Rosa Valletta

Effect of 2-Hydroxyethyl Methacrylate on Human Pulp Cell Survival Pathways ERK and AKT

Previous investigations have revealed that dental monomers could affect intracellular pathways leading to cell survival or cell death. Mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) might mediate cell responses as well as cell survival and apoptosis. The purpose of this study was to evaluate the effects of 2-hydroxyethyl methacrylate (HEMA) on the ERK1/2 and AKT pathways in human primary pulp fibroblasts (HPCs). HPCs were treated with various concentrations of HEMA, after which viability and reactive oxygen species levels were determined by flow cytometry with Annexin V-PI staining and 2,7-dichlorofluorescine diacetate, respectively. Whole-cell extracts were immunoblotted with anti-P-Akt or anti-P-ERK1/2. Cell viability decreased in a dose-dependent manner after HEMA exposure, showing a significant decrease with 10 mmol/L HEMA (p < .05). HEMA treatment resulted in a 4-fold increase in reactive oxygen species formation (p < .05). A short HEMA exposure (30-90 minutes) increased ERK1/2 phosphorylation, whereas a decrease in the AKT phosphorylation was observed. Selective inhibitors of the ERK (PD98059) and AKT (LY294002) pathways amplified HPC cell damage after HEMA exposure. Our findings demonstrated that HEMA exposure modulates the ERK and AKT pathways in different manners, and that in turn, they function in parallel to mediate pro-survival signaling in pulp cells subjected to HEMA cytotoxicity.

Evaluation of surface roughness of orthodontic wires by means of atomic force microscopy

OBJECTIVE To compare the surface roughness of different orthodontic archwires. MATERIALS AND METHODS Four nickel-titanium wires (Sentalloy(®), Sentalloy(®) High Aesthetic, Titanium Memory ThermaTi Lite(®), and Titanium Memory Esthetic(®)), three β-titanium wires (TMA(®), Colored TMA(®), and Beta Titanium(®)), and one stainless-steel wire (Stainless Steel(®)) were considered for this study. Three samples for each wire were analyzed by atomic force microscopy (AFM). Three-dimensional images were processed using Gwiddion software, and the roughness average (Ra), the root mean square (Rms), and the maximum height (Mh) values of the scanned surface profile were recorded. Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Tukeys post hoc test (P < .05). RESULTS The Ra, Rms, and Mh values were expressed as the mean ± standard deviation. Among as-received archwires, the Stainless Steel (Ra  =  36.6 ± 5.8; Rms  =  48 ± 7.7; Mh  =  328.1 ± 64) archwire was less rough than the others (ANOVA, P < .05). The Sentalloy High Aesthetic was the roughest (Ra  =  133.5 ± 10.8; Rms  =  165.8 ± 9.8; Mh  =  949.6 ± 192.1) of the archwires. CONCLUSIONS The surface quality of the wires investigated differed significantly. Ion implantation effectively reduced the roughness of TMA. Moreover, Teflon(®)-coated Titanium Memory Esthetic was less rough than was ion-implanted Sentalloy High Aesthetic.

N-acetyl cysteine directed detoxification of 2-hydroxyethyl methacrylate by adduct formation

Cytotoxicity of the dental resin monomer 2-hydroxyethyl methacrylate (HEMA) and the protective effects of N-acetyl cysteine (NAC) on monomer-induced cell damage are well demonstrated. The aim of our study was to analyze the hypothesis that the protection of NAC from HEMA cytotoxicity might be due to direct NAC adduct formation. To this end, using HPLC we first measured the actual intracellular HEMA concentrations able to cause toxic effects on 3T3-fibroblasts and then determined the decrease in intracellular and extracellular HEMA levels in the presence of NAC. In addition, by capillary electrophoresis coupled with mass spectrometry analysis (CE-MS), we evaluated NAC-HEMA adduct formation. HEMA reduced 3T3 cell vitality in a dose- and time-dependent manner. The concentration of HEMA inside the cells was 15-20 times lower than that added to the culture medium for cell treatment (0-8 mmol/L). In the presence of 10 mmol/L NAC, both intracellular and extracellular HEMA concentrations greatly decreased in conjunction with cytotoxicity. NAC-HEMA adducts were detected both in the presence and absence of cells. Our findings suggest that the in vitro detoxification ability of NAC against HEMA-induced cell damage occurs through NAC adduct formation. Moreover, we provide evidence that the actual intracellular concentration of HEMA able to cause cytotoxic effects is at least one magnitude lower than that applied extracellularly.

