In vitro study of BSA gel/polyelectrolite complexes core shell microcapsules encapsulating doxorubicin for antitumoral targeted treatment

Abstract

The aim of this study is to develop core shell microcapsules of bovine serum albumin (BSA) gel with a complex polyelectrolite multilayer shell of natural polysaccharides with opposite charges, pectin (P), chitosan (Chi), and hyaluronic acid (HA) respectively, encapsulating Doxorubicin (Dox) as a carrier for targeted anti-tumoral treatment of hepatic cell carcinoma (HCC). A sacrificial CaCO₃ template method was used in order to obtain microcapsules with a BSA gel core and a layer-by-layer (Lbl) deposition technique of polyelectrolite complexes formed between P/Chi in the inner layers and HA/Chi in the outer shell layers. The preformed microcapsules, BSA gel/P/Chi/HA, noted as ms, have been applied for Dox encapsulation (ms-Dox). Dox encapsulation and release in different pH media were studied in order to elucidate the interactions between pH dependently charged species involved in the Dox loading/releasing processes. The structure characterization of ms/ms-Dox was evaluated by FTIR and UV-Vis spectroscopy, X-ray diffraction, thermal analy sis, optical microscopy, confocal laser scanning microscopy, and scanning electron microscopy. The in vitro study for citotoxicity assessment on normal and tumoral cells of both ms and ms-Dox was performed using mesenchymal stem cells (MSCs) and Hep2G HCC cell lines. Results of physical-chemical analyses confirm the successful encapsulation of Dox in ms, and the in vitro biological study recommends ms-Dox as a candidate for future in vivo research as a targeted anti-tumoral treatment modality applications.     

Chitosan/polyethylene glycol–alginate microcapsules for oral delivery of hirudin

A mild chitosan/calcium alginate microencapsulation process, as applied to encapsulation of biological macromolecules such as albumin and hirudin, was investigated. The polysaccharide chitosan was reacted with sodium alginate in the presence of calcium chloride to form microcapsules with a polyelectrolyte complex membrane. Hirudin-entrapped alginate beads were further surface coated with polyethylene glycol (PEG) via glutaraldehyde functionalities. It was observed that approximately 70% of the content is being released into Tris-HCl buffer, pH 7.4 within the initial 6 h and about 35% release of hirudin was also observed during treatment with 0.1 M HCl, pH 1.2 for 4 h. But acid-treated capsules had released almost all the entrapped hirudin into Tris-HCl, pH 7.4 media within 6 h. From scanning electron microscopic and swelling studies, it appears that the chitosan and PEG have modified the alginate microcapsules and subsequently the protein release. The microcapsules were also prepared by adding dropwise albumin-containing sodium alginate mixture into a PEG– CaCl₂ system. Increasing the PEG concentration resulted in a decrease rate of albumin release. The results indicate the possibility of modifying the formulation to obtain the desired controlled release of bioactive peptides (hirudin), for a convenient gastrointestinal tract delivery system. © 1998 John Wiley & Sons, Inc. J Appl Polym Sci 70: 2143–2153, 1998     

Design of intelligent chitosan/heparin hollow microcapsules for drug delivery

In this study, a novel strategy has been developed for the assembly of polyelectrolyte multilayer (PEM) on CaCO₃ templates in acidic pH solutions, where consecutive polyelectrolyte layers (heparin/poly(allylamine hydrochloride) or heparin/chitosan) were deposited on PEM hollow microcapsules established previously on CaCO₃ templates. The PEM build‐up, hollow capsule characterization and successful encapsulation of fluorescein 5(6)‐isothiocyanate (FITC)‐Dextran by coprecipitation with CaCO₃ are demonstrated. Improvement by the removal of CaCO₃ core was achieved while the depositions. In the course of the release profile, high retardation for encapsulated FITC‐Dextran was observed. The combined shell capsules system is a significant trait that has potential use in tailoring functional layer‐by‐layer capsules as intelligent drug delivery vehicles where the preliminary in vitro tests showed the responsiveness on the enzymes. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44425.     

Structure and control release of chitosan/carboxymethyl cellulose microcapsules

Microcapsules of chitosan/sodium carboxymethyl cellulose (NaCMC) were successfully prepared using a novel method of emulation phase separation. Their structure and morphology were characterized by infrared spectroscopy (IR), scanning electron microscopy (SEM), and X-ray diffraction. Bovine serum albumin (BSA) was encapsulated in the microcapsules to test their release behavior. The swelling behavior, encapsulation efficiency, and release behavior of the microcapsules with different chitosan contents and pH conditions were investigated. The results indicated that the microcapsules have a high encapsulation efficiency (75%) and a suitable size (20–50 μm). The BSA in the microcapsules was speedily released at pH 7.2, namely, in intestinal fluid. The BSA release was reduced with increase of the chitosan content from 17 to 38% in the microcapsules. Acid-treated microcapsules have a compact structure, owing to a strong electrostatic interaction caused by —NH₂ groups of chitosan and —COOH groups of CMC, and the encapsulated BSA was hardly released at pH 1.0, namely, in gastric juice. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 82: 584–592, 2001     

Biodegradable Nanoparticles-Loaded PLGA Microcapsule for the Enhanced Encapsulation Efficiency and Controlled Release of Hydrophilic Drug

Recently, nano- and micro-particulate systems have been widely utilized to deliver pharmaceutical compounds to achieve