Cytotoxicity effects and differentiation potential of ormocer-based and nanohybrid composite resins on human dental pulp stem cells\.

OBJECTIVE: to compare conventional nanohybrid (Ceram.x Spectra) and ormocer-based (Admira fusion) dental composite resins effects on human dental pulp stem cells (hDPSCs) in terms of cytotoxicity, self-renewal, migration and osteogenic differentiation. METHODS: hDPSCs were cultured in presence of different dilutions (undiluted, form 1:2 to 1:100) of CeramX (CX) and Admira fusion (AD) eluates and viability assay in standard or osteogenic conditions were performed. Samples and eluates were prepared according to ISO 10993-12. In addition, apoptosis, self-renewal and migration activity evaluations were carried out. Osteogenic differentiation potential was tested by Alkaline Phosphatase Activity, alizarin red staining and gene expression of specific markers (ALP, RUNX2, OCN, OPN and COL1α1). Statistical analysis was performed by means of a One-way analysis of variance (One-way ANOVA) followed by a Tukey's test for multiple comparison; results were presented as mean ± standard error of mean (SEM). RESULTS: Admira Fusion demonstrated to be highly biocompatible and showed positive effects on hDPSCs proliferation and differentiation; on the contrary, conventional nanohybrid composite showed to be more cytotoxic and without any notable effect on stem cells differentiation. Moreover, the obtained results were further corroborated by a significant upregulation of osteogenic differentiation markers obtained in presence of ormocer-based composite resin eluate. Specifically, in AD 1:50 group expression levels of ALP, Runx2, Col1α1 were double than control (ALP, p = 0.045; Runx2, p = 0.003; Col1α1, p = 0.001) and CX 1:50 (ALP, p = 0.006; RUNX2, p = 0.029; Col1α1, p = 0.005). Moreover, in the same group, OPN and OCN resulted about 5 times more expressed as compared to control (OPN, p = 0.009; OCN, p = 0.0005) and CX 1:50 (OPN, p = 0.012; OCN, p = 0.0006). SIGNIFICANCE: The less cytotoxicity obtained by AD than conventional nanohybrid composite may be attributed to a reduced monomers release in the oral environment, supporting the hypothesis of limited adverse effect and enhanced healing potential, mainly when the material is positioned in close contact with pulp tissue.

Oral-Gut-Estrobolome Axis May Exert a Selective Impact on Oral Cancer.

A subset of bacterial species that holds genes encoding for ß-glucuronidase and ß-galactosidase, enzymes involved in the metabolism of conjugated estrogens, is called the "estrobolome." There is an emerging interest embracing this concept, as it may exert a selective impact on a number of pathologies, including oral cancer. Although the estrobolome bacteria are typically part of the gut microbiota, recent experimental pieces of evidence have suggested a crosstalk among oral and gut microbiota. In fact, several oral bacterial species are well represented also in the gut microbiota, and these microbes can effectively induce the estrobolome activation. The main pathways used for activating the estrobolome are based on the induction of the expression patterns for 2 bacterial enzymes: ß-glucuronidase and aromatase, both involved in the increase of estrogen released in the bloodstream and consequently in the salivary compartment. Mechanistically, high estrogen availability in saliva is responsible for an increase in oral cancer risk for different reasons: briefly, 1) estrogens directly exert biological and metabolic effects on oral mucosa cells; 2) they can modulate the pathological profile of some bacteria, somewhere associated with neoplastic processes (i.e., Fusobacterium spp., Parvimonas ssp.); and 3) some oral bacteria are able to convert estrogens into carcinogenic metabolites, such as 4-hydroxyestrone and 16α-hydroxyestrone (16α-OHE), and can also promote local and systemic inflammation. Nowadays, only a small number of scientific studies have taken into consideration the potential correlations among oral dysbiosis, alterations of the gut estrobolome, and some hormone-dependent cancers: this lack of attention on such a promising topic could be a bias affecting the full understanding of the pathogenesis of several estrogen-related oral pathologies. In our article, we have speculated on the activity of an oral-gut-estrobolome axis, capable of synergizing these 2 important microbiotas, shedding light on a pilot hypothesis requiring further research.

Oral health status and Unmet Restorative Treatment Needs (UTN) in disadvantaged migrant and not migrant children in Italy.

