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.

Effect of different molecular weight PLGA on flurbiprofen nanoparticles: formulation, characterization, cytotoxicity, and in vivo anti-inflammatory effect by using HET-CAM assay.

Objective: The effect of polymers used in nanoparticle (NP) production on the formulation properties is one of the few studied issues. Therefore, this study aims to formulate flurbiprofen (FLB) loaded NPs with different molecular weight (Mw) poly lactic-co-glycolic acid (PLGA) and investigate the effect of Mw on NP character. One of the most important objectives is to provide a high anti-inflammatory effect with a low dose and the anti-inflammatory efficacy of the selected optimal formulation is to be determined by in vivo hen's egg test on Chorioallantoic Membrane (HET-CAM) analysis that a new, popular and in vivo animal experiment alternative method.Significance: To determine the anti-inflammatory efficacy of the optimum formulation by HET-CAM analysis. To the best of our knowledge, this is the first report on the in vivo anti-inflammatory evaluation of FLB-loaded PLGA NP using the in vivo HET-CAM assay.Methods: Blank and FLB-loaded PLGA NPs were prepared using a nanoprecipitation technique. The cell viability test for all formulation was performed with MTT in the NIH-3T3 mouse embryonic fibroblast cell line. The anti-inflammatory activity of optimum formulation (A6) was examined using the in vivo HET-CAM assay.Results: The particle sizes (PSs) of the FLB-loaded PLGA NPs were between 175 and 198 nm. The encapsulation efficiency (EE%) was a range of 82-93%. In vitro release of NPs showed extended-release up to 144 h. The release kinetics were fitted to the Peppas-Sahlin and Weibull models. The results showed that PS, PDI, EE%, and release rates of NPs were directly related to the Mw of PLGA. There is no statistically significant difference in cell viability study was observed between blank and FLB-loaded PLGA NPs. The in vivo anti-inflammatory activity results indicated that A6 coded formulation was showed significantly good anti-inflammatory potential at low dose.Conclusions: It could be concluded that FLB-loaded NPs seem to be a promising extended-release drug delivery system for oral administration with a low dose and high efficiency.

New approaches to tumor therapy with siRNA-decorated and chitosan-modified PLGA nanoparticles.

Objective: In this study, we aimed to develop a candidate modifited polymeric nanoparticle (NP) system that will kill cancer cells by facilitated to apoptosis and also reduce pain. Significance: The primary goal of treatment, especially for metastatic cancers, is to control the growth of the cancer and to alleviate the symptoms. Pain is one of the commonest symptoms of cancer. In cancer treatment, directing cancer cells to death while simultaneously relieving pain will be a new approach. Methods: Chitosan-modified PLGA NPs were prepared using an nanoprecipitation technique. The NPs were loaded with flurbiprofen and decorated with folic acid. STAT3-siRNA was adsorbed to these polymeric NPs using antisense technology. Results: The NPs were small in size (176.9-220.3 nm) with positive zeta potential (+14.1 mV to +27.2 mV). They had high loading capacity and prolonged release properties over 144 hours. Cytotoxicity studies performed with siRNA showed effective electrostatic interaction due to the positively charged NPs. Folic acid facilitated entry into cancer cells and helped to kill them. Conclusion: The formulation we developed is a potential carrier system for both treatment of cancer and prevention of pain, especially for metastatic cancers.

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.

Evaluation of Cell Proliferation and Wound Healing Effects of Vitamin A Palmitate-Loaded PLGA/Chitosan-Coated PLGA Nanoparticles: Preparation, Characterization, Release, and Release Kinetics.

In this study, vitamin A palmitate (VAP)-loaded poly(lactic-co-glycolic acid) (PLGA)/chitosan-coated PLGA nanoparticle (NP) systems were prepared by the nanoprecipitation technique. The prepared systems were characterized by parameters such as particle size, polydispersity index (PDI), ζ-potential, encapsulation efficiency, in vitro dissolution, and release kinetic study. Then, the cytotoxicity and wound healing profiles of the designed NP formulations in HaCaT (human keratinocyte skin cell lines) were determined. The particle size of VAP-loaded NPs was obtained between 196.33 ± 0.65 and 669.23 ± 5.49 nm. PDI data proved that all NPs were prepared as high quality and monodisperse. While negative ζ-potential values of Blank-NP-1 and NP-1 encoded PLGA NP formulations were obtained, positive ζ-potential was obtained in chitosan-coated NPs. In vitro release studies of NPs observed rapid dissolution in the first 1-6 h, but prolonged dissolution of VAP after rapid dissolution. As a result of cell culture studies and wound healing activity studies, it was determined that NP-7 was the most effective. It was thought that the reason for this was that the NP-7 coded formulation was a chitosan-coated PLGA nanoparticle with the smallest particle size, and it was concluded that the efficiency of VAP was increased with its nanoparticle structure. This study demonstrated the similar wound healing effects of VAP-loaded nanoparticle systems, in particular NP-7, which increases keratinocyte cell proliferation at lower concentrations (10 µg·mL-1) than vitamin A alone (100 µg·mL-1). VAP-loaded nanocarriers that can be used in the pharmaceutical industry have been successfully produced and the results obtained have been evaluated as promising for this industry.

