RESUMEN
The present study aimed to evaluate the effect of photobiomodulation therapy (PBM) on different stages of osteogenesis in vitro. For this, osteoblastic-like cells (Saos-2 cell lineage) were irradiated in two different periods: during the Proliferation phase (PP; from the second to the fourth day) and during the Differentiation phase (DP; from the seventh to the ninth day). The energy density used in the study was 1.5 J/ cm2. The following parameters were evaluated: 1) quantification of collagen type 1 (COL 1), osteopontin (OPN), and bone morphogenetic protein 2 (BMP-2); 2) quantification of alkaline phosphatase (ALP) activity; and 3) quantification of extracellular matrix (ECM) mineralization. Non-irradiated cultures were used as controls. The data were analyzed using the Student's t-test or one-way ANOVA, considering a significance level of 5%. The results indicated that COL 1 and BMP-2 quantification was higher in Saos-2 irradiated during the DP in relation to the control group at day 10 (p < 0.05). No differences were observed for other comparisons at this time point (p > 0.05). OPN expression was greater in PP compared with the other experimental groups at day 10 (p < 0.05). Irradiation did not affect ALP activity in Saos-2 regardless of the exposure phase and the time point evaluated (p > 0.05). At day 14, ECM mineralization was higher in Saos-2 cultures irradiated during the DP in relation to the PP (p < 0.05). In conclusion, the results suggested that the effects of PBM on osteoblastic cells may be influenced by the stage of cell differentiation.
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Fosfatasa Alcalina , Proteína Morfogenética Ósea 2 , Diferenciación Celular , Proliferación Celular , Colágeno Tipo I , Terapia por Luz de Baja Intensidad , Osteoblastos , Osteogénesis , Osteopontina , Osteogénesis/efectos de la radiación , Humanos , Proteína Morfogenética Ósea 2/metabolismo , Fosfatasa Alcalina/metabolismo , Osteopontina/metabolismo , Diferenciación Celular/efectos de la radiación , Colágeno Tipo I/metabolismo , Osteoblastos/efectos de la radiación , Osteoblastos/citología , Osteoblastos/metabolismo , Proliferación Celular/efectos de la radiación , Matriz Extracelular/metabolismo , Matriz Extracelular/efectos de la radiaciónRESUMEN
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.
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Diseño de Implante Dental-Pilar , Implantes Dentales , Torque , Pilares Dentales , Ensayo de Materiales , Estrés Mecánico , Tornillos Óseos , Análisis del Estrés DentalRESUMEN
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.
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Sustitutos de Huesos , Fibrina Rica en Plaquetas , Elevación del Piso del Seno Maxilar , Sustitutos de Huesos/uso terapéutico , Trasplante Óseo/métodos , Implantación Dental Endoósea/métodos , Humanos , Maxilar/cirugía , Seno Maxilar/patología , Seno Maxilar/cirugía , Osteogénesis , Elevación del Piso del Seno Maxilar/métodosRESUMEN
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.
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Encía , Acero Inoxidable , Adhesión Celular , Humanos , Polietilenglicoles/farmacología , Propiedades de Superficie , TitanioRESUMEN
OBJECTIVES AND BACKGROUND: Sodium ascorbyl phosphate (SAP) is a hydrophilic and stable L-ascorbic acid derivative, being converted by the cell phosphatases into free ascorbic acid (AA), which allows its sustained release in the medium. AA participates in the maintenance and healing of the periodontium. It presents a regulatory role of the osteoblastic activity, stimulating the deposition of collagen extracellular matrix followed by the induction of genes associated with the osteoblastic phenotype. It also acts in the elimination of reactive oxygen species, abundantly produced by defense cells in periodontal disease. The aim of this study was to evaluate the effect of SAP on osteoblast viability and differentiation. METHODS: Mouse preosteoblastic cells of the MC3T3-E1 strain were used. Cell viability was assessed by the trypan blue dye exclusion assay and the expression of genes related to osteoblast differentiation by quantitative PCR. Collagen I secretion was evaluated by ELISA, and mineralized matrix formation was assayed by Alizarin red S staining. RESULTS: The results showed that SAP at concentrations from 50 to 500 µmol/L does not influence preosteoblast cell viability, but stimulates their differentiation, observed by the induction of RUNX2, COL1A1, and BGLAP2; by the higher secreted levels of collagen I; and also by the increase in the mineralization of the extracellular matrix in cells exposed to this agent at 200 or 400 µmol/L, compared with those not exposed. CONCLUSION: By its stability and capacity to induce preosteoblastic cell differentiation, our results indicate that the incorporation of SAP into local release devices, membranes/scaffolds or biomaterials, could favor bone tissue formation and therefore periodontal healing.
