Chemodiversity and Antibacterial Activity of the Essential Oil of Leaves of Croton argyrophyllus.

The Croton argyrophyllus leaf essential oil has anti-inflammatory, antioxidant, cytotoxic among other activities. However, there are chemical composition variations in the literature. This work reports the first study of the intraspecific chemical variation of C. argyrophyllus population from Pernambuco state, Brazil. The essential oils of nine specimens (OCA1 to OCA9) were analyzed by GC/MS and NIR to identify the chemical compositions and to verify the similarities between the analyzed samples. These analyses resulted in the identification of bicyclogermacrene (mean 38.42 %), (Z)-caryophyllene (mean of 14.06 %), epi-longipinanol (mean of 9.78 %) and germacrene B (mean of 9.1 %) as the major constituents, as well as the same chemical markers for all oil samples. However, these are different to those that were previously registered in the literature for C. argyrophyllus essential oil. The data obtained from the analysis by NIR spectroscopy were treated by PCA and HCA and showed similarities in the chemical samples' profile. By statistical analyses three clusters were obtained: OCA1-6, OCA7-8 and OCA9. All these groups were potentially active against Staphylococcus aureus. However, the OCA7-8 group was the most active.

Chitosan/Essential Oils Formulations for Potential Use as Wound Dressing: Physical and Antimicrobial Properties.

Film-forming emulsions and films, prepared by incorporating different concentrations of clove essential oil (CEO) and melaleuca essential oil (MEO) into chitosan (CS) were obtained and their properties were evaluated. Film-forming emulsions were characterized in terms of qualitative assessment, hydrogen potential and in vitro antibacterial activity, that was carried by the agar diffusion method, and the growth inhibition effects were tested on the Gram-positive microorganism of Staphylococcus aureus, Gram-negative microorganisms of Escherichia coli, and against isolated fungi such as Candida albicans. In order to study the impact of the incorporation of CEO and MEO into the CS matrix, the appearance and thickness of the films were evaluated. Furthermore, Fourier transform infrared spectroscopy (FTIR), contact angle measurements, a swelling test, scanning electron microscopy and a tensile test were carried out. Results showed that the film-forming emulsions had translucent aspect with cloudy milky appearance and showed antimicrobial properties. The CEO had the highest inhibition against the three strains studied. As regards the films' properties, the coloration of the films was affected by the type and concentration of bioactive used. The chitosan/CEO films showed an intense yellowish coloration while the chitosan/MEO films presented a slightly yellowish coloration, but in general, all chitosan/EOs films presented good transparency in visible light besides flexibility, mechanical resistance when touched, smaller thicknesses than the dermis and higher wettability than chitosan films, in both distilled water and phosphate-buffered saline (PBS). The interactions between the chitosan and EOs were confirmed by. The chitosan/EOs films presented morphologies with rough appearance and with EOs droplets in varying shapes and sizes, well distributed along the surface of the films, and the tensile properties were compatible to be applied as wound dressings. These results revealed that the CEO and MEO have a good potential to be incorporated into chitosan to make films for wound-healing applications.

N-Acetyl-D-Glucosamine-Loaded Chitosan Filaments Biodegradable and Biocompatible for Use as Absorbable Surgical Suture Materials.

The aim of this study was to prepare chitosan (CS) filaments incorporated with N-acetyl-D-Glucosamine (GlcNAc), using the wet spinning method, in order to combine the GlcNAc pharmacological properties with the CS biological properties for use as absorbable suture materials. The filaments were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), uniaxial tensile testing, in vitro biodegradation, and through in vitro drug release and cytotoxicity studies. It was observed that the addition of GlcNAc did not alter the morphology of the filaments. The CS and CS/GlcNAc filaments presented diameters 145 µm and 148 µm, respectively, and the surfaces were homogeneous. Although the mechanical resistance of the chitosan filaments decreased with the incorporation of the GlcNAc drug, this property was greater than the mean values indicated in the U.S. Pharmacopeia (1.7 N) for suture number 6-0 (filament diameter of 100-149 µm). The biodegradation of the CS filaments was accelerated by the addition of GlcNAc. After 35 days, the CS/GlcNAc filaments degradability was at its total, and for the CS filaments it was acquired in 49 days. The in vitro kinetic of the release process was of the zero-order and Hopfenberg models, controlled by both diffusion and erosion process. The in vitro cytotoxicity data of the CS and CS/GlcNAc filaments toward L929 cells showed that these filaments are nontoxic to these cells. Thus, the GlcNAc-loaded CS filaments might be promising as absorbable suture materials. In addition, this medical device may be able to enhance healing processes, relieve pain, and minimize infection at the surgery site due the prolonged release of GlcNAc.

