Biomineralization potential and biological properties of a new tantalum oxide (Ta2O5)-containing calcium silicate cement.

OBJECTIVE: The present study evaluated the biological effects and biomineralization potential of a new tantalum oxide (Ta2O5)-containing material designed for vital pulp therapy or perforation repair (NeoMTA 2), compared to NeoMTA Plus and Bio-C Repair. MATERIAL AND METHODS: Human dental pulp stem cells (hDPSCs) were exposed to different eluates from NeoMTA Plus, NeoMTA 2, and Bio-C Repair. Ion release from each material was determined using inductively coupled plasma-optical emission spectrometry (ICP-MS). The biological experiments performed were MTT assays, apoptosis/necrosis assays, adhesion assays, migration assays, morphology evaluation, and reactive oxygen species (ROS) production analysis. Biomineralization was assessed by Alizarin red S staining. Finally, osteo/odontogenic gene expression was determined by real-time quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR). Data were analyzed using one-way ANOVA followed by Tukey's multiple comparison test. RESULTS: NeoMTA 2 displayed a significantly higher calcium release compared to the other materials (p < 0.05). When hDPSCs were cultured in presence of the different material eluates, all groups exhibited similar hDPSC viability and migration rates when compared to untreated cells. Substantial cell attachment and spreading were observed in all materials' surfaces, without significant differences. hDPSCs treated with NeoMTA 2 displayed an upregulation of ALP, Col1A1, RUNX2 (p < 0.001), ON, and DSPP genes (p < 0.05), and showed the highest mineralization potential compared to other groups (p < 0.001). Finally, the more concentrated eluates from these materials, specially NeoMTA Plus and NeoMTA 2, promoted higher ROS production in hDPSCs compared to Bio-C Repair and control cells (p < 0.001), although these ROS levels did not result in increased cell death. CONCLUSIONS: The new tantalum oxide (Ta2O5)-containing material shows an adequate cytocompatibility and the ability to promote biomineralization without using chemical osteogenic inducers, showing great potential as a new material for vital pulp therapy. CLINICAL RELEVANCE: NeoMTA 2 seems to be a promising material for vital pulp therapy. Further studies considering its biocompatibility and biomineralization potential are necessary.

In Vitro Biocompatibility of CPP-ACP and Fluoride-containing Desensitizers on Human Gingival Cells.

OBJECTIVES: To analyze the biocompatibility of different desensitizers containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and fluoride in their composition: MI Varnish (MV), Clinpro White Varnish (3M Oral Care), Profluorid Varnish (VOCO), Duraphat (Colgate) and Embrace Varnish (Pulpdent) on human gingival fibroblast cells (hGF). METHODS AND MATERIALS: Human gingival fibroblast (hGF) cells were exposed to several desensitizer extracts at different concentrations (0.1%, 1%, and 4% eluates). Then, in vitro biocompatibility was studied by analyzing the IC50 value, cell proliferation (MTT assay and cell cycle), cell migration (wound healing assay), cell morphology and F-actin content (immunocytofluorescence), and induction of apoptosis/necrosis (flow cytometry). Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey test. RESULTS: The lowest cell viability and IC50 were observed in all concentrations of Embrace Varnish-treated hGFs (p<0.001), whereas the highest were exhibited by those treated with Clinpro White Varnish. Similar effects were evidenced when induction of apoptosis/necrosis and cell migration assays were assessed. Finally, MI Varnish, Profluorid Varnish, Duraphat, and Embrace Varnish extracts showed lower numbers of attached cells, some of them with an unusual fibroblastic morphology when cultured with 4% concentration of the varnishes, while Clinpro White Varnish exhibited a similar number of cells with an evident actin cytoskeleton compared to the control group. CONCLUSIONS: The results obtained in this study indicate that hGFs show better in vitro biocompatibility after exposure to Clinpro White Varnish, even at the highest concentration employed, making it the most eligible for topical applications. In contrast, Embrace Varnish exhibited a high cytotoxicity towards hGFs that could potentially delay the healing process and regeneration of the oral mucosa, although more studies are needed to confirm this hypothesis.