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.

Quercetin, Rutin And Quercetin-Rutin Incorporated Hydroxypropyl ß-Cyclodextrin Inclusion Complexes.

Quercetin (Q) and rutin (R) are well known and most studied flavonoids due to their activities in reduction of inflammation, oxidative damage, platelet aggregation and inhibition of cancer proliferation. Despite their remarkable potentials they have limited oral bioavailability due to the low water solubility. Therefore in this study inclusion complexes of Q and R with hydroxypropyl-ß-cyclodextrin (HP-ß-CD) were formulated to improve the aqueous solubility, antiproliferative efficacy and also antioxidant activity of the flavonoids. According to the analyses results, aqueous solubilities of Q and R were increased up to ∼630 fold and ∼55 fold, respectively. ZP values were ranged between -21.7±0.3 mV and -6.1±0.8 mV showing the anionic structure of the complexes. 1H-NMR analyses revealed the complex formation considering the shifts of the protons of the APIs as well as HP-ß-CD. The in vitro release analyses revealed that the cumulative release of Q was decreased from 22.9 % to 18.1 and 15.2 for T9 and T 24 formulations respectively while the cumulative release of R increased from 26.8 % up to 64.5 % and 75.8 % with T14 and T24 formulations respectively. According MTT analyses results, Q showed higher antiproliferative effect in MDA-MB-231 and A549 cell lines compared to NIH-3T3 cell lines while R showed remarkable effect only on MDA-MB-231 cell lines at the end of 48 h of incubation period. A synergistic effect was observed in the formulation of combined flavonoid (Q/R) inclusion complexes and an antiproliferative effect was ordered as MDA-MB-231 > A549 > NIH-3T3. The selected complexes T9 (Q), T14 (R) and T24 (Q/R) have shown the highest antioxidant activity with 93.8 %, 65.3 % and 93.1 % respectively with DPPH analyses. In conclusion incoporation of Q, R and Q/R to HP-ß-CD based inclusion complexes have great potentials with enhanced in vitro dissolution characteristics and antiproliferative effects on different types of cancer cell lines for efficient treatment of severe disorders.

Nano Spray-Dried Drugs for Oral Administration: A Review.

Spray drying is an important technology that is fast, simple, reproducible, and scalable. It has a wide application range, that is, in food, chemicals, and encapsulation of pharmaceuticals. The technology can be divided into conventional spray drying and nano spray drying. The key advantage of nano spray drying is the production of drug-loaded nanosized particles for various drug delivery applications. The recent developments in nano spray dryer technology and the market launch of the Nano Spray Dryer B-90 by Büchi Labortechnik AG in 2009 enabled the production of submicron spray-dried particles. This review focuses on nanosized drug delivery systems intended for oral administration produced by nano spray drying. First, the nano spray drying concept, the basic technologies implemented in the equipment, and the effects of the various process parameters on the final dry submicron powder properties are presented. Then, the topics of new formulation strategies of oral drugs are highlighted with examples that have entered the research literature in recent years. Next, the subjects of direct conversion of poorly water-soluble drugs, encapsulation of drugs, and drying of preformed nanoparticles are considered. Finally, topics such as morphology, particle size, size distribution, surface analysis, bioavailability, drug release, release kinetics, and solid-state characterization (by differential scanning calorimetry, X-ray diffraction, Fourier transform infrared spectroscopy, nuclear magnetic resonance) of oral drug delivery systems produced by nano spray drying are discussed. The review attempts to provide a comprehensive knowledge base with current literature and foresight to researchers working in the field of pharmaceutical technology and nanotechnology and especially in the field of nano spray drying.

Levocetirizine Dihydrochloride-Loaded Chitosan Nanoparticles: Formulation and In Vitro Evaluation.

