Development of Chlorhexidine Digluconate and Lippia sidoides Essential Oil Loaded in Microemulsion for Disinfection of Dental Root Canals: Substantivity Profile and Antimicrobial Activity.

The dental intracanal disinfection is crucial to achieve the success of endodontic treatment, avoiding the maintenance of endodontic infections. Chlorhexidine digluconate can act as an irrigating agent for it. However, it can cause tissue irritation in high concentrations. Therefore, combinations with other antimicrobial agents and more efficient therapeutic alternatives are studied, which make it possible to administer drugs more safely and with minimal adverse effects. Thus, the objective of this study was the development of a microemulsion containing chlorhexidine digluconate and essential oil of Lippia sidoides to be used for disinfection of dental root canals and to evaluate its profile of substantivity and antimicrobial activity. The microemulsions were obtained through phase diagrams, using the spontaneous formation method. We completed a physical-chemical characterization and evaluate the stability of the microemulsions, in addition to the substantivity profile in a bovine root dentin model, and in vitro antibacterial effect on Enterococcus faecalis. A method for quantifying chlorhexidine was developed using UV-Vis spectroscopy. The microemulsions showed acid pH, conductivity above 1.3 µScm-1, and dispersion index similar to water. The microemulsions showed antimicrobial inhibition halos similar to the commercial gel conventionally used, but with four times more substantivity to dentinal tissues. Microemulsions were obtained as a therapeutic alternative to formulations available on the market, presenting themselves as a system with great potential for the administration of drugs for disinfection of root canals.

In Situ Synthesis of Silver Nanoparticles in a Hydrogel of Carboxymethyl Cellulose with Phthalated-Cashew Gum as a Promising Antibacterial and Healing Agent.

Silver nanoparticles have been shown to possess considerable antibacterial activity, but in vivo applications have been limited due to the inherent, but low, toxicity of silver. On the other hand, silver nanoparticles could provide cutaneous protection against infection, due to their ability to liberate silver ions via a slow release mechanism, and their broad-spectrum antimicrobial action. Thus, in this work, we describe the development of a carboxymethyl cellulose-based hydrogel containing silver nanoparticles. The nanoparticles were prepared in the hydrogel in situ, utilizing two variants of cashew gum as a capping agent, and sodium borohydride as the reducing agent. This gum is non-toxic and comes from a renewable natural source. The particles and gel were thoroughly characterized through using rheological measurements, UV-vis spectroscopy, nanoparticles tracking analysis, and transmission electron microscopy analysis (TEM). Antibacterial tests were carried out, confirming antimicrobial action of the silver nanoparticle-loaded gels. Furthermore, rat wound-healing models were used and demonstrated that the gels exhibited improved wound healing when compared to the base hydrogel as a control. Thus, these gels are proposed as excellent candidates for use as wound-healing treatments.

Characterization and Biological Activities of Ocellatin Peptides from the Skin Secretion of the Frog Leptodactylus pustulatus.

Eight new peptides were isolated from the skin secretion of the frog Leptodactylus pustulatus and their amino acid sequences determined by de novo sequencing and by cDNA cloning. Structural similarities between them and other antimicrobial peptides from the skin secretion of Leptodactylus genus frogs were found. Ocellatins-PT1 to -PT5 (25 amino acid residues) are amidated at the C-terminus, while ocellatins-PT6 to -PT8 (32 amino acid residues) have free carboxylates. Antimicrobial activity, hemolytic tests, and cytotoxicity against a murine fibroblast cell line were investigated. All peptides, except for ocellatin-PT2, have antimicrobial activity against at least one Gram-negative strain. Ocellatin-PT8 inhibited the growth of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Salmonella choleraesuis strains with MICs in the 60-240 µM range. No significant effect was observed in human erythrocytes and in a murine fibroblast cell line after exposure to the peptides at MICs. A comparison between sequences obtained by both direct HPLC-MS de novo sequencing and cDNA cloning demonstrates the secretion of mature peptides derived from a pre-pro-peptide structure.

