5-Fluorouracil-Loaded BSA Nanoparticles: Formulation Optimization and In Vitro Release Study

Response surface methodology (RSM) was used to optimize the process of preparing bovine serum albumin (BSA) nanoparticles by desolvation, then the resulting BSA nanoparticles (BSANPs) were conjugated with folate to produce a drug carrier system that can specifically target tumors. The anticancer drug, vinblastine sulfate (VBLS), was loaded to this tumor-specific drug carrier system for the purpose of overcoming the nonspecific targeting characteristics and side effects of the drug. A central composite design was applied for modeling the process, which was composed of four independent variables, namely BSA concentration, the rate of adding ethanol (ethanol rate), ethanol amount, and the degree of crosslinking. The mean particle size and residual amino groups of the BSANPs were chosen as response variables. The interactive effects of the four independent variables on the response variables were studied. The characteristics of the nanoparticles; such as amount of folate conjugation, drug entrapment efficiency, drug-loading efficiency, surface morphology and release kinetics in vitro were investigated. Optimum conditions for preparing desired BSANPs, with a mean particle size of 156.6 nm and residual amino groups of 668.973 nM/mg, were obtained. The resulting folate-conjugated BSANPs (FA-BSANPs) showed a drug entrapment efficiency of 84.83% and drug-loading efficiency of 42.37%, respectively, and the amount of folate conjugation was 383.996 microM/g BSANPs. The results of this study indicate that using FA-BSANPs as a drug carrier system could be effective in targeting VBLS-sensitive tumors in the future.

Preparation of Bovine Serum Albumin (BSA) nanoparticles by desolvation using a membrane contactor: A new tool for large scale production

Objective To study the clinical application of local drug control-release,modified bovine serum albumin (BSA) nanoparticles was used as controlled release carrier for local drug delivery to the inner ear.Methods Modified BSA nanoparticles,with or without Rhodamine B (RhB),were prepared by improved desolvation method and be confirmed by electron microscope and particle size analyzer.Drug loading,encapsulation efficiency and drug release were analyzed.L929 cells were used for cell biocompatibility assay.Nanoparticles with RhB was used for in vivo imaging to investigate the penetration of particles into the round window membrane (RWM).Results The BSA nanoparticles were smooth-surfaced solid-spheres with average diameter of 476 nm.The Zetapotential of blank nanoparticle was 15.4 mV.RhB,a model drug,was proved to be adsorbed on the nanoparticles and to be release control.Glutaraldehyde cross-linked BSA nanoparticles had certain cytotoxicity,while heat denatured BSA nanoparticles had good cell compatibility.Nanoparticles with RhB was proved to be adsorbed on and penetrated through the RWM.Conclusion The nanoparticles have complete structure and carry drug well for control-release.The preparation is simple and non-toxic.The outcome provides the basis for the study of injectable cross RWM control-release nanogel. Key words: Bovine serum albumin; Nanoparticle; Controlled release carrier, Round window membrane; Inner ear

Chemical coupling of polyethylene glycol (PEG) to proteins or particles (PEGylation), prolongs their circulation half-life by greater than 50-fold, reduces their immunogenicity, and also promotes their accumulation in tumors due to enhanced permeability and retention effect. Herein, phase separation method was used to prepare bovine serum albumin (BSA) nanoparticles. PEGylation of BSA nanoparticles was performed by SPA activated mPEG through their free amino groups. Effect of process variables on PEGylation efficiency of BSA nanoparticles was investigated and optimized through response surface methodology with the amount of free amino groups as response. Optimum conditions was found to be 32.5 g/l of PEG concentration, PEG-nanoparticle incubation time of 10 min, incubation temperature of 27°C, and pH of 7 for 5 mg of BSA nanoparticles in 1 mL phosphate buffer. Analysis of data showed that PEG concentration had the most noticeable effect on the amount of PEGylated amino groups, but pH had the least. Mean diameter and zeta potential of PEGylated nanoparticles under these conditions were 217 nm and -14 mV, respectively. In conclusion, PEGylated nanoparticles demonstrated reduction of the negative surface charge compared to the non modified particles with the zeta potential of -31.7 mV. Drug release from PEGylated nanoparticles was almost slower than non-PEGylated ones, probably due to existence of a PEG layer around PEGylated particles which makes an extra resistance in opposition to drug diffusion.

Preparation of size-controlled bovine serum albumin (BSA) nanoparticles by a modified desolvation method

The aim of this study was to design hirudin-loaded bovine serum albumin (BSA) nanoparticles to control release and improve antithrombotic effect of hirudin. BSA nanoparticles were designed as carriers for delivery of hirudin. Hirudin–BSA nanoparticles were prepared by a desolvation procedure and cross linked on the wall material of BSA. The hirudin–BSA nanoparticles were characterised by particle size distribution, zeta potential, entrapment efficiency, differential scanning calorimetry (DSC), and powder X-ray diffractometry (PXRD). The in vitro release characteristics and pharmacological availability were investigated. The morphology of hirudin–BSA nanoparticles was approximately spherical. The mean particle size was 164.1 ± 5.40 nm and the zeta potential was −20.41 ± 0.64 mV. The mean entrapment efficiency and drug loading were 85.14% ± 4.79% and 66.38% ± 3.54%, respectively. Results from DSC and PXRD revealed that hirudin in BSA existed in an amorphous state. The release behaviours of hirudin from BSA nanoparticles in phosphate buffer solution were fitted to the bioexponential model. The in vivo result obtained after intravenous injection of hirudin–BSA nanoparticles in normal rats demonstrated that BSA nanoparticles could prolong the antithrombotic effect of hirudin in comparison with hirudin solution. These results suggest that hirudin–BSA nanoparticles may be a promising drug delivery system for thrombosis and disseminated intravascular coagulation therapy.