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Ácido Ascórbico/análogos & derivados , Osteoblastos , Animales , Ácido Ascórbico/farmacología , Diferenciación Celular , Ratones , Osteoblastos/efectos de los fármacos , OsteogénesisRESUMEN
BACKGROUND: There has been great interest recently in the mechanisms of cell-to-cell communication through microvesicles (MV). These structures are produced by many different cell types and can modulate cellular activity by induction of epigenetic alterations. These vesicles may promote tumor mass increase either by stimulating cell proliferation via growth factors or by inhibiting apoptosis, which reinforces the role of such vesicles as important modulators of tumor progression. METHODS: The present in vitro study aimed to characterize MV derived from malignant neoplastic epithelial cell cultures (EP) and their effect on the expression of apoptosis/autophagy and invasion related genes of benign myoepithelial (Myo) cell cultures. RESULTS: The results revealed round structures with a mean size of 153.6 (±0.2) nm, with typical MV morphology. CD63 quantification indicated that EP cell culture at 70%-80% confluence secreted 3.088 × 108 MV/mL. Overall, Myo exposed to MVs derived from EP showed both up- and downregulation of tumorigenesis promoting genes. MVs from EP cells promoted cell death of Myo cells and positively modulate BAX, SURVIVIN, LC3B, MMP-2, and MMP-9 expression. Furthermore, an increasing of MMP-2 and MMP-9 secretion by Myo was observed after MV exposure. CONCLUSIONS: These findings suggest that MVs from EP modulate autophagy of Myo cells, which may, in part, explain the disappearance of these cells in in situ areas of invasive carcinoma ex-pleomorphic adenoma. Additionally, the overexpression of MMPs contributes to the development of an invasive phenotype of Myo cells, which could favor the dissolution of the basement membrane during tumorigenesis process.
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Adenoma Pleomórfico , Carcinoma de Células Escamosas , Autofagia , Carcinoma de Células Escamosas/genética , Muerte Celular , Células Epiteliales , HumanosRESUMEN
PURPOSE: To apply titanium tetrafluoride (TiF4) in an aqueous solution or incorporated into the primer of a self-etching adhesive (Clearfil SE Bond) as dentin pre-treatment and evaluate its antimicrobial effect, determine the minimum bactericidal concentraion (MBC) against Streptococcus mutans and Lactobacillus casei and analyse its potential to inhibit the development of carious lesions at the restoration interface. MATERIALS AND METHODS: For MBC, an aqueous solution or primer with different concentrations (in %) of TiF4 were used (from 0.0 to 4.0). Also, 50 cavities were prepared at the enamel/dentin junction of third molars and received the following dentin pre-treatments (n = 10): Clearfil SE Bond (CL); aqueous solution of 2.5% TiF4 + CL (T2.5%); aqueous solution of 4% TiF4 + CL (T4%); 2.5% TiF4 incorporated into the primer (P2.5%); 4% TiF4 incorporated into the primer (P4%). Cavities were restored and submitted to pH cycling to create artificial caries lesions. Microhardness tests were performed after sectioning the restorations to assess the demineralisation at margins. RESULTS: ANOVA and Tukey's tests showed that TiF4 in aqueous solution presented MBC against S. mutans and L. casei of over 2.0%. TiF4 in the primer of a self-etching adhesive presented MBC of over 1% for L. casei. For enamel, CL showed no significant differences in microhardness between the depths. CONCLUSIONS: The aqueous solution had an antimicrobial effect against Streptococcus mutans and Lactobacillus casei of over 2.0%. Pretreatment with the aqueous solution or primer did not inhibit demineralisation at enamel or dentin restoration interfaces.