Antibacterial, antibiofilm and cytotoxic activities of Terminalia fagifolia Mart. extract and fractions.

BACKGROUND: The methicillin resistance of bacteria from the genus Staphylococcus and its ability to form biofilms are important factors in pathogenesis of these microorganisms. Thus, the search for new antimicrobials agents, especially from plants, has been intensified. In this context, Terminalia species have been the subject of research for many pharmacological activities. In this study we evaluated the antibacterial, antibiofilm and cytotoxic activities of the ethanol extract (EtE) from Terminalia fagifolia stem bark as well as that of three fractions of the extract (AqF, HaF and WSF). METHODS: We determined the minimum inhibitory concentration (MIC) by microdilution in 96-well plates, where the strains were exposed to serial dilutions of the ethanol extract and fractions, ranging from 12.5 to 400 µg/mL. We then determined the minimum bactericidal concentration (MBC), seeding the inoculum (10 µL) with concentrations equal to or greater than the MIC in Mueller-Hinton agar. To test the antibiofilm activity biofilm formation was induced in the presence of concentrations equivalent to 1/2, 1/4 and 1/8 of the MIC extract or fraction tested. In addition, the effect of the EtE and the fractions on cell viability was tested by the MTT assay on human MCF-7 breast cancer and mouse fibroblast NIH/3T3. To obtain high-resolution images of the effect of the aqueous fraction on the bacterial morphology, atomic force microscopy (AFM) imaging of treated S. aureus cells was performed. RESULTS: We observed antibacterial activity of EtE and fractions with MICs ranging from 25-200 µg/mL and MBCs ranging from 200-400 µg/mL. Regarding antibiofilm activity, both the EtE as the AqF, HaF and WSF fractions showed significant inhibition of the biofilm formation, with inhibition of biofilms formation of over 80% for some strains. The EtE and fractions showed a moderate cytotoxicity in cell line NIH/3T3 viability and potential antitumoral activity on human breast cancer cell line MCF-7. The microscopic images obtained revealed morphological changes to the S. aureus ATCC 29213 surface caused by AqF, as well as significant size alterations. CONCLUSIONS: The results show potential antibacterial, antibiofilm and antitumoral activities of the ethanol extract and fractions of T. fagifolia.

Chemical Characterization and Effect of a Lactobacilli-Postbiotic on Streptococcus mutans Biofilm In Vitro.

Postbiotic is the term used to define the soluble factors, metabolic products, or byproducts released by live probiotic bacteria or after its lysis. The objective of this study was to carry out the chemical characterization of the postbiotic of Lacticaseibacillus rhamnosus LR-32 and to evaluate its in vitro effect on the development of the Streptococcus mutans biofilm. After the cultivation of the probiotic strain, the postbiotic was extracted by centrifuging the culture and filtering the supernatant. This postbiotic was characterized by using gas chromatography coupled with mass spectrometry (GC-MS), and then it was used to determine the growth inhibition of S. mutans in its planktonic form; additionally, its effects on the following parameters in 48 h biofilm were evaluated: viable bacteria, dry weight, and gene expression of glucosyltransferases and VicR gene. The control group consisted of the biofilm without any treatment. A paired t-test was performed for statistical analysis, with the p-value set at 5%. Seventeen compounds of various chemical classes were identified in the postbiotic, including sugars, amino acids, vitamins, and acids. The treatment with the postbiotic led to an inhibition of the growth of S. mutans in its planktonic form, as well as a decrease in the number of viable bacteria, reduction in dry weight, and a negative regulation of the gene expression of gtfB, gtfC, gtfD, and vicR in its biofilm state, compared with the nontreated group (p < 0.05). The postbiotic of L. rhamnosus impaired the development of S. mutans biofilm.

