Release Profiles Of Bovine Serum Albumin Research Articles

Protein encapsulated in electrospun nanofibrous scaffolds for tissue engineering applications

AbstractThe main purpose of tissue engineering is the preparation of fibrous scaffolds with similar structural and biochemical cues to the extracellular matrix in order to provide a substrate to support the cells. Controlled release of bioactive agents such as growth factors from the fibrous scaffolds improves cell behavior on the scaffolds and accelerates tissue regeneration. In this study, nanofibrous scaffolds were fabricated from biocompatible and biodegradable poly(lactic‐co‐glycolic acid) through the electrospinning technique. Nanofibers with a core–sheath structure encapsulating bovine serum albumin (BSA) as a model protein for hydrophilic bioactive agents were prepared through emulsion electrospinning. The morphology of the nanofibers was evaluated by field‐emission scanning electron microscopy and the core–sheath structure of the emulsion electrospun nanofibers was observed by transmission electron microscopy. The results of the mechanical properties and X‐ray diffraction are reported. The scaffolds demonstrated a sustained release profile of BSA. Biocompatibility of the scaffolds was evaluated using the MTT (3(4,5‐ dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide) assay for NIH‐3T3 fibroblast cells. The results indicated desirable biocompatibility of the scaffolds with the capability of encapsulation and controlled release of the protein, which can serve as tissue engineering scaffolds. © 2013 Society of Chemical Industry

A nanocomposite contact lens for the delivery of hydrophilic protein drugs.

To improve the efficiency of topical ocular drug administration, we developed a nanocomposite contact lens to deliver hydrophilic protein drugs over a prolonged period of time. Here, an in situ route was used to encapsulate the hydrophilic protein drug bovine serum albumin (BSA) within gelatin nanoparticles (NPs), 180 ± 20 nm in diameter, which were then grafted onto the lens material, a copolymer of 2-hydroxyethyl methacrylate and 2-aminoethyl methacrylate p(HEMA-co-AEMA), through photopolymerization. The thickness of the nanocomposite lens was controlled at 150 μm. The release kinetics of BSA from plain p(HEMA-co-AEMA), gelatin NPs, and gelatin NP-grafted p(HEMA-co-AEMA) in phosphate buffer saline (PBS) at pH = 7.4 were studied. The release profile of BSA encapsulated within gelatin NPs could be monitored for 7 days, three times longer than that of BSA soaked in p(HEMA-co-AEMA). Our findings indicate that use of the nanocomposite contact lens, i.e. BSA-loaded gelatin NPs incorporated into p(HEMA-co-AMEA), can prolong the release profile of BSA to 12 days. The swelling behavior and interior strain of p(HEMA-co-AEMA) with and without grafted NPs (1000 : 1 w/w) were further investigated. The nanocomposite lens shows higher swelling behavior than the plain p(HEMA-co-AEMA) lens does. The addition of gelatin NPs to hydrogels leads to a relatively uniform interior strain with lower stiffness. Thus, the prolonged release might be due to a combination of effects. Internal diffusion of the nanocomposite lens materials may significantly contribute to the prolonged release of protein drugs. Furthermore, the nanocomposite lens materials had no cytotoxicity. This new biocompatible nanocomposite might be further developed as an alternative tool for continuous topical ocular drug delivery over a prolonged period of time.

Calcium Alginate Nanoparticles Synthesized Through a Novel Interfacial Cross-Linking Method as a Potential Protein Drug Delivery System

The goal of this research work was to develop a novel technique to synthesize calcium alginate nanoparticles using pharmaceutically relevant microemulsions. Stable microemulsion-based reactors were prepared using aqueous sodium alginate, aqueous calcium chloride, dioctyl sodium sulfosuccinate (DOSS), and isopropyl myristate. The reactor microemulsions were characterized via conductivity and dynamic light scattering (DLS) experiments. The conductivity data indicated composition- and reagent-dependent variations in electrical conductivity when the aqueous phase containing reagents were present at or above a Wo (Wo = [DOSS]/[water]) value of 14. The reactor microemulsions were of approximately 6 nm sized droplets. When the reactor microemulsions were mixed and sonicated for 1 h approximately, 350-nm-sized calcium alginate nanoparticles were produced, as indicated by DLS measurements. The particles were isolated and characterized via low-vacuum scanning electron microscopy. The electron micrographs corroborate the DLS results. The nanoparticles were evaluated as a drug delivery system by incorporating bovine serum albumin (BSA) and performing in vitro release and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) studies. The BSA release profile was characterized by an initial burst release followed by a sustained-release phase. SDS-PAGE studies indicated that the incorporated protein did not suffer covalent aggregation or degradation via fragmentation.

