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1.
Adv Mater ; : e2001736, 2020 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-32700332

RESUMO

Biomaterial scaffolds have served as the foundation of tissue engineering and regenerative medicine. However, scaffold systems are often difficult to scale in size or shape in order to fit defect-specific dimensions, and thus provide only limited spatiotemporal control of therapeutic delivery and host tissue responses. Here, a lithography-based 3D printing strategy is used to fabricate a novel miniaturized modular microcage scaffold system, which can be assembled and scaled manually with ease. Scalability is based on an intuitive concept of stacking modules, like conventional toy interlocking plastic blocks, allowing for literally thousands of potential geometric configurations, and without the need for specialized equipment. Moreover, the modular hollow-microcage design allows each unit to be loaded with biologic cargo of different compositions, thus enabling controllable and easy patterning of therapeutics within the material in 3D. In summary, the concept of miniaturized microcage designs with such straight-forward assembly and scalability, as well as controllable loading properties, is a flexible platform that can be extended to a wide range of materials for improved biological performance.

2.
Lasers Med Sci ; 35(3): 767-775, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31407100

RESUMO

Papacarie Duo™ is clinically used and has proven effectiveness; however, it is necessary to improve its antimicrobial action. The combined treatment of Papacarie Duo™ with Urucum (Bixa Orellana) could create a potential tool for dental caries treatment; its extract obtained from the seeds' pericarp contains a water-soluble primary pigment (cis-bixin) with smaller amounts of other carotenoids. The dicarboxylic acid salts of cis-norbixin and trans-norbixin occur in heated alkaline solutions. To analyze the absorption spectra and cytotoxicity (with human dermal fibroblasts) in different concentrations of Urucum, associated or not with Papacarie Duo™, we performed this in vitro study. The effects of pure Urucum, Papacarie Duo™, and PapaUrucum™ on the microstructure of collagen were also analyzed. The application of papain-based gel with Urucum did not present cytotoxicity, its exhibit UV absorption spectrum peak around 460 ± 20 nm. Also, it showed that the compound used did not alter the chemical structure of collagen. Consequently, this product could be used as a chemomechanical method to remove dentin caries as well as being a potential product for antimicrobial photodynamic therapy (aPDT) application.

3.
Lab Chip ; 20(2): 405-413, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31854401

RESUMO

The tooth has a unique configuration with respect to biomaterials that are used for its treatment. Cells inside of the dental pulp interface indirectly with biomaterials via a calcified permeable membrane, formed by the dentin matrix and several thousands of dentinal tubules (∼2 µm in diameter). Although the cytotoxic response of the dental pulp to biomaterials has been extensively studied, there is a shortage of in vitro model systems that mimic the dentin-pulp interface and enable an improved understanding of the morphologic, metabolic and functional influence of biomaterials on live dental pulp cells. To address this shortage, here we developed an organ-on-a-chip model system which integrates cells cultured directly on a dentin wall within a microfluidic device that replicates some of the architecture and dynamics of the dentin-pulp interface. The tooth-on-a-chip is made out of molded polydimethylsiloxane (PDMS) with a design consisting of two chambers separated by a dentin fragment. To characterize pulp cell responses to dental materials on-chip, stem cells from the apical papilla (SCAPs) were cultured in odontogenic medium and seeded onto the dentin surface, and observed using live-cell microscopy. Next, to evaluate the tooth-on-a-chip as a platform for materials testing, standard dental materials used clinically (2-hydroxyethylmethacrylate - HEMA, phosphoric acid - PA, and Adper-Scotchbond - SB) were tested for cytotoxicity, cell morphology, and metabolic activity on-chip, and compared against standardized off-chip controls. All dental materials had cytotoxic effects in both on-chip and off-chip systems in the following order: HEMA > SB > PA (p < 0.05), and cells presented consistently higher metabolic activity on-chip than off-chip (p < 0.05). Furthermore, the tooth-on-a-chip enabled real-time tracking of gelatinolytic activity in a model hybrid layer (HL) formed in the microdevice, which suggests that dental pulp cells may contribute to the proteolytic activity in the HL more than endogenous proteases. In conclusion, the tooth-on-a-chip is a novel platform that replicates near-physiologic conditions of the pulp-dentin interface and enables live-cell imaging to study dental pulp cell response to biomaterials.

