAL AND METHODS: A total of 120 dentin specimens and 120 yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) (BruxZir) cylindrical specimens were used. Zirconia and dentin specimens were randomly divided into 8 study groups (n=15) based on 2 luting cement types (a bioceramic cement or glass ionomer cement as control), 2 airborne-particle abrasion protocols (50 µm or 110 µm), and 2 water storage durations (24 hours or 30 days). After the shear bond strength test using a universal machine at a crosshead speed of 1 mm/min, fracture patterns were evaluated under a stereomicroscope and a scanning electron microscope. Strength values were statistically analyzed with a 3-factor ANOVA model (α=.05). Bioactivity was evaluated in simulated body fluid (SBF). RESULTS: The control glass ionomer cement achieved significantly greater shear bond strength compared with the tested bioceramic cement. Mean bond strength values ranged from 2.52 MPa to 5.23 MPa for the bioceramic cement tested and from 4.20 MPa to 6.61 MPa for the control cement. The duration of water storage played a significant role in the bond strength, with groups stored for 30 days reaching higher bond strength values, whereas the particle size of airborne-particle abrasion did not have a significant effect. Failure types were primarily mixed. No apatite formation was recorded on the surface of the specimens even after 30 days of immersion in SBF. CONCLUSIONS: The evaluated cement did not develop apatite in SBF, and its bond strength values were below the control glass ionomer cement.

Assessment of cytotoxicity and antibacterial effects of silver nanoparticle-doped titanium alloy surfaces.

OBJECTIVES: This study aimed to develop silver nanoparticle (AgNP)-doped Ti6Al4V alloy surfaces and investigate their antibacterial properties against representative periopathogens and potential cytotoxicity on osteoblastic cells. METHODS: AgNPs of different size distributions (5 and 30nm) were incorporated onto the Ti6Al4V surfaces by electrochemical deposition, using colloid silver dispersions with increasing AgNP concentrations (100, 200 and 300ppm). The time-course silver release from the specimen surfaces to cell culture media was assessed by Atomic Absorption Spectroscopy (AAS). Cell attachment, viability and proliferation were investigated by SEM, live/dead staining MTT and BrdU assays. The antibacterial effects were assessed against P. gingivalis and P. intermedia by serial dilution spotting assays. RESULTS: A time- and concentration-dependent silver release from the experimental surfaces was observed. Overall, cell viability and attachment on the AgNP-doped surfaces, suggested adequate cytocompatibility at all concentrations. A transient cytotoxic effect was detected at 24h for the 5nm-sized groups that fully recovered at later time-points, while no cytotoxicity was observed for the 30nm-sized groups. A statistically significant, concentration-dependent decrease in cell proliferation rates was induced at 48h in all AgNP groups, followed by recovery at 72h in the groups coated with 5nm-sized AgNPs. A statistically significant, concentration-dependent antibacterial effect up to 30% was confirmed against both periopathogens. SIGNIFICANCE: This study sheds light to the optimal size-related concentrations of AgNP-doped Ti6Al4V surfaces to achieve antibacterial effects, without subsequent cytotoxicity. These results significantly contribute to the development of antibacterial surfaces for application in oral implantology.

Evaluation of the biological behaviour of various dental implant abutment materials on attachment and viability of human gingival fibroblasts.

OBJECTIVE: This study aimed to investigate the biological effects of yttria-stabilized zirconia (Y-TZP) compared to other dental implant abutment materials, i.e. lithium disilicate (LS2) and titanium alloy (Ti), as well as the effects of aging of Y-TZP on viability/proliferation and attachment properties of Human Gingival Fibroblasts (HGFs). METHODS: Cylindrical specimens of each material were prepared as per manufacturer's instructions. Y-TZP specimens were divided into three groups: 1. no aging (Zr0), 2. aging for 5 h, 134 °C, 2 bars, 100% humidity (Zr5), 3. aging for 10 h under the same conditions (Zr10). Surface roughness was evaluated by optical profilometry; cell metabolic activity/viability by MTT assay, morphological changes by Scanning Electron Microscopy (SEM) and ratio of live/dead cells by confocal microscopy. RESULTS: Results showed statistically significant reduction of HGF metabolic activity/viability in contact with Y-TZP after aging. Nevertheless, non-aged zirconia showed no significant differences compared with LS2, Ti and control cultures. In contrast, significant stimulation of cell metabolic activity/viability was observed in HGFs exposed to LS2 eluates. Differential morphological patterns were observed for HGF in contact with different materials/treatments, with obviously increased number of dead cells and sparser distribution of HGFs cultured on Zr10 specimens. These effects were not correlated with surface topography, since Y-TZP aging did not alter surface micro-roughness. SIGNIFICANCE: These findings indicate that Y-TZP shows comparable biological properties to Ti and LS2 as implant abutment material. Nevertheless, Y-TZP aging might influence gingival cell attachment and proliferation properties, providing an alert to a potentially negative effect on the long-term maintenance of gingival architecture.