Nanoencapsulation of Protein Drug for Controlled Release

In this paper, gelatin nanoparticles (NPs) have been studied as a carrier to encapsulate protein drug for the controlled release with a prolonged manner. Bovine serum albumin (BSA), a hydrophilic protein drug, was loaded within gelatin NPs through an in situ two-step esolvation method. The average diameter of the NPs is estimated at 180 ± 10 nm by using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Gelatin NPs show the mesoporous structure with average pore size of 2.82 nm. To confirm the encapsulation of BSA in gelatin NPs, fluorescent-labelled BSA was encapsulated in gelatin NPs, which were identified by confocal laser scanning microscopy CLSM). The release kinetics of BSA from gelatin NPs in phosphate buffer saline (PBS) was studied through UV-Vis spectrometry. The release profile of BSA from nanoparticulate system could be monitored for 7 days. In addition, the rate of BSA release from gelatin NPs decreases with increasing the concentration of the cross-linker. The release profile of BSA from the gelatin NPs follows a diffusion-controlled release mechanism. Our results also indicate that the acidic condition can delay the release profile of protein from gelatin NPs.

Superporous polyacrylate/chitosan IPN hydrogels for protein delivery

In this study, poly(acrylamide), poly(AAm), and poly(acrylamide-co-acrylic acid), poly(AAm-co-AA) superporous hydrogels (SPHs) were synthesized by radical polymerization in the presence of gas blowing agent, sodium bicarbonate. In addition, ionically crosslinked chitosan (CH) superporous hydrogels were synthesized to form interpenetrating superporous hydrogels, i.e. poly(AAm)-CH and poly(AAm-co-AA)-CH SPH-IPNs. The hydrogels have a structure of interconnected pores with pore sizes of approximately 100-150 μm. Although the extent of swelling increased when AA were incorporated to the poly(AAm) structure, the time to reach the equilibrium swelling (~30 s) was not affected so much. In the presence of chitosan network mechanical properties significantly improved when compared with SPHs, however, equilibrium swelling time (~30 min) was prolonged significantly as due to the lower porosities and pore sizes of SPH-IPNs than that of SPHs. Model protein bovine serum albumin (BSA) was loaded into SPHs and SPH-IPNs by solvent sorption in very short time (<1 h) and very high capacities (~30-300 mg BSA/g dry gel) when compared to conventional hydrogels. BSA release profiles from SPHs and SPH-IPNs were characterized by an initial burst of protein during the first 20 min followed by a completed release within 1 h. However, total releasable amount of BSA from SPH-IPNs was lower than that of SPHs as due to the electrostatic interactions between chitosan and BSA.

Morphology and Release Kinetics of Protein-Loaded Porous Poly(L-Lactic Acid) Spheres Prepared by Freeze-Drying Technique

Freeze-drying a biodegradable polymer, poly(L-lactic acid) (PLLA), from 1,4-dioxane solutions provided very porous spherical particles of ca. 3 mm in radius with specific surface area of 8–13 m2 g−1. The surface of the particle was found to be less porous compared with its interior. To apply the freeze-dried PLLA (FDPLLA) to drug delivery system, its morphology and drug releasing kinetics were investigated, bovine serum albumin (BSA) being used as a model drug compound. Immersion of FDPLLA into a BSA aqueous solution gave BSA-loaded FDPLLA, where mass fraction of the adsorbed BSA reached up to 79%. Time-dependent release profile of BSA in water suggested a two-step mechanism: (1) very rapid release of BSA deposited on and near the particle surface, which results in an initial burst, and (2) leaching of BSA from the interior of the particle by the diffusion process. It was suggested that the latter process is largely governed by the surface porosity. The porosity of both the interior and surface was found to decrease remarkably as the concentration of the original PLLA/1,4-dioxane solution increases, C 0. Thus, C 0 is a key parameter that controls the loading and releasing of BSA.

