High Incorporation Efficiency Research Articles

Formulation of Controlled Release Gellan Gum Macro Beads of Amoxicillin

Gellan gum has been reported to have wide pharmaceutical applications such as tablet binder, disintegrant, gelling agent and as a controlled release polymer. Multiparticulate delivery systems spread out more uniformly in the gastrointestinal tract and reduce the local irritation. The purpose of this study is to explore possible applicability of gellan macro beads as an oral controlled release system of a sparingly soluble drug, amoxicillin. Gellan gum beads were prepared by ionotropic gelation with calcium ions. The effect of drug loading, stirring time, polymer concentration, electrolyte (CaCl2) concentration, curing time etc. influencing the preparation of the gellan gum macro beads and the drug release from gellan gum beads were investigated in this study. Optimal preparation conditions allowed very high incorporation efficiency for amoxicillin (91%) The release kinetics of amoxicillin from gellan beads followed the diffusion model for an inert porous matrix in the order: 0.1 N HCl > phosphate buffer > distilled water. Change in curing time did not significantly affect the release rate constant, but drug concentration, polymer concentration and electrolyte concentration significantly affect the release rate of amoxicillin from the beads. The gellan macro beads may be suitable for gastro retentive controlled delivery of amoxicillin.

Development of a binary lipid nanoparticles formulation of itraconazole for parenteral administration and controlled release

The principal aim of this study was to develop an intravenous formulation of itraconazole (ITZ) using lipid nanoparticles based on binary mixture of liquid and solid lipids. Lipid nanoparticles were developed to provide the controlled release of ITZ as well as to improve the solubility of ITZ. Lipid nanoparticles were prepared with tristearin as a solid lipid, triolein as a liquid lipid, and a surfactant mixture of eggPC, Tween 80 and DSPE-PEG 2000. ITZ was incorporated at the concentration of 20 mg/g. Lipid nanoparticles were manufactured by high-pressure homogenization method. The particle size and polydispersity index (PI) of lipid nanoparticles were below 280 nm and 0.2, respectively. Zeta potentials and incorporation efficiencies of lipid nanoparticles were around −30 mV and above 80%, respectively. Lipid nanoparticles containing 1% of liquid lipid showed the smallest particles size and the highest incorporation efficiency. Results from SEM, DSC and PXRD revealed that ITZ in lipid nanoparticles exists in an amorphous state. Release rates were increased as the amount of liquid lipid in lipid core increased, demonstrating that the release of ITZ from lipid nanoparticles could be controlled by modulation of the amount of liquid lipid in lipid core. Pharmacokinetic studies were performed after intravenous administration of lipid nanoparticles in rats at the dose of 5 mg/kg. The plasma concentration of ITZ was prolonged after intravenous administration of lipid nanoparticles. It is concluded that binary lipid nanoparticles could control the release and pharmacokinetic parameters of ITZ.

Preparation of microparticles of naproxen with Eudragit RS and Talc by spherical crystallization technique

Microparticles of naproxen with Eudragit RS and talc were prepared by the spherical crystallization technique, i.e. quasi-emulsion solvent diffusion method. The obtained microparticles were evaluated by micromeritic properties, yield, encapsulation efficiency, drug physical state and dissolution rate of drug. The influence of formulation factors and preparation condition (drug: polymer ratio, talc: polymer ratio, SLS concentration, stirring speed) on the properties of the microparticles were also examined. The resultant microparticles were finely spherical and uniform with high incorporation efficiency and yield. Greater encapsulation efficiency was obtained by increasing the drug: polymer ratio and talc: polymer ratio and by reducing the SLS %. The dissolution rate of naproxen from microparticles was enhanced significantly with increasing the ratio of drug: polymer and stirring rate, and sustained by increasing SLS % in crystallization medium. The results of X-ray diffraction and differential scanning calorimeter analysis indicated that naproxen was highly dispersed in microparticles, so as amorphous state. Studies carried out to characterize the micromeritic properties of formulations, such as flowability and packability showed that microparticles were suitable for further pharmaceutical manipulation (e.g. capsule filling). Hence, the spherical crystallization technique can be successfully used for obtaining spherical microparticles, generating a heterogeneous matrix system and providing sustained drug release.

