Surface topography of resorbable porcine collagen membranes, and their effect on early osteogenesis: An in vitro study.

OBJECTIVE: Guided tissue regeneration (GTR) is based on the use of different membranes that function as sealants and barriers in specific clinical situations. Among the several tissue production methods and origins, resorbable porcine-derived membranes are the most commonly used. Because these membranes are so diverse, and have several different clinical applications, doubts linger as to their effect in stimulating osteogenesis. The objective of this study was to make an in vitro evaluation of the viability and differentiation of osteoblastic cells cultured on the surface of the following collagen membranes: Jason® (Botiss Biomaterials), Collprotect® (Botiss Biomaterials), and Bio-Gide® (Geistlich). MATERIAL AND METHODS: Fragments of the 3 resorbable collagen membranes (5 × 5 mm) were used, and pre-osteoblastic SAOS-2 cells (ATCC, USA) were plated on their porous surfaces. Evaluation of the membranes was performed at 3, 5 and 7 days, considering the following parameters: (1) topographic analysis of the different surfaces by scanning electron microscope; (2) cellular viability by MTT, (3) quantification of type I collagen and osteopontin by Elisa. The quantitative analyses were carried out using a significance level of 5%. RESULTS: Collprotect® and Jason® membranes presented a rough surface with an irregular aspect on both sides, while double-layered Bio-Gide® had one layer with a smooth surface and the other with a rough surface along each respective length. The viability assays revealed that the cells cultured the cells grown on Collprotect® showed higher viability than those grown in Bio-Gide® or Jason®, especially after 5 and 7 days. After 3 and 5 days, evaluation of type I collagen showed that the cells plated on the Jason® and Collprotect® surfaces had greater collagen secretion than those plated on BioGide®. After 7 days, an increase in osteopontin levels was observed when the cells were plated on all the experimental membranes, compared with the control group. CONCLUSION: All the tested membranes were suitable for use in GTR clinical procedures. Their indication in specific regenerative cases depends on the mechanical and biological properties of their originating tissues, thus enabling better results and assertive choices by dental professionals.

Conditioned Media from Human Pulp Stem Cell Cultures Improve Bone Regeneration in Rat Calvarial Critical-Size Defects.

The aim of this study was to test whether lyophilized conditioned media from human dental pulp mesenchymal stem cell cultures promote the healing of critical-size defects created in the calvaria of rats. Prior to the surgical procedure, the medium in which dental pulp stem cells were cultured was frozen and lyophilized. After general anesthesia, an 8 mm diameter bone defect was created in the calvaria of twenty-four rats. The defects were filled with the following materials: xenograft alone (G1) or xenograft associated with lyophilized conditioned medium (G2). After 14 or 42 days, the animals were euthanized, and the specimens processed for histologic and immunohistochemical analysis. Bone formation at the center of the defect was observed only in the G2 at 42 days. At both timepoints, increased staining for VEGF, a marker for angiogenesis, was observed in G2. Consistent with this, at 14 days, G2 also had a higher number of blood vessels detected by immunostaining with an anti-CD34 antibody. In conclusion, conditioned media from human dental pulp mesenchymal stem cell cultures had a positive effect on the regenerative process in rat critical-size bone defects. Both the formation of bone and enhancement of vascularization were stimulated by the conditioned media.

Sema4D is diminished in leukocyte platelet-rich fibrin and impairs pre-osteoblastic MC3T3-E1 cells' functionality.

OBJECTIVE: Semaphorin 4D (Sema4D) is a coupling factor expressed on osteoclasts that may hinder osteoblast differentiation. Since the leukocyte platelet-rich fibrin (L-PRF) membrane promotes growth factor concentration, this study aims to quantify the amount of Sema4D in L-PRF membranes, and analyze the impact of Sema4D on osteoblast cell function in vitro. DESIGN: Enzyme-linked immunosorbent assay (ELISA) was used to quantify the levels of Sema4D in both L-PRF and whole blood (serum). To analyze the impairment of Sema4D on osteoblasts, MC3T3-E1 cells were induced to osteogenic differentiation and exposed to Sema4D ranging from 10 to 500 ng/ml concentrations. The following parameters were assayed: 1) cell viability by MTT assay after 24, 48, and 72 h; 2) matrix mineralization by Alizarin Red staining after 14 days, 3) Runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), osteonectin (ONC), bone sialoprotein (BSP) and alkaline phosphatase (ALP) gene expression by qPCR. For all data, the significance level was set at 5%. RESULTS: The amount of Sema4D in the whole blood (serum) was higher than in L-PRF. Osteoblasts exposed to Sema4D at all tested concentrations exhibited a decrease in matrix mineralization formation as well in RUNX-2, OCN, ONC, BSP, and ALP gene expression (p < 0.05). CONCLUSION: The presence of Sema4D, a molecule known for suppressing osteoblast activity, diminishes within L-PRF, enhancing its ability to facilitate bone regeneration.