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Antiinfecciosos , Recubrimiento Dental Adhesivo , Resinas Compuestas , Esmalte Dental , Dentina , Recubrimientos Dentinarios , Fluoruros , Ensayo de Materiales , TitanioRESUMEN
PURPOSE: Etidocaine (EDC) is a long lasting local anesthetic, which alleged toxicity has restricted its clinical use. Liposomes can prolong the analgesia time and reduce the toxicity of local anesthetics. Ionic gradient liposomes (IGL) have been proposed to increase the upload and prolong the drug release, from liposomes. METHODS: First, a HPLC method for EDC quantification was validated. Then, large unilamellar vesicles composed of hydrogenated soy phosphatidylcholine:cholesterol with 250 mM (NH4)2SO4 - inside gradient - were prepared for the encapsulation of 0.5% EDC. Dynamic light scattering, nanotracking analysis, transmission electron microscopy and electron paramagnetic resonance were used to characterize: nanoparticles size, polydispersity, zeta potential, concentration, morphology and membrane fluidity. Release kinetics and in vitro cytotoxicity tests were also performed. RESULTS: IGLEDC showed average diameters of 172.3 ± 2.6 nm, low PDI (0.12 ± 0.01), mean particle concentration of 6.3 ± 0.5 × 1012/mL and negative zeta values (-10.2 ± 0.4 mV); parameters that remain stable during storage at 4°C. The formulation, with 40% encapsulation efficiency, induced the sustained release of EDC (ca. 24 h), while reducing its toxicity to human fibroblasts. CONCLUSION: A novel formulation is proposed for etidocaine that promotes sustained release and reduces its cytotoxicity. IGLEDC can come to be a tool to reintroduce etidocaine in clinical use.
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Anestésicos Locales/administración & dosificación , Anestésicos Locales/toxicidad , Supervivencia Celular/efectos de los fármacos , Preparaciones de Acción Retardada/química , Etidocaína/administración & dosificación , Etidocaína/toxicidad , Liposomas/química , Anestésicos Locales/farmacocinética , Línea Celular , Colesterol/química , Liberación de Fármacos , Etidocaína/farmacocinética , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Humanos , Iones/química , Fosfatidilcolinas/químicaRESUMEN
The aim of this study was to measure and record the universal transmucosal abutment height, and then evaluate whether it influenced loosening of the abutment screw by analyzing the torque and detorque values after mechanical cycling. Thirty-six implants, model CM Unitite, with internal conical connections (3.5 × 10 mm) and respective universal prosthetic abutments (n = 36, 3.25 × 6 mm), were divided into three groups (n = 12 each) with respective transmucosal heights of 0.8, 3.5, and 5.5 mm. Insertion torque of 20 Ncm was used in accordance with the manufacturer's specifications. Afterward, the samples were submitted to fatigue tests consisting of 500,000 cycles at a frequency of 2Hz, a dynamic compressive load of 120N, and an angle of 30°. The detorque values were measured with a digital torque meter and tabulated to perform statistical analyses; a level of significance of 5% was adopted. The mean detorque values (SD) obtained were 22.83 (6.30), 22.5 (5.45), and 19.41 (4.69) Ncm for transmucosal abutments with heights of 0.8, 3.5, and 5.5 mm, respectively, and showed no statistically significant difference ( P = .262). The authors of this study concluded that the transmucosal height of prosthetic abutments submitted to mechanical fatigue did not influence the detorque values.