In VitroEvaluation of Macroporous Hydrogels to Facilitate Stem Cell Infiltration, Growth, and Mineralization

Hydrogels have gained acceptance as biomaterials in a wide range of applications, including pharmaceutical formulations, drug delivery, and tissue sealants. However, exploiting the potential of hydrogels as scaffolds for cell transplantation, tissue engineering, and regenerative medicine still remains a challenge due to, in part, scaffold design limitations. Here, we describe a highly interconnected, macroporous poly(ethylene glycol) diacrylate hydrogel scaffold, with pores ranging from 100 to 600 microm. The scaffold exhibits rapid cell uptake and cell seeding without the need of any external force or device with high incorporation efficiency. When human mesenchymal stem cells are seeded within the porous scaffolds, the scaffolds were found to promote long-term stem cell viability, and on exposure to osteogenic medium, elicit an mineralization response as evaluated by an increased alkaline phosphatase activity (per cell) and calcium and phosphate content within the constructs. The atomic composition of the mineralized matrix was further determined by energy dispersive spectroscopy and found to be similar to calcium-deficient hydroxyapatite, the amorphous biological precursor of bone. The macroporous design of the hydrogel appears advantageous over similar porous hydrogel scaffolds with respect to ease of synthesis, ease of stem cell seeding, and its ability to support long-term stem cell survival and possible differentiation.

Preparation of aminoglycoside-loaded chitosan nanoparticles using dextran sulphate as a counterion

Purpose. To prepare aminoglycoside (AG) (streptomycin, gentamicin and tobramycin) loaded chitosan nanoparticles with high drug incorporation efficiency and test the in vivo oral efficacy of streptomycin (SM) loaded chitosan nanoparticles in a Mycobacterium tuberculosis (TB) chronic infection mouse model.Method. Dextran sulphate (a polyanion) was used to shield the positive charge of AG and increase the drug incorporation in the chitosan nanoparticle. By varying the concentration of each component, the formulation of SM-loaded chitosan nanoparticle was optimized by monitoring the drug incorporation efficacy and particle size. The mechanism of the nanoparticle formation was suggested and the preparation method was applied to two other aminoglycosides (AG): gentamicin (GM) and tobramycin (TM). The resulting nanoparticles were characterized by particle diameter, drug incorporation efficacy, drug loading efficacy and zeta potential. The in vitro drug release from these nanoparticles was carried out in pH 1.2 and pH 7.4 buffer. Preliminary in vivo oral efficacy studies of SM-loaded chitosan nanoparticles was performed in a Mycobacterium tuberculosis (TB) chronic infection mouse model.Results. The optimal concentration of streptomycin (SM)/dextran sulphate/chitosan/tripolyphosphate (TPP) for SM nanoparticles preparation was 2/1.2/2/0.8 mg mL−1. Through calculation, the optimal concentrations of dextran sulphate are 2.5 mg mL−1 and 2.4 mg mL−1 for 2 mg mL−1 gentamicin and tobramycin, respectively (). The resulting AG chitosan nanoparticles had a high drug incorporation efficacy with particle sizes in the nanometre range. The in vitro drug release studies showed that more than 60% drug is retained inside the nanoparticles in pH 1.2 buffer after 6 h. The preliminary in vivo results indicated that oral SM chitosan nanoparticles induced one log 10 reduction (p < 0.01) in growth of the bacilli and were as effective as subcutaneously injected aqueous SM solution at the same concentration (100 mg kg−1).Table 1. The optimal amount of dextran sulphate for AG (2 mg) chitosan nanoparticles based upon charge-neutralization calculation.Aminoglycoside (AG)StreptomycinGentamicinTobramycinFormulaC21H39N7O12C19H39N5O7C18H37N5O9Number of associated H2SO43/211Total MW728547565.5Number of positive charges355Calculated dextran amount (mg)1.22.52.4Conclusion. Dextran sulphate can significantly increase AG incorporation into the chitosan nanoparticles. The concentration of each component was critical in preparing AG-loaded chitosan nanoparticles. The chitosan nanoparticles designed in this study may provide a promising oral drug delivery formulation for AG which usually, in tuberculosis treatment, is administrated as an injectible preparation.