AIM: Despite a global decline in caries experience in children, dental caries remains a common and persistent public health problem, being more prevalent among socioeconomically disadvantaged groups. The aim of this survey is to evaluate DMFT index, caries prevalence and Unmet Restorative Treatment Needs (UNT) index in migrant and not migrant children with low income, in Campania region (Italy), in order to plan and build up strategies for promoting dental health in children and adequate health care for vulnerable groups. MATERIALS AND METHODS: A sample of 553 children (age range: 12-14 years old) were recruited from volunteer patients at a charitable foundation in Naples (Italy) and were categorised into 2 subgroups, according to their status of "migrant" or "not migrant". Parents completed a questionnaire in order to investigate demographic and oral health behaviour. The participants were healthy and from low family income levels. DMFT scores and caries prevalence were calculated; UNT index was also calculated [D?(D + F)]%. The mean DMFT values related to the examined variables were calculated using one-way ANOVA. RESULTS: Mean DMFT and UTN index were 3.92±2.92 and 86.3% for migrants, and 3.29±3.21 and 68.4% for not migrants, respectively. The difference in the mean DMFT between the two groups was statistically significant (p=.027). Caries prevalence was 77.5% and 55.9% in the migrant group and in the not migrant group, respectively. High DMFT values were statistically associated to history of previous dental visit, sugar intake, brushing teeth once a day, and low mothers' educational level for both groups. CONCLUSION: Economically disadvantaged children showed high levels of dental caries. Furthermore, DMFT values, caries prevalence and unmet restorative treatment needs index among migrant children were higher than that of not migrant. In order to reduce inequalities in dental caries experience, there is a need to design policies aimed at primary prevention through health promotion programmes.

Thermafil: A New Clinical Approach Due to New Dimensional Evaluations.

BACKGROUND: There are a lot of techniques to obturate the root canals, but lateral condensation of gutta-percha is the most used one. An important aspect of thermafil is the error margin tolerated by the manufacturer in the production of plastic carriers. In literature, there is no evidence about discrepancy percentage between different carriers. It is demonstrated that the error margin of gutta-percha is 0.5% and is 0.2% for metal files (ISO standards). OBJECTIVE: The aim of this study was to evaluate the real dimensions of thermafil plastic carriers observed by the stereo microscope measuring the dimensional discrepancy between them. METHODS: For this study, 80 new thermafil (Dentsply Maillefer) have been selected. 40 thermafil 0.25 and 40 thermafil 0.30. Through 60X stereo microscope, the dimensions of the plastic carrier tips have been measured. The dimensions of the plastic carrier were also measured after a heating cycle. ZL GAL 11TUSM (Zetaline stereo evolution) microscope was used to observe the samples. Measurements were made through a dedicated software (Image Focus). All samples were analysed at 60X. RESULTS: A non-parametric paired test (Wilcoxon test) was used to compare baseline and after heating values; p-values ≤ 0.05 were assumed to be statistically significant. CONCLUSION: The samples we measured showed a mean value of the diameters in Thermafil 25 that was 0.27 mm, for Thermafil 30 the mean value was 0.33 mm.We have measured a dimensional variable of 8% in the 25 group while in group 30 the maximum possible variation found was 4%, that's why we propose a new protocol of obturation with thermafil. We can also conclude that a single heating process does not affect clinically the plastic carrier dimensions.

Comparison of views on the need for continuing education on oral cancer between general dentists and oral medicine experts: A Delphi survey.

OBJECTIVES: The role of dental professionals in screening for oral cancer has been limited. The purpose of this study was to investigate and compare the educational priorities of oral medicine specialists, general dental practitioners, and doctors of dental surgery with regards to the diagnosis and management of oral cancers and potential malignant disorders. STUDY DESIGN: This was a longitudinal survey. MATERIALS AND METHODS: A Delphi survey was directed to a panel of 25 oral medicine specialists asking them to identify the major difficulties in diagnosing and managing patients with oral cancer or suspected malignancy. In a second phase, two groups of generalists were asked to express their ratings on the issues identified by experts. RESULTS: The response rate of the experts to the survey was 84%, while only 44% of the generalists participated. Although the three groups agreed on most of the issues, there were significant differences of opinions on 10 of the items proposed by specialists (P < 0.05 from the Kruskal-Wallis test), which were observed mainly between experts and general dental practitioners (P < 0.017 from the Mann-Whitney U test). The opinion of the participants about future investments in the field of education resulted in similar results (P > 0.05 from the Chi-square test), with the specialists ranking highest on mandatory annual thematic courses, while the generalists prioritizing more interactive and extensive pre-graduation courses on oral cancer detection. CONCLUSION: This study confirms a clear need to improve the educational foundation on oral cancer by a didactic process starting with pre-graduation courses that should involve National Health Care Services, National Dental Associations, and academia.