Synthesis and characterization of star-shaped poly (lactide-co-glycolide) and its drug-loaded microspheres

The star-shaped poly (lactide-co-glycolide) (PLGA) was synthesized via the ring-opening polymerization of d,l-lactide and glycolide, with pentaerythritol as a multifunctional initiator and stannous 2-ethyl hexanoate as a catalyst. The structures of these polymers were characterized by 13C-NMR spectroscopy, while the molecular weight and polydispersity index (PDI) were determined by gel permeation chromatography (GPC). The glass transition temperature (Tg) of copolymer was determined by differential scanning calorimetry (DSC). Bovine serum albumin (BSA) loaded microspheres were fabricated using star-shaped PLGA by a W/O/W double emulsion solvent evaporation method. The results of characterization demonstrated that the particle size of the PLGA microspheres were about 80–150 μm, the maximum loading capacity and encapsulation efficiency of BSA-loaded microspheres were 67.51 μg/(mg microspheres) and 78.39%, respectively, which were better than linear PLGA. The in vitro release profiles of BSA in phosphate buffer saline (PBS) lasted for 37 h. Drug release profiles can be affected by polymer molecular weight and the ratio of polymer to drug. The maximum release percentage was 80%.

A novel pH-sensitive hydrogel based on dual crosslinked alginate/N-α-glutaric acid chitosan for oral delivery of protein

A novel pH-sensitive hydrogel based on dual crosslinked alginate/N-α-glutaric acid chitosan (GAC) was prepared. The homogeneous alginate/GAC solution was dropped into calcium chloride solution and crosslinked by Ca2+ ions. Sequentially, the crosslinked beads were suspended in sodium sulfate solution for forming dual crosslinked beads. The swelling behaviors of dual crosslinked beads were investigated in simulated gastric fluid (SGF), simulated intestinal fluid (SIF) and simulated colonic fluid (SCF). Bovine serum albumin (BSA) was loaded in beads with the high loading efficiency. The sustained release profiles of BSA loaded beads were studied at different pH environment for simulating gastrointestinal condition. The amount of BSA released from the beads at pH 1.2 was relatively low (below 18%), while almost 100% of BSA could be released at pH 7.4. The preliminary results clearly suggested that the dual crosslinked alginate/GAC hydrogel may be a potential polymeric carrier for oral delivery of protein drugs.

Protein micro and nanoencapsulation within glycol-chitosan/Ca2+/alginate matrix by spray drying

Encapsulation of therapeutic peptides and proteins into polymeric micro and nanoparticulates has been proposed as a strategy to overcome limitations to oral protein administration. Particles having diameter less than 5 µm are able to be taken up by the M cells of Peyer’s patches found in intestinal mucosa. Current formulation methodologies involve organic solvents and several time consuming steps. In this study, spray drying was investigated to produce protein loaded micro/nanoparticles, as it offers the potential for single step operation, producing dry active-loaded particles within the micro to nano-range. Spherical, smooth surfaced particles were produced from alginate/protein feed solutions. The effect of operational parameters on particle properties such as recovery, residual activity and particle size was studied using subtilisin as model protein. Particle recovery depended on the inlet temperature of the drying air, and mean particle size ranged from 2.2 to 4.5 µm, affected by the feed rate and the alginate concentration in the feed solution. Increase in alginate:protein ratio increased protein stability. Presence of 0.2 g trehalose/g particle increased the residual activity up to 90%. Glycol-chitosan-Ca2+alginate particles were produced in a single step operation, with resulting mean diameter of 3.5 μm. Particles showed fluorescein isothiocyanate labeled bovine serum albumin (BSA)-protein entrapment with increasing concentration toward the particle surface. Similar, limited release profiles of BSA, subtilisin and lysozyme were observed in gastric simulation, with ultimate full release of the proteins in gastrointestinal simulation.

Controlling Bovine Serum Albumin Release From Biomimetic Calcium Phosphate Coatings

Biomimetic calcium phosphate (CaP) coating has been used successfully for protein delivery, but the release of protein from CaP coating is mainly dependent on the limited dissolution of the CaP coating and the passive diffusion of the protein in the CaP coating. In the present work, our aim is to improve the release behavior of protein from CaP coating and make it more controllable. By using bovine serum albumin (BSA) as a model protein, our strategy is to tailor BSA release profiles by controlling the distribution of BSA in CaP coatings. To achieve this aim, BSA was added to a modified simulated body fluid (m-SBF) at different stages of coating formation to obtain tailored BSA release profiles. Sustained BSA release was obtained when BSA was added to m-SBF at the initial stage of the coating where the BSA was incorporated into the lattice structure of the coating. In contrast, a relatively faster release was observed when BSA was added during the later stage of coating formation where BSA was mainly adsorbed to the coating surface. As a result, the BSA release efficiency could be tailored by adding BSA into m-SBF at different coating formation stages. More importantly, the coating composition was not altered with the change of BSA adding times and all the beneficial properties of the biomimetic coating were reserved. Therefore, the BSA release from CaP coatings can be tailored by adjusting its distribution in the coating to achieve a more satisfactory release profile.