Photodynamic inactivation of viruses by immobilized chlorin-containing liposomes

The viral safety of blood derived products relies in properly chosen inactivation procedures. In this way, it has been reported that some photosensitizers are useful products for blood sterilization. The data presented here show the high incorporation efficiency of the chlorin 3-phorbinepropanol, 9,14-diethyl-4,8,13,18-tetramethyl-20-(3S-trans) (CHL) into anionic unilamellar liposomes, give a protocol for the steric immobilization of chlorin-containing liposomes in a chromatographic support and provide the studies of photodynamic inactivation of bovine viral diarrhea virus (BVDV) and encephalomyocarditis virus (EMCV) with chlorin-containing liposomes, free in solution and immobilized on Sephacryl S-1000 beads. The study demonstrates the successful inactivation of the enveloped virus BVDV by both preparations in culture medium and the resistance of the non-enveloped virus EMCV. The effectiveness of CHL-containing liposomes, in solution and immobilized in the chromatographic support, decreased when the culture media was replaced with human blood plasma. Moreover, the reduction factor of the virus titer after irradiation was smallest when immobilized liposomes were used. Nevertheless, the reduction factor for the virus titers of enveloped viruses after irradiation of human blood plasma samples with immobilized chlorin-containing liposomes increased with the reduction of the sample thickness. The more outstanding aspect of this paper is the design of a system useful for blood sterilization that can be easily removed after photodynamic treatment and, therefore, able to be applied in the manufacturing processes.

Calcium phosphate‐DNA nanoparticle gene delivery from alginate hydrogels induces in vivo osteogenesis

There is a significant need for improved therapy for bone regeneration. The delivery of recombinant bone morphogenetic proteins has been approved for clinical use to promote osteogenesis, but still has limitations such as expense, degradation of the proteins in vivo and difficulties retaining protein at the site of injury. Localized gene delivery is a promising alternative therapy, as it would allow sustained expression of specific osteoinductive growth factors by cells near the damaged site. We have engineered an injectable system for localized, sustained nonviral gene delivery from alginate hydrogels containing preosteoblastic cells and calcium phosphate-DNA nanoparticles. The nanoparticles utilized in this report are stable, on the order of 100 nm, and have a high DNA incorporation efficiency (>66%). When the nanoparticles were incorporated in alginate hydrogels, sustained release of DNA was observed. Furthermore, MC3T3-E1 preosteoblast cells exhibited the capacity to form bony tissue in as little as two and half weeks when mixed with DNA nanoparticles encoding for BMP-2 into the alginate hydrogels and injected subcutaneously in the backs of mice. This injectable, minimally invasive gene delivery system may be efficacious in bone regeneration applications.

Physicomechanical properties of naproxen-loaded microparticles prepared from Eudragit l100.

Microparticles of naproxen with Eudragit L100 and Aerosil were prepared by the emulsion solvent diffusion method in order to avoid local gastrointestinal irritation, one of the major side effects of nonsteroidal anti-inflammatory drugs after oral ingestion. The process of preparation involved the use of ethanol as good solvent, dichloromethane as a bridging liquid, water as poor solvent, Aerosil as anti-adhesion agent, and sodium dodecyl sulfate to aid in the dispersion of the drug and excipients into the poor solvent. The obtained microparticles were evaluated for micromeritic properties, yield, encapsulation efficiency, drug physical state, and drug release properties. The influence of formulation factors and preparation condition (polymer/naproxen ratio, Aerosil/polymer ratio, and the initial difference of temperature between the solvent and nonsolvent) on the properties of the microparticles were also examined. The resultant microparticles were finely spherical and uniform with high incorporation efficiency (>79%) and yield (>71%). The incorporation efficiency was enhanced with increasing the ratio of excipients to drug and the initial difference of temperature between the solvent and nonsolvent. The mean diameter of the microparticles was influenced by all of the manufacturing parameters. Studies carried out to characterize the micromeritic properties of formulations, such as flowability and packability, showed that microparticles were suitable for further pharmaceutical manipulation (e.g., capsule filling). Drug release studies of the microparticles confirmed the gastroresistance, and mathematical studies showed that the drug released followed a Hixon and Crowell kinetic. These microparticles represent a simple method for the preparation of drug-loaded enteric microparticles with desired micromeritic properties and gastroresistance release.