Microbiological analysis of bacterial sealing of internal conical implants with different taper angles.

The present study evaluated the effect of the taper angle of different internal conical connection implants and cyclic loading on the implant-abutment bacterial seal. A total of 96 implant-abutment sets were divided into eight groups. Four groups of different taper degrees with cyclic mechanical loading of 500,000 cycles per sample, with a 120-N load at 2 Hz before analysis [16DC (16-degree, cycled), 11.5DC (11.5-degree, cycled), 3DC (3- degree, cycled) and 4DC (4- degree, cycled)] were compared to four control groups without cyclic loading [16D (16-degree), 11.5D (11.5-degree), 3D (3-degree), and 4D (4-degree)]. Microbiological analysis was performed by immersing all samples in a suspension containing Escherichia coli and incubating them at 37°C. After 14 days, the presence of bacterial seals was evaluated. Fisher-Freeman-Halton exact tests and binomial tests were performed (5% significance level). The groups showed significant differences in bacterial seal, and mechanical load cycling improved the bacterial seal in the 3DC group. In all other groups, no significant differences in bacterial seal were found between cycled and uncycled samples. To conclude, the internal conical connection with a 3-degree taper angle showed better results than the other connection with different angles when subjected to load cycling. However, none of the angles tested were fully effective in sealing the implant-abutment interface.

Short implants: Influence of different diameters, torques, and transmucosal heights of straight prosthetic abutments on screw preload loss after cycling.

This study evaluated the influence of different implant diameters, insertion torques, and transmucosal heights on the loosening of abutments installed on short implants, after mechanical cycling. The Morse taper connection implants (n = 96) tested were 5 mm high, divided according to the platform diameter: 4 or 6 mm. A universal abutment was coupled to each implant (with different transmucosal heights: 1 or 5 mm). The sets were subdivided into 20- and 32-Ncm torque. After the cycle fatigue test, the detorque values were measured with a digital torque indicator. After mechanical cycling, the mean detorque values obtained for the abutment with 20-Ncm insertion torque were lower than for implants with 32-Ncm insertion torque, regardless of the platform diameter or transmucosal height. In the 20-Ncm torque group, there was no statistically significant difference in the detorque values between platform diameters or transmucosal heights. Otherwise, for 32-Ncm sets, a smaller platform diameter (4 mm), and a longer transmucosal height (5 mm) showed the lowest detorque values. In conclusion, implants placed with 32-Ncm insertion torque and abutments with 1 mm transmucosal height and a 6 mm implant diameter demonstrated the highest detorque values.

Pulp-Dentin Complex Regeneration with Cell Transplantation Technique Using Stem Cells Derived from Human Deciduous Teeth: Histological and Immunohistochemical Study in Immunosuppressed Rats.

The aim of this study was to histologically verify the performance of pulp-derived stem cells used in the pulp-dentin complex regeneration. Maxillary molars of 12 immunosuppressed rats were divided into two groups: the SC (stem cells) group, and the PBS (just standard phosphate-buffered saline) group. After pulpectomy and canal preparation, the teeth received the designated materials, and the cavities were sealed. After 12 weeks, the animals were euthanized, and the specimens underwent histological processing and qualitative evaluation of intracanal connective tissue, odontoblast-like cells, intracanal mineralized tissue, and periapical inflammatory infiltrate. Immunohistochemical evaluation was performed to detect dentin matrix protein 1 (DMP1). In the PBS group, an amorphous substance and remnants of mineralized tissue were observed throughout the canal, and abundant inflammatory cells were observed in the periapical region. In the SC group, an amorphous substance and remnants of mineralized tissue were observed throughout the canal; odontoblasts-like cells immunopositive for DMP1 and mineral plug were observed in the apical region of the canal; and a mild inflammatory infiltrate, intense vascularization, and neoformation of organized connective tissue were observed in the periapical region. In conclusion, the transplantation of human pulp stem cells promoted partial pulp tissue neoformation in adult rat molars.