Novel Scaffold Based on Chitosan Hydrogels/Phthalated Cashew Gum for Supporting Human Dental Pulp Stem Cells.

Hydrogels are structures that have value for application in the area of tissue engineering because they mimic the extracellular matrix. Naturally obtained polysaccharides, such as chitosan (CH) and cashew gum, are materials with the ability to form polymeric networks due to their physicochemical properties. This research aimed to develop a scaffold based on chitosan and phthalated cashew tree gum and test it as a support for the growth of human mesenchymal stem cells. In this study, phthalation in cashew gum (PCG) was performed by using a solvent-free route. PCG-CH scaffold was developed by polyelectrolyte complexation, and its ability to support adherent stem cell growth was evaluated. The scaffold showed a high swelling rate. The pore sizes of the scaffold were analyzed by scanning electron microscopy. Human dental pulp stem cells (hDPSCs) were isolated, expanded, and characterized for their potential to differentiate into mesenchymal lineages and for their immunophenotypic profile. Isolated mesenchymal stem cells presented fibroblastoid morphology, plastic adhesion capacity, and differentiation in osteogenic, adipogenic, and chondrogenic lineages. Mesenchymal stem cells were cultured in scaffolds to assess cell adhesion and growth. The cells seeded on the scaffold showed typical morphology, attachment, and adequate distribution inside the matrix pores. Thus, cells seeded in the scaffold may improve the osteoinductive and osteoconductive properties of these biomaterials.

Antibacterial application of natural and carboxymethylated cashew gum-based silver nanoparticles produced by microwave-assisted synthesis.

This study presents a green synthesis route to silver nanoparticles (AgNPs) stabilized with cashew gum (CG) or carboxymethylated cashew gum (CCG) using microwave-assisted synthesis and evaluates their antibacterial activity. The antimicrobial activity was measured by determining the minimum inhibitory concentration (MIC) with Staphylococcus aureus and Escherichia coli. In both cases of the presence of CG and CCG, it was found that higher pH lead to more efficient conversion of silver nitrate to AgNPs with well dispersed, spherical and stable particles as well as low crystallinity. CCG-capped AgNPs were slightly smaller (137.0 and 96.3 nm) than those coated with non-modified gum (144.7 and 100.9 nm). The samples presented promising antibacterial activity, especially on Gram-negative bacteria, resulting in significant membrane damage on treated bacteria in comparison to the untreated control, observed by atomic force microscopy. Thus, a quick and efficient synthesis route was applied to produce CGAgNPs and CCGAgNPs with antimicrobial potential.

Solvent-free production of phthalated cashew gum for green synthesis of antimicrobial silver nanoparticles.

This work describes a solvent-free method for the chemical modification of cashew gum (Anacardium occidentale L.) using phthalic anhydride in different proportions with different reaction times. Four biopolymers were synthesized and characterized by FTIR, NMR, and elemental analysis. A computational chemistry study was conducted to understand better the reaction. Phthalated cashew gum was used in preparation of silver nanoparticles (AgNPs) by a conventional route, using sodium borohydride (NaBH4) as reducing agent, and for green route. AgNPs were evaluated for antimicrobial activity and characterized by UV-Vis spectroscopy, FTIR, nanoparticle tracking analysis, Zeta Potential analysis, and atomic force microscopy. AgNPs produced by the green route had an average size of 51.9 nm and Zeta Potential of -55.8 mV, and AgNPs produced by the conventional method had an average size of 47.7 nm and Zeta Potential of -39.3 mV. AgNPs synthesized using phthalated cashew gum showed antimicrobial activity against Staphylococcus aureus and Escherichia coli.

Thaulin-1: The first antimicrobial peptide isolated from the skin of a Patagonian frog Pleurodema thaul (Anura: Leptodactylidae: Leiuperinae) with activity against Escherichia coli.