4.
Sci Rep ; 9(1): 10860, 2019 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-31350423

RESUMO

Direct visualization of the spatial relationships of the dental pulp tissue at the whole-organ has remained challenging. CLARITY (Clear Lipid-exchanged Acrylamide Tissue hYdrogel) is a tissue clearing method that has enabled successful 3-dimensional (3D) imaging of intact tissues with high-resolution and preserved anatomic structures. We used CLARITY to study the whole human dental pulp with emphasis on the neurovascular components. Dental pulps from sound teeth were CLARITY-cleared, immunostained for PGP9.5 and CD31, as markers for peripheral neurons and blood vessels, respectively, and imaged with light sheet microscopy. Visualization of the whole dental pulp innervation and vasculature was achieved. Innervation comprised 40% of the dental pulp volume and the vasculature another 40%. Marked innervation morphological differences between uni- and multiradicular teeth were found, also distinct neurovascular interplays. Quantification of the neural and vascular structures distribution, diameter and area showed that blood vessels in the capillary size range was twice as high as that of nerve fibers. In conclusion whole CLARITY-cleared dental pulp samples revealed 3D-morphological neurovascular interactions that could not be visualized with standard microscopy. This represents an outstanding tool to study the molecular and structural intricacies of whole dental tissues in the context of disease and treatment methods.

5.
Trials ; 19(1): 590, 2018 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-30373650

RESUMO

BACKGROUND: Halitosis is an unpleasant breath odour that can interfere with the professional life, social life and quality of life of people who suffer from it. A modality of treatment that has been increasing in dentistry is antimicrobial photodynamic therapy (aPDT). Bixa orellana, popularly known as "urucum" is a plant native to Brazil. The seeds are used to produce a dye that is largely used in the food, textile, paint and cosmetic industries. The aim of this study is to verify whether aPDT with Bixa orellana extract and blue light-emitting diodes (LEDs) is effective in reducing halitosis. This method will also be compared with tongue scraping, the most commonly used conventional method for tongue coating removal, and the association of both methods will be evaluated. METHODS/DESIGN: A randomized clinical trial will be conducted at the dental clinic of the Universidade Nove de Julho. Thirty-nine patients will be divided by block randomization into three groups (n = 13) according to the treatment to be performed. In Group 1, tongue scraping will be performed by the same operator in all patients for analysis of the immediate results. Patients will also be instructed on how to use the scraper at home. Group 2 will be treated with aPDT with Bixa orellana extract and the LED light curing device: Valo Cordless Ultradent®. Six points in the tongue dorsum with a distance of 1 cm between them will be irradiated. The apparatus will be pre-calibrated at wavelength 395-480 nm for 20 s and 9.6 J per point. In Group 3, patients will be submitted to the tongue scraping procedure, as well as to the previously explained aPDT. Oral air collection with the Oral Chroma™ and microbiological collections of the tongue coating shall be done before, immediately after and 7 days after treatment for comparison. DISCUSSION: Halitosis treatment is a topic that still needs attention. The results of this trial could support decision-making by clinicians regarding aPDT using blue LEDs for treating halitosis on a daily basis, as most dentists already have this light source in their offices. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03346460 . Registered on 17 November 2017.


Assuntos
Bixaceae , Luzes de Cura Dentária , Halitose/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Extratos Vegetais/uso terapêutico , Língua/efeitos dos fármacos , Adolescente , Adulto , Bixaceae/química , Brasil , Luzes de Cura Dentária/efeitos adversos , Feminino , Halitose/diagnóstico , Halitose/microbiologia , Humanos , Masculino , Fotoquimioterapia/efeitos adversos , Fármacos Fotossensibilizantes/efeitos adversos , Fármacos Fotossensibilizantes/isolamento & purificação , Extratos Vegetais/efeitos adversos , Extratos Vegetais/isolamento & purificação , Ensaios Clínicos Controlados Aleatórios como Assunto , Fatores de Tempo , Língua/microbiologia , Resultado do Tratamento , Adulto Jovem
6.
Lasers Med Sci ; 33(9): 1925-1931, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29931589

RESUMO

In this work, we present the efficacy of photodynamic therapy against yeast cells in an animal model. We tested two photosensitizers, methylene blue and protoporphyrin IX. Thirty-seven female BALB-c mice with a body mass of 20-25 g were used. To achieve persistent vaginitis, estrus was induced by subcutaneous injection of 0.1 mg/mL estradiol valerate applied weekly. Three days after pseudo-estrus, intravaginal inoculation with Candida albicans was performed. Mice were anesthetized with ketamine (80 mg/kg) and xylazine (10 mg/kg) by intraperitoneal injection before inoculation, and antimicrobial photodynamic therapy (aPDT) was performed 5 days after fungal inoculation. Two photosensitizers were tested, methylene blue (MB; 100 µM) and protoporphyrin IX (PpNetNI; 10 µM). Two custom-made LEDs emitting light at 660 and 630 nm at approximately 800 mW each were used for irradiation. The aPDT treatment reduced the fungal colony-forming units (CFUs) by one order of magnitude for the MB (p = 0.020) and PpNetNI (p = 0.018) photosensitizers. Seven days after the treatment, there were significantly fewer CFUs compared to the control group (p = 0.041 and p = 0.035 for MB and PpNetNI, respectively), but this was not increased compared to the initial number immediately after aPDT. Using aPDT as a therapeutic option to decrease fungal infection in a vaginal candidiasis model resulted in a significant reduction in the C. albicans population. Both photosensitizers were effective for preventing reinfection within 7 days. The aPDT also had no effect on the vaginal mucosa at the ultrastructural level. In addition to the fungicide effect, we observed reduced swelling and lack of the formation of abscesses, microabscesses coating the cornified epithelial layer, and the accumulation of neutrophils in the submucosa.


