Translation of nanoscale discoveries from the laboratory to the clinic promises new diagnostic tools, drug targeting modalities, gene therapy platforms, and tissue constructs for patients. Tissue morphogenesis during fetal development is highly dependent on spatial and temporal expression of multiple morphogens targeting different progenitor cells. During fetal development, mesenchymal stem cells (MSCs) and endothelial colony forming cells (ECFCs) are implicated in bone formation coupled by paracrine signalling between the two progenitor cells. The invading ECFCs secrete osteogenic morphogens (BMP2) to stimulate cell differentiation and mineralization whereas the differentiating MSCs release vasculogenic morphogens (VEGF) to further stimulate capillary formation for the metabolically active osteoblasts. I will present in my seminar novel drug delivery strategies and tissue models for timed and localized release of morphogens using self-assembled nanogels in a micro-patterned co-culture system to stimulate paracrine signalling and coupling osteogenesis to vasculogenesis.
Crystallization of pharmaceutical ingredients, primarily those that possess several polymorphic forms, particle size and morphology critical properties, are among the most serious and least understood manufacturing procedure. Many processes and product failures can be traced to a poor understanding or lack control of the crystallization procedures. Clearly the pharmaceutical industry requires to getting more competitive and robust process by the knowledge of molecular complexity and solid form challenges, due to the impact of material properties for production efficiency related to the solid implication on drug product formulation.The content of this presentation is focused on reporting real case examples from the lab and development scale to production, throughout in-process crystallization measurements and control by means also of PAT approaches. Process understanding using in-process techniques in development scale, such as automated batch reactor vessels equipped with Reaction Calorimetry, ATR/FT-IR Spectroscopy, Focus Beam Reflectance (FBRM) probes plus temperature and pH sensors, were suitable methods to reach the desired active ingredient requirements, and then define a suitable tool in control the Critical Quality Attributes with as well benefits in process cycle time reduction. A variety of in situ analytical methods applied, combined with chemometric tools for the analysis of multivariate process information, have provided a basis for future improvement in modelling, simulation and control of crystallization procedures. These on-line recorded data together with chemical properties parameters assessed by off-line controlling techniques, are the starting point for intended processes for high quality products.
Over the last several years, mesoporous silica nanoparticles (MSNs) have been extensively developed in the field of biomedicine as nanocarriers for delivering anti-cancer drugs, due to their biocompatibility and ease of surface functionalization. However, it has certain limitations for being applied in pharmaceuticals, for example this type of material usually has low drug loading capacity. In this study, hollow mesoporous silica nanoparticles (HMSNs) were fabricated in order to increase the drug loading capacity of nanosilica materials. The synthesized HMSNs possessed inner hollow cores that could remarkably raise the total pore volume and thus improve the capacity for cargo loading. HMSNs were synthesized according to the hard-template method with three main steps: (1) forming of solid SiO2 nanoparticles as templates, (2) forming of core-shell structure by coating MSN layers onto the templates, and (3) forming of hollow core structure by etching away the solid template. The HMSNs product was characterized by TEM, XRD, TGA, FTIR and BET. In addition, drug loading capacity of the material was evaluated with doxorubicin (DOX) as model drug. The results indicated remarkable improvement in drug loading capacity, compared to MSN sample. More importantly, MTT assay data showed that the prepared HMSNs were biocompatible nanocarriers and furthermore, the incorporation of DOX into these HMSNs has been proven successful in reducing the toxicity of DOX. In the interest of the system’s biodistribution, the imaging and measurement on mice with tumor cell lines were conducted and the obtained results indicated excellent accumulation of the desired materials at tumor sites. As a part of histological investigation on HMSNs nano-drug-carrier system, long-term toxicity study was also managed by H&E organ staining. Base on the condition of the cells’ components, for examples cellular shrinkage in liver cells, condensation of chromatin or rupture of cell membrane, etc., the conclusion about toxic features from the administration of HMSNs system can be made, which in this report, showed minor in vivo toxicity. These results demonstrated the potential of HMSNs in the delivery of anticancer agents.