Clinical and Spectrophotometric Evaluation of LED and Laser Activated Teeth Bleaching.

BACKGROUND: Auxiliary power sources (LED and laser) are used in in-office teeth bleaching techniques to accelerate the redox reaction of the whitening gel to increase ease of use, to improve comfort and safety, and to decrease the procedure time. OBJECTIVE: The aim this study is to evaluate the efficiency of the teeth whitening procedures performed with hydrogen peroxide and carbamide peroxide, LED or Laser activated. METHOD: 18 patients, affected by exogenous dyschromia, were treated with a bleaching agent composed by 35% hydrogen peroxide and 10% carbamide peroxide. They were divided into two groups: in the first group the bleaching agent was activated by a LED lamp; in the second group it was activated by a Laser diode lamp. Both groups were subjected to 3 bleaching cycle of 15' each. The chromatic evaluations were performed before and after one week from the treatment, using a chromatic scale and a spectrophotometer. The mean value of pre, post bleaching and follow-up were analyzed using a T-test, with results statistically significant for P<0,05. RESULTS: Results showed that the variations in brightness, chroma and hue are significantly influenced by the interaction between the whitening agent and the original colour of the teeth. Laser-activation has marginally improved the bleaching effectiveness. All patients treated with laser activation complained an increase in dental sensitivity. CONCLUSION: The use of laser-activating systems did not improve the efficacy of bleaching.

Relation between conversion degree and cytotoxicity of a flowable bulk-fill and three conventional flowable resin-composites.

OBJECTIVE: The aim of this study is to evaluate if the cytotoxic effects of the Surefil SDR flow, bulk fill flowable composite resin and three conventional flowable materials (Venus Diamond Flow, Filtex Supreme XTE Flowable and Enamel plus HRi Flow) correlated with the conversion degree (DC); hardness and depth of cure are also assessed. MATERIALS AND METHODS: Disks of each materials--cured using LED lamp--are utilized to evaluate DC (by FT-IR technique), amount of leached monomers (by HPLC technique), hardness (by Vickers hardness tester) and cytotoxicity (by MTT test). RESULTS: All tested materials show light cytotoxic effects, independently from DC values. Both the latter parameter and the hardness, in fact, change in function of thickness and type of material. HPLC results show that the monomers amount leached from each specimen is influenced by thickness but it is always very low which justifies the absence of any cytotoxic effect. CONCLUSIONS: Our findings suggest that there are not statistically significant differences in cytotoxicity in all experimental conditions, notwithstanding the differences in hardness and in degree of conversion.

Calcium silicate/calcium phosphate biphasic cements for vital pulp therapy: chemical-physical properties and human pulp cells response.

OBJECTIVES: The aim was to test the properties of experimental calcium silicate/calcium phosphate biphasic cements with hydraulic properties designed for vital pulp therapy as direct pulp cap and pulpotomy. METHODS: CaSi-αTCP and CaSi-DCDP were tested for ion-releasing ability, solubility, water sorption, porosity, ability to nucleate calcium phosphates, and odontoblastic differentiation­alkaline phosphatase (ALP) and osteocalcin (OCN) upregulation­of primary human dental pulp cells (HDPCs). RESULTS: The materials showed high Ca and OH release, high open pore volume and apparent porosity, and a pronounced ability to nucleate calcium phosphates on their surface. HDPCs treated with CaSi-αTCP showed a strong upregulation of ALP and OCN genes, namely a tenfold increase for OCN and a threefold increase for ALP compared to the control cells. Conversely, CaSi-DCDP induced a pronounced OCN gene upregulation but had no effect on ALP gene regulation. CONCLUSIONS: Both cements showed high biointeractivity (release of Ca and OH ions) correlated with their marked ability to nucleate calcium phosphates. CaSi-αTCP cement proved to be a potent inducer of ALP and OCN genes as characteristic markers of mineralization processes normally poorly expressed by HDPCs. CLINICAL RELEVANCE: Calcium silicate/calcium phosphate cements appear to be attractive new materials for vital pulp therapy as they may provide odontogenic/dentinogenic chemical signals for pulp regeneration and healing, and dentin formation in regenerative endodontics.