Zn- and Mg-doped hydroxyapatite nanoparticles for controlled release of protein.

Bovine serum albumin (BSA) protein incorporated with hydroxyapatite (HA) nanoparticles (NPs) were synthesized by an in situ precipitation process. 2 mol % Zn(2+) and Mg(2+) were used as dopants to synthesize Zn(2+)/Mg(2+)-doped HA-BSA NPs. In our study we used BSA as a model protein. The amount of BSA uptake by doped and undoped HA NPs and subsequent release of BSA from NPs were investigated. Zn-doped HA NPs showed the highest amount of BSA uptake, whereas the amount of BSA loaded in undoped HA NPs was the lowest. A two-stage BSA release profile from doped and undoped HA NPs was observed in phosphate buffer solution (PBS) at pH 7.2 +/- 0.2. The initial burst release was due to the desorption of BSA from the HA surface. The later stage of slow release was controlled by the dissolution of BSA incorporated HA NPs. The BSA release rate from Zn-doped HA NPs was found to be the highest, whereas undoped HA NPs released BSA at the slowest rate. Our study showed that the protein release rate from HA NPs can be controlled by the addition of suitable dopants, and doped HA-based NP systems can be used in bone growth factor and drug release study.

Preparation and in vitro evaluation of new pH‐sensitive hydrogel beads for oral delivery of protein drugs

AbstractNew biodegradable pH‐responsive hydrogel beads based on chemically modified chitosan and sodium alginate were prepared and characterized for the controlled release study of protein drugs in the small intestine. The ionotropic gelation reaction was carried out under mild aqueous conditions, which should be appropriate for the retention of the biological activity of an uploaded protein drug. The equilibrium swelling studies were carried out for the hydrogel beads at 37°C in simulated gastric (SGF) and simulated intestinal (SIF) fluids. Bovine serum albumin (BSA), a model for protein drugs was entrapped in the hydrogels and the in vitro drug release profiles were established at 37°C in SGF and SIF. The preliminary investigation of the hydrogel beads prepared in this study showed high entrapment efficiency (up to 97%) and promising release profiles of BSA. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010

Phospholipid-based ultrasonic microbubbles for loading protein and ultrasound-triggered release

Background: Ultrasonic microbubbles are used as ultrasound-triggered delivery carriers for protein drugs. Aim: This work was to prepare stabilized protein-loaded phospholipid-based ultrasonic microbubbles (PUM) and to determine its value as a protein delivery system. Method: Bovine serum albumin (BSA) was used as a model protein drug. BSA-containing PUM were prepared by dissolving lyophilized PUM powder in BSA solution. The particle size and microbubble concentration of BSA-containing PUM were measured. The BSA encapsulation efficiency as a function of BSA concentration was determined. Contrast enhancement of BSA-containing PUM in vivo was detected. The release profile of BSA from PUM was also investigated. Results: The mean particle size and microbubble concentration of PUM were unchanged by the presence of BSA for at least 30 minutes after preparation. The net amount of BSA entrapped in PUM was maintained unchanged with increasing BSA concentration. BSA-containing PUM were shown easily to be visible in in vivo rabbit kidney. There was no difference in echogenicity between the loaded and unloaded PUM. Ultrasound duration had a positive relationship with BSA release. Ultrasound of 30 seconds stimulated 94.1% and 93.3% of BSA release from PUM solutions containing 0.3% and 1.5% BSA, respectively. Conclusions: Protein-loaded PUM exhibited satisfactory physical characteristics and were potent for using in ultrasound-triggered delivery.