Impact of surface modification in BSA nanoparticles for uptake in cancer cells

The glutathione-conjugated bovine serum albumin (BSA) nanoparticles were constructed in the present exploration as a novel biodegradable carrier for brain-specific drug delivery with evaluation of its in vitro and in vivo delivery properties. BSA nanocarriers were activated and conjugated to the distal amine functions of the glutathione via carbodiimide chemistry using EDAC as a mediator. These nanoparticles were characterized for particle shape, average size, SPAN value, drug entrapment and in vitro drug release. Further, presence of glutathione on the surface of BSA nanoparticles was confirmed by Ellman’s assay, which has suggested that approximately 750 units of glutathione were conjugated per BSA nanoparticle. To evaluate the brain delivery properties of the glutathione-conjugated BSA nanoparticles fluorescein sodium was used as a model hydrophilic compound. Permeability and neuronal uptake properties of developed formulations were evaluated against the MDCK-MDR1 endothelial and neuro-glial cells, respectively. The permeability of glutathione-conjugated BSA nanoparticles across the monolayer of MDCK-MDR1 endothelial tight junction was shown significantly higher than that of unconjugated nanoparticles and fluorescein sodium solution. Similarly, glutathione-conjugated nanoparticles exhibited considerably higher uptake by neuro-glial cells which was inferred by high fluorescence intensity under microscope in comparison to unconjugated nanoparticles and fluorescein sodium solution. Following an intravenous administration, nearly three folds higher fluorescein sodium was carried to the rat brain by glutathione-conjugated nanoparticles as compared to unconjugated nanoparticles. The significant in vitro and in vivo results suggest that glutathione-conjugated BSA nanoparticles is a promising brain drug delivery system with low toxicity.

Adsorption of lactoferrin and bovine serum albumin nanoparticles on pellicular two-layer agarose-nickel at reactive blue 4 in affinity chromatography

Chitosan-coated bovine serum albumin nanocarriers mediate efficient delivery of methotrexate in breast cancer therapeutics

Robust size control of bovine serum albumin (BSA) nanoparticles by intermittent addition of a desolvating agent and the particle formation mechanism

The widely used bovine serum albumin (BSA) nanoparticles (NPs) were modified with poly(allylamine hydrochloride) (PAH)/sodium poly(4-styrene sulfonate) (PSS) multilayers and aptamers to improve their suspension stability and targeting ability. For this to occur, a PAH-g-poly(ethylene glycol) (PAH-g-PEG–COOH) layer was further adsorbed onto the (PAH/PSS)2 multilayer-coated BSA NPs and used to covalently bond the aptamer AS1411, which is known to specifically bind the over-expressed nucleolin on cancer cell membranes. The PEGylated multilayer-coated BSA NPs showed good suspension stability in diverse media, in particular in a serum containing medium. By a mechanism of spontaneous deposition, doxorubicin (DOX) was effectively loaded into the pre-formed BSA NPs with both good encapsulation efficiency (98.6%) and loading percentage (9%). The loaded drug showed a pH-dependent release behaviour, i.e. faster at pH 5.5 than at pH 7.4. The multilayer coating did not significantly influence either both drug loading or release. In vitro cell culture demonstrated that the as-prepared BSA NPs could be specifically delivered to liver cancer cells, leading to higher cellular uptake and cytotoxicity.

Tuning the binding, release and cytotoxicity of hydrophobic drug by Bovine Serum Albumin nanoparticles: Influence of particle size

Preparation, characterization and biodistribution of the lactone form of 10-hydroxycamptothecin (HCPT)-loaded bovine serum albumin (BSA) nanoparticles

Albumin based nanocarriers have been widely used in drug delivery studies. Here, we developed a water‐in‐ionic liquid (IL) emulsion‐like method to prepare bovine serum albumin (BSA) nanoparticles as alternative to the traditional organic solvents containing techniques. Conformational changes of albumin induced by the imidazolium based ILs at the water‐IL interface triggers the BSA nanoparticle formation. The albumin nanoparticle formation are dependent on the experimental parameters and the hydophobicity of the IL. At pH 9.0, using 1.3%wt of BSA in water/1‐butyl‐3‐methyl imidazolium tetrafluoroborate (BmimBF 4 ) (50/50 mol%) and TX‐100/butanol surfactant mixture yields uniformly distributed 200 nm average sized BSA nanoparticles. Different than BmimBF 4 , using a more hydrophilic IL, EmimBF 4 yielded albumin aggregates. Instead, using a more hydrophobic IL, HmimBF 4 produced albumin nanoparticles but a non‐uniform size distribution was obtained. These results indicate that the ionic liquids called green and designer solvents can be also used to synthesize albumin nanoparticles.