Liposomes are the most versatile carriers for the delivery of a large variety of both lipophilic and hydrophilic drug molecules, being biocompatible and biodegradable. Despite their great advantages, their tendency to degrade and aggregate in biological fluids as well as in storage conditions drove the research towards new preparative approaches for liposomes stabilization, essentially based on superficial coatings or inclusion in polymeric materials. However the most used techniques, such as spraying, layering, sometimes by exploiting supercritical fluids, require complex and expensive apparatuses. Therefore, this work was developed with the idea to overcome typical liposomes stabilization limits, by choosing the consolidated wet granulation process as a method to obtain granules containing liposomes in a single step, by spraying on powder the liposomal suspension, previously produced by a novel continuous and rapid simil-microfluidic method. Literature highlights, although with only few works, the use of wet granulation as method to stabilize polymeric nanoparticles suspensions, by adding them in the binder phase. Instead, there are not experimental evidences about the use of liposomes suspensions as binder phase in wet granulation for both their stabilization and incorporation in pharmaceutical solid forms. Thus, this novel combination between wet granulation and the use of liposomes suspension spray as binder phase, allowed to reach both easy and cheap liposomes stabilization and the production of smart solid multiparticulate dosage forms, which could be ideal candidates for a combined fast/slow release of active ingredients, enhancers, fortifiers.
While vaccines represent the strongest weapons against a contagious disease, several shortcomings still limit their optimal function. This can largely be attributed to the lack of a proper delivery system as well as route of administration for effective activation of the immune system [1-3]. The intradermal route has the potential to be an outstanding route of vaccination due to the localization of potent antigen present cell (APC) and other immune cells in the dermis . MNPs are tiny needles, small enough (100 –1,000 μm) that perforate superficial layers of the skin to deliver a therapeutic agent into the dermis in a relatively painless and invasive manner. MNPs are generally preferred by patients as compared to traditional hypodermic injections. they take advantage of the potent intradermal immune system, which can create stronger responses than what is typical of the muscle or can generate equivalent responses from lower doses. The majority of works with microneedles has involved bolus delivery of drugs and vaccines using either coated or dissolvable microneedles, but introducing microparticles into microneedles in order to sustained release has received less attention . Given these objectives and challenges, we designed a multi-compartment dissolvable and degradable polymeric microneedle patches for the sustained release of fd-OVA.
A central issue in the understanding of the pathogenesis of nonalcoholic fatty liver disease is the problem of the underlying mechanisms which are not fully understood. In the setting of excessive central adiposity, insulin resistance is the major underlying cause of fat accumulation in hepatocytes. Because of the difficulties with human trials, several animal models have been developed for this purpose mainly characterized as follows: genetically disturbed or murine fatty liver, methionine-choline deficient diet fed or murine steatohepatitis, and high-fat or sucrose diet fed models. Although these animal models have provided useful information, none of them accurately reflect genetic, metabolic and biochemical characteristics of the human disease.
Nitrile derivatives are known to cause toxicities like neurotoxicity, hepatotoxicity, cardiovascular, renal and gastrointestinal disorders. Nitriles collates a broad range of chemicals including Acetonitrile, Acrylonitrile, Butyronitrile, Valeronitrile etc. widely utilized in different manufacturing industries and materials. Mitochondria and free radical generation are indicated to play a crucial role in the mechanism and uplifting the biological effects of induced toxicities. Alpha lipoic acid (ALA) is reported to be efficient enough to inhibit and prevent free radical generation persuaded toxicity because of its antioxidative activity. In this study, we investigated the effects of nitrile derivatives and Alpha lipoic acid (ALA) on hippocampal cellular and mitochondrial enzyme activities such as SOD (superoxide dismutase), GPx(glutathione peroxidase), CAT (catalase) and GSH (reducedglutathione) and MnSOD (manganese Superoxide dismutase), GPx,GST (glutathione-s-transferase) and GSH respectively. We also studied the effects and alterations caused by nitrile derivatives administration on the enzymes of TCA cycle and mitochondrial membrane. In this study, damage in mitochondrial membrane and significant reduction in the enzymatic activities of TCA Cycle such as ICDH (isocitrate dehydrogenase), SDH (succinate dehydrogenase), α-KDH (Alpha ketoglutarate dehydrogenase) and MDH (malate dehydrogenase) was observed. Considerable downfall in the levels of these enzymes was observed on subsequent treatment with Alpha Lipoic Acid (ALA). Activities of ETC enzymes NADH dehydrogenase and Cytochrome C oxidase were also studied, where nitrile derivatives were found to downregulate the enzymatic action. It is hereby concluded that whilenitrile derivatives demonstrated toxicities by altering enzymatic activities and damaging mitochondrial membrane, Alpha lipoic acid (ALA) restored the altered enzymatic action and efficiently preserved the hippocampal cellular and mitochondrial damage induced by nitrile derivatives.