Preparation, characterization and in-vitro evaluation of sustained release protein-loaded nanoparticles based on biodegradable polymers

Controlled drug delivery technology of proteins/peptides from biodegradable nanoparticles has emerged as one of the eminent areas to overcome formulation associated problems of the macromolecules. The purpose of the present investigation was to develop protein-loaded nanoparticles using biodegradable polymer poly L-lactide-co-glycolidic acid (PLGA) with bovine serum albumin (BSA) as a model protein. Despite many studies available with PLGA-based protein-loaded nanoparticles, production know-how, process parameters, protein loading, duration of protein release, narrowing polydispersity of particles have not been investigated enough to scale up manufacturing of protein-loaded nanoparticles in formulations. Different process parameters such as protein/polymer ratio, homogenizing speed during emulsifications, particle surface morphology and surface charges, particle size analysis and in-vitro protein release were investigated. The in-vitro protein release study suggests that release profile of BSA from nanoparticles could be modulated by changing protein-polymer ratios and/or by varying homogenizing speed during multiple-emulsion preparation technique. The formulation prepared with protein-polymer ratio of 1:60 at 17,500 rpm gave maximum protein-loading, minimum polydispersion with maximally sustained protein release pattern, among the prepared formulations. Decreased (10,000 rpm) or enhanced (24,000 rpm) homogenizing speeds resulted in increased polydispersion with larger particles having no better protein-loading and -release profiles in the present study.

In vitro release of bovine serum albumin from alginate/HPMC hydrogel beads

In recent years, the use of swelling polymeric matrices for the encapsulation and controlled release of protein drugs has received significant attention. The purpose of the present study was to investigate the release of albumin, a model protein from alginate/hydroxypropyl-methylcellulose (HPMC) gel beads. A hydrogel system comprised of two natural, hydrophilic polymers; sodium alginate and HPMC was studied as a carrier of bovine serum albumin (BSA) which was used as a model protein. The morphology, bead size and the swelling ratio were studied in different physical states; fully swollen, dried and reswollen using scanning electron microscopy and image analysis. Finally the effect of different alginate/HPMC ratios on the BSA release profile in physiological saline solution was investigated. Swelling experiments revealed that the bead diameter increases with the viscosity of the alginate solution while the addition of HPMC resulted in a significant increase of the swelling ratio. The BSA release patterns showed that the addition of HPMC increased the protein-release rate while the release mechanism fitted the Peppas model. Alginate/HPMC beads prepared using the ionic gelation exhibited high BSA loading efficiency for all formulations. The presence of HPMC increased the swelling ability of the alginate beads while the particle size remained unaffected. Incorporation of HPMC in the alginate gels also resulted in improved BSA release in physiological saline solution. All formulations presented a non-Fickian release mechanism described by the Peppas model. In addition, the implementation of non-parametric tests showed significant differences in the release patterns between the alginate/HPMC and the pure alginate beads, respectively.

Water-free microencapsulation of proteins within PLGA microparticles by spray drying using PEG-assisted protein solubilization technique in organic solvent

Poly( d, l-lactic- co-glycolic acid) (PLGA) microparticles encapsulating therapeutic proteins were prepared under a water-free formulation condition. Bovine serum albumin (BSA) and recombinant human growth hormone (rhGH) were homogeneously solubilized as nano-scale complexes in methylene chloride phase by using polyethylene glycol (PEG) as a complex-forming agent. The organic phase containing dissolved PLGA and PEG/protein complexes was directly spray dried to obtain PLGA microparticles encapsulating proteins. They exhibited sustained release profiles of BSA and rhGH up to 30 days with reduced initial bursts. The released protein molecules from the microparticles maintained structural integrity without aggregation, suggesting that the current single-step protein microencapsulation method without using water could be potentially applied for sustained delivery of a wide range of therapeutic protein drugs that are not soluble in organic solvents.

Porous Devices Derived from Co-Continuous Polymer Blends as a Route for Controlled Drug Release

In this study we examine the release profile of bovine serum albumin (BSA) from a porous polymer matrix derived from a co-continuous polymer blend. The porosity is generated through the selective extraction of one of the continuous phases. This is the first study to examine the approach of using morphologically tailored co-continuous polymer blends as a template for generating porous polymer materials for use in controlled release. A method for the preparation of polymeric capsules is introduced, and the effect of matrix pore size and surface area on the BSA release profile is investigated. Furthermore, the effect of surface charge on release is examined by surface modification of the porous substrate using layer-by-layer deposition techniques. Synthetic, nonerodible polymer, high-density polyethylene (HDPE), was used as a model substrate prepared by melt blending with two different styrene-ethylene-butylene copolymers. Blends with HDPE allow for the preparation of porous substrates with small pore sizes (300 and 600 nm). A blend of polylactide (PLA) and polystyrene was also used to prepare porous PLA with a larger pore size (1.5 microm). The extents of interconnectivity, surface area, and pore dimension of the prepared porous substrates were examined via gravimetric solvent extraction, BET nitrogen adsorption, mercury porosimetry, and image analysis of scanning electron microscopy micrographs. With a loading protocol into the porous HDPE and PLA involving the alternate application of pressure and vacuum, it is shown that virtually the entire porous network was accessible to BSA loading, and loading efficiencies of between 80% and 96% were obtained depending on the pore size of the carrier and the applied pressure. The release profile of BSA from the microporous structure was monitored by UV spectrophotometry. The influence of pore size, surface area, surface charge, and number of deposited layers is demonstrated. It is shown that an effective closed-cell structure in porous PLA can be prepared, effectively eliminating all short-term BSA release.

