Treatment of oxidative stress-induced pain and inflammation with dexketoprofen trometamol loaded different molecular weight chitosan nanoparticles: Formulation, characterization and anti-inflammatory activity by using in vivo HET-CAM assay.

Angiogenesis is a fundamental process of wound healing, embryogenesis etc. but occurs in cancer and chronic inflammation pathologically. HET-CAM assay is a useful, well established and animal alternative test to screen anti-inflammatory potentials of pharmaceutical products as well as nano-formulations. Dexketoprofen trometamol (DT) belongs to the nonsteroidal anti-inflammatory drug (NSAID) group which is a rapidly acting analgesic ingredient. Because DT has a short half-life, high and frequent dosing is used in treatment. The need of design and producing a new oral prolonged-release dosage form containing DT is the major aim of the study with low dose and low side effects. Chitosan (CS) has been widely used in the pharmaceutical area because of its favorable biological properties. In this study, DT loaded CS nanoparticles (CS-NPs) were produced by spray drying method for oral drug delivery. Structures of CS-NPs were elucidated by particle size, zeta potential, SEM, DSC, FT-IR and 1H NMR. High encapsulation efficiency was obtained (73-84%) for the prepared formulations. In vitro release was examined in pH 1.2 buffer and pH 6.8 buffer. DT-loaded CS-NPs showed prolonged release, particularly at pH 6.8. Weibull kinetic model was found to fit best to DT release from CS-NPs in both release medium. The anti-inflammatory activity of optimum formulation (M-DT) was examined using the in vivo HET-CAM assay. The anti-inflammatory activity results indicated that M-DT coded NPs formulation showed significantly good anti-inflammatory potential with closer inhibition value to the standard anti-inflammatory DT at one fifth lower dosage. According to the proposed method and results it can be successfully applicable to the NP preparation containing DT and it could be concluded that DT loaded CS-NPs seem to be a promising prolonged release drug delivery system for oral administration with low dose and high efficiency.

Diclofenac sodium loaded PLGA nanoparticles for inflammatory diseases with high anti-inflammatory properties at low dose: Formulation, characterization and in vivo HET-CAM analysis.

The development of a new drug active substance is not only time-consuming and expensive, but also a chain of operations that often fails. However, increasing the bioavailability, effectiveness, safety, or targeting the drugs used in clinic by various methods, such as nanoparticles (NPs), may be a more effective way of using them in clinic. In addition, NP formulations are becoming increasingly popular in modern medical treatments. Angiogenesis, formation of new capillaries from a pre-existing one, fundamentally occurs in physiological processes such as wound healing, embryogenesis and menstrual cycle, also has a vital role in pathology of cancer, psoriasis, diabetic retinopathy and chronic inflammation. The Hen's Egg Test on the Chorioallantoic Membrane (HET-CAM) assay is a useful, well established and animal alternative in vivo procedure for evaluation of anti-inflammatory potentials and anti-irritant properties of nano drug delivery systems. In this study, diclofenac sodium (DS) loaded PLGA NPs were prepared and characterized. The particle size (PS) of DS-loaded PLGA NPs was between 114.7 and 124.8 nm and all NPs were monodisperse with negative zeta potential values. The encapsulation efficiency was in range of 41.4-77.8%. In vitro dissolution studies of NPs showed up to 24 h of DS release after the first 3 h of burst effect. The 3 h burst effect and 24 h release kinetics studied with DDSolver were found to be predominantly driven not only by one mechanism, by a combined mechanism of Fickian and non-Fickian. Solid state structures of formulations were clarified by DSC and FT-IR analysis. PS, EE% and release rates were found to be affected by the amount of DS added to the formulations. Increasing the amount of DS added to the formulations increased PS, while the EE% decreased. The release rates were affected by PS and the formulation with the lowest PS value showed slower release. The anti-inflammatory activity of optimum formulation (NP-1) was examined using in vivo HET-CAM assay. The anti-inflammatory activity results indicated that NP-1 coded NP formulation showed significantly good anti-inflammatory potential at low dose. As a result, a low dose high anti-inflammatory effect was achieved with the NP structure of DS. To the best of our knowledge this is the first study on in vivo anti-inflammatory activities of DS loaded PLGA NPs by HET-CAM.

Anti-angiogenic effect of a Palladium(II)-Saccharinate Complex of Terpyridine in vitro and in vivo.