Complexation hydrogels for intestinal delivery of interferon β and calcitonin

Recent studies have suggested that complexation hydrogels poly(methacrylic acid-g-ethylene glycol) (henceforth designated as P(MAA-g-EG)) exhibit high insulin incorporation efficiency, rapid insulin release in the intestine based on their pH-dependent complexation properties, enzyme-inhibiting effects and mucoadhesive characteristics. Therefore, they are promising carriers for insulin delivery via an oral route. As we designed these hydrogels as carriers suitable for oral administration of various peptide/protein drugs, in this study we aimed at investigating the applicability of P(MAA-g-EG) hydrogels to improving the intestinal absorption of various peptide/protein drugs. High loading efficiency into hydrogels was observed for insulin, calcitonin, and interferon β. In addition, polymer microparticles loaded with calcitonin and interferon β exhibited complexation/decomplexation and pH-sensitive release behavior. The molecular weight and chemical structure appeared to affect the efficiency of loading and release depending on the peptides and proteins. Furthermore, a drastic reduction of plasma calcium concentration accompanied by calcium absorption and a dose-dependent enhancement of plasma interferon β concentration were observed after the administration of particles loaded with calcitonin or interferon β into closed rat ileal segments. These findings indicate that P(MAA-g-EG) hydrogels are promising carriers for administration of various peptides and proteins via an oral route.

High loading efficiency and tunable release of plasmid DNA encapsulated in submicron particles fabricated from PLGA conjugated with poly-L-lysine

Poly(lactic- co-glycolic acid) (PLGA) particles have been widely explored as vehicles for delivery of plasmid DNA to mammalian cells both in vitro and in vivo. Achieving high incorporation efficiencies and control over release kinetics are significant challenges in encapsulating hydrophilic molecules such as DNA within submicron particles fabricated from PLGA. This study explored two modifications in the preparation of submicron particles to specifically address these challenges. Firstly, we compared homogenization and sonication as energy sources for emulsification. It was demonstrated that particles prepared with homogenization resulted in higher encapsulation efficiency and a linear release profile of DNA as compared to particles prepared with sonication, which exhibited lower encapsulation efficiency and a burst release. Also investigated was conjugation of poly-L-lysine to PLGA (PLGA–PLL) to create an electrostatically favorable interaction between the carrier material and the DNA. Particles fabricated with high weight percentages of PLGA–PLL/PLGA resulted in remarkably increased loading (> 90%). Additionally, the release profile could be dictated by the quantity of PLGA–PLL incorporated into the particles. Particles incubated in vitro on COS-7 cells were able to transfect cells. These results demonstrated that DNA encapsulation and release were modulated by the method of fabrication.

Preparation and Characterization of 10-Hydroxycamptothecin Loaded Nanostructured Lipid Carriers

