Use of liquid-based cytology samples reveals genomic instability and cell death in patients undergoing orthodontic treatment.

PURPOSE: To examine the use of liquid-based exfoliative cytology to determine the presence of genomic instability and cell death in the oral mucosa of patients with orthodontic appliances. METHODS: Fifty-four oral mucosa samples were collected from 18 patients and divided into three stages: T0, before fixation of orthodontic appliances; T1, 25 days after appliance fixation; T2, 90 days after appliance fixation. All samples were Papanicolaou-stained and observed by microscopy (1,000 cells/sample) to ascertain the frequency of micronucleated cells (MN) and nuclear abnormalities (nuclear buds (NB), binucleated (BN), condensed chromatin (CC), karyorrhexis (KR), and karyolysis (KL)). Differences were analyzed statistically using the Mann-Whitney, Wilcoxon, Kruskal-Wallis and chi-squared tests. RESULTS: After placement of orthodontic appliances, significant differences were observed for genomic instability biomarkers (MN and NB) and cell death (CC, KR and KL) (P < 0.05). Female patients and older patients exhibited a higher frequency of MN. CONCLUSION: Liquid-based cytology has revealed that orthodontic appliances induce genomic instability and cell death in epithelial tissue of the oral mucosa, facilitating sample preservation and yielding more than one preparation per sample. Future studies should investigate whether such cell damage can be reversed through cell repair or whether cell alterations evolve and lead to disease.

Enhancement of fluoride release in glass ionomer cements modified with titanium dioxide nanoparticles.

BACKGROUND: Several efforts have been made to improve the glass ionomer cements (GICs) properties with nanotechnology. Fluoride release in once of most beneficial properties of GICs. The purpose of this study was to evaluate the fluoride release, recharge, and cytotoxicity in GICs reinforced with titanium dioxide nanoparticles (TiO2N). OBJECTIVE: Evaluate the fluoride release, recharge, and cytotoxicity in GICs reinforced with TiO2N. METHODS: Four GICs, FUJI IX EXTRA (G1c), KETAC MOLAR (G2c), IONOFILL MOLAR (G3c), and FUJI IX (G4c) were combined with TiO2N (G1e, G2e, G3e, and G4e) and divided into blocks of 5-mm width and 1-mm thickness 10 each. A total of 80 samples were arranged as follows: GICs alone as negative control (n = 40) and GICs + TiO2N as experimental groups (n = 40). The fluoride release was determined for periods of 1, 2, 6, 10, 31, 90, 180, 240, and 300 days. On days 30 and 179, samples were recharged by submerging in 1 mL of 20,000 ppm sodium fluoride gel. Cytotoxic activity was carried out with gingival fibroblasts, using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide cell viability assay. RESULTS: The experimental groups obtained the highest and more constant fluoride released when compared to control groups. After the first recharge, experimental groups (G1e, G3e, and G4e) showed statistically significant results (P = .001, 0.010, and 0.001 respectively) enhancing their recharge ability regarding control groups. The second recharge showed better results in G1e concerning the rest of the groups. No cytotoxic activity was observed in all experimental groups, although significant differences were observed in G3e and G4e regarding control group. CONCLUSION: The incorporation of TiO2N enhance the fluoride release in glass ionomers with a noncytotoxic effect on human gingival fibroblasts.

TcVac1 vaccine delivery by intradermal electroporation enhances vaccine induced immune protection against Trypanosoma cruzi infection in mice.

The efforts for the development and testing of vaccines against Trypanosoma cruzi infection have increased during the past years. We have designed a TcVac series of vaccines composed of T. cruzi derived, GPI-anchored membrane antigens. The TcVac vaccines have been shown to elicit humoral and cellular mediated immune responses and provide significant (but not complete) control of experimental infection in mice and dogs. Herein, we aimed to test two immunization protocols for the delivery of DNA-prime/DNA-boost vaccine (TcVac1) composed of TcG2 and TcG4 antigens in a BALB/c mouse model. Mice were immunized with TcVac1 through intradermal/electroporation (IDE) or intramuscular (IM) routes, challenged with T. cruzi, and evaluated during acute phase of infection. The humoral immune response was evaluated through the assessment of anti-TcG2 and anti-TcG4 IgG subtypes by using an ELISA. Cellular immune response was assessed through a lymphocyte proliferation assay. Finally, clinical and morphopathological aspects were evaluated for all experimental animals. Our results demonstrated that when comparing TcVac1 IDE delivery vs IM delivery, the former induced significantly higher level of antigen-specific antibody response (IgG2a + IgG2b > IgG1) and lymphocyte proliferation, which expanded in response to challenge infection. Histological evaluation after challenge infection showed infiltration of inflammatory cells (macrophages and lymphocytes) in the heart and skeletal tissue of all infected mice. However, the largest increase in inflammatory infiltrate was observed in TcVac1_IDE/Tc mice when compared with TcVac1_IM/Tc or non-vaccinated/infected mice. The extent of tissue inflammatory infiltrate was directly associated with the control of tissue amastigote nests in vaccinated/infected (vs. non-vaccinated/infected) mice. Our results suggest that IDE delivery improves the protective efficacy of TcVac1 vaccine against T. cruzi infection in mice when compared with IM delivery of the vaccine.