Shell Cross-Linked Hyaluronic Acid/Polylysine Layer-by-Layer Polyelectrolyte Microcapsules Prepared by Removal of Reducible Hyaluronic Acid Microgel Cores

Shell cross-linked hollow polyelectrolyte microcapsules composed of hyaluronic acid (HA) and poly- l-lysine (PLL) were prepared by layer-by-layer (LBL) adsorption and subsequent core removal by a reductive agent. Disulfide cross-linked HA microgels were used as template core materials for the LBL deposition on the surface and removed by treatment of dithiothreitol at neutral pH condition. HA/PLL polyelectrolyte multilayers on the shell were chemically cross-linked via carbodiimide chemistry, and their physicochemical properties and drug release behaviors were investigated. Shell cross-linked HA/PLL polyelectrolyte microcapsules exhibited far enhanced physical stability against freeze-thaw cycles and acidic pH conditions compared to the un-cross-linked ones. The cross-linked HA/PLL multilayer shell also demonstrated pH responsive permeability, which became more permeable at low pH than at neutral pH. When bovine serum albumin (BSA), as a model protein drug, was loaded inside using the pH-dependent permeability, BSA release profiles from the microcapsules could be readily modulated by varying medium pH values or adding an HA digesting enzyme (hyaluronidase) in the incubation medium.

Preparation of nanoparticles composed of chitosan and its derivatives as delivery systems for macromolecules

AbstractThree different kinds of nanoparticles for paracellular transport were prepared using a simple and mild ionic‐gelation method. Sodium tripolyphosphate (TPP) as crosslinking agent was added into three kinds of solutions, which were chitosan solution, physical blending solution of chitosan, and glycidyl trimethylammonium chloride (GTMAC), and O‐(2‐hydroxyl) propyl‐3‐trimethyl ammonium chitosan chloride (O‐HTCC) solution respectively. O‐HTCC was synthesized by coupling of GTMAC to chitosan whose functional groups of the NH2 groups were protected. The nanoparticles were characterized by transmission electron microscopy, atomic force microscopy, photon correlation spectroscopy, and zeta potential measurement. The results showed that increasing TPP concentration promoted the size of chitosan nanoparticles, a decrease in the size of O‐HTCC nanoparticles incurred on the contrary. The size of O‐HTCC nanoparticles is slightly bigger than that of pure chitosan nanoparticles, and smaller than that of physical blending nanoparticles (PBN). Bovine serum albumin (BSA), as a model protein drug, was incorporated into the nanoparticles. Compared with chitosan nanoparticles and PBN, high BSA loading efficiency (87.5%) and loading capacity (99.5%) are achieved by quaternized chitosan (O‐HTCC) nanoparticles, and the release profile of BSA from nanoparticles has an obvious burst effect and a slowly continuous release phase followed. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2007

Incorporation of a model protein into chitosan–bile salt microparticles

In order to develop a mucosal delivery system based on biocompatible polymers, a new methodology for production of protein-loaded microparticles is developed. Chitosan anionic precipitation/coacervation is accomplished by the addition of sodium deoxycholate (DCA). These microparticles were prepared under mild conditions, where bovine serum albumin (BSA) and DCA were simply dipped into a chitosan solution under stirring. Platelet-like and/or spherical microparticles, having high protein loading efficiency and relatively low protein external exposure, are obtained. To achieve a better compaction of the microparticle matrix, block copolymers and other non-ionic surfactants are added to the formulation. BCA analysis and fluorescence quenching were used to assess the degree of protein exposure. BSA release profiles for chitosan–DCA formulations in PBS pH 7.4 and HCl 0.1N revealed, in most cases, an initial burst release, but more than 55% of the BSA remains protected inside the microparticles. It is also observed that in acidic environment (HCl 0.1N) the protein is better shielded from the environment. Some of the formulations show good properties for mucosal protein delivery, and one of those here developed is now being tested in vivo, for mucosal administration of an adenovirus vaccine.