Efficacy of the Sander bite-jumping appliance in growing patients with mandibular retrusion: a randomized controlled trial

OBJECTIVES The efficacy of functional appliances remains highly debated. This randomized controlled trial investigated the skeletal and dentoalveolar effects determined by the Sander bite-jumping appliance (BJA). The null hypothesis to be tested was that the appliance would not induce supplementary mandibular growth compared to untreated controls. SETTING AND SAMPLE POPULATION This study was carried out at the Section of Orthodontics, University of Naples Federico II, Italy. Forty-six patients receiving a clinical diagnosis of skeletal and dental class II due to mandibular retrusion were either allocated to a treatment (23 patients;15 boys, 8 girls; mean age ± SD: 10.9 ± 1.3 years) or to an untreated control group (23 patients;11 boys, 12 girls; mean age ± SD: 10.5 ± 1.2 years), by using a balanced block randomization. METHODS Lateral cephalograms were taken before and after treatment and used for comparisons. Measurements were analyzed by descriptive statistics, univariate and multivariate statistical tests. RESULTS Treated individuals had a significant increase in mandibular length (6.4 ± 2.3 vs. 3.5 ± 2.5 mm; p < 0.001), overjet reduction (-5.0 ± 2.9 vs. 0.3 ± 1.2 mm; p < 0.001) and molar relationship improvement (-5.3 ± 2.4 vs. 0.1 ± 1.1 mm; p < 0.001) compared to controls. The use of the appliance did not significantly affect jaw divergence. Proclination of lower incisors was slightly greater (3.0°, p = 0.023) in treated patients than in controls. The increase in mandibular length was not significantly influenced by cervical stage (p = 0.40). CONCLUSION The BJA can effectively correct class II malocclusions by a combination of dentoalveolar and skeletal effects. The long-term stability of the correction needs to be evaluated.

Effect of Nickel Chloride on Cell Proliferation

Objective: Metal alloys used in dentistry and in other biomedical fields may release nickel ions in the oral environment. The release of nickel might influence the normal biological and physiological processes, including tissue wound healing, cell growth and proliferation. The aim of this study was to evaluate in vitro the effects of nickel ions on cell cycle, viability and proliferation. Materials and Methods: Human osteosarcoma cells (U2OS) and human keratinocytes (HaCat) were exposed to different nickel chloride (NiCl2) concentrations (0 - 5mM) for various periods exposure. The viability of cultured cells was estimated by flow cytometry using Annexin V-FITC and Propidium Iodide (PI). Cell proliferation was evaluated by using carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometry. Finally, the effects of NiCl2 on cell cycle were assessed and quantified by flow cytometry. Statistical analysis was performed by means of ANOVA followed by Tukey’s test. Results: NiCl2 induced a dose and time dependent decrease in cell viability. After 24h, 1mM NiCl2 caused a similar and significant reduction of viability in U2OS and HaCat cells, while higher NiCl2 concentrations and longer exposure times showed a reduced cytotoxic effect in HaCat as compared to U2OS cells. Exposure to NiCl2 caused a dose- and time-dependent inhibition of cell proliferation in both cell lines tested, with a prominent effect on U2OS cells. Furthermore, both cell lines exposed to NiCl2 exhibited significant changes in cell cycle distribution after 24h exposure 2mM NiCl2, as compared to untreated cells (p<0.05). Conclusion: Our results indicate that release of nickel ions may affect cell proliferation. The inhibition of cell growth by NiCl2 is mediated by both cell cycle arrest and by induction of cell death.

The influence of Ni(II) on surface antigen expression in murine macrophages.

Biomedical alloys may release nickel ions during corrosion phenomena and, in addition to their interaction with oral tissues, these ions may also influence characteristic properties of the immune system cells. The aim of this study was to evaluate the effect of nickel chloride on the expression of functionally distinct surface antigens in murine RAW macrophages. The expression of the surface antigens CD14, CD40, MHC class I, MHC class II, CD80, CD86, CD54 was analyzed by flow cytometry. The bacterial endotoxin lipopolysaccharide (LPS) was used as a positive control to induce antigen expression. Cells were stimulated with NiCl(2) (0.1 and 0.5mm) in the presence and absence of LPS (0.1 or 25 microg/ml). After exposure periods of 6, 24 and 48 h, LPS caused a time- and dose-dependent increase in the expression of all surface antigens. CD14 expression was up-regulated by 0.1 microg/ml LPS by about 10-fold after 24h and 100-fold after 48 h. After 48 h, NiCl(2) alone up-regulated the expression of all surface antigens between 2- and 4-fold, while in cells stimulated by LPS, 0.1mm NiCl(2) was effective only on CD14, CD40 and MHC class I. Moreover, 0.5mm NiCl(2) even inhibited the LPS-induced expression of all surface antigens, except for CD54, which was still significantly up-regulated. These results show that nickel chloride is able to induce an up-regulation of surface antigen expression, but a high concentration may impair essential functions of macrophages stimulated by LPS.

Behaviour of human mesenchymal stem cells on chemically synthesized HA-PCL scaffolds for hard tissue regeneration.