Synthesis and Preparation of Chitosan/Clay Microspheres: Effect of Process Parameters and Clay Type.

This work aimed to prepare chitosan/clay microspheres, by the precipitation method, for use in drug carrier systems. The influence of the process parameters, particularly two airflows of the drag system (2.5 and 10 L·min-1) on the microspheres physical dimensions and properties, such as microstructure, degree of swelling and porosity were evaluated. The samples were characterized by optical microscopy (OM), scanning electron microscopy (SEM) and X-ray diffraction (XRD). Water absorption and porosity tests were also performed. The results showed that the process parameters affected the size of the microspheres. The diameter, volume and surface area of the chitosan/clay microspheres decreased when they were prepared with the higher airflow of the drag system. The microspheres presented a porous microstructure, being the pore size, percentage of porosity and degree of swelling affected not only by the process parameters but also by the type of clay. Hybrids (chitosan/clay) with intercalated morphology were obtained and the hybrid prepared with montmorillonite clay at higher airflows of the drag system presented the greatest interlayer spacing and a more disordered morphology. Thus, it is certain that the chitosan/clay nanocomposite microspheres prepared with montmorillonite (CL clay) at higher airflows of the drag system can have good drug-controlled release properties.

Preparation and Characterization of Chitosan Obtained from Shells of Shrimp (Litopenaeus vannamei Boone).

The main source of commercial chitosan is the extensive deacetylation of its parent polymer chitin. It is present in green algae, the cell walls or fungi and in the exoskeleton of crustaceans. A novel procedure for preparing chitosan from shrimp shells was developed. The procedure involves two 10-minutes bleaching steps with ethanol after the usual demineralization and deproteinization processes. Before deacetylation, chitin was immersed in 12.5 M NaOH, cooled down and kept frozen for 24 h. The obtained chitosan was characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), UV, X-ray diffraction (XRD) and viscosimetry. Samples of white chitosan with acetylation degrees below 9 % were obtained, as determined by FTIR and UV-first derivative spectroscopy. The change in the morphology of samples was followed by SEM. The ash content of chitosan samples were all below 0.063 % . Chitosan was soluble in 1 % acetic acid with insoluble contents of 0.62 % or less. XRD patterns exhibited the characteristic peaks of chitosan centered at 10 and 20 degrees in 2 θ . The molecular weight of chitosan was between 2.3 and 2.8 × 10 5 g/mol. It is concluded that the procedure developed in the present work allowed obtaining chitosans with physical and chemical properties suitable for pharmaceutical applications.

Anabolic Effect of Insulin Therapy on the Bone: Osteoprotegerin and Osteocalcin Up-Regulation in Streptozotocin-Induced Diabetic Rats.

Type 1 diabetes mellitus (T1DM) is associated with several skeletal alterations, particularly in conditions of poor glycaemic control. Insulin therapy is the major conservative treatment for T1DM; however, the effects of this hormone on bone markers of T1DM rats are limited, and the regulatory mechanisms remain elusive. Therefore, the evaluation of molecular and non-molecular parameters in a chronic animal model of T1DM-induced bone loss, treated with and without insulin, may help in elucidating the insulin mechanisms. Male Wistar rats were assigned into three groups: control, T1DM (T1DM rats induced with streptozotocin [STZ] at 40 mg/kg intravenously) and T1DM plus insulin therapy (T1DMI). After 8 weeks, we evaluated the serum biochemical, tibia histomorphometric and biomechanical parameters, as well as the gene expression of the receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG) and osteocalcin (OC) of femur mRNA. Compared with T1DM, the T1DMI group showed less bone loss, which was revealed by the increased trabecular width (TbWi, p < 0.001) and trabecular bone area (BAr, p < 0.01), reduced trabecular separation (TbSp, p < 0.01) and increased Young's modulus (p < 0.05). Moreover, molecular analyses indicated that the expression of OPG and OC was up-regulated (p < 0.001 and p < 0.05, respectively). In summary, the up-regulation of OPG and OC in the T1DMI group supports an anabolic effect of insulin, which was demonstrated by the maintenance of bone architecture and flexibility. These results suggest that insulin therapy may prevent T1DM-induced bone loss via the effects on the bone formation.