DoE development of ionic gradient liposomes: A successful approach to improve encapsulation, prolong anesthesia and decrease the toxicity of etidocaine.

Etidocaine (EDC) is a long-acting local anesthetic of the aminoamide family whose use was discontinued in 2008 for alleged toxicity issues. Ionic gradient liposomes (IGL) are nanostructured carriers for which an inner/outer gradient of ions increases drug upload. This work describes IGLEDC, a formulation optimized by Design of Experiments, composed of hydrogenated soy phosphatidylcholine:cholesterol:EDC, and characterized by DLS, NTA, TEM/Cryo-TEM, DSC and 1H NMR. The optimized IGL showed significant encapsulation efficiency (41 %), good shelf stability (180 days) and evidence of EDC interaction with the lipid bilayer (as seen by DSC and 1H NMR results) that confirms its membrane permeation. In vitro (release kinetics and cytotoxicity) tests showed that the encapsulation of EDC into the IGL promoted sustained release for 24 h and decreased by 50 % the intrinsic toxicity of EDC to Schwann cells. In vivo IGLEDC decreased the toxicity of EDC to Caenorhabditis elegans by 25 % and extended its anesthetic effect by one hour, after infiltrative administration, at clinically used (0.5 %) concentration, in rats. Thus, this novel drug delivery system is a promise for the possible reintroduction of EDC in clinics, aiming at the control of operative and postoperative pain.

Effect of platelet-rich fibrin (PRF) membranes on the healing of infected skin wounds.

Tissue engineering focuses on wound healing and tissue regeneration. Platelet-rich fibrin (PRF) is a fibrin matrix containing cytokines, growth factors and cells that are gradually released into the wound over time. This study aimed to evaluate the effect of PRF membranes on wound repair and microbial control in infected wounds. Skin wounds were performed on the dorsum of rats using a 6 mm diameter metal punch. The defects were randomly assigned into four groups (n = 12/each) accordingly to the treatment: G1, noninfected wound filled only with clot; G2, noninfected wound with PRF; G3, infected wound (S. aureus) without PRF; G4, infected wound (S. aureus) with PRF. After 7 and 14 days, macroscopic and histological analyses of the wounds were performed. Furthermore, the quantification of ß-defensin in PRF was measured by ELISA. At 14 days, the groups with PRF (G2 and G4) had wound sizes significantly smaller than the original defects (6 mm) (p < 0.05) and significantly smaller than those not treated with PRF, in both the infected and noninfected groups (p < 0.05). Furthermore, the groups with infected wounds (G3 and G4) demonstrated a significantly lower inflammation score in the PRF group than in the noninfected groups (p < 0.05). In vitro analysis of ß-defensin was performed in all PRF membrane groups, and the median value was 1.444 pg/mL. PRF in the wounds of both control and infected rats played an important role in the modulation of tissue healing, most notably in infected sites.

In vivo evaluation of a collagen membrane in bone neoformation: A morphological and histomorphometric study.

OBJECTIVE: Guided bone regeneration (GBR) is a technique that involves the placement of mechanical barriers to protect the blood clot, and create an isolated space to prevent competition from epithelial and connective tissues in bone augmentation treatments. Collagen membranes stand out from other materials available for performing regenerative surgeries, and are widely used because of their ability to promote cell adhesion and homeostasis, and their biocompatibility, ease of handling, and low immunogenicity. In this context, researchers have investigated xenogenic membranes/barriers that cost less and have slower resorption rates. The current study aimed to assess the osteogenic potential induced by a crosslinked, synthesized xenogenic membrane 100 µm thick when applied in vivo to critical defects in rat calvaria. MATERIAL AND METHODS: Critical size defects were created in the calvaria of thirty male Wistar rats, and randomly divided into the following two groups: G1 - clot covered with a commercial xenogenic membrane (Lumina-Coat®, Criteria, Brazil), and G2 - clot covered with a synthesized xenogenic membrane. The animals were euthanized after 7, 15 and 30 days, and samples of calvaria were processed to perform morphometric evaluations to measure bone neoformation in the defect region. In addition, ultrastructural characterization of the collagen membranes was performed by scanning electron microscope. The quantitative analyses were carried out by adopting a significance level of 5%. RESULTS: The ultrastructural characterization revealed that the synthesized membrane had thicker collagen fibers and a more cohesive surface, compared with the Lumina-Coat® collagen membrane (G1). There was no significant difference in bone neoformation between the membranes (p>0.05), at any of the time periods analyzed. The bone quantification area increased significantly over time for both membranes (p<0.05). CONCLUSION: The synthesized membrane exhibited morphological characteristics similar to those of the commercial membrane evaluated, allowed potentially active participation in the bone neoformation process, and served as a low-cost alternative for GBR procedures.