Two types of 10-hydroxycamptothecin (10-HCPT) loaded nanostructured lipid carriers (NLC-F 68 and NLC-Brij 35) intended for use as the alternative formulation of 10-HCPT for parenteral administration were prepared using an emulsification-ultrasonication method and fully characterized from physicochemical and in vitro release standpoint. The particle size was measured by laser diffraction, being 108 nm and 126 nm for NLC-F 68 and NLC-Brij 35, respectively. Zeta potentials of two NLCs were −28.5 mV and −32.1 mV analyzed by photon correlation spectroscopy. The incorporated efficiency was more than 85%. It is observed that NLCs are homogeneous and spherical in shape by transmission electron microscopy. Differential scanning calorimetry analysis of NLCs showed that 10-HCPT was dispersed within NLC in an amorphous state. The combination of trehalose and mannitol as cryoprotectant was most suitable for HCPT-NLC lyophilization. The in vitro release behavior for two types of NLC was similar and displayed biphasic drug release pattern with rapid release at the initial stage and prolonged release afterwards. These results suggest that NLC could be exploited as a carrier of 10-HCPT with high incorporation efficiency and controlled release and that NLC may serve as the alternative delivery system for parenteral administration of 10-HCPT.

Analysis of tellurium as n-type dopant in GaInP: Doping, diffusion, memory effect and surfactant properties

The use of tellurium as n-type dopant for GaAs and InP has several advantages, including a high incorporation efficiency, the very high doping levels achievable and a low diffusion coefficient. However, its use to dope Ga x In 1− x P is not straightforward, since it shows several problems like a remarkable memory effect and an acute inertia of the material to become Te-doped, which gives rise to gradual doping profiles. In this paper, all these phenomena are studied and quantified using secondary ion mass spectroscopy (SIMS) and electrochemical CV profiling (ECV) measurements. Concerning the gradual doping profiles, their origin is linked to the interaction of Te and In in the gas phase and on the growth surface. A phenomenological explanation is given for this effect although the exact physical processes behind remain to be defined.

Gut passage of phosphorus-limited algae through Daphnia: do they take up nutrients in the process?

Nutrient-limited algae are known to be a food source of inferior quality for zooplankters. Three factors are thought to determine this poor quality: direct elemental limitations of the algae, biochemical limitations and an increased resistance to digestion because of an increase in cell wall thickness. Thus far, most studies have concentrated on the effect of the algae on the daphniids. It has recently been hypothesized, however, that while going through the digestive tract of herbivorous zooplankters the digestion resistant nutrient-limited algae might actually take-up nutrients, in a similar way as it has been described for gelatinous alga such as Sphaerocystis. In this study, we present results of different experiments investigating whether nutrient-limited algae are indeed more resistant to digestion, and whether nutrient-limited algae take-up the limiting nutrient in the guts of their predators. We observed that digestion resistance is not very important, and that it can only be observed at high food levels. As a result, we could not find any evidence for nutrient uptake of these algae when they pass through the daphniids. We did find that animals adapted to low-P environments have a higher incorporation efficiency for P, and conclude that digestion resistance in nutrient stressed algae is of very limited ecological relevance.

Nickel Stabilization Efficiency of Aluminate and Ferrite Spinels and Their Leaching Behavior

Stabilization efficiencies of spinel-based construction ceramics incorporating simulated nickel-laden waste sludge were evaluated and the leaching behavior of products investigated. To simulate the process of immobilization, nickel oxide was mixed alternatively with gamma-alumina, kaolinite, and hematite. These tailoring precursors are commonly used to prepare construction ceramics in the building industry. After sintering from 600 to 1480 degrees C at 3 h, the nickel aluminate spinel (NiAl204) and the nickel ferrite spinel (NiFe204) crystallized with the ferrite spinel formation commencing about 200-300 degrees C lower than for the aluminate spinel. All the precursors showed high nickel incorporation efficiencies when sintered at temperatures greater than 1250 degrees C. Prolonged leach tests (up to 26 days) of product phases were carried out using a pH 2.9 acetic acid solution, and the spinel products were invariably superior to nickel oxide for immobilization over longer leaching periods. The leaching behavior of NiAl2O4 was consistent with congruent dissolution without significant reprecipitation, but for NiFe2O4, ferric hydroxide precipitation was evident. The major leaching reaction of sintered kaolinite-based products was the dissolution of cristobalite rather than NiAl2O4. This study demonstrated the feasibility of transforming nickel-laden sludge into spinel phases with the use of readily available and inexpensive ceramic raw materials, and the successful reduction of metal mobility under acidic environments.