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Pilares Dentales , Diseño de Implante Dental-Pilar , Implantes Dentales , Análisis del Estrés Dental , Estrés Mecánico , TorqueRESUMEN
BACKGROUND: Polymorphous low-grade adenocarcinoma (PLGA) remains a diagnostic challenge for most pathologists due to its large spectrum of histological patterns. In this study, the expression of two new markers recently described for salivary gland tumors was studied in PLGA. METHODS: The morphology of 33 cases of PLGA was carefully evaluated using hematoxylin-and-eosin-stained sections and confirmed by immunohistochemistry for cytokeratin 7, vimentin, and S-100. Periodic acid-Schiff with diastase digestion was also used. The expression of mammaglobin and DOG-1 was carried out using the EnVision System. Mammaglobin was assessed according to the percentage of positively stained tumor cells, while DOG-1 was evaluated according to its presence and site. For MCM-2 and Ki-67, markers of proliferation, the labeling index of cell nuclei positivity was evaluated using total cell number. The ETV6-NTRK3 fusion was examined by fluorescence in situ hybridization analysis. RESULTS: The histological patterns of the tumor were classified as lobular or non-lobular. For the non-lobular pattern, tubular, cribriform, glomeruliform, trabecular, and papillary patterns were observed. Mammaglobin was present in all PLGA cases, and its expression was stronger (P = 0.01) in the lobular than in the non-lobular pattern. The expression of DOG-1 was present in the apical portion and cytoplasm of the cells. Proliferation markers were low for all cases independent of histological pattern. CONCLUSIONS: Polymorphous low-grade adenocarcinoma has been confirmed to originate from the intercalated duct and to feature high expression of mammaglobin in its lobular pattern resembling that of mammary secretory analogue carcinoma, except for the ETV6 gene rearrangement.
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Adenocarcinoma/metabolismo , Anoctamina-1/metabolismo , Biomarcadores de Tumor/metabolismo , Mamoglobina A/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias de las Glándulas Salivales/metabolismo , Adenocarcinoma/patología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Neoplasias de las Glándulas Salivales/patologíaRESUMEN
PURPOSE: The goal from this in vitro study was to evaluate osteoblast protein expression of collagen type 1 (col 1) and osteopontin (OPN) on titanium surface treated with double etching compared with a zirconia implant surface. MATERIALS AND METHODS: Discs were selected on both surfaces with respective treatments. Mouse MC3T3-E1 osteoblastic cells were tested for cell viability using the MTT assay. Subsequently, the quantification of col 1 and OPN secreted by osteoblastic cells plated on different surfaces was evaluated by enzyme linked immunosorbent assay (ELISA). RESULTS: Regarding the cell proliferation, statistical analysis showed no significant effect of surface-time interaction and viable cell count on the surfaces of titanium versus zirconia. No statistical differences between the surfaces of titanium and zirconia on cell viability was found. The zirconia surface expression of OPN was significantly higher, which occurred in all times. Furthermore, zirconia demonstrated significantly greater in col 1 expression in 48 and 96 hours compared with titanium. CONCLUSION: In this study, the zirconia surface demonstrated OPN and col 1 expression significantly higher as compared with titanium.
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Osteoblastos/citología , Titanio/química , Circonio/química , Animales , Adhesión Celular , Técnicas de Cultivo de Célula , Diferenciación Celular , Proliferación Celular , Supervivencia Celular , Células Cultivadas , Colágeno Tipo I/metabolismo , Materiales Dentales/química , Ensayo de Inmunoadsorción Enzimática , Técnicas In Vitro , Ensayo de Materiales , Ratones , Osteoblastos/metabolismo , Osteopontina/metabolismo , Propiedades de SuperficieRESUMEN
INTRODUCTION: In this study, we sought to assess the perception of changes in soft-tissue profile after Herbst appliance treatment by comparing facial profile silhouettes before treatment, immediately after treatment, and 2 years after treatment, as examined by orthodontists, general dentists, and laypersons. METHODS: The sample comprised 21 patients (mean age, 9.5 ± 0.5 years), who were treated with the Herbst appliance for a mean period of 12 (±1.1) months. Three lateral cephalograms were obtained at different times: baseline, immediately after removal of the Herbst appliance, and 2 years after removal of the appliance. The 63 resulting profile silhouettes were evaluated by 120 examiners divided into 3 groups: orthodontists, general dentists, and laypersons. The examiners were instructed to choose their preferred profile and note how much change they perceived across profiles using a visual analog scale. RESULTS: All groups of examiners preferred the posttreatment profiles. However, on quantitative assessment, the magnitudes of changes in the profile were found to be variable and rather small, with laypersons quantifying the greatest magnitude of change. CONCLUSIONS: Early Herbst appliance treatment produced positive changes in the facial profile that were visually appreciable both immediately after removal of the appliance and 2 years after treatment.