Effect of ozone therapy on the modulation of inflammation and on new bone formation in critical defects of rat calvaria filled with autogenous graft.

OBJECTIVE: The aim of this study was to evaluate the effect of ozone therapy on new bone formation and inflammation modulation in defects of rat calvaria filled with autogenous bone. MATERIAL AND METHODS: Critical size defects were created in the calvaria of 24 male Wistar rats. The animals were randomly divided into four groups according to the treatment: G1: clot; G2: clot and covered with xenogenic membrane; G3: particulate autogenous bone graft; G4: autogenous bone graft and application of 3 mL O2/O3 gas mixture (10 µg/ml). The defects were filled immediately after surgery with a bilateral retroauricular application, in the region immediately above the incision. After 21 days, the animals were euthanized, and the samples were processed for morphometric evaluations designed to measure both the intensity of the inflammatory infiltrate, and the presence of new bone formation in the defect. RESULTS: The results showed a lower inflammation score and higher mean of newly formed bone in the region of the defect for the group associated with ozone therapy (G4). The bone formed in the region of the defect could be observed as being more lamellar and mineralized in the case of associated ozone therapy. CONCLUSION: Ozone therapy represents a promising adjuvant therapy to accelerate tissue regeneration.

Microbiological analysis of bacterial sealing of internal conical implants with different taper angles

Abstract The present study evaluated the effect of the taper angle of different internal conical connection implants and cyclic loading on the implant-abutment bacterial seal. A total of 96 implant-abutment sets were divided into eight groups. Four groups of different taper degrees with cyclic mechanical loading of 500,000 cycles per sample, with a 120-N load at 2 Hz before analysis [16DC (16-degree, cycled), 11.5DC (11.5-degree, cycled), 3DC (3- degree, cycled) and 4DC (4- degree, cycled)] were compared to four control groups without cyclic loading [16D (16-degree), 11.5D (11.5-degree), 3D (3-degree), and 4D (4-degree)]. Microbiological analysis was performed by immersing all samples in a suspension containing Escherichia coli and incubating them at 37°C. After 14 days, the presence of bacterial seals was evaluated. Fisher-Freeman-Halton exact tests and binomial tests were performed (5% significance level). The groups showed significant differences in bacterial seal, and mechanical load cycling improved the bacterial seal in the 3DC group. In all other groups, no significant differences in bacterial seal were found between cycled and uncycled samples. To conclude, the internal conical connection with a 3-degree taper angle showed better results than the other connection with different angles when subjected to load cycling. However, none of the angles tested were fully effective in sealing the implant-abutment interface.

Evaluation of the dosimetry and centralization of scout-view function in CBCT.

This study evaluated the centralization of the region of interest (ROI) in acquisition of the CBCT images, when the freely positionable scout-view (SV) function is applied. Additionally, the dosimetry of the acquired images was assessed in the SV function alone as well as in complete tomographic image in two different fields of view (FOV) (50x50 and 78x150mm). A three-location device was created to accommodate the dosimeters and the specimens, in the right, middle and left location during image acquisition. For dose assessment, thermoluminescent dosimeters were irradiated within the FOV and analyzed in a portable reader. For ROI evaluation, three specimens of gutta-percha stick were placed on the same device and the CT scans were acquired (CBCT OP 300 Maxio device, 90kV, 13mA, 85 µm voxel size, FOV of 50X50mm), with and without the SV, in three positions (3-9, 1-7 and 5-11 o'clock), simulating different regions of the mouth. Two image evaluations were performed, an objective and subjective. There was a slight percentage increase (1.36% to 1.40%) of the radiation dose with the use of SV. The distances were significantly greater in the images acquired without SV (p < 0.05). Every image obtained with SV was classified as being at the FOV's center. In conclusion, the results demonstrated that SVs function is effective to centralize the ROI in the FOV, increasing the scan precision and avoiding repetitions due to positioning errors.