Microstructural composition, ion release, and bioactive potential of new premixed calcium silicate-based endodontic sealers indicated for warm vertical compaction technique.

OBJECTIVE: The aim of this study was to evaluate the microstructural composition, ion release, cytocompatibility, and mineralization potential of Bio-C Sealer ION+ (BCI) and EndoSequence BC Sealer HiFlow (BCHiF), compared with AH Plus (AHP), in contact with human periodontal ligament cells (hPDLCs). MATERIALS AND METHODS: The sealers' ionic composition and release were assessed using energy-dispersive spectroscopy (EDS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. For the biological assays, hPDLCs were isolated from third molars, and sealer extracts were prepared (undiluted, 1:2, and 1:4 ratios). An MTT assay, wound-healing assay, and cell morphology and adhesion analysis were performed. Activity-related gene expression was determined using RT-qPCR, and mineralization potential was assessed using Alizarin Red staining (ARS). Statistical analyses were performed using one-way ANOVA and Tukey's post hoc test (α < 0.05). RESULTS: The three sealers exhibited variable levels of silicon, calcium, zirconium, and tungsten release and in their composition. Both BCI and BCHiF groups showed positive results in cytocompatibility assays, unlike AHP. The BCHiF group showed an upregulation of CAP (p < 0.01), CEMP1, ALP, and RUNX2 (p < 0.001) compared with the negative control, while the BCI group showed an upregulation of CEMP1 (p < 0.01), CAP, and RUNX2 (p < 0.001). Both groups also exhibited a greater mineralization potential than the negative and positive controls (p < 0.001). CONCLUSIONS: The calcium silicate-based sealers considered in the present in vitro study exhibited a high calcium ion release, adequate cytocompatibility, upregulated osteo/cementogenic gene expression, and increased mineralized nodule formation in contact with hPDLCs. CLINICAL RELEVANCE: From a biological perspective, BCI and BCHiF could be clinically suitable for root canal filling.

Prevalence of Apical Periodontitis in patients with Multiple Myeloma.

BACKGROUND: Aim Previous reports have been analyzed the prevalence/association of apical periodontitis (AP) with systemic diseases. The present study aims to analyze the prevalence of healthy/diseased periapex and endodontic treatments in patients with Multiple Myeloma (MM) and compare the results with those of control subjects. MATERIAL AND METHODS: Methodology Panoramic radiographs of 50 individuals with MM were evaluated and compared with 50 controls that were sex and age matched exactly with the diseased group. Radiographic analysis was performed by 2 two experienced endodontists under standardized conditions. The periapical status (presence or not of AP) was assessed using the periapical index (PAI). Data included systemic health, technical quality of root fillings, total number of teeth, quality of restoration, and periapical status. Statistical evaluation of differences between groups included used chi-squared tests and Fisher's exact tests. RESULTS: The prevalence of root canal-treated teeth was 10.11% in the MM group and 12.05% in the control group (p=0.90). The average root canal-treated teeth in the test group was 2,34 and 2.48 in the control group, where the difference was statistically significant (p=0.05). AP in 1 or more teeth was found in 86 % and in 78% of the patients in the MM and the control groups, respectively. When analyzed by subject, there was no statistically significant difference in the prevalence of AP (p>0.72). Similarly there was also no statistically significant difference in the prevalence of PA (p=0.85), when analyzed by tooth, AP was found in 63.2% and 62.9% in MM and control groups. CONCLUSIONS: The presence of AP and endodontic treatment was not significantly different in individuals with MM compared with control subjects. Future studies are needed to elucidate and confirm the association between MM and AP.

Cytocompatibility, bioactivity potential, and ion release of three premixed calcium silicate-based sealers.