Assuntos
Anti-Infecciosos/uso terapêutico , Candidíase Vulvovaginal/tratamento farmacológico , Fotoquimioterapia , Animais , Candida albicans/efeitos dos fármacos , Candidíase Vulvovaginal/microbiologia , Feminino , Humanos , Inflamação/patologia , Azul de Metileno/farmacologia , Azul de Metileno/uso terapêutico , Camundongos Endogâmicos BALB C , Neutrófilos/metabolismo , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Protoporfirinas/farmacologia , Protoporfirinas/uso terapêutico , Vagina/efeitos dos fármacos , Vagina/microbiologia , Vagina/patologia
7.
Biomed Pharmacother ; 96: 1147-1153, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29191696

RESUMO

AIMS: The purpose of this study was toclarify the effect of low intensity pulsed ultrasound (LIPUS) on the phenotype of the inflammatory infiltrate in muscle tissue following acute injury. MAIN METHODS: Forty Wistar rats were submitted to cryoinjury of the tibialis anterior muscle and only half were treated with LIPUS. After 1, 2, 3 and 7 days macrophages and neutrophils were quantified. KEY FINDINGS: With one day, LIPUS led to reductions in the number of neutrophils and M1 macrophages. After two days, muscles treated with LIPUS had fewer total macrophages and M1 macrophages, but a greater number of M2 macrophages. Muscles treated with LIPUS showed fewer macrophages after three and seven days. SIGNIFICANCE: As the permanence of cells with pro-inflammatory and anti-inflammatory action can lead to the perpetuation of inflammation with consequent tissue damage and tissue fibrosis, respectively, the ability of LIPUS to modulate the occurrence of these cells demonstrates the therapeutic potential of this resource.


Assuntos
Macrófagos/efeitos da radiação , Músculo Esquelético/lesões , Músculo Esquelético/efeitos da radiação , Neutrófilos/efeitos da radiação , Fenótipo , Ondas Ultrassônicas , Animais , Inflamação/patologia , Inflamação/terapia , Macrófagos/patologia , Masculino , Músculo Esquelético/patologia , Neutrófilos/patologia , Ratos , Ratos Wistar
8.
PLoS One ; 12(11): e0187945, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29145431

RESUMO

To evaluate whether periodontitis modulates lung inflammation in an experimental model of asthma as well as the photodynamic therapy (PDT) is associated with a reduction of lung inflammation. Seventy-two BALB/c male mice (~2 months) were randomly divided into 8 groups (n = 9): Basal, Periodontitis (P), P+PT, P+PT+PDT, Asthma (A), A+P, A+P+PT, and A+P+PT+PDT. Periodontitis was induced by using the ligature technique and asthma was induced by ovalbumin (OVA). PT was performed with curettes and PDT with methylene blue (0.005%), λ = 660nm, with a radiant exposure of 318J/cm2. After 43 days, euthanasia was carried out prior to lung and mandible morphological analyzes. All of the manipulations of the animals were performed by only one operator. The total and differential cell counts and cytokines IL-4, IL-5, IL-10, IFN-γ, TNF-α, IL-1ß, and IL-6 were evaluated in the bronchoalveolar lavage (BAL) and in the serum. Mucus and alkaline phosphatase were also quantified. Statistical analyzes were performed by a blinded statistician. One-way analysis of variance (ANOVA) was employed, followed by the Student-Newman-Keuls test. Periodontitis group (P) increased alkaline phosphatase and bone resorption (p<0.05), validating the experimental model of periodontitis. The A group and the P group increased the total amount of cells (p <0.05) in the BAL. However, in the A+P group, there was a decrease in these cells, except for in the A+P+PT+PDT group (p<0.05). The asthma group increased the Th2 cytokines and P group increased the Th1 cytokine profile, and A+P+PT+PDT group increased IL-10 cytokine. Mucus was increased for the A and P groups. In conclusion, periodontitis in the asthmatic mice reduced the inflammatory migrated cells in the BAL (eosinophils, lymphocytes, macrophages). In addition, it reduced the levels of the IL-4 and TNF-α cytokines, which was also accompanied by a decreased mucus production. After PDT treatment the total cell count increased however, this increase was not accompanied by a pro-inflammatory cytokines release. Only in PDT group the anti-inflammatory IL-10 was increased. Further studies are needed to understand this mechanism of action.