Biologic Behavior of Pressed Lithium Disilicate Ceramic and Zirconia on Human Gingival Fibroblasts: An In Vitro Study.

This study evaluated in vitro the biologic profile of manually polished surfaces of pressed lithium disilicate (LD) ceramic compared to zirconia (Zir) in human gingival fibroblasts. Samples with a 10-mm diameter and 3-mm thickness were used. After manual polishing, the average roughness (Ra) of the samples was measured. The cell proliferation and viability of gingival fibroblasts on the surfaces were assessed at 24, 48, and 96 hours. Additionally, the morphology, cell adhesion, and type III collagen (COLIII) and vimentin (VIM) expression by fibroblasts plated onto these surfaces was analyzed. Polystyrene (Pol) was used as control for all assays. The mean Ra was 0.261 ± 0.053 µm for Zir and 0.345 ± 0.130 µm for LD. Both surfaces presented similar cell proliferation and viability (P > .05). The cell morphology demonstrated that, for both surfaces, the cells were occasionally spindle-shaped, parallel to the direction of the grooves. Compared to Pol, an upregulation of COLIII and VIM gene expression was observed by fibroblasts cultured on Zir and LD at all time points (P < .05). The characteristics presented by LD and Zir surfaces after manual polishing protocol were similar and had biologically favorable performances, thus suggesting LD as a suitable alternative to Zir in the peri-implant region for esthetic purposes.

Evaluation of the dosimetry and centralization of scout-view function in CBCT

Abstract This study evaluated the centralization of the region of interest (ROI) in acquisition of the CBCT images, when the freely positionable scout-view (SV) function is applied. Additionally, the dosimetry of the acquired images was assessed in the SV function alone as well as in complete tomographic image in two different fields of view (FOV) (50x50 and 78x150mm). A three-location device was created to accommodate the dosimeters and the specimens, in the right, middle and left location during image acquisition. For dose assessment, thermoluminescent dosimeters were irradiated within the FOV and analyzed in a portable reader. For ROI evaluation, three specimens of gutta-percha stick were placed on the same device and the CT scans were acquired (CBCT OP 300 Maxio device, 90kV, 13mA, 85 µm voxel size, FOV of 50X50mm), with and without the SV, in three positions (3-9, 1-7 and 5-11 o'clock), simulating different regions of the mouth. Two image evaluations were performed, an objective and subjective. There was a slight percentage increase (1.36% to 1.40%) of the radiation dose with the use of SV. The distances were significantly greater in the images acquired without SV (p < 0.05). Every image obtained with SV was classified as being at the FOV's center. In conclusion, the results demonstrated that SVs function is effective to centralize the ROI in the FOV, increasing the scan precision and avoiding repetitions due to positioning errors.

Resumo Este estudo avaliou a centralização da região de interesse (ROI) na aquisição das imagens de TCFC, quando a função scout-view (SV) posicionável livremente é aplicada. Adicionalmente, a dosimetria das imagens adquiridas foi avaliada isoladamente na presença da função SV, bem como após aquisição de imagem tomográfica completa em dois diferentes campos de visão (FOV) (50x50 e 78x150mm). Um dispositivo de três localizações foi criado para acomodar os dosímetros e os espécimes, na localização direita, central e esquerda, durante a aquisição das imagens. Para avaliação da dose, dosímetros termoluminescentes foram irradiados dentro dos campos de visão e analisados em leitor portátil. Para avaliação da ROI, três espécimes de guta percha foram colocados no mesmo aparelho e as tomografias foram adquiridas (CBCT OP 300 Maxio, 90kV, 13mA, 85 μm tamanho de voxel, FOV de 50X50mm), com e sem a SV, em três posições (3-9, 1-7 e 5-11 horas), simulando diferentes regiões da boca. Foram realizadas duas avaliações de imagem, uma objetiva e outra subjetiva. Houve um leve aumento percentual (1,36% para 1,40%) da dose de radiação com o uso de SV. As distâncias foram significativamente maiores nas imagens adquiridas sem SV (p < 0,05). Todas as imagens obtidas com SV foram classificadas como sendo do centro do FOV. Em conclusão, os resultados do presente estudo demonstraram que a função scout view é eficaz para centralizar a ROI no FOV, aumentando a precisão do escaneamento e evitando repetições devido a erros de posicionamento.