Marginal Leakage of Class V Composite Restorations Assessed Using Microcomputed Tomography and Scanning Electron Microscope.

OBJECTIVE: The aim of the study was to compare in Class V composite restorations marginal leakage measurements obtained with microcomputed tomography (micro-CT) and scanning electron microscopy (SEM) observations. METHODS: Class V cavities were prepared on 10 human molars and restored using Optibond FL (Kerr, Orange, CA, USA) and Premise Flowable (Kerr). Sealing ability was evaluated by assessing silver-nitrate penetration depth along enamel and dentin margins. Leakage was quantified using a scoring system. Micro-CT analysis provided 502 cross-sectional images for each tooth. Microleakage evaluation was performed first on three cross-sections corresponding to the sections examined by SEM, then on all 502 of the obtained micro-CT images. SEM observations were performed first at 20× magnification, then, if showing a zero score, at 80× magnification. Enamel and dentin microleakage scores assigned to corresponding sections through micro-CT and SEM (20×) were compared (Wilcoxon signed-rank test, α=0.05). RESULTS: No statistically significant difference in leakage scores emerged between micro-CT and 20×-magnification SEM. Eight tooth sections that were given a zero score under SEM at 20× magnification showed to be infiltrated at the higher magnification (80×). For five teeth a higher score was assigned following scanning of 502 cross-sections than based on the observation of three sections. CONCLUSIONS: Micro-CT presents as a valid, nondestructive in vitro method to quantitatively evaluate marginal leakage of adhesive restorations.

Effect of autoclaving on the surfaces of TiN -coated and conventional nickel-titanium rotary instruments.

AIM: To evaluate the effects of repeated autoclave sterilization cycles on surface topography of conventional nickel-titanium ( NiTi ) and titanium nitride ( TiN )-coated rotary instruments. METHODOLOGY: A total of 60 NiTi rotary instruments, 30 ProTaper (Dentsply Maillefer) and 30 TiN -coated AlphaKite (Komet/Gebr. Brasseler), were analysed. Instruments were evaluated in the as-received condition and after 1, 5 and 10 sterilization cycles. After sterilization, the samples were observed using scanning electron microscope (SEM), and surface chemical analysis was performed on each instrument with energy dispersive X-ray spectroscopy (EDS). Moreover, the samples were analysed by atomic force microscopy (AFM), and roughness average (Ra) and the root mean square value (RMS) of the scanned surface profiles were recorded. Data were analysed by means of anova followed by Tukey's test. RESULT: Scanning electron microscope observations revealed the presence of pitting and deep milling marks in all instruments. EDS analysis confirmed that both types of instruments were composed mainly of nickel and titanium, whilst AlphaKite had additional nitride. After multiple autoclave sterilization cycles, SEM examinations revealed an increase in surface alterations, and EDS values indicated changes in chemical surface composition in all instruments. Ra and RMS values of ProTaper significantly increased after 5 (P = 0.006) and 10 cycles (P = 0.002) with respect to the as-received instruments, whilst AlphaKite showed significant differences compared with the controls after 10 cycles (P = 0.03). CONCLUSIONS: Multiple autoclave sterilization cycles modified the surface topography and chemical composition of conventional and TiN -coated NiTi rotary instruments.

Effects of sodium hypochlorite and ethylenediaminetetraacetic acid on rotary nickel-titanium instruments evaluated using atomic force microscopy.