OBJECTIVE: Compositional modifications may alter the biological and physicochemical characteristics of calcium silicate-based sealers (CSBS) and, ultimately, their bioactivity. The main objective of this study was to evaluate the biological properties of three CSBS: EndoSequence BC Sealer, Ceraseal, and Endoseal mineral trioxide aggregate. MATERIALS AND METHODS: Human periodontal ligament stem cells (hPDLSCs) were exposed to several eluates of CSBS. The ion release profile and pH were determined, and metabolic activity and cell migration were assessed using the MTT and wound healing assays. hPDLSCs were cultured in direct contact with the surface of each material, and cell morphology and attachment were analyzed by scanning electron microscopy (SEM). Bioactivity potential was assessed by RT-qPCR and mineralization assays. Statistical differences between biomaterials were assessed using one- or two-way ANOVA (α < 0.05). RESULTS: All materials showed an alkaline pH, although Endoseal exhibited a significantly higher pH compared with the other CSBS (p < 0.05). Ceraseal released significantly more Ca2+ (p < 0.05) than EndoSequence BC Sealer and Endoseal. Interestingly, Endoseal induced a significant reduction in cell viability and cell migration compared with the control (p < 0.001). Moreover, SEM showed abundant cells adhering to EndoSequence BC Sealer and Ceraseal surfaces, whereas very few round cells were detected on the surface of Endoseal. Finally, Ceraseal and EndoSequence induced ALP, CAP, and CEMP-1 expression and a significantly higher mineralization capacity than Endoseal (***p < 0.001). CONCLUSIONS: The eluates from EndoSequence BC Sealer and Ceraseal displayed higher cell viability, cell attachment, cell migration rates, and ion release rates than Endoseal. Ceraseal and EndoSequence BC Sealer exhibited significantly more gene expression and mineralization capacity than Endoseal. CLINICAL RELEVANCE: The results obtained in the present work suggest that EndoSequence BC Sealer and Ceraseal possess biological properties that make them suitable materials for root canal treatment.

Decreased lipid metabolism but increased FA biosynthesis are coupled with changes in liver microRNAs in obese subjects with NAFLD.

BACKGROUND/OBJECTIVE: Many controversies regarding the association of liver miRNAs with obesity and nonalcoholic fatty liver diseases (NAFLD) call for additional validations. This study sought to investigate variations in genes and hepatic miRNAs in a sample of obese patients with or without NAFLD and human hepatocytes (HH). SUBJECTS/METHODS: A total of 60 non-consecutive obese women following bariatric surgery were recruited. Subjects were classified as NAFLD (n=17), borderline (n=24) and controls (n=19) with normal enzymatic profile, liver histology and ultrasound assessments. Profiling of 744 miRNAs was performed in 8 obese women with no sign of hepatic disease and 11 NAFLD patients. Additional validation and expression of genes related to de novo fatty acid (FA) biosynthesis, uptake, transport and ß-oxidation; glucose metabolism, and inflammation was tested in the extended sample. Induction of NAFLD-related genes and miRNAs was examined in HepG2 cells and primary HH treated with palmitic acid (PA), a combination of palmitate and oleic acid, or high glucose, and insulin (HG) mimicking insulin resistance in NAFLD. RESULTS: In the discovery sample, 14 miRNAs were associated with NAFLD. Analyses in the extended sample confirmed decreased miR-139-5p, miR-30b-5p, miR-122-5p and miR-422a, and increased miR-146b-5p in obese subjects with NAFLD. Multiple linear regression analyses disclosed that NAFLD contributed independently to explain miR-139-5p (P=0.005), miR-30b-5p (P=0.005), miR-122-5p (P=0.021), miR-422a (P=0.007) and miR-146a (P=0.033) expression variance after controlling for confounders. Decreased miR-122-5p in liver was associated with impaired FA usage. Expression of inflammatory and macrophage-related genes was opposite to decreased miR-30b-5p, miR-139-5p and miR-422a, whereas increased miR-146b-5p was associated with FABP4 and decreased glucose metabolism and FA mobilization. In partial agreement, PA (but not HG) led to decreased miR-139-5p, miR-30b-5p, miR-422a and miR-146a in vitro, in parallel with increased lipogenesis and FA transport, decreased glucose metabolism and diminished FA oxidation. CONCLUSION: This study confirms decreased liver glucose and lipid metabolism but increased FA biosynthesis coupled with changes in five unique miRNAs in obese patients with NAFLD.