Patagonia's biodiversity has been explored from many points of view, however, skin secretions of native amphibians have not been evaluated for antimicrobial peptide research until now. In this sense, Pleurodema thaul is the first amphibian specie to be studied from this large region of South America. Analysis of cDNA-encoding peptide in skin samples allowed identification of four new antimicrobial peptides. The predicted mature peptides were synthesized and all of them showed weak or null antimicrobial activity against Klebsiella pneumoniae, Staphylococcus aureus and Escherichia coli with the exception of thaulin-1, a cationic 26-residue linear, amphipathic, Gly- and Leu-rich peptide with moderate antimicrobial activity against E. coli (MIC of 24.7µM). AFM and SPR studies suggested a preferential interaction between these peptides and bacterial membranes. Cytotoxicity assays showed that thaulin peptides had minimal effects at MIC concentrations towards human and animal cells. These are the first peptides described for amphibians of the Pleurodema genus. These findings highlight the potential of the Patagonian region's unexplored biodiversity as a source for new molecule discovery.

Descrição da morfologia dos frênulos labiais superiores em escolares de Teresina / Description of the Morphology of Upper Labial Frenulum in Students from Teresina

O objetivo desta pesquisa foi o de fazer um levantamento da prevalência dos diferentes tipos de frênulos labiais superiores, além de suas inserções em escolares de Teresina - PI. Para tanto, foram realizados exames clínicos em 300 escolares com idades entre 7 e 10 anos. As crianças foram escolhidas ao acaso, e o exame consistiu na distensão do lábio superior com o auxílio dos dedos de um único examinador, envoltos por gaze,sendo os achados anotados em ficha padronizada. Foi utilizada a classificação dos frênulos labiais superiores proposta por Sewerin (1971) e das inserções proposta por Placek et al. (1974). Os dados foram processados através do programa de computação SPSS, sendo a análise estatística do tipo descritiva. O frênulo labial de maior prevalência foi o simples, seguido do teto-labial persistente, simples com nódulo, coincidência de duasou mais variações ou anormalidades, simples com apêndice, com recesso, bífido e duplo. O tipo de inserção mais prevalente foi a inserção na mucosa alveolar, seguida da gengiva inserida, papila interdental e penetrantena papila

The objective of this paper was to estimate the prevalence and attachment of different kinds of upper lip frenulum in students from Teresina - PI. A sample of 300 students aged 7 to 10 years were clinicallyexamined. The children were chosen at random and the examiner distended their upper lip just using his fingers wrapped up in gauze. The findings were recorded on a standard chart. The classification for upperlabial frenulum proposed by Sewerin (1971) and that of the attachments proposed by Placek et al. (1974) were used The data were processed using the computation program SPSS and descriptive statistical analysis. The simple upper labial frenulum showed the largest prevalence, followed by the persistent tectolabial, simple with nodule, coincidence of two or more variations or abnormalities, simple with appendix, with recess, bifid and double. The attachment to alveolar mucous was the most prevalent, followed by inserted gum, interdental papilla and penetrating in the papilla attachments.

Parêmetros clínicos e atividade eletromiográfica em pacientes com disfunção temporomandibular / Clinical Parameters and Electromyographic Activity in Patients with Temporomandibular Disorder

Este estudo compara os parâmetros clínicos da palpação muscular e a auto percepção do lado de mastigaçãopreferido com a atividade eletromiográfica dos músculos masseter (feixe superficial) e temporal (feixe anterior).Seis pacientes dentados com diagnóstico de disfunção temporomandibular foram selecionados para ogrupo experimental; o grupo controle era formado por quatro voluntários sadios. Os músculos masseter etemporal direitos de ambos os grupos apresentaram maior atividade eletromiográfica. Os músculos masseteresdos pacientes com disfunções temporomandibulares apresentaram uma maior atividade eletromiográfica dosque os músculos temporais. A atividade elétrica indica uma tendência de hiperfunção, preferência de lado demastigação e dor articular e está primariamente associada com a atividade dos músculos masseteres