Assuntos
Asma/complicações , Periodontite/complicações , Fotoquimioterapia , Animais , Asma/metabolismo , Citocinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Periodontite/metabolismo
9.
Sci Rep ; 7(1): 13687, 2017 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-29057895

RESUMO

We previously reported that cholinergic stimulation with pyridostigmine (PY) induces anti-inflammatory cell recruitment soon after myocardial infarction (MI). In this study, we evaluated the anti-inflammatory effects of PY during the proliferative phase of cardiac repair by analyzing the infiltration of macrophages, Treg lymphocytes, oxidative stress and inflammatory cytokines. Wistar rats underwent control sham surgery or ligation of the left coronary artery and were randomly allocated to remain untreated (untreated infarcted group, I) or to receive PY (30 mg·kg(-1)·day(-1)) in the supplied water (infarcted treated group, I + PY). Blood pressure and heart rate variability were registered at day 5 post-MI. The animals were euthanized 7 days after thoracotomy, when the hearts were removed and processed for immunohistochemistry (CD68, CD206, FOXP3), cytokines (IL-1ß, IL-6, IL-10, TNF-α) and oxidative stress (superoxide dismutase, catalase, glutathione peroxidase, lipidic and protein peroxidation). PY treatment increased parasympathetic modulation, M2 macrophages and the anti-oxidant enzyme activity but reduced protein oxidation (carbonyls) and the concentration of IL-1ß, IL-6, TNF-α and IL-10. Cholinergic stimulation induces parasympathetic neuro-immune modulation and anti-inflammatory cell enrollment as well as prevents oxidative stress and cytokine production after MI.


Assuntos
Cardiotônicos/farmacologia , Inibidores da Colinesterase/farmacologia , Inflamação/tratamento farmacológico , Infarto do Miocárdio/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Brometo de Piridostigmina/farmacologia , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Citocinas/metabolismo , Modelos Animais de Doenças , Hemodinâmica/efeitos dos fármacos , Inflamação/metabolismo , Inflamação/patologia , Masculino , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Estresse Oxidativo/fisiologia , Distribuição Aleatória , Ratos Wistar
10.
Rev Assoc Med Bras (1992) ; 63(1): 29-34, 2017 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-28225874

RESUMO

OBJECTIVE: To analyze morphological characteristics and organization of the collagen fibers of third degree burns from scalding compared to laser therapy and silver sulfadiazine, the latter considered as the gold standard. METHOD: Were selected 12 animals (Rattus norvegicus) also divided into three groups (control group [CG] - untreated burns; sulfadiazine group [SG] - burns were treated with silver sulfadiazine at 1%; laser group [LG] - burns were treated with photobiomodulation). The scald burns were carried out by using PVC mold, and the material collected on the 14th day after burn was prepared for morphological and optical retardation analysis for evaluation of inflammatory infiltrates and collagen organization, respectively. RESULTS: On the 14th day, the laser and sulfadiazine groups had mild inflammatory response, while the control group showed an intense inflammatory process, with statistical significance between laser and control groups, but not between sulfadiazine and control groups. Laser and sulfadiazine groups no longer had granulation tissue, opposite to what was seen in the control group. The presence of hair follicles and ulcer did not significantly differ between groups. The optical retardation of collagen fibers was higher in sulfadiazine group, followed by laser and control groups. As for systemic effect, we were able to identify it by simply analyzing the presence or absence of granulation tissue. CONCLUSION: Morphologically, the laser or silver sulfadiazine treatments were similar and both provided better organization of collagen fibers in relation to the untreated group. However, the sulfadiazine group modulated the deposition of collagen fibers more efficiently than the laser group.


Assuntos
Queimaduras/tratamento farmacológico , Queimaduras/radioterapia , Terapia com Luz de Baixa Intensidade/métodos , Sulfadiazina de Prata/administração & dosagem , Animais , Queimaduras/patologia , Colágeno/efeitos dos fármacos , Colágeno/efeitos da radiação , Modelos Animais de Doenças , Feminino , Ratos , Fatores de Tempo , Cicatrização/fisiologia
11.
Rev. Assoc. Med. Bras. (1992) ; 63(1): 29-34, Jan. 2017. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-842517