The utilization of polymer-grafted liposomes for the delivery of anticancer drug, particularly Paclitaxel (PTX) to the tumor target sites is a new and advanced method in the branch of chemotherapy. Liposome in the pharmaceutical view is the possessor of superb properties for drug delivery, but it is held back by the fact that liposome has a very short circulation time. The grafting of methylated-polyethylene glycol on the surface of liposome is the key to solve and eliminate the only drawback for liposome as a drug delivery system. In this study, liposome formulated from natural soy lecithin (SLP) was prepared and partially surface modified with methoxy polyethylene glycol (mPEG) for the delivery of PTX. The PTX/SLP@mPEG was successfully developed with nanometric particle size, high entrapment efficiency, and long-lasting drug release. Moreover, in vitro cytotoxicity assays demonstrated that SLP@mPEG is nontoxic and cytocompatible. PTX/SLP@mPEG also showed remarkable suppress the proliferation of cancer cells. In addition, the in vivo antitumor effect of PTX/SLP@mPEG was also observed in a mouse model that had been subcutaneously inoculated with breast cancer cells by virtue of higher passive accumulation in tumor tissue via the enhanced permeability retention effect. Overall, these obtained results provide insights into the potential of SLP@mPEG as a platform for the development of more effective therapies against cancers.
Diabetes is severe chronic metabolic disease characterized by high blood glucose levels leads to generation of reactive oxygen species that causes oxidative damage in kidney, liver and pancreas.The aim and purpose of present research work was to find out the antidiabetic and hypolipdimic potential of commonly available and economical medicinal plant i.e Musa paradisiaca L bract and tracheal fluid methanolic extract. For this purpose, diabetes was induced in rats with alloxon, two standard drugs (Glibenclamide 2.5 mg/kg orally, insulin 4 U/200 mg/dl blood sugar subcutaneous injection) and different doses (100 mg/kg, 250 mg/kg and 500 mg/kg) of morphological parts (bract, and tracheal fluid) methanolic extract were administered orally to different groups for 20 days to the diabetic rats. Fasting blood glucose level and weight of rat was measures every fifth day and TG, TC, HDL-C, LDL-C, VLDL, TC/HDL and LDL/HDL was find out at the end of experiment. Histopathology of pancreas was performed. Results of fasting blood glucose level, change in weight, TG, TC, HDL-C, LDL-C, VLDL, TC/HDL and LDL/HDL was analyzed by using ANOVA followed by turkey test, compared with positive and negative control and graph was drawn by using the 5th version of graph pad prism. It is concluded from the results that tracheal fluid has significant antidiabetic and hypo-lipidimic activity whereas bract have non-significant activity.
New antimicrobial hydrogels were prepared via reaction of functionalized-Tragacanth Gum biopolymer by glycidyltrimethylammonium chloride (QTG) with acrylamide (AM). Characterization of the QTG hydrogels with AM (QTG-AM) were carried out by thermogravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FTIR), and 1HNMR. Swelling behavior of the QTG hydrogels were investigated on the pH, medium, and temperature, immersion time. Loading and in-vitro drug release from the final hydrogels were studied by using quercetin as a model drug. Five type of kinetic models were applied, and the release of drug from the drug-loaded hydrogels occurred through non-Fickian diffusion mechanism. The water uptake by hydrogels decreased in 0.9% NaCl solution and increased in pH 9.2. The antimicrobial effect of the prepared hydrogels QTG-AM was investigated against Candida albicans, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa. The antimicrobial results demonstrated that the hydrogels possess good antimicrobial activity against the all tested microorganism.
Toxic substances and secretions of animals such as snakes, spiders and scorpions have been used in traditional medicine for the treatment of several diseases. Despite of the inherent difficulty and high economic risk in the discovery and development of new drugs from natural products, the development and rapid advancement of new technologies such as mass spectrometry-based proteomic analysis has allowed researchers to advance in a more focused characterization for the determination of key molecular targets in several diseases. In order to characterize and find potential key molecular species in cancer or disease linked to central nervous system (CNS) that may be targets in snake venom therapeutics, we have tested the Bothrops jararaca snake venom and Tityus serrulatus scorpion venom on several cancer cell lines and Crotalus durissus terrificus rattle snake venom on mice cerebellum and screened for proteome differences with and without the venom treatment. We have observed several up- and down-regulated proteins that play important roles related to cancer, such as cell proliferation, invasion, metastasis, apoptosis and stress response and proteins related to synapses inhibition and oxidative stress that are key processes in some CNS-related diseases. These data show that venom or some of their components may have potential usage for cancer or CNS-related disease therapy.
