1-monooleoyl-rac-glycerol and Disease-Models--Animal

The possibility of using the RNA interference (RNAi) mechanisms in gene therapy was one of the scientific breakthroughs of the last century. Despite the extraordinary therapeutic potential of this approach, the need for an efficient gene carrier is hampering the translation of the RNAi technology to the clinical setting. Although a diversity of nanocarriers has been described, liposomes continue to be one of the most attractive siRNA vehicles due to their relatively low toxicity, facilitated siRNA complexation, high transfection efficiency and enhanced pharmacokinetic properties. This review focuses on RNAi as a therapeutic approach, the challenges to its application, namely the nucleic acids' delivery process, and current strategies to improve therapeutic efficacy. Additionally, lipid-based nanocarriers are described, and lessons learned from the relation between biophysical properties and biological performance of the dioctadecyldimethylammonium:monoolein (DODAX: MO) system are explored. Liposomes show great potential as siRNA delivery systems, being safe nanocarriers to protect nucleic acids in circulation, extend their half-life time, target specific cells and reduce off-target effects. Nevertheless, several issues related to delivery must be overcome before RNAi therapies reach their full potential, namely target-cell specificity and endosomal escape. Understanding the relationship between biophysical properties and biological performance is an essential step in the gene therapy field.

Topics: Animals; Disease Models, Animal; Genetic Therapy; Glycerides; Humans; Liposomes; Nanoparticles; Quaternary Ammonium Compounds; RNA Interference; RNA, Small Interfering

Skin dendritic cells (DCs) such as Langerhans cells and dermal dendritic cells have a pivotal role in inducing antigen-specific immunity; therefore, transcutaneous cancer vaccines are a promising strategy to prophylactically prevent the onset of a variety of diseases, including cancers. The largest obstacle to delivering antigen to these skin DC subsets is the barrier function of the stratum corneum. Although reverse micellar carriers are commonly used to enhance skin permeability to hydrophilic drugs, the transcutaneous delivery of antigen, proteins, or peptides has not been achieved to date because of the large molecular weight of drugs. To achieve effective antigen delivery to skin DCs, we developed a novel strategy using a surfactant as a skin permeation enhancer in a reverse micellar carrier. In this study, glyceryl monooleate (MO) was chosen as a skin permeation enhancer, and the MO-based reverse micellar carrier enabled the successful delivery of antigen to Langerhans cells and dermal dendritic cells. Moreover, transcutaneous vaccination with the MO-based reverse micellar carrier significantly inhibited tumor growth, indicating that it is a promising vaccine platform against tumors.

Topics: Administration, Cutaneous; Animals; Cancer Vaccines; Cell Line, Tumor; Dendritic Cells; Disease Models, Animal; Drug Carriers; Female; Glycerides; Humans; Melanoma; Melanoma-Specific Antigens; Mice; Mice, Inbred C57BL; Micelles; Skin; Skin Neoplasms; Tumor Burden; Vaccination

In this study, the use of cubosomes for topical delivery of the antimicrobial peptide (AMP) LL-37 was investigated. Topical delivery of AMPs is of great interest for treatment of skin infections caused by bacteria, such as Staphylococcus aureus. AMP containing cubosomes were produced by three different preparation protocols and compared: (i) pre-loading, where LL-37 was incorporated into a liquid crystalline gel, which thereafter was dispersed into nanoparticles, (ii) post-loading, where LL-37 was let to adsorb onto pre-formed cubosomes, and (iii) hydrotrope-loading, where LL-37 was incorporated during the spontaneously formed cubosomes in an ethanol/glycerol monooleate mixture. Particle size and size distribution were analyzed using dynamic light scattering (DLS), liquid crystalline structure by small angle x-ray scattering (SAXS) and release of LL-37 by a fluorescamine assay. Proteolytic protection of LL-37 as well as bactericidal effect after enzyme exposure was investigated. The skin irritation potential of cubosomes was examined by an in vitro epidermis model. Finally, the bacterial killing property of the cubosomes was examined by an ex vivo pig skin wound infection model with Staphylococcus aureus. Data showed that a high loading of LL-37 induced formation of vesicles in case of cubosomes prepared by sonication (pre-loading). No release of LL-37 was observed from the cubosomes, indicating strong association of the peptide to the particles. Proteolysis studies showed that LL-37 was fully protected against enzymatic attacks while associated with the cubosomes, also denoting strong association of the peptide to the particles. As a consequence, bactericidal effect after enzyme exposure remained, compared to pure LL-37 which was subjected to proteolysis. No skin irritation potential of the cubosomes was found, thus enabling for topical administration. The ex vivo wound infection model showed that LL-37 in pre-loaded cubosomes killed bacteria most efficient.