RESUMO

Summary Objective: To analyze morphological characteristics and organization of the collagen fibers of third degree burns from scalding compared to laser therapy and silver sulfadiazine, the latter considered as the gold standard. Method: Were selected 12 animals (Rattus norvegicus) also divided into three groups (control group [CG] - untreated burns; sulfadiazine group [SG] - burns were treated with silver sulfadiazine at 1%; laser group [LG] - burns were treated with photobiomodulation). The scald burns were carried out by using PVC mold, and the material collected on the 14th day after burn was prepared for morphological and optical retardation analysis for evaluation of inflammatory infiltrates and collagen organization, respectively. Results: On the 14th day, the laser and sulfadiazine groups had mild inflammatory response, while the control group showed an intense inflammatory process, with statistical significance between laser and control groups, but not between sulfadiazine and control groups. Laser and sulfadiazine groups no longer had granulation tissue, opposite to what was seen in the control group. The presence of hair follicles and ulcer did not significantly differ between groups. The optical retardation of collagen fibers was higher in sulfadiazine group, followed by laser and control groups. As for systemic effect, we were able to identify it by simply analyzing the presence or absence of granulation tissue. Conclusion: Morphologically, the laser or silver sulfadiazine treatments were similar and both provided better organization of collagen fibers in relation to the untreated group. However, the sulfadiazine group modulated the deposition of collagen fibers more efficiently than the laser group.


Resumo Objetivo: Analisar características morfológicas e organização das fibras colágenas de queimaduras de terceiro grau provocadas por escaldo em relação à terapia com laser e àquela considerada padrão-ouro, a sulfadiazina de prata. Método: Foram selecionados 12 animais (Rattus norvegicus), divididos igualmente em três grupos (grupo controle [GC] - queimaduras não tratadas; grupo sulfadiazina [GS] - queimaduras tratadas com sulfadiazina de prata 1%; grupo laser [GL] - queimaduras tratadas com fotobiomodulação). As queimaduras foram realizadas por escaldo com a utilização de molde de PVC, e o material coletado no 14º dia pós-queimadura foi preparado para análise morfológica e de retardo óptico, para avaliação do infiltrado inflamatório e da organização do colágeno, respectivamente. Resultados: No 14º dia, os grupos laser e sulfadiazina apresentaram resposta inflamatória leve, enquanto o grupo controle apresentou processo inflamatório intenso, havendo significância estatística entre os grupos laser e controle, mas não entre os grupos sulfadiazina e controle. Enquanto os grupos laser e sulfadiazina não apresentavam mais tecido de granulação, o grupo controle ainda apresentava. A presença de folículo piloso e de úlcera não diferiu significantemente entre os grupos. O retardo óptico das fibras colágenas foi maior no grupo sulfadiazina, seguido dos grupos laser e controle. Apenas a análise da presença ou ausência de tecido de granulação permitiu identificar o efeito sistêmico. Conclusão: Morfologicamente, os tratamentos com laser ou sulfadiazina de prata foram similares e ambos proporcionaram maior organização das fibras colágenas em relação ao grupo não tratado. Entretanto, o grupo sulfadiazina modulou a deposição das fibras colágenas mais eficientemente que o grupo laser.

12.
Clin Oral Investig ; 21(5): 1681-1685, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27600723

RESUMO

OBJECTIVES: The aim of this study is to investigate whether patients with multiple sclerosis (MS) have more orofacial dysfunctions than the general population, using the Nordic Orofacial Test-Screening (NOT-S). MATERIALS AND METHODS: The NOT-S instrument was applied in 34 patients with MS, who went to the MS Reference Center, Universidade Metropolitana de Santos and 34 healthy patients, matched for gender and age. NOT-S results were compared between patients with MS and control subjects. Disability and disease duration were assessed among the patients, in order to establish whether these parameters might affect the results from NOT-S. RESULTS: There was no significant difference in orofacial function between patients with MS and control subjects. There was no statistically significant correlation between disability and NOT-S or between disease duration and NOT-S. However, the correlation between disease duration and the degree of disability was statistically significant, thus suggesting that the results are in accordance with what would be expected regarding MS. CONCLUSIONS: These results indicate that there was no correlation between orofacial dysfunction and MS, although there were some differences in the affected domains. CLINICAL RELEVANCE: This study points out the orofacial dysfunctions which health professionals should be aware in this population.