Anti-angiogenic activity of palladium (Pd)(II)-based complexes is unknown despite their quite powerful anticancer activity. This study was therefore carried out to evaluate both in vivo anti-angiogenic effect and in vitro cytotoxic activity of a Pd(II)-based complex. ([Pd(sac)(terpy)](sac)·4H2O(sac=saccharinate and terpy=2,2':6',2″-terpyridine)) on HUVEC cells. The anti-angiogenic activity of the complex was evaluated in vivo using the chick embryo chorioallantoic membrane (CAM) assay, tube formation assay and the cytotoxicity was screened using the MTT viability assays. The CAM treated with the complex (50µg/pellet) showed a strikingly high anti-angiogenic effect (score 1.1±0.2) compared to the positive controls cortisone, prednisone and (±)-thalidomide (e.g. (±)-thalidomide score 0.9±0.2) tested at the same concentration. Furthermore, the complex showed neither membrane toxicity nor irritation at the tested concentration. According to the MTT assays, the human umbilical vein endothelial cell (HUVEC) viability was inhibited in a dose-dependent manner at tested concentrations (1.56-100µM). Pd(II) complex also reduced the tube network at the lower dose than the compared with thalidomide. These results suggest that the Pd(II)-complex has strong anti-angiogenic activity, which adds an important feature to the previously-described anticancer activity of the complex.

Preparation, Characterization, Antioxidant Activities, and Determination of Anti-Alzheimer Effects of PLGA-Based DDSs Containing Ferulic Acid.

Nanoparticle (NP) systems have attracted the attention of researchers in recent years due to their advantages, such as modified release features, increased therapeutic efficacy, and reduced side effects. Ferulic acid (FA) has therapeutic effects such as anti-inflammatory, anti-Alzheimer's, antioxidant, antimicrobial, anticancer, antihyperlipidemic, and antidiabetic. In this study, FA-loaded PLGA-based NPs were prepared by a nanoprecipitation method and the effect of varying concentrations of Poloxamer 188 and Span 60 on NP properties was investigated. FA-loaded A-FA coded formulation was chosen as optimum. High encapsulation efficiency has been achieved due to the low affinity of FA to the water phase and, therefore, its lipophilic nature, which tends to migrate to the organic phase. It was determined that the release of FA from the A-FA was slower than pure FA and prolonged release in 24 h. Antioxidant and anti-Alzheimer's effects of A-FA coded NP formulation were investigated by biological activity studies. A-FA coded NP formulation showed strong DPPH free radical scavenging, ABTS cation decolorizing, and reducing antioxidant activity. Since it has both AChE inhibitor and antioxidant properties according to the results of its anti-Alzheimer activity, it was concluded that the formulation prepared in this study shows promise in the treatment of both oxidative stress-related diseases and Alzheimer's.

A Novel Phytotherapy Application: Preparation, Characterization, Antioxidant Activities and Determination of Anti-inflammatory Effects by in vivo HET-CAM Assay of Chitosan-based DDSs Containing Endemic Helichrysum pamphylicum P.H. Davis & Kupicha Methanolic Extract.

BACKGROUND: Numerous pharmaceutical applications for chitosan, a polysaccharide made from the shells of crustaceans by deacetylating chitin that occurs naturally, are currently being researched. Chitosan, a natural polymer, is successfully used to prepare many drug-carrier systems, such as gel, film, nanoparticle, and wound dressing. OBJECTIVE: Preparing chitosan gels without external crosslinkers is less toxic and environmentally friendly. METHODS: Chitosan-based gels containing Helichrysum pamphylicum P.H.Davis & Kupicha methanolic extract (HP) were produced successfully. RESULTS: The F9-HP coded gel prepared with high molecular weight chitosan was chosen as the optimum formulation in terms of pH and rheological properties. The amount of HP was found to be 98.83 %± 0.19 in the F9-HP coded formulation. The HP release from the F9-HP coded formula was determined to be slower and 9 hours prolonged release compared to pure HP. It was determined that HP release from F9-HP coded formulation with the DDSolver program was by anomalous (non-fickian) diffusion mechanism. The F9-HP coded formulation significantly showed DPPH free radical scavenger, ABTS•+ cation decolorizing and metal chelating antioxidant activity while weakly reducing antioxidant potential. According to the HET-CAM scores, strong anti-inflammatory activity was obtained by the F9-HP coded gel at a dose of 20 µg.embryo-1 (p<0.05 compared with SDS). CONCLUSION: In conclusion, it can be said that chitosan-based gels containing HP, which can be used in both antioxidant and anti-inflammatory treatment, were successfully formulated and characterized.