Bryophyllum pinnatum is a succulent perennial plant traditionally used in the treatment of premature labour, first in anthroposophic hospitals and, recently, in conventional settings as an add-on medication. Experimental evidence obtained with uterus strips supports this use and suggests that bufadienolides might be responsible for the tocolytic effect. Previous work with myometrial cells showed that B. pinnatum leaf press juice (BPJ) inhibits the increase of intracellular free calcium concentration ([Ca2+]i) induced by oxytocin (OT), a hormone known to trigger myometrium contractions. However, it is not known which compounds in BPJ contribute to this inhibition and whether BPJ affects other OT-induced signalling pathways. Here we compare the inhibitory effects of BPJ, a bufadienolide-enriched fraction (BEF), a flavonoid-enriched fraction (FEF), the corresponding flavonoid aglycon mixture (A-Mix), bersaldegenin-1,3,5-orthoacetate (BO), the combination of BEF and FEF, and the OT-receptor antagonist atosiban on the secondary intracellular effects triggered by OT, such as changes in intracellular calcium levels and phosphorylation of mitogen activated protein kinases (MAPKs). To study the effects on OT-triggered [Ca2+]i increase, human myometrial hTERT-C3 and PHM1-41 cells that had been loaded with a calcium specific fluorescent probe (Fura-2-AM) were pre-incubated with test substances or vehicle controls. Then, cells were stimulated with OT to induce a rapid and transient increase in [Ca2+]i. [Ca2+]i was measured by real-time fluorescence spectrophotometry. To examine the phosphorylation of MAPKs by Western-blotting, hTERT-C3 cells were incubated for 30 min with BPJ and then stimulated with OT. BPJ led to concentration-dependent decrease of the OT-induced increase of [Ca2+]i in both cell lines (p<0.0001), achieving ca. 75% inhibition at a 20 µg/mL concentration. Atosiban also promoted a concentration-dependent effect on [Ca2+]i in both cell lines (p<0.0001). BEF, FEF, BO, A-Mix, and the combination of BEF and FEF led to concentration-dependent decrease of the OT-induced increase of [Ca2+]i in hTERT-C3 cells (p<0.05). BEF (2.2 µg/mL), FEF (17.4 µg/mL), BO (0.04 µg/mL), and A-Mix (0.7 µg/mL), at concentrations corresponding to 20 µg/mL BPJ led to ca. 25% decrease of the OT-induced increase of [Ca2+]i. The combination of BEF plus FEF led to a decrease of 55.3%. In addition, BPJ significantly reduced OT-induced phosphorylation of MAPKs p38 and JNK at its maximum (5 min incubation). We have demonstrated that BPJ not only promotes a specific and concentration-dependent effect on the OT-induced increase of [Ca2+]i, but also inhibit the OT-induced activation of important MAPKs. Compounds present in BEF and in FEF seem to have a synergistic effect on the inhibition of the OT-induced increase of [Ca2+]i, which is comparable to the effect of BPJ.
Design and development of biodegradable water-based poly (urethane-urea) dispersions, with poly(ε-caprolactone)–D-glucose polyester polyol as a central core for combining properties of end-functionalized star-shaped polymers, block copolymers and advantages of the presence of both hydrophobic and hydrophilic segments in amphiphilic star copolymers. Herein, A green Star- liked poly(ε-caprolactone) (PCL)–glucose was synthesized in one-pot in the presence of tin(II) 2-ethylhexanoate (Sn(Oct)2) catalyst via ring-opening polymerization(ROP) of CL by D-glucose as the initiator. The prepared pentol was reacted with hexamethylene diisocyanate (HDI), dimethylol propionic acid (DMPA), triethylamine (TEA), and ethyldiamine (EDA) as a chain extender to afford the WPCL-b-PUU star-block copolymer through the acetone process. The chemical structure of polymers was characterized by Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (1H NMR). Also, the degree of polymerization (DPn: 14), average molecular weight (Mn: about 8,000 g mol−1), OH value and functionality of oligopolyol (4-5) were determined using 1HNMR spectroscopy. The polydispersity indices (PDI) were (1.69) for star polyol and (1.797) for star-block copolymers. The glass transition temperatures, melting points, crystallinities, and decomposition temperatures of the copolymers were measured by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) with a heating rate of 10° C/min under nitrogen. X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to analyze the crystallinity and morphology of synthesized polymers. In vitro cytocompatibility studies performed over 72 h indicated that WPCL-b-PUU star-block copolymer was nontoxic to mouse fibroblast cells (L929). The in vitro degradability of this was investigated with evaluating the morphological changes, and gravimetric measurements after soaking the copolymer in PBS (pH 7.4, Invitrogen, CA, USA) at 37 °. Therefore, these analyses indicate That the WPCL-b-PUU star-block copolymer is a biocompatible, biodegradable polymer and can be considered as a suitable carrier for designing novel drug delivery systems.