Assuntos
Transtornos de Deglutição/fisiopatologia , Avaliação da Deficiência , Dor Facial/fisiopatologia , Esclerose Múltipla/fisiopatologia , Sensação/fisiologia , Distúrbios da Fala/fisiopatologia , Xerostomia/fisiopatologia , Adolescente , Adulto , Estudos de Casos e Controles , Expressão Facial , Feminino , Humanos , Masculino , Mastigação/fisiologia , Pessoa de Meia-Idade , Respiração
13.
Sci Rep ; 6: 33270, 2016 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-27641507

RESUMO

This study intended to evaluate the effects of a papain-gel with a red-light absorbing pigment (methylene blue - MB) to mediate photodynamic therapy (PDT) against Streptococcus mutans biofilms. The PapaMBlue was compared with free MB to generate reactive oxygen species using fluorescence probes (SOSG and HPF). PDT (660-nm light) was carried out against S. mutans biofilms grown on either plastic dishes or on collagen membrane and assayed by CFU, live-dead staining using confocal microscopy, transmission electron microscopy and H&E staining for collagen films. Cytotoxicity and subcellular localization was studied in human fibroblasts. Sponges of bioabsorbable type I collagen membrane were exposed to papain based gel, irradiated with laser and analyzed about their integrity by ATR-FTIR. The PapaMBlue produced higher amounts of singlet oxygen and hydroxyl radicals than free MB, possibly due to better disaggregation of the dye in solution. The PapaMBlue antimicrobial effects on biofilms proved to be capable of reducing the S. mutans. Both MTT and PrestoBlue assays showed higher cell viability and metabolism scores in fibroblasts treated with PapaMBlue and MB, possibly due to stimulation of mitochondrial activity and that collagen triple helix is unaffected. The PapaMBlue is equally effective as MB in destroying S. mutans biofilms growing on plastic or collagen without affecting fibroblasts.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Cárie Dentária/tratamento farmacológico , Azul de Metileno/farmacologia , Papaína/farmacologia , Streptococcus mutans/efeitos dos fármacos , Cárie Dentária/microbiologia , Géis , Humanos , Radical Hidroxila/metabolismo , Radical Hidroxila/farmacologia , Azul de Metileno/metabolismo , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Papaína/metabolismo , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Oxigênio Singlete/metabolismo , Oxigênio Singlete/farmacologia , Streptococcus mutans/fisiologia , Streptococcus mutans/ultraestrutura
14.
J Biophotonics ; 9(11-12): 1337-1343, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27248838

RESUMO

Halitosis can exert a negative influence on the social relations of adolescents and affect one's self-image. The aim of this study was to evaluate the clinical and microbiological effect of antimicrobial photodynamic therapy (aPDT) on halitosis in adolescents. Forty-six individuals aged 12 to 19 years were randomly allocated: Group 1 - treatment with photodynamic therapy; Group 2 - treatment with a tongue scraper and Group 3 - treatment with a tongue scraper and photodynamic therapy. The count of bacterial colony-forming units per milliliter was used for the microbiological analysis. Statistical analysis involved the Kruskal-Wallis test followed by the Student-Newman-Keuls test. ANOVA was used for the determination of colony-forming units after treatment. The level of significance for all statistical tests was 5% (p < 0.05). After treatment, a statistically significant reduction in total volatile sulfur compounds was found in all groups (p < 0.001), with the largest reduction (median: 0) found in Group 3 (tongue scraper and photodynamic therapy). Moreover, a statistically significant difference was found between treatment with aPDT and a tongue scraper alone (p < 0.001). The present findings demonstrate an option for the treatment of halitosis in adolescents, with an immediate effect and without the mechanical aggression to the toungue. Clinical Trials: NCT02007993.


Assuntos
Halitose/tratamento farmacológico , Fotoquimioterapia , Adolescente , Criança , Feminino , Humanos , Masculino , Compostos de Enxofre/análise , Língua/microbiologia , Adulto Jovem
16.
Am J Physiol Regul Integr Comp Physiol ; 310(8): R697-706, 2016 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-26791829

RESUMO

We tested the hypothesis that an increase in the anti-inflammatory cholinergic pathway, when induced by pyridostigmine (PY), may modulate subtypes of lymphocytes (CD4+, CD8+, FOXP3+) and macrophages (M1/M2) soon after myocardial infarction (MI) in rats. Wistar rats, randomly allocated to receive PY (40 mg·kg(-1)·day(-1)) in drinking water or to stay without treatment, were followed for 4 days and then were subjected to ligation of the left coronary artery. The groups-denominated as the pyridostigmine-treated infarcted (IP) and infarcted control (I) groups-were submitted to euthanasia 3 days after MI; the heart was removed for immunohistochemistry, and the peripheral blood and spleen were collected for flow cytometry analysis. Noninfarcted and untreated rats were used as controls (C Group). Echocardiographic measurements were registered on the second day after MI, and heart rate variability was measured on the third day after MI. The infarcted groups had similar MI areas, degrees of systolic dysfunction, blood pressures, and heart rates. Compared with the I Group, the IP Group showed a significant higher parasympathetic modulation and a lower sympathetic modulation, which were associated with a small, but significant, increase in diastolic function. The IP Group showed a significant increase in M2 macrophages and FOXP3(+)cells in the infarcted and peri-infarcted areas, a significantly higher frequency of circulating Treg cells (CD4(+)CD25(+)FOXP3(+)), and a less extreme decrease in conventional T cells (CD25(+)FOXP3(-)) compared with the I Group. Therefore, increasing cholinergic modulation with PY induces greater anti-inflammatory cell recruitment soon after MY in rats.