OBJECTIVES: The aim of the present study was to formulate levocetirizine hydrochloride (LCD)-loaded chitosan nanoparticles at submicron level with high entrapment efficiency and prolonged effect for optimizing the plasma drug concentration enhancing bioavailability. MATERIALS AND METHODS: LCD was successfully incorporated into chitosan nanoparticles by spray drying for the purpose of oral application. In vitro characteristics were evaluated in detail. RESULTS: LCD was successfully loaded into the polymeric matrices by spray drying. Characterization of the nanoparticles including encapsulation efficiency, particle size, zeta potential, morphology, polydispersity index, solid-state characterizations, and LCD quantification by high performance liquid chromatography was performed. The release pattern of LCD from the nanoparticles was determined using a dialysis tube in simulated intestinal fluid (pH 6.8). In vitro release profiles indicated prolonged release of LCD from the nanoparticles that followed the Korsmeyer-Peppas kinetic model. CONCLUSION: Chitosan-based LCD-loaded polymeric nanoparticles appear to be a promising drug delivery system for the active agent.

Clarithromycin-Loaded Poly (Lactic-co-glycolic Acid) (PLGA) Nanoparticles for Oral Administration: Effect of Polymer Molecular Weight and Surface Modification with Chitosan on Formulation, Nanoparticle Characterization and Antibacterial Effects.

Clarithromycin (CLR) is a member of the macrolide antibiotic group. CLR has low systemic oral bioavailability and is a drug of class II of the Biopharmaceutical Classification System. In many studies, using nanoparticles (NPs) as a drug delivery system has been shown to increase the effectiveness and bioavailability of active drug substances. This study describes the development and evaluation of poly (lactic-co-glycolic acid) (PLGA) NPs and chitosan (CS)-coated PLGA NPs for oral delivery of CLR. NPs were obtained by nanoprecipitation technique and characterized in detail, and the effect of three molecular weights (Mw1: 7.000-17.000, Mw2: 38.000-54.000, Mw3: 50.000-190.000) of PLGA and CS coating on particle size (PS), zeta potential (ZP), entrapment efficiency (EE%), and release properties etc. were elucidated. Gastrointestinal stability and cryoprotectant effect tests were performed on the NPs. The PS of the prepared NPs were in the range of 178 to 578 nm and they were affected by the Mw and CS coating. In surface-modified formulations with CS, the ZP of the NPs increased significantly to positive values. EE% varied from 62% to 85%, depending upon the Mw and CS coating. In vitro release studies of CLR-loaded NPs showed an extended release up to 144 h. Peppas-Sahlin and Weibull kinetic model was found to fit best for CLR release from NPs. By the broth microdilution test method, the antibacterial activity of the formulations was determined on Staphylococcus aureus (ATCC 25923), Listeria monocytogenes (ATCC 1911), and Klebsiella pneumoniae (ATCC 700603). The structures of the formulations were clarified by thermal (DSC), FT-IR, and 1H-NMR analysis. The results showed that PS, ZP, EE%, and dissolution rates of NPs were directly related to the Mw of PLGA and CS coating.

Supervised Machine Learning Algorithms for Evaluation of Solid Lipid Nanoparticles and Particle Size.

AIMS AND OBJECTIVES: Solid Lipid Nanoparticles (SLNs) are pharmaceutical delivery systems that have advantages such as controlled drug release, long-term stability etc. Particle Size (PS) is one of the important criteria of SLNs. These factors affect drug release rate, bio-distribution etc. In this study, the formulation of SLNs using high-speed homogenization technique has been evaluated. The main emphasis of the work is to study whether the effect of mixing time and formulation ingredients on PS can be modeled. For this purpose, different machine learning algorithms have been applied and evaluated using the mean absolute error metric. MATERIALS AND METHODS: SLNs were prepared by high-speed homogenizaton. PS, size distribution and zeta potential measurements were performed on freshly prepared samples. In order to model the formulation of the particles in terms of mixing time and formulation ingredients and evaluate the predictability of PS depending on these parameters, different machine learning algorithms were applied on the prepared dataset and the performances of the algorithms were also evaluated. RESULTS: PS of SLNs obtained was in the range of 263-498nm. The results present that PS of SLNs can be best estimated by decision tree based methods, among which Random Forest has the least mean absolute error value with 0.028. As a result, the estimation of machine learning algorithms demonstrates that particle size can be estimated by both decision rule-based machine learning methods and function fitting machine learning methods. CONCLUSION: Our findings present that machine learning methods can be highly useful for determining formulation parameters for further research.