Spinel Formation for Stabilizing Simulated Nickel-Laden Sludge with Aluminum-Rich Ceramic Precursors

The feasibility of stabilizing nickel-laden sludge from commonly available Al-rich ceramic precursors was investigated and accomplished with high nickel incorporation efficiency. To simulate the process, nickel oxide was mixed alternatively with gamma-alumina, corundum, kaolinite, and mullite and was sintered from 800 to 1480 degrees C. The nickel aluminate spinel (NiAl2O4) was confirmed as the stabilization phase for nickel and crystallized with efficiencies greater than 90% for all precursors above 1250 degrees C and 3-h sintering. The nickel-incorporation reaction pathways with these precursors were identified, and the microstructure and spinel yield were investigated as a function of sintering temperature with fixed sintering time. This study has demonstrated a promising process for forming nickel spinel to stabilize nickel-laden sludge from a wide range of inexpensive ceramic precursors, which may provide an avenue for economically blending waste metal sludges via the building industry processes to reduce the environmental hazards of toxic metals. The correlation of product textures and nickel incorporation efficiencies through selection of different precursors also provides the option of tailoring property-specific products.

Inhibition of liver metastasis by all- trans retinoic acid incorporated into O/W emulsions in mice

All- trans retinoic acid (ATRA) was incorporated into lipid emulsions in an attempt to alter its distribution characteristics and improve its inhibition of liver cancer metastasis. Lipid emulsions composed of egg phosphatidylcholine, cholesterol, and soybean oil were the optimized carriers for ATRA delivery, as shown by the submicron particle size and high incorporation efficiency. The particle size and zeta potential of ATRA incorporated into emulsions were about 133 nm and −11 mV, respectively. In vitro drug release study demonstrated that the release of ATRA from emulsions was sustained in the absence and present of bovine serum albumin, suggesting that ATRA was stable when incorporated in emulsions. After intravenous administration in mice, [ 3H]cholesteryl hexadecyl ether incorporated into emulsion, which is the inherent distribution of emulsions, accumulated gradually mainly in the liver. The blood concentration and hepatic accumulation of [ 3H]ATRA incorporated into emulsion was significantly higher than that of serum dissolving [ 3H]ATRA, which represent the original distribution characteristic of free ATRA. In a murine liver metastasis model by colon adenocarcinoma, the liver metastasis number and liver weight were significantly reduced and the survival time of mice was prolonged following intravenous injection of ATRA incorporated into emulsions.

Simultaneous Detection of Precore/Basal Core Promoter Mutations in Hepatitis B Virus Using Arrayed Primer Extension

Hepatitis B is a major disease that causes serious public health problems worldwide. The loss of HBeAg expression due to point mutations or single nucleotide polymorphisms (SNPs) in the precore/basal core promoter region of the hepatitis B virus (HBV) is associated with hepatocellular cirrhosis and carcinoma. Simultaneous screening for these mutations is strongly advocated for monitoring disease development in HBV-infected patients. The aim of this study is to apply arrayed primer extension (APEX) for the detection of HBV SNPs at the precore/basal core promoter. We optimized APEX for simultaneous detection of eight potential sites of SNPs in the precore/basal core promoter region of HBV. The precore/basal core promoter regions of HBV from 36 HBV-infected patients were amplified by PCR. After purification and DNA fragmentation, the short, single-stranded HBV DNA fragments were allowed to hybridize with the oligonucleotides corresponding to the sites of SNPs immobilized on glass slides, followed by incorporation of different fluorescently labeled dideoxynucleotides. This allows fast and unequivocal discrimination between wild-type and mutant genotypes with high dideoxy-nucleotide incorporation efficiency, sensitivity, and specificity. The coexistence of both genotypes was also detected; this was undetected by DNA sequencing. The simultaneous detection of SNPs in HBV precore/basal core promoter by APEX enables large-scale diagnostic analysis, which can be extended to the whole HBV genome.