Assuntos
Anti-Inflamatórios/farmacologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Quimiotaxia de Leucócito/efeitos dos fármacos , Neurônios Colinérgicos/efeitos dos fármacos , Inibidores da Colinesterase/farmacologia , Macrófagos/efeitos dos fármacos , Infarto do Miocárdio/tratamento farmacológico , Miocárdio/imunologia , Brometo de Piridostigmina/farmacologia , Linfócitos T Reguladores/efeitos dos fármacos , Animais , Biomarcadores/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Neurônios Colinérgicos/metabolismo , Neurônios Colinérgicos/patologia , Modelos Animais de Doenças , Fatores de Transcrição Forkhead/metabolismo , Frequência Cardíaca/efeitos dos fármacos , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Infarto do Miocárdio/imunologia , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miocárdio/metabolismo , Miocárdio/patologia , Fenótipo , Ratos Endogâmicos WKY , Baço/efeitos dos fármacos , Baço/imunologia , Baço/metabolismo , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Fatores de Tempo , Função Ventricular Esquerda/efeitos dos fármacos
17.
Lasers Med Sci ; 31(1): 41-7, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26510574

RESUMO

UNLABELLED: Light with or without chemical agents has been used to induce therapeutic and antimicrobial effects. With photodynamic therapy, the antimicrobial effect is confined to areas covered by a photosensitive dye and irradiated with light. The aim of the present study was to evaluate the effect of photodynamic therapy for the treatment of halitosis in adolescents through the analysis of volatile sulfur compounds, especially sulfide. A controlled, clinical trial was conducted with 45 adolescents randomly allocated to three groups: group 1, photodynamic therapy administered to the dorsum of the tongue; group 2, treatment with a tongue scraper; and group 3, treatment with a tongue scraper combined with photodynamic therapy. The diagnosis of halitosis was performed using gas chromatography before and after treatment. Comparisons were made using the Kruskal-Wallis test followed by the Student-Newman-Keuls test, with the level of significance set at 5 % (p < 0.05). After treatment, a statistically significant reduction in halitosis was found in all groups (p < 0.001). The greatest reduction in total sulfides (median = 0) occurred with the combination of tongue scraper and photodynamic therapy. The present study describes a novel option for the treatment of halitosis in adolescents with an immediate effect that does not involve the mechanical aggression of the lingual papillae that occurs with conventional treatment. TRIAL REGISTRATION: Photodynamic Therapy in Adolescents Halitosis ( https://clinicaltrials.gov/ct2/show/NCT02007993?term=NCT02007993&rank=1 )Number: NCT02007993FUNDING:FAPESPNumber: 2013/13032-8.


Assuntos
Halitose/tratamento farmacológico , Fotoquimioterapia , Adolescente , Feminino , Halitose/metabolismo , Humanos , Masculino , Sulfetos/metabolismo , Língua , Resultado do Tratamento
18.
Lasers Surg Med ; 48(1): 45-51, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26627532

RESUMO

BACKGROUND AND OBJECTIVE: Diabetes alters innate and specific immunity, causing an imbalanced tissue repair process. Very active neutrophils and macrophages are found for a long time in chronic wounds in those individuals. The aim of this study was to evaluate the response of the main effector cells of immunity (neutrophils, macrophages, and T lymphocytes) and to compare the effects of two laser therapy regimens in the postoperative treatment of excision wounds. METHOD: Diabetes was induced in female Wistar rats and a punch was used to cause wounds in the dorsum of each individual. The animals were randomly allocated to a control group (CG), in which the wound was untreated, a single-dose laser group (SLG), in which the wound was submitted to single dose of laser therapy at wavelength of 660 nm, output power of 30 mW, energy density of 4 J/cm(2), and 26-second exposure time, and a fractionated-dose laser group (FLG), submitted to 1 J/cm(2) of laser therapy on Days 1, 3, 8, and 10. Euthanasia was performed on five animals from each group Days 1, 3, 8, 10, 15, and 22. The wound was removed and routinely processed for immunohistochemistry against elastase, CD3, CD68, and CD206 antibodies. The samples were photographed and labeled cells were counted by a blinded observer. The Kruskal-Wallis test was used for the statistical analysis. RESULTS: Neutrophils were predominant in the SLG on Day 1, whereas these cells were mostly found in the CG on Day 3 (P < 0.05). The T lymphocyte count was similar in all groups in the throughout the experiment. On Day 3, the SLG exhibited a greater number of total macrophages (CD68+) (P < 0.05), whereas the macrophage count was similar among the different groups on the other evaluation days. The CD206 + cell counts revealed that the SLG had more M2 macrophages than the CG on Day 8 (P < 0.05), whereas the FLG exhibited more M2 macrophages than the CG on Day 10 (P < 0.05). CONCLUSION: The present findings demonstrate that laser therapy can alter the composition of inflammatory infiltrate in diabetic wounds, leading to a more balanced response transiting from a rapid neutrophil infiltration through to M2 macrophage polarization, especially with a single application of 4 J/cm(2) in the immediate postoperative period.