Topics: Administration, Topical; Animals; Anti-Infective Agents; Antimicrobial Cationic Peptides; Cathelicidins; Disease Models, Animal; Drug Delivery Systems; Drug Liberation; Epidermis; Escherichia coli; Ethanol; Glycerides; Humans; Liquid Crystals; Microbial Sensitivity Tests; Nanoparticles; Scattering, Small Angle; Skin Irritancy Tests; Staphylococcal Skin Infections; Staphylococcus aureus; Swine; Treatment Outcome; Wound Infection; X-Ray Diffraction

Alzheimer's disease (AD) is one of the most patient devastating central nervous system diseases with no curative therapy. An effective oral therapy with brain-targeting potential is required that is hampered by blood-brain barrier. Piperine (PIP) is a natural alkaloid with memory enhancing potentials. Oral PIP delivery suffers from its hydrophobicity and first-pass metabolism. In this study, novel Tween-modified monoolein cubosomes (T-cubs) were elaborated as bioactive nanocarriers for brain-targeted oral delivery of PIP. Seven liquid crystalline nanoparticles (cubosomes) were prepared testing different bioactive surfactants (Tween 80, poloxamer, and Cremophor). Full in vitro characterization was carried out based on particle size, zeta potential, polydispersity index, entrapment efficiency, and in vitro release. Morphological examination and structure elucidation were performed using transmission and polarizing microscopes. Sporadic dementia of Alzheimer's type was induced in 42 male Wistar rats on which full behavioral and biochemical testing was conducted. Brain toxicity was assessed based on Caspase-3 assay for apoptosis and tumor necrosis factor-α for inflammation. Liver and kidney toxicity studies were conducted as well. Among others, T-cubs exhibited optimum particle size (167.00±10.49 nm), polydispersity index (0.18±0.01), and zeta potential (-34.60±0.47 mv) with high entrapment efficiency (86.67%±0.62%). Cubs could significantly sustain PIP in vitro release. In vivo studies revealed T-cubs potential to significantly enhance PIP cognitive effect and even restore cognitive function to the normal level. Superiority of T-cubs over others suggested brain-targeting effect of Tween. Toxicological studies contended safety of cubs on kidney, liver, and even brain. T-cubs exhibited potential anti-inflammatory and anti-apoptotic activity of loaded PIP, indicating potential to stop AD progression that was first suggested in this article. Novel oral nanoparticles elaborated possess promising in vitro and in vivo characteristics with high safety for effective chronic treatment of AD.

Topics: Administration, Oral; Alkaloids; Alzheimer Disease; Animals; Benzodioxoles; Blood-Brain Barrier; Brain; Caspase 3; Disease Models, Animal; Drug Delivery Systems; Glycerides; Humans; Inflammation; Kidney; Liquid Crystals; Liver; Male; Nanomedicine; Nanoparticles; Oxidative Stress; Particle Size; Piperidines; Poloxamer; Polyethylene Glycols; Polysorbates; Polyunsaturated Alkamides; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

In this paper a novel copolymer, chitosan graft glyceryl monooleate (CS-GO) was synthesized and its potential as the nanocarrier for enhancing the peroral delivery of enoxaparin was studied systemically. The successful synthesis was characterized by (1)H NMR. Enoxaparin nanocomplexes were prepared by self-assembly. Mucoadhesive properties of the nanocomplexes were evaluated using mucin particle method. Uptake and transport of the nanocomplexes were investigated in Caco-2 cells. In vivo absorption was studied in rats. The therapeutic effects of the nanocomplexes were evaluated using pulmonary thromboembolism model in mice. This study demonstrated that compared to chitosan based system, hydrophobic modification of CS with GO enhanced the oral absorption of enoxaparin significantly, which is in good agreement with the enhanced mucoadhesion, cellular internalization and transport in cell culture. Cellular uptake of CS-GO based enoxaparin nanocomplexes was incubation time, enoxaparin concentration and incubation temperature dependent. The uptake mechanism was assumed to be adsorptive endocytosis via clathrin- and caveolae-mediated process. Its therapeutic efficacy was further demonstrated by pharmacodynamic study with pulmonary thromboembolism inhibition percentage 47.1%. In conclusion, CS-GO copolymer is a promising nanocarrier for enhancing the oral absorption of enoxaparin.

Topics: Adhesiveness; Administration, Oral; Animals; Anticoagulants; Caco-2 Cells; Chitosan; Disease Models, Animal; Drug Carriers; Drug Compounding; Enoxaparin; Glycerides; Humans; Male; Mice, Inbred Strains; Molecular Structure; Particle Size; Pulmonary Embolism; Rats, Sprague-Dawley; Surface Properties