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Cara/anatomía & histología , Maloclusión Clase II de Angle/terapia , Aparatos Ortodóncicos Funcionales , Ortodoncia Correctiva/métodos , Cefalometría , Niño , Estética Dental , Femenino , Humanos , Masculino , Resultado del TratamientoRESUMEN
OBJECTIVE: The aim of this in vitro study was to evaluate the mechanical behavior and bacterial microleakage at the implant/abutment-tapered interface following mechanical cycling. MATERIALS AND METHODS: Two groups of screwless (Morse taper) implants (G1 and G2) and two groups of prosthetic screwed implants (G3 and G4) were tested. One group from each model (G2 and G4) were submitted to mechanical cycling, 500,000 cycles per sample, at a load of 120 N at 2 Hz prior to analysis. Microbiological analysis was performed via immersion of all samples in an Escherichia coli-containing suspension, incubated at 37 °C. After 14 days, the abutments were removed from their respective implants, registering the removal force (G1 and G2) or reverse torque (G3 and G4), and the presence of bacterial leakage was evaluated. Scanning electron microscopy (SEM) was performed to analyze the tapered surfaces of the selected samples. The Student t, binomial, and G tests were used for statistical analysis at a 5 % significance level. RESULTS: The results showed no significant difference between removal force, reverse torque, and contamination values when comparing implants of the same type. However, when the four groups were compared, contamination differed significantly (p = 0.044), with G1 having the least number of contaminated samples (8.3 %). SEM analysis showed superficial defects and damage. CONCLUSIONS: The abutment removal force or torque was not affected by mechanical cycling. Bacterial sealing of the implant/abutment tapered interface was not effective for any condition analyzed. Imprecise machining of implant parts does not allow a sufficient contact area between surfaces to provide effective sealing and prevent bacterial leakage. CLINICAL RELEVANCE: The microscopic gap caused by unsatisfactory implant/abutment adaptation, surface irregularities, and plastic deformation of all parts enabled bacterial contamination of the oral implants.
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Pilares Dentales/microbiología , Implantes Dentales/microbiología , Filtración Dental , Diseño de Implante Dental-Pilar , Análisis del Estrés Dental , Remoción de Dispositivos , Escherichia coli , Técnicas In Vitro , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Propiedades de Superficie , TorqueRESUMEN
Myoepithelial cells play a central role in glandular tumors, regulating the progression of in situ to invasive neoplasias, with the tumor microenvironment being shown to be involved in both initiation and progression. This study aimed to analyze the in vitro effects of fibroblast growth factor-2 (FGF-2) and hepatocyte growth factor (HGF) in myoepithelial cells under the influence of the fibronectin matrix extracellular protein. Benign myoepithelial cells were obtained from pleomorphic adenoma and cultured on a fibronectin substratum. FGF-2 and HGF were supplemented at different concentrations and time intervals, in order to evaluate cell proliferation, morphology and immunophenotype. Individually, FGF-2 and HGF supplementation did not alter myoepithelial cell proliferation, morphology or immunophenotype. The fibronectin substratum provoked an increase in cell proliferation and immunopositivity for α-smooth muscle actin and FGF-2. The myoepithelial cell morphology changed when the fibronectin substratum and FGF-2 acted together, highlighting the importance of the fibronectin extracellular matrix protein on the behavior of these cells.