Analysis of the Viability and Morphology of Gingival Cells on Materials Used in Novel Prosthetic Components: In Vitro Study.

AIM: The objective of this in vitro study was to evaluate the viability and morphology of human fibroblasts and keratinocytes cells, both grown on stainless steel (steel) (18Cr14Ni2.5Mo), and polyether-ether-ketone (PEEK) surfaces, hypothesizing the use of these surfaces as novel materials for prosthetic components. MATERIALS AND METHODS: Gingival human keratinocytes and gingival human fibroblasts lines were grown on discs made by steel (n = 36), PEEK (n = 36), and titanium (Ti) (Ti6A14V) (n = 36)-control. For viability assay, cultures were grown at 24 hours (TV1), 48 hours (TV2), and 72 hours (TV3) times and evaluated by the colorimetric tetrazolium assay (MTT). For morphology and cell adhesion assays, after 24 hours (TM1), 48 hours (TM2), and 96 hours (TM3) of cell culture, cells were examined by scanning electron microscopy (SEM) and analyzed at magnifications with 500×, 1,000×, and 2,500×. RESULTS: Regarding the viability, the keratinocytes did not present statistical difference on the different materials, in TV1 and TV3 times of culture. Their growth rate increased on all materials, being more expressive in steel; the fibroblasts did not present statistical difference on the different materials, in TV2 and TV3 times of culture. The growth rate of these decreased on all materials, being more expressive in PEEK. The morphology analyses show increase in cell numbers, adequate spreading, and adhesion at all cultivation times (TM1, TM2, and TM3) in both cell lines, on all materials. CONCLUSION: All materials tested are suitable for use in the manufacture of prosthetic components for implant-supported rehabilitations, considering the limitations of this study. CLINICAL SIGNIFICANCE: This work analyzes the cellular response of cells present in the human gingiva, as a way to simulate the peri-implant tissue response around novel angular prosthetic components made of stainless steel and PEEK.

Histomorphometric evaluation of different graft associations for maxillary sinus elevation in wide antral cavities: a randomized controlled clinical trial.

OBJECTIVES: Pneumatization of the maxillary sinus can make it difficult, if not impossible, to install osseointegrated implants, and undertake their eventual functional rehabilitation, which may ultimately require regenerative techniques to achieve. This randomized controlled study proposed conducting a histological evaluation of the behavior of different graft materials in wide maxillary sinuses, at a height of 8 to 10 mm from the alveolar ridge, combined with bone remnants less than 3mm. MATERIALS AND METHODS: Thirty-six patients underwent a sinus elevation procedure through the lateral window. The sinuses were randomly filled with the following materials (n=12/group): group 1, xenogenic bone + autogenous bone (ratio 70:30, respectively); group 2, xenogenic bone + L-PRF; and group 3, xenogenic bone. At 8 months, bone biopsies of engrafted sites were harvested and analyzed histomorphometrically in order to quantify newly formed bone tissue. RESULTS: The results showed a greater area of newly formed bone for G1, averaging 2678.37 (1116.40) µm2, compared with G2 at 984.87 (784.27) µm2, and G3 at 480.66 (384.76) µm2 (p < 0.05). Additionally, fewer xenogenic bone particles and a large amount of connective tissue were observed in G2. CONCLUSIONS: In maxillary sinuses with large antral cavities, autogenous bone combined with xenogenic bone seems to demonstrate better graft remodeling and improve bone formation, compared with the addition of L-PRF. CLINICAL RELEVANCE: L-PRF produces few advantages regarding new bone formation in the wide maxillary sinuses. TRIAL REGISTRATION: Brazilian Clinical Trials Registry (REBEC) number RBR-2pbbrvg.

In vitro analysis of prosthetic abutment and angulable frictional implant interface adaptation: Mechanical and microbiological study.