Thyroid hormone responsive Spot 14 increases during differentiation of human adipocytes and its expression is down-regulated in obese subjects.

CONTEXT: Very limited information is available regarding the function of human thyroid hormone responsive Spot 14 (human S14, hS14) in adipogenesis and human adiposity. OBJECTIVE: To evaluate hS14 levels during differentiation of human pre-adipocytes, in human fat depots and isolated fat cells. DESIGN: This was a cross-sectional study. SUBJECTS: A total of 161 omental (OM) and 87 subcutaneous (SC) adipose tissue samples obtained during elective surgical procedures from a population who varied widely in terms of obesity. MEASUREMENTS: hS14 gene expression and protein levels during adipogenesis were assessed by RT-PCR, western blot, and using an automated confocal imaging approach. RESULTS: hS14 gene expression levels were decreased in OM adipose tissue from overweight (-42.0%) and obese subjects (-56.5%) compared with lean subjects (P<0.05 and P<0.0001, respectively). hS14 mRNA (but not hS14-related) was inversely associated with obesity measures such as body mass index (P=0.001), percent fat mass (P=0.001), waist-to-hip ratio (P=0.020), and systolic blood pressure (P=0.031). hS14 gene expression and protein levels were up-regulated at the early stages of differentiation of human pre-adipocytes as well as for 3T3-L1 cells. That observation was most prominent in those individual cells exhibiting the more marked differentiation features. hS14 gene expression levels increased by approximately 45 000-fold in mature adipocytes. Increased hS14 levels were also found in stromal-vascular cells/pre-adipocytes (3.8-fold, P<0.05) and in adipose tissue samples (1.9-fold, P<0.0001) from SC compared with OM fat depots. CONCLUSIONS: These results suggest that hS14 is involved in human adipogenesis, but inversely related to obesity and OM fat accumulation.

Evaluation of the influence of the addition of biodegradable polymer matrices in the formulation of self-curing polymer systems for biomedical purposes.

The solid phase of self-curing formulations of poly(methyl methacrylate) was modified by different biodegradable polymer matrices, such as poly(l-lactic acid), poly(beta-hydroxybutyrate) and thermoplastic starches (TPSs). The aim of this modification was the acquisition of a short- to medium-term drug delivery system to release bisphosphonates for hard tissue treatment. Different physico-chemical characterization techniques were used in order to determine the influence of these matrices and their mechanical capacity, in vitro behaviour, curing parameters, residual monomer content and surface topography for the preparation of the self-curing formulations. The incorporation of the polyesters did not induce an increase in water uptake capacity of the system due to their apolar aliphatic character. On the other hand, TPSs exhibited values of water absorption up to 15.3%, related with their hydrophilic chemical structure, dependent on the commercial formulation and the particle size distribution of the powder. The modifications of the solid phase led in all cases to a decrease in the mechanical behaviour of the material, although the formulations modified with TPS were in the range of accepted values according to standard specifications. The immersion of TPS formulations in a simulated physiological environment (phosphate buffer solution, pH 7.4, 37 degrees C) conducted to a surface porosity related with release of plasticizers of the domains of the biodegradable component of the formulation. Finally drug release capacity was studied by loading the material with Ibandronate, observing high dependence with the kind of TPS added, as well as its particle size.

Surgically treated perforations of the gastrointestinal tract caused by ingested foreign bodies.