The main objective of this study was to improve the safety and oxidative stability of glycerol monooleate (GMO)-based dry-emulsion (DE) formulation containing cyclosporine A (CsA) for inhalation therapy. GMO or highly purified GMO (hpGMO) was used as surfactant for the DE formulations (GMO/DE or hpGMO/DE), the toxicological and physicochemical properties of which were characterized with a focus on oxidative stability, in vitro/in vivo toxicity, and dissolution property. Incubation of GMO at oxidation accelerating conditions for 10 days at 60°C resulted in the formation of lipid peroxides as evidenced by increased malondialdehyde (111 μmol/mg); however, hpGMO samples exhibited increase of only 20.7 μmol/mg in malondialdehyde level. No significant acute cytotoxicity was observed in rat alveolar L2 cells exposed to hpGMO (0.28mM), and intratracheal administration of hpGMO powder in rats did not cause an increase of the plasma LDH level. The hpGMO/DE exhibited marked improvement in dissolution behavior of CsA, and stable fine micelles with a mean diameter of 320 nm were formed when suspended in water. A respirable powder formulation of hpGMO/DE (hpGMO/DE-RP) was newly prepared, and its in vitro inhalation property and in vivo efficacy were also evaluated. The hpGMO/DE-RP exhibited high dispersibility in laser diffraction analysis and significantly improved potency to attenuate recruitment of inflammatory cells into airway and thickening of airway wall in an animal model. Thus, the strategic use of hpGMO would improve oxidative stability and local toxicity compared with a GMO-based DE formulation, and its application to RP formulation could be a promising approach for effective inhalation therapy.

Topics: Animals; Anti-Inflammatory Agents; Antigens; Asthma; Cell Line; Cyclosporine; Disease Models, Animal; Emulsions; Glycerides; Male; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Rats; Rats, Sprague-Dawley; Respiratory Therapy; Surface-Active Agents

The purpose of this work was to evaluate the new bone tissue, comparing two different carriers for rhBMP-2, monoolein and chitosan gels, using the decortication and nondecorticatication surgical technique in rat mandibles, evaluated by histomorphometrical method. It was used 56 male Wistar rats (300 g), divided into 8 groups according to the rhBMP-2 carrier used, monoolein or chitosan gels; surgical technique, bone decortication or nondecortication; and period of time, 3 or 6 weeks until the sacrifice by perfusion. Results obtained in this study showed that the rhBMP-2/monoolein and rhBMP-2/chitosan used in this experimental model was able to induce osteogenesis, contributing to the bone healing process. The bone repair process was time dependent, so that at 6 weeks there was an improved amount of new bone in relation to 3 weeks, considered each analyzed group, and the decortication was able to expose the bone marrow and speed up the bone healing process, which was showed by histomorphometrical methods. Both of carriers were capable to adapt to the bone surgical area, according to the clinical observations, and had favorable properties in relation to protein releasing, revealed by the amount of new bone tissue found in the histological analysis.

Topics: Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Chitosan; Disease Models, Animal; Drug Carriers; Gels; Glycerides; Humans; Male; Mandible; Osteogenesis; Random Allocation; Rats; Rats, Wistar; Recombinant Proteins; Transforming Growth Factor beta

The number of femoral artery catheterizations will increase over the next decade to more than 9 million worldwide. Accordingly, a new era of access site management with vascular closure techniques utilizing biologics are being developed and implemented. Glycerol mono-oleate (GMO) is one such biologic - a biodegradable compound that changes from a solid phase to a bioadhesive swollen semisolid phase when exposed to aqueous solutions and heat. We assessed whether GMO would: 1) achieve hemostasis more effectively than control when injected into a swine liver biopsy tract; and 2) inhibit common percutaneous procedure pathogens.. During the hemostasis experiment, seven swine anticoagulated with heparin (ACT > 250) underwent 10 open-liver biopsies with a 14 gauge cutting needle; 5 injected with GMO (treatment) and 5 injected with nothing (control). Thirty seconds, 2 minutes, 5 minutes and 10 minutes after the procedure, bleeding was objectively graded; 0 = no bleeding (success) and 1 = bleeding (failure). During the bacteria experiment, GMO was injected into plates containing culture media for 4 common percutaneous pathogens (Enteroccocis faecalis, Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae). When injected, GMO converted to a semisolid phase with definitive margins in the culture media. Each bacterium was then coated over their respective media and GMO.. The results showed a significant treatment effect (p < 0.017) on each success/failure bleeding outcome at 30 seconds (p < 0.0001), 2 minutes (p < 0.0001) and 5 minutes (p = 0.0038) based on a multiple logistic regression analysis controlling for initial bleeding, pig and side-of-liver biopsy (medial or lateral lobe). At 10 minutes, the bleeding results were not significant (p = 0.0917), likely explained by a pig's innate ability to clot at this time period. For the bacteria experiment, there was no growth of bacteria on the GMO for any of the plates. Specifically, the Staphylococcus aureus plate displayed a 200 micron halo containing no bacterial growth surrounding the GMO.. In conclusion, these results illustrate a significant hemostatic effect post liver biopsy at multiple time points using GMO. Furthermore, GMO displays bacterial deterrent properties.

Topics: Absorbable Implants; Animals; Biocompatible Materials; Biopsy, Needle; Blood Loss, Surgical; Disease Models, Animal; Glycerides; Hemorrhage; Hemostatic Techniques; Hemostatics; Liver; Probability; Random Allocation; Sensitivity and Specificity; Swine; Tissue Adhesives