Our goal was to characterize the response of human mesenchymal stem cells (hMSCs) to a novel composite scaffold for bone tissue engineering. The hydroxyapatite–polycaprolactone (HA–PCL) composite scaffolds were prepared by a sol–gel method at room temperature and the scaffold morphology was investigated by scanning electron microscopy (SEM)/energy‐dispersive spectroscopy (EDS) to validate the synthesis process. The response of two different lines of hMSCs, bone‐marrow‐derived human mesenchymal stem cells (BMSCs) and dental pulp stem cells (DPSCs) in terms of cell proliferation and differentiation into the osteoblastic phenotype, was evaluated using Alamar blue assay, SEM, histology and alkaline phosphatase activity. Our results indicate that tissue engineering by means of composite HA–PCL scaffolds may represent a new therapeutic strategy to repair craniofacial bone defects. Copyright

Effects of intraoral aging on surface properties of coated nickel-titanium archwires

OBJECTIVE To evaluate the effects of intraoral aging on surface properties of esthetic and conventional nickel-titanium (NiTi) archwires. MATERIALS AND METHODS Five NiTi wires were considered for this study (Sentalloy, Sentalloy High Aesthetic, Superelastic Titanium Memory Wire, Esthetic Superelastic Titanium Memory Wire, and EverWhite). For each type of wire, four samples were analyzed as received and after 1 month of clinical use by an atomic force microscope (AFM) and a scanning electronic microscope (SEM). To evaluate sliding resistance, two stainless steel plates with three metallic or three monocrystalline brackets, bonded in passive configuration, were manufactured; four as-received and retrieved samples for every wire were pulled five times at 5 mm/min for 1 minute by means of an Instron 5566, recording the greatest friction value (N). Data were analyzed by one-way analysis of variance and by Students t-test. RESULTS After clinical use, surface roughness increased considerably. The SEM images showed homogeneity for the as-received control wires; however, after clinical use esthetic wires exhibited a heterogeneous surface with craters and bumps. The lowest levels of friction were observed with the as-received Superelastic Titanium Memory Wire on metallic brackets. When tested on ceramic brackets, all the wires exhibited an increase in friction (t-test; P < .05). Furthermore, all the wires, except Sentalloy, showed a statistically significant increase in friction between the as-received and retrieved groups (t-test; P < .05). CONCLUSION Clinical use of the orthodontic wires increases their surface roughness and the level of friction.

Dental and skeletal effects of palatal expansion techniques: a systematic review of the current evidence from systematic reviews and meta-analyses

The aim was to assess the quality and to summarise the findings of the Systematic Reviews (SRs) and Meta-Analyses (MAs) on the dental and skeletal effects of maxillary expansion. Electronic and manual searches have been independently conducted by two investigators, up to February 2015. SRs and MAs on the dentoalveolar and skeletal effects of fixed expanders were included. The methodological quality was assessed using the AMSTAR (A Measurement Tool to Assess Systematic Reviews). The design of the primary studies included in each SR/MA was assessed with the LRD (Level of Research Design scoring). The evidence for each outcome was rated applying a pre-determined scale. Twelve SRs/MAs were included. The AMSTAR scores ranged from 4 to 10. Two SRs/MAs included only RCTs. The current findings from SRs/MAs support with high evidence a significant increase in the short-term of maxillary dentoalveolar transversal dimensions after Rapid Maxillary Expansion (RME). The same effect is reported with moderate evidence after Slow Maxillary Expansion (SME). However, there is moderate evidence of a non-significant difference between the two expansion modalities concerning the short-term dentoalveolar effects. With both RME and SME, significant increase of skeletal transversal dimension in the short-term is reported, and the skeletal expansion is always smaller than the dentoalveolar. Even though dental relapse to some extent is present, long-term results of the dentoalveolar effects show an increase of the transversal dimension, supported by moderate evidence for RME and low evidence for SME. Skeletal long-term effects are reported only with RME, supported by very low evidence.

Effect of pH on in vitro biocompatibility of orthodontic miniscrew implants

BackgroundAlthough the clinical use of miniscrews has been investigated on a large scale, little is known about their biocompatibility. Since low pH can affect corrosion resistance, the aim of this study was to evaluate the cytotoxic effect of orthodontic miniscrews in different pH conditions.MethodsFour orthodontic miniscrews of stainless steel and grade IV and grade V titanium were immersed in a pH 7 and pH 4 saline solution for 1, 7, 14, 21, 28, and 84 days. Human osteogenic sarcoma cells (U2OS), permanent human keratinocytes (HaCat), and primary human gingival fibroblasts (HGF) were exposed to eluates, and the mitochondrial dehydrogenase activity was measured after 24 h to assess the cytoxicity. The results were analyzed using the Mann-Whitney U test (P < 0.05).ResultsWhen exposed to pH 7-conditioned eluates, the cell lines showed an even greater viability than untreated cells. On the contrary, the results revealed a statistically significant decrease in U2OS, HaCat, and HGF viability after exposure to eluates obtained at pH 4. Among the cell lines tested, HGF showed the most significant decrease of mitochondrial activity. Interestingly, grade V titanium miniscrews caused highest toxic effects when immersed at pH 4.ConclusionsThe results suggested that at pH 7, all the miniscrews are biocompatible while the eluates obtained at pH 4 showed significant cytotoxicity response. Moreover, different cell lines can produce different responses to miniscrew eluates.