OBJECTIVE: Intestinal perforation due to foreign body (FB) ingestion is rare (1%). We describe our experience in treating these lesions surgically. METHOD: From 1995 to 2006, data were collected prospectively in 33 patients (18 women and 15 men; mean age 64 years) operated on for intestinal perforation due to an ingested FB. The type of object, preoperative diagnosis, perforation site, treatment, morbidity and mortality were reviewed. RESULTS: Foreign body ingestion was predominantly involuntary (88%). The mean time from ingestion to perforation was 10.4 days. The most frequently ingested objects were dietary FB (n = 21) and toothpicks (n = 6). The most frequent predisposing factors were dentures or an orthodontic appliance (73%). The most common preoperative diagnoses were acute abdomen of uncertain origin (n = 7), acute appendicitis (n = 7) and acute diverticulitis (n = 5). Pneumoperitoneum was observed in 10 cases. The diagnosis was reached during laparotomy in 30 (91%) cases. The most frequent perforation site was the colorectal region (n = 18, 54.5%), followed by the terminal ileum (n = 7, 21.2%); intraperitoneal perforation was the most common (n = 30, 91%). All cases had abdominal contamination and 22 (66.7%) had diffuse peritonitis. Treatment was always by surgery and antibiotics. Thirteen patients required a colostomy. Morbidity was 57.6% (n = 19) and mortality 6.1% (n = 2). CONCLUSION: Intestinal perforation by a foreign body is rare and normally affects the sigmoid colon, rectum or distal ileum. Dentures are a common risk factor. Patients are rarely aware of foreign body ingestion. Dietary FB and toothpicks are the most commonly ingested objects. Treatment consists of surgery and antibiotics. Appendicitis and acute diverticulitis should be considered in the differential diagnosis.

Chemical modification of jute fibers for the production of green-composites.

Natural fiber reinforced composites is an emerging area in polymer science. Fibers derived from annual plants are considered a potential substitute for non-renewable synthetic fibers like glass and carbon fibers. The hydrophilic nature of natural fibers affects negatively its adhesion to hydrophobic polymeric matrices. To improve the compatibility between both components a surface modification has been proposed. The aim of the study is the chemical modification of jute fibers using a fatty acid derivate (oleoyl chloride) to confer hydrophobicity and resistance to biofibers. This reaction was applied in swelling and non-swelling solvents, pyridine and dichloromethane, respectively. The formation of ester groups, resulting from the reaction of oleoyl chloride with hydroxyl group of cellulose were studied by elemental analysis (EA) and Fourier Transform infrared spectroscopy (FTIR). The characterization methods applied has proved the chemical interaction between the cellulosic material and the coupling agent. The extent of the reactions evaluated by elemental analysis was calculated using two ratios. Finally electron microscopy was applied to evaluate the surface changes of cellulose fibers after modification process.

Population-based incidence and survival of gastrointestinal stromal tumours (GIST) in Girona, Spain.

BACKGROUND: Gastrointestinal stromal tumours (GIST) are rare malignancies characterised by their association with KIT oncogene mutations. Until now, population-based reports of the incidence or survival of kit-confirmed GIST have been rare, and none have originated in Southern Europe. MATERIALS AND METHODS: We used the Girona Cancer Registry to identify malignant mesenchymal tumours of the digestive tract between 1994 and 2001, and performed c-kit testing in the tumour samples. Age-adjusted incidence rates and survival rates were calculated, and they were also analysed by sex and NIH risk categories. RESULTS: Forty-six cases were categorised as GIST. Fifty percent were localised in the stomach, 43.5% in small intestine, 4.3% in the omentum, and 2.2% in colon. Thirty-seven percent were classified as high risk of an aggressive behaviour, 30.4% as intermediate risk and 32.6% as low or very low risk. Only one patient received treatment with imatinib mesilate. The annual incidence by 100,000 inhabitants in crude rate, European age-standardised rate and world age-standardised rate was, respectively, 1.09, 0.90 and 0.65 cases. The relative 5-year survival rate was 74.7% for the entire cohort, and it was markedly lower in the high-risk cases (20.3%). CONCLUSIONS: We report the first population-based study of GIST incidence and survival in Southern Europe. The incidence rate is low and comparable with that of cancer registries from Northern Europe. Survival was favourable in our pre-imatinib population although it was low in high risk cases. Prognostic discrimination of the cases with intermediate, low, or very low risk is inadequate, and these categories should be considered jointly in the future. Our results will help researchers in establishing baseline values against which they can compare, in the future, the impact of imatinib and other Kit tyrosine inhibitors on survival.