Dental pulp stem cells in chitosan/gelatin scaffolds for enhanced orofacial bone regeneration.

OBJECTIVE: Biomimetic chitosan/gelatin (CS/Gel) scaffolds have attracted great interest in tissue engineering of several tissues. However, limited information exists regarding the potential of combining CS/Gel scaffolds with oral cells, such as dental pulp stem cells (DPSCs), to produce customized constructs targeting alveolar/orofacial bone reconstruction, which has been the aim of the present study. METHODS: Two scaffold types, designated as CS/Gel-0.1 and CS/Gel-1, were fabricated using 0.1 and 1% (v/v) respectively of the crosslinker glutaraldehyde (GTA). Scaffolds (n=240) were seeded with DPSCs with/without pre-exposure to recombinant human BMP-2. In vitro assessment included DPSCs characterization (flow cytometry), evaluation of viability/proliferation (live/dead staining, metabolic-based tests), osteo/odontogenic gene expression analysis (qRT-PCR) and structural/chemical characterization (scanning electron microscopy, SEM; energy dispersive X-ray spectroscopy, EDX; X-ray powder diffraction, XRD; thermogravimetry, TG). In vivo assessment included implantation of DPSC-seeded scaffolds in immunocompromised mice, followed by histology and SEM-EDX. Statistical analysis employed one/two-way ANOVA and Tukey's post-hoc tests (significance for p<0.05). RESULTS: Both scaffolds supported cell viability/proliferation over 14 days in culture, showing extensive formation of a hydroxyapatite-rich nanocrystalline calcium phosphate phase. Differential expression patterns indicated GTA concentration to significantly affect the expression of osteo/odontogenic genes, with CS/Gel-0.1 scaffolds being more effective in upregulating DSPP, IBSP and Osterix. In vivo analysis demonstrated time-dependent production of a nanocrystalline, mineralized matrix at 6, 8 and 10 weeks, being more prominent in constructs bearing rhBMP-2 pre-treated cells. The latter showed higher amounts of osteoid and fully mineralized bone, as well as empty space reduction. SIGNIFICANCE: These results reveal a promising strategy for orofacial bone tissue engineering.

Fibro/chondrogenic differentiation of dental stem cells into chitosan/alginate scaffolds towards temporomandibular joint disc regeneration.

Tissue engineering (TE) may provide effective alternative treatment for challenging temporomandibular joint (TMJ) pathologies associated with disc malpositioning or degeneration and leading to severe masticatory dysfunction. Aim of this study was to evaluate the potential of chitosan/alginate (Ch/Alg) scaffolds to promote fibro/chondrogenic differentiation of dental pulp stem cells (DPSCs) and production of fibrocartilage tissue, serving as a replacement of the natural TMJ disc. Ch/Alg scaffolds were fabricated by crosslinking with CaCl2 combined or not with glutaraldehyde, resulting in two scaffold types that were physicochemically characterized, seeded with DPSCs or human nucleus pulposus cells (hNPCs) used as control and evaluated for cell attachment, viability, and proliferation. The DPSCs/scaffold constructs were incubated for up to 8 weeks and assessed for extracellular matrix production by means of histology, immunofluorescence, and thermomechanical analysis. Both Ch/Alg scaffold types with a mass ratio of 1:1 presented a gel-like structure with interconnected pores. Scaffolds supported cell adhesion and long-term viability/proliferation of DPSCs and hNPCs. DPSCs cultured into Ch/Alg scaffolds demonstrated a significant increase of gene expression of fibrocartilaginous markers (COLI, COL X, SOX9, COM, ACAN) after up to 3 weeks in culture. Dynamic thermomechanical analysis revealed that scaffolds loaded with DPSCs significantly increased storage modulus and elastic response compared to cell-free scaffolds, obtaining values similar to those of native TMJ disc. Histological data and immunochemical staining for aggrecan after 4 to 8 weeks indicated that the scaffolds support abundant fibrocartilaginous tissue formation, thus providing a promising strategy for TMJ disc TE-based replacement.