AIM: To use atomic force microscopy (AFM) to evaluate the effects of sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA) on the surface characteristics of ProTaper rotary nickel-titanium instruments. METHODOLOGY: A total of twenty ProTaper (Dentsply Maillefer, Ballaigues, Switzerland) instruments (S1, S2, F1, F2) were divided into five groups: no immersion, immersion in 5.25% NaOCl for 5 or 10min and immersion in 17% EDTA for 5 or 10min. Twenty surface areas along 3-mm sections at the tip of the files (perfect squares of 1x1µm) were analysed by AFM operating in contact mode under ambient conditions. Three-dimensional images (400x400 lines) were processed using Gwyddion software, and the roughness average (Ra) and the root mean square value (RMS) of the scanned surface profiles were recorded. Data were analysed by means of anova and paired samples t-test. RESULTS: Three-dimensional AFM images of the surface of ProTaper instruments, including new and those immersed in NaOCl and EDTA solutions, revealed topographic irregularities at the nanometric scale. RMS and Ra values of instruments treated with NaOCl and EDTA solutions were statistically higher than that of the new ones (P<0.05). CONCLUSIONS: Atomic force microscopy three-dimensional images and roughness values indicated that short-term contact between NaOCl and EDTA endodontic irrigants and ProTaper instruments caused alterations in the surface of instruments.

Inhibition of TEGDMA and HEMA-induced genotoxicity and cell cycle arrest by N-acetylcysteine.

OBJECTIVES: Dental resin monomers like triethylene glycol dimethacrylate (TEGDMA) and 2-hydroxyethyl methacrylate (HEMA) are able to cause an imbalance of the redox state in mammalian cells. The resulting oxidative stress originating from reactive oxygen species (ROS) has been associated with cytotoxicity. We hypothesized that ROS might contribute to the generation of genotoxicity by TEGDMA and HEMA as well. Therefore, we examined the formation of micronuclei in V79 cells by both resin monomers in the presence of the antioxidant N-acetylcysteine (NAC), which scavenges ROS. In addition, we analyzed the effects of TEGDMA and HEMA on the normal cell cycle in the presence of NAC. METHODS: V79 fibroblasts were exposed to increasing concentrations of TEGDMA and HEMA in the presence and absence of NAC for 24h. Genotoxicity was indicated by the formation of micronuclei. The modification of the normal cell cycle was analyzed by flow cytometry (FACS). RESULTS: A dose-related increase in the number of micronuclei in V79 cells-induced by TEGDMA and HEMA indicated genotoxicity of both chemicals. However, the formation of micronuclei was reduced in the presence of 10 mmol/L NAC, indicating its protective role. A cell cycle delay in G2 phase caused by TEGDMA was absent when cells were co-treated with NAC. Similarly, the presence of NAC led to a reversion of the cell cycle delay in HEMA-treated cell cultures. SIGNIFICANCE: Our results suggest that genotoxic effects and the modification of the cell cycle caused by TEGDMA and HEMA are mediated, at least in part, by oxidative stress.

Genetic and cellular toxicology of dental resin monomers.

Monomers are released from dental resin materials, and thus cause adverse biological effects in mammalian cells. Cytotoxicity and genotoxicity of some of these methacrylates have been identified in a vast number of investigations during the last decade. It has been well-established that the co-monomer triethylene glycol dimethacrylate (TEGDMA) causes gene mutations in vitro. The formation of micronuclei is indicative of chromosomal damage and the induction of DNA strand breaks detected with monomers like TEGDMA and 2-hydroxyethyl methacrylate (HEMA). As a consequence of DNA damage, the mammalian cell cycle was delayed in both G1 and G2/M phases, depending on the concentrations of the monomers. Yet, the mechanisms underlying the genetic and cellular toxicology of resin monomers have remained obscure until recently. New findings indicate that increased oxidative stress results in an impairment of the cellular pro- and anti-oxidant redox balance caused by monomers. It has been demonstrated that monomers reduced the levels of the natural radical scavenger glutathione (GSH), which protects cell structures from damage caused by reactive oxygen species (ROS). Depletion of the intracellular GSH pool may then significantly contribute to cytotoxicity, because a related increase in ROS levels can activate pathways leading to apoptosis. Complementary, cytotoxic, and genotoxic effects of TEGDMA and HEMA are inhibited in the presence of ROS scavengers like N-acetylcysteine (NAC), ascorbate, and Trolox (vitamin E). Elevated intracellular levels of ROS can also activate a complex network of redox-responsive macromolecules, including redox-sensitive transcription factors like nuclear factor kappaB (NF-kappaB). It has been shown that NF-kappaB is activated probably to counteract HEMA-induced apoptosis. The induction of apoptosis by TEGDMA in human pulp cells has been associated with an inhibition of the phosphatidylinositol 3-kinase (PI3-K) cell-survival signaling pathway. Although the details of the mechanisms leading to cell death, genotoxicity, and cell-cycle delay are not completely understood, resin monomers may be able to alter the functions of the cells of the oral cavity. Pathways regulating cellular homeostasis, dentinogenesis, or tissue repair may be modified by monomers at concentrations well below those which cause acute cytotoxicity.