Unilamellar Liposomes with Enhanced Boron Content

A new type of boron-rich, DSPC-free, unilamellar liposomes was formed using the novel dual-chain, ionic, nido-carborane lipid, K[nido-7-(C16H33OCH2)2CHOCH2-7,8-C2B9H11] (DAC-16), and cholesterol for encapsulation of an aqueous buffer core. Since DSPC was not necessary for the formation of stable DAC-16 liposomes, the boron concentration of these vesicles was increased dramatically to approximately 8.8 wt % in the dry lipid; these liposomes had a high bilayer boron incorporation efficiency of 98%. DSPC-free liposomes exhibited a size distribution pattern of 40-60 nm, which was in the range normally associated with selective tumor uptake. This size distribution was maintained throughout storage at room temperature for several months. Additionally, optimized liposome formulations incorporating DAC-16, DSPC, and cholesterol were identified having stable size distribution patterns after storage for more than two months at a variety of temperatures. Although animal studies indicate that DAC-16 liposomes are toxic, this new ionic nido-carborane lipid allows the formation of liposomes of high boron content for in vitro applications that require the delivery of large amounts of boron.

Genospheres: self-assembling nucleic acid-lipid nanoparticles suitable for targeted gene delivery

We describe the assembly of a cationic lipid-nucleic acid nanoparticle from a liquid monophase containing water and a water miscible organic solvent where both lipid and DNA components are separately soluble prior to their combination. Upon removal of the organic solvent, stable and homogenously sized (70-100 nm) lipid-nucleic acid nanoparticles (Genospheres) were formed. The low accessibility (<15%) of the nanoparticle-encapsulated DNA to a DNA intercalating dye indicated well-protected nucleic acids and high DNA incorporation efficiencies. It was demonstrated that Genospheres could be stably stored under a variety of conditions including a lyophilized state where no appreciable increase in particle size or DNA accessibility was observed following reconstitution. Finally, Genospheres were made target-specific by insertion of an antibody-lipopolymer (anti-HER2 scFv (F5)-PEG-DSPE) conjugate into the particle. The target specificity (>100-fold) in HER2 overexpressing SK-BR-3 breast cancer cells was dependent on the degree of PEGylation, where the incorporation of high amounts of PEG-lipid on the particle surface (up to 5 mol%) had only a minor effect on the transfection activity of the targeted Genospheres. In summary, this work describes a novel, readily scalable method for preparing highly stable immunotargeted nucleic acid delivery vehicles capable of achieving a high degree of specific transfection activity.

Novel oral insulin delivery systems based on complexation polymer hydrogels: Single and multiple administration studies in type 1 and 2 diabetic rats

Insulin-loaded polymer microparticles (ILP) composed of crosslinked poly(methacrylic acid) and poly(ethylene glycol) are multi-functional carriers showing high insulin incorporation efficiency, a rapid insulin release in the intestine based on their pH-dependent complexation properties, enzyme-inhibiting effects and mucoadhesive characteristics. Thus, they are potential carriers for insulin delivery via an oral route. Recent studies suggest that the polymer composition and particle size of ILP strongly influenced insulin bioavailability. Therefore, the present study aimed at finding an optimal formulation and designing carriers for oral insulin delivery using in vivo experiments. Various types of ILPs were prepared and administered orally to healthy and type 1 and 2 diabetic rats. The most promising formulation was subsequently used for in vivo multiple oral administration studies using diabetic rats. The microparticles of diameters of < 53 μm (SS-ILP) composed of a 1 : 1 molar ratio of methacrylic acid / ethylene glycol units showed the most pronounced hypoglycaemic effects following oral administration to healthy rats, achieving a 9.5% pharmacological availability compared to subcutaneous insulin injection. Their usefulness was also confirmed with both type 1 and 2 diabetic rat groups. In a multiple administration study, SS-ILP significantly suppressed the postprandial rise in blood glucose and showed continuous hypoglycaemic effects following 3 times/day oral administration to both diabetic rat groups in the presence of foods. These results indicate that the blood glucose levels of diabetic rats can be effectively controlled by oral SS-ILP administration, and thus SS-ILP would be a promising delivery carrier of insulin via the oral route.