Protection against T1DM-Induced Bone Loss by Zinc Supplementation: Biomechanical, Histomorphometric, and Molecular Analyses in STZ-Induced Diabetic Rats.

Several studies have established an association between diabetes and alterations in bone metabolism; however, the underlying mechanism is not well established. Although zinc is recognized as a potential preventive agent against diabetes-induced bone loss, there is no evidence demonstrating its effect in chronic diabetic conditions. This study evaluated the effects of zinc supplementation in a chronic (90 days) type 1 diabetes-induced bone-loss model. Male Wistar rats were distributed in three groups: control, type 1 diabetes mellitus (T1DM), and T1DM plus zinc supplementation (T1DMS). Serum biochemical analysis; tibia histomorphometric, biomechanical, and collagen-content analyses; and femur mRNA expression were evaluated. Relative to T1DM, the zinc-supplemented group showed increased histomorphometric parameters such as TbWi and BAr and decreased TbSp, increased biomechanical parameters (maximum load, stiffness, ultimate strain, and Young's modulus), and increased type I collagen content. Interestingly, similar values for these parameters were observed between the T1DMS and control groups. These results demonstrate the protective effect of zinc on the maintenance of bone strength and flexibility. In addition, downregulation of OPG, COL1A, and MMP-9 genes was observed in T1DMS, and the anabolic effects of zinc were evidenced by increased OC expression and serum ALP activity, both related to osteoblastogenesis, demonstrating a positive effect on bone formation. In contrast, T1DM showed excessive bone loss, observed through reduced histomorphometric and biomechanical parameters, characterizing diabetes-associated bone loss. The bone loss was also observed through upregulation of OPG, COL1A, and MMP-9 genes. In conclusion, zinc showed a positive effect on the maintenance of bone architecture and biomechanical parameters. Indeed, OC upregulation and control of expression of OPG, COL1A, and MMP-9 mRNAs, even in chronic hyperglycemia, support an anabolic and protective effect of zinc under chronic diabetic conditions. Furthermore, these results indicate that zinc supplementation could act as a complementary therapy in chronic T1DM.

Avaliação da bioatividade de ligas de NiTi tratadas a laser para aplicação odontológica / Bioactivity evaluation of laser treated NiTi alloys for dental application

As ligas de NiTi são amplamente utilizadas em disposi-tivos da área odontológica, como na Implantodontia, devido as suas propriedades de biocompatibilidade, memória de forma e superelasticidade. Objetivo: Ava-liar a bioatividade e as características morfológicas de liga de NiTi (45,3% de Ni e 54,7% de Ti), tratadas a laser com e sem recobrimento de apatita. Materiais e método: As ligas de NiTi foram obtidas pela técnica Plasma Skull Push Pull, tratadas superficialmente. As amostras foram tratadas a laser (Yb:YAG) e pelo método biomimético em SBF-0 e SBF-6 (Simulated Body Fluid), ambas a 1,5M. A análise de bioatividade foi realizada por meio da pesagem, espectroscopia por energia dis-persiva de raios X (EDS) e microscopia eletrônica de varredura (MEV). Resultados: As espectroscopias reve-laram aumento na concentração de oxigênio e carbono após a ablação, favorecendo a difusão de átomos de oxigênio e formação de óxidos de titânio. Após a de-posição da apatita pelo método biomimético em SBF-0 e 6, o recobrimento apresentou uma razão molar de Ca/P de aproximadamente 1,0, caracterizando a forma-ção de apatitas de baixa cristalinidade. Conclusão: A associação da irradiação a laser e deposição química de apatitas pelo método biomimético em SBF-6, embo-ra tenha demonstrado um melhor desempenho quanto à bioatividade e topografia das ligas, o ganho de massa não foi estatisticamente significativo ao nível de 5%. Tais propriedades podem contribuir para maior inte-ração do biomaterial com o meio biológico e possível redução no tempo de osseointegração.