[Lateral popliteal block: a modification of anatomical references]. / Bloqueo poplíteo por vía lateral: una modificación de las referencias anatómicas.

OBJECTIVE: To study the efficacy of a lateral approach in performing a sciatic nerve block at the popliteal fossa using modified anatomical references: the intersection of the groove between the lateral vastus and biceps femoris muscles and the axis of the femur, which passes through the upper vertex of the popliteal fossa. PATIENTS AND METHOD: The described lateral approach was used in a series of 50 patients scheduled for foot surgery. We located the nerve by stimulation and infused 40 mL of 1.5% mepivacaine. Several technical features, efficacy and complications were analyzed. RESULTS: The mean patient age was 55 years, mean weight was 67 +/- 12.6 Kg, and mean height was 159.8 +/- 6.7 cm. Only 1 puncture was necessary in 90% of the cases; 2 punctures were needed in 10%. The point of puncture was proximal to the lateral condyle of the femur at a mean 9.1 +/- 2.1 cm. Depth ranged from 3.8 to 9 cm. The mean latency time to onset of the surgical block was 28.3 +/- 8 minutes. The block was considered excellent in 42 cases and adequate in 6; the technique failed in 2 cases. No major complications were observed. CONCLUSIONS: The proposed approach is easy to implement, involves no remarkable complications, and is particularly useful in patients who have difficulty taking a prone decubitus position.

[Changes in HIV-related attitudes and knowledge of teenagers following the Aulasida intervention in 1996-1997]. / Cambios en las actitudes y conocimientos de los adolescentes sobre la infección por VIH tras la intervención escolar Aulasida, 1996-1997.

BACKGROUND: Schools being the ideal setting for carrying out Health Education activities, the aim of this study was that of pinpointing and quantifying the changes in attitudes and knowledge on the part of teenagers enrolled in school in Algemesí (Valencia) following an educational intervention regarding HIV infection. METHOD: Eleven schools at which a total of 2,599 teenagers (ages 12-19) were enrolled in eleven different years of study (Secondary Education, Secondary Ed. and School Leaving Certificate, College Preparation Course and Vocational Training) were invited to take part. The Aulasida intervention carried out during the 1996-1997 school year consisted of an informative lecture-panel discussion and student involvement activities in small groups using educational materials. The gauging instrument was a questionnaire. This questionnaire was designed in a before-and-after cross-sectional study. An analysis was made divided into age and educational level strata. The averages were compared with the Student "t" test and the percentages of change with ji square. RESULTS: A total of nine schools accepted taking part. 1575 students answered the "before" test (47.4% males and 52.1% females), the average age being 15.2 (1.96) years old. The average number of correct answers to the "before" test was 13.5 (2.8). The "after" test showed an overall increase of up to 14.7 (3.0) correct answers (p < 0.01). By educational levels, this increase was highly appreciable in the younger age group. The most common sources of information on HIV were: television (80.8%); Aulasida (76.8%), teachers (60.9%), pamphlets (58.4%) and films (53.7%9. CONCLUSIONS: Educational interventions are useful tools for increasing knowledge and improving attitudes regarding HIV infection. Secondary schools are the best environment for this purpose, it being necessary to carry out interventions among younger groups, as a greater impact is thus achieved.