Assuntos
Terapia a Laser , Macrófagos/imunologia , Macrófagos/efeitos da radiação , Neutrófilos/imunologia , Neutrófilos/efeitos da radiação , Linfócitos T/imunologia , Linfócitos T/efeitos da radiação , Cicatrização/imunologia , Cicatrização/efeitos da radiação , Animais , Diabetes Mellitus Experimental , Feminino , Distribuição Aleatória , Ratos , Ratos Wistar
19.
J Lasers Med Sci ; 7(3): 159-162, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28144435

RESUMO

Introduction: The aim of the present study was to determine the effective transmission of 660 and 780 nm lasers through mandibular and maxillary alveolar bones in the buccal-lingual/ palatal direction. Methods: The laser probe was positioned in direct contact with the surface of the anterior, middle and posterior regions of each bone (5 dried maxillae and 5 mandibles) and the power meter was positioned on the bone wall opposite to the radiated wall for the measure of the remaining energy passing through the bone tissue. Ten measurements were performed with each laser at each irradiated point. Results: Transmitted power was significantly higher in bones irradiated with 780 nm laser. Tendencies toward greater average power transmitted in the anterior region of both bones at both wavelengths were also observed. Conclusion: Dosimetry and the choice of light source may be adjusted according to the anatomic region of the alveolar bone to be treated.

20.
PLoS One ; 10(8): e0134858, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26267811

RESUMO

Immune escape and metastasis are the hallmarks of several types of cancer including bladder cancer. One of the mechanisms involved in these processes has been linked to indoleamine 2,3-dioxygenase (IDO). Although IDO is classically recognized for its immunomodulatory property, it has presented nonimmunological effects in some tumors. TGF-ß1 is believed to contribute to carcinoma development by modulating immunossupressive molecules, including IDO. In addition, TGF-ß1 induces the epithelial-mesenchymal transition (EMT), which is a critical step in the tumor invasiveness and metastasis. We investigated the role of MT and IDO modulation in the induction of EMT by TGF-ß1 in T24 human bladder carcinoma cells. When T24 cells were incubated with the IDO inhibitor (MT, 1-methyl-D-tryptophan), with TGF-ß1, and with MT+TGF-ß1, a significant decrease of IDO expression and activity was observed. In addition, downregulation of e-cadherin and upregulation of n-cadherin and EMT transcription factors were induced by the treatments, confirming the induction of EMT. siRNA-mediated knockdown of IDO decreased e-cadherin expression, but had no effect on EMT transcription factors. In the scratch-wound assay, the heightened migration process was intensified when the cells were incubated with MT+TGF-ß1. These effects were associated with a robust inhibition of Akt activation. After inoculation of T24 cells under the kidney capsule of Balb/c nude, the cells were positive for IDO in the center of the cell infiltrate, being negative in the periphery, where EMT is high. In conclusion, inhibition of IDO by TGF-ß1 and MT is associated with EMT in T24 human bladder carcinoma cells. MT has potentiating effect in TGF-ß1-induced EMT, independently of IDO. This nonimmunological effect of MT should be considered if IDO is the target to avoid immune escape in bladder cancer.


Assuntos
Indolamina-Pirrol 2,3,-Dioxigenase/biossíntese , Fator de Crescimento Transformador beta1/metabolismo , Triptofano/administração & dosagem , Neoplasias da Bexiga Urinária/genética , Animais , Caderinas/biossíntese , Linhagem Celular Tumoral , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Indolamina-Pirrol 2,3,-Dioxigenase/antagonistas & inibidores , Indolamina-Pirrol 2,3,-Dioxigenase/genética , Camundongos , Invasividade Neoplásica/genética , Invasividade Neoplásica/imunologia , Metástase Neoplásica , RNA Interferente Pequeno , Fator de Crescimento Transformador beta1/administração & dosagem , Fator de Crescimento Transformador beta1/imunologia , Triptofano/análogos & derivados , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/imunologia , Neoplasias da Bexiga Urinária/patologia , Ensaios Antitumorais Modelo de Xenoenxerto
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