NF-kappaB protection against apoptosis induced by HEMA.

The cytotoxicity of dental monomers has been widely investigated, but the underlying mechanisms have not been elucidated. We studied the molecular mechanisms involved in cell death induced by HEMA. In human primary fibroblasts, HEMA induced a dose-dependent apoptosis that was confirmed by the activation of caspases-8, -9, and -3. We found an increase of reactive oxygen species (ROS) and NF-kappaB activation after HEMA exposure. Blocking of ROS production by anti-oxidants had no direct influence on apoptosis caused by HEMA, but inhibition of NF-kappaB increased the fraction of apoptotic cells. Accordingly, mouse embryonic fibroblasts (MEF) from p65-/- mice were more susceptible to HEMA-induced apoptosis than were wild-type controls. Our results indicate that exposure to HEMA triggers apoptosis and that this mechanism is not directly dependent upon redox signaling. Nevertheless, ROS induction by HEMA activates NF-kappaB, which exerts a protective role in counteracting apoptosis.

Inhibition of phosphatidylinositol 3-kinase amplifies TEGDMA-induced apoptosis in primary human pulp cells.

Cytotoxicity of triethylene glycol dimethacrylate (TEGDMA), a co-monomer of dental resinous restorative materials, is firmly established in vitro, but the molecular mechanisms are unknown. Here we examined apoptosis and necrosis induced by TEGDMA in human primary pulp cells. The levels of apoptotic and necrotic cell populations differentially increased after exposure to increasing concentrations of TEGDMA. A two-fold increase in the percentage of apoptotic cells was induced by 1 mmol/L TEGDMA. However, a population shift among cells in apoptosis and necrosis was detected when cell cultures were exposed to 2 mmol/L TEGDMA. Inhibition of the MAP Kinase/ERK pathway had no influence on cell survival, but inhibition of phosphatidylinositol 3 kinase (PI3-Kinase; Akt/protein kinase B) by LY294002 amplified TEGDMA-induced apoptosis. Moreover, Akt phosphorylation was inhibited in the presence of TEGDMA. These results suggest that depression of PI3K signaling may be a primary target in TEGDMA-induced apoptosis.

Evaluation of circulating levels and renal clearance of natural amino acids in patients with Cushing's disease.

Although the hypercortisolism-induced impairment of protein homeostasis is object of several studies, a detailed evaluation of the complete amino acid profile of patients with Cushing's syndrome (CS) has never been performed. The aim of the current open transversal controlled study was to evaluate serum and urinary concentrations as well as renal clearance of the complete series of natural amino acids and their relationship with glucose tolerance in patients with Cushing's disease (CD). Twenty patients with CD (10 active and 10 cured) and 20 sex- and age-matched healthy controls entered the study. Measurement of serum and urinary levels of the complete series of natural amino acids was performed in all patients analyzed by cationic exchange high performance liquid cromatography (HPLC) after 2 weeks of a standardized protein intake regimen. The renal clearance (renal excretion rate) of each amino acid was calculated on the basis of the serum and urinary concentrations of creatinine and the specific amino acid. Fasting glucose and insulin levels, glucose and insulin response to standard glucose load, insulinogenic and homeostasis model insulin resistance (Homa-R) indexes were also evaluated and correlated to the circulating levels and renal clearances of each amino acid. Significantly higher serum (p<0.01) and urinary (p<0.05) levels of alanine and cystine, lower serum and higher urinary levels of leucine, isoleucine and valine (p<0.05) and higher renal excretion rates of leucine, isoleucine and valine (p<0.01) were found in patients with active CD than in patients cured from the disease and in controls. No difference was found between cured patients and controls. Creatinine clearance was similar in active and cured patients and in controls. In patients with active CD, urinary cortisol levels were significantly correlated to urinary cystine levels (r=0.85; p<0.01) and renal excretion rate of leucine (r=-0.76; p<0.05), isoleucine (r=-0.76; p<0.05) and valine (r=-0.66; p<0.05). Fasting blood glucose levels were significantly correlated to serum alanine levels (r=0.70; p<0.05). Although Homa-R was significantly correlated to BMI in active patients (r=0.74 p<0.05), it was not correlated to amino acid levels. In conclusion, the results of the current study demonstrate that patients with CD have significant changes in serum and urinary concentration of several amino acids and changes in renal clearance of some specific amino acids. Normalization of cortisol levels restored the amino acid profile.