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Adenoma Pleomórfico/patología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Fibronectinas/farmacología , Factor de Crecimiento de Hepatocito/farmacología , Neoplasias de las Glándulas Salivales/patología , Actinas/inmunología , Adenoma Pleomórfico/inmunología , Proliferación Celular , Células Cultivadas , Células Epiteliales/efectos de los fármacos , Células Epiteliales/inmunología , Células Epiteliales/patología , Humanos , Fenotipo , Neoplasias de las Glándulas Salivales/inmunologíaRESUMEN
During tumor development, benign neoplastic cells are influenced by the expression of cytokines, growth factors, and proteases present in the tumor microenvironment. Epidermal growth factor (EGF) is the most studied growth factor and is considered important for cell proliferation and migration. Metalloproteinases (MMPs) are also involved in tumor progression. The present study aimed to analyze the proliferation, viability and migration index of pleomorphic adenoma myoepithelial cells, in addition to the secretion of MMPs with EGF supplementation. Benign myoepithelial cells were cultured with two different EGF doses (5 and 10 ng/ml), and the influence of EGF on cell proliferation and viability, using trypan blue and MTT assays, respectively, after 24, 48, and 72 h, was evaluated. To analyze cellular morphology, hematoxylin-eosin staining and indirect immunofluorescence using the anti-vimentin antibody, was performed. In vitro migration assays were performed in Transwell chambers with an 8-µm pore covered with Matrigel and supplemented with 5 or 10 ng/ml of EGF, after 96 h. After 4 days of cell culture, ELISA was performed to determine the MMP-2 and MMP-13 levels. One-way analysis of variance (ANOVA) with post hoc Tukey test was applied, with a significance level of 0.05. The results revealed that EGF influences myoepithelial cell morphology, without alteration of proliferation and viability. The migration assay showed that EGF increased the mean index from 16 % in the control group to 40 and 76 % for 5 and 10 ng/ml of EGF, respectively. ELISA revealed that when the cells were supplemented with either of the EGF doses, an increase in MMP-2 levels was observed when compared with the control group (C). This study concludes that EGF aids in the production of MMP-2, which favors the dissolution of the basement membrane, contributing to cell migration and tumor progression, hence permitting contact between the myoepithelial cells and stroma.
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Adenoma Pleomórfico/metabolismo , Movimiento Celular , Factor de Crecimiento Epidérmico/fisiología , Metaloproteinasa 13 de la Matriz/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Neoplasias de las Glándulas Salivales/metabolismo , Adenoma Pleomórfico/enzimología , Adenoma Pleomórfico/patología , Proliferación Celular , Forma de la Célula , Células Epiteliales/enzimología , Células Epiteliales/metabolismo , Humanos , Mioepitelioma/enzimología , Mioepitelioma/metabolismo , Mioepitelioma/patología , Neoplasias de las Glándulas Salivales/enzimología , Neoplasias de las Glándulas Salivales/patología , Transducción de Señal , Células Tumorales CultivadasRESUMEN
The aim of this study is to validate a minimally invasive surgical procedure to harvest palate periosteum as a source of tissue for mesenchymal stromal/stem cells. We performed a standardized procedure to harvest the palate periosteum in ten subjects, which consisted of a 3 mm disposable punch and a Molt periosteal elevator to harvest a small full-thickness fragment of soft tissue at the hard palate area, between the upper bicuspids, 3 to 4 mm apical to the cement enamel junction. The one-third inner portion was fragmented, and following standard cell culture procedures, the adherent cells were cultured for three passages, after obtaining 70-90% confluence. Cell morphology analysis, flow cytometry analysis, and viability and osteogenic differentiation assays were performed. In all 10 cases, uneventful healing was observed, with no need for analgesic intake. The evaluation of cell morphology showed elongated spindle-shaped cells distributed in woven patterns. A high viability range was verified as well as an immunophenotype compatible with mesenchymal stem cell lineage. The differentiation assay showed the potential of the cells to differentiate into the osteogenic lineage. These results demonstrate that the minimally invasive proposed surgical technique is capable of supplying enough periosteum source tissue for stem cell culture and bone tissue engineering.
RESUMEN
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.
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Colágeno , Osteogénesis , Ratas , Masculino , Humanos , Animales , Ratas Wistar , Colágeno/química , Colágeno/farmacología , Regeneración Ósea , Cráneo/cirugíaRESUMEN
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.