The aim of this study was to evaluate, in vitro, the microbiological sealing at the implant and different angles frictional prosthetic abutment interface, submitted or not to mechanical cycling, as well as the deactivation force and evaluation of the implant-abutment interface by scanning electron microscopy. For this study, the sealing capacity of eighty sets of abutments/implants of each angle, with and without mechanical cycling, with internal conical connection (locking tapper) (4.3 mm × 9.0 mm) constituted in Titanium alloy (Ti6Al4V), and stainless steel angled prosthetic abutment was evaluated (18Cr14Ni2.5Mo) according to ASTM F138-13a (Arcsys, FGM, Joinville, Brazil), 6 mm high and 4.2 mm in diameter at the coronary portion, and 3.5 mm high transmucosal, in 4 different angles (0, 5, 10 and 20°). After in vitro tests, 100% biological sealing was observed at the implant / prosthetic abutment interface within cycled and non-cycled conditions, for the straight, 5, 10 and 20° inclination groups. There was no statistically significant difference in the removal force of the prosthetic abutments at different angles, under non-cycled conditions; however, under mechanical loading, the deactivation force was significantly higher for straight prosthetic abutments than with 10 and 20° of angulation. Surface analysis revealed good adaptation between implants and abutments, and the presence of wear areas, independently of mechanical loading. It is concluded that the analysis of implant and prosthetic abutment interface revealed good adaptation between the parts, for all analyzed samples.

Low concentrations of caffeic acid phenethyl ester stimulate osteogenesis in vitro.

OBJECTIVE: The aim of this study was to evaluate the effects of caffeic acid phenethyl ester (CAPE) on osteoblast-like cell cultures (SAOS-2). METHODS: SAOS-2 were exposed to CAPE at 1 nM, 10 nM, 100 nM, 1 µM, and 10 µM. Non-exposed cultures were used as control. The following parameters were assayed: 1) cell viability at 1, 3, and 7 days; 2) alkaline phosphatase (ALP) activity at 5 and 10 days; 3) matrix mineralization at 14 days; and 4) Runt-related transcription factor 2 (RUNX2), ALP, osteopontin (SPP1), and osteocalcin (BGLAP) gene expression at 5 and 10 days. The data were analyzed by ANOVA two-way or Kruskal-Wallis (α = 5%). RESULTS: At day 1, cell viability was similar among all groups (p > 0.05). At days 3 and 7, cultures exposed to CAPE at 10 µM exhibited a significant reduction in cell viability compared with the others groups (p < 0.05). At day 5, ALP activity was similar among all experimental groups; at day 10, however, the stain intensity was higher in cultures exposed to CAPE at 100 nM and 10 nM in comparison with the other groups (p < 0.05). At days 5 and 10, RUNX2, ALP, SPP1, and BGLAP gene expression was greater in cultures exposed to CAPE in comparison with the control (p < 0.05). At day 14, matrix mineralization was similar in cultures exposed to CAPE at 1 nM and 10 nM (p > 0.05), but superior to those ones observed in the other experimental groups (p < 0.05). CONCLUSION: CAPE at low concentrations can positively module the osteogenesis in vitro.

Titanium dioxide nanotubes added to glass ionomer cements affect S. mutans viability and mechanisms of virulence.

This in vitro study evaluated the impact of TiO2 nanotubes (n-TiO2) incorporated into glass ionomer cement (GIC) on Streptococcus mutans (S. mutans) characteristics at cellular and molecular levels. n-TiO2, synthesized by the alkaline method (20 nm in size), was added to Ketac Molar EasyMix® at 0%, 3%, 5%, and 7% by weight. S. mutans strains were cultured on GIC disks with addition or not of n-TiO2 for 1, 3, and 7 days and the following parameters were assessed: inhibition halo (mm) (n=3/group); cell viability (live/dead) (n=5/group); cell morphology (SEM) (n=3/group); and gene expression by real-time PCR (vicR, covR, gtfB, gtfC, and gtfD) (n=6/group). The data were analyzed by the Kruskal-Wallis test, repeated-measures ANOVA or two-way ANOVA, and Tukey's and Dunn's post-hoc tests (α=0.05). The agar diffusion test showed a higher antibacterial property for 5% n-TiO2 compared with 3% and 7% (p<0.05) with no effect of time (1, 3, and 7 days). The cell number was significantly affected by all n-TiO2 groups, while viability was mostly affected by 3% and 5% n-TiO2, which also affected cell morphology and organization. Real-time PCR demonstrated that n-TiO2 reduced the expression of covR when compared with GIC with no n-TiO2 (p<0.05), with no effect of time, except for 3% n-TiO2 on vicR expression. Within-group and between-group analyses revealed n-TiO2 did not affect mRNA levels of gtfB, gtfC, and gtfD (p>0.05). Incorporation of n-TiO2 at 3% and 5% potentially affected S. mutans viability and the expression of key genes for bacterial survival and growth, improving the anticariogenic properties of GIC.