Diagnosis and follow-up of cystinuria: use of proton magnetic resonance spectroscopy.

Proton Nuclear Magnetic Resonance (NMR) Spectroscopy of urine (as well as of other biological fluids) is a very powerful technique enabling multi-component analysis useful in both diagnosis and follow-up of a wide range of inherited metabolic diseases. Among these pathologies, cystinuria is characterised by accumulation in urine of four dibasic amino acids, namely lysine, arginine, ornithine and cystine; the last one, being only slightly water soluble, generates urolithiasis. The mentioned aminoacids can be detected in the urine NMR spectrum of cystinuric patients, the most abundant being the lysine (5 mM and over are often detected), whose typical signals become very high; arginine and ornithine are also usually detectable, although pathologic concentrations are lower (usually below 2mM). The proposed NMR technique is also suitable in monitoring the therapy with alpha-mercaptopropionylglycine (MPG), providing quantitation of several metabolites of interest in the follow-up of the pathology, like cystine, creatinine and citrate.

Macroporous hydroxyapatite as alloplastic material for dental applications.

In the dentistry field, synthetic hydroxyapatite can be conveniently used as semiabsorbable alloplastic material to solve a number of clinical problems. The hydroxyapatite structure influences bone ingrowth as well as its resorption. In particular, pore size must exceed 100 microm to allow new bone ingrowth. Because of its brittleness, highly porous hydroxyapatite is difficult to handle without causing damage, and, therefore, its mechanical performance has to be improved placing the material on a dense hydroxyapatite substrate. Dense/porous hydroxyapatite laminates can be obtained by slip casting technology. The method starts with the preparation of a ceramic powder-binder-solvent system. This slurry produces, after solvent evaporation, a soft green tape, that is successively baked at 300 degrees C to remove all organic component, and finally sintered at high temperature (1200 degrees C). The material porosity cannot be significantly modified by changes in the slurry composition and sintering temperature; therefore, the macroporous hydroxyapatite layer must be obtained using a porous hydroxyapatite powder. The powder is prepared by grinding of green tape pieces and the resulting coarse material is successively baked at 300 degrees C. Such material is mixed with a polymer solution and cast on a green tape substrate. Layer fractures are not observed in both film bulks and interface, because during sintering the consolidation of two layers happens simultaneously. In the resulting material, the first layer consisted of macroporous hydroxyapatite with high osteoconductive properties, and the second layer was a dense hydroxyapatite substrate able to improve the laminate mechanical properties.

European multicentre evaluation of the Super Aution SA-4220 urinalysis analyser.

A multicentre evaluation of the urine test strip analyser Super Aution-4220 was carried out in six laboratories. The analytical performance of the instrument with regard to imprecision, linearity, detection limit, drift, carry-over and method comparison was studied. Using the Aution stick 8 test strip the pH, glucose, protein, ketones, bilirubin, blood, urobilinogen and leukocyte esterase were analysed. Specific gravity measurements were performed by refractive index method. Within-run and between-run imprecision determined at three levels of analyte were good. No carry-over was observed. Obtained results were linear through all the described analytical range. No significant drift was detected. Method comparison with some quantitative methods was performed and showed a good correlation with most of the analytes. The study of interferences showed minor interferences by common therapeutic drugs with the measurement of some analytes. During the assessment period of about 6 months no breakdown occurred in any laboratory. The Super Aution urine analyser appeared to be a highly automated analyser of urinary test strips. The operation was simple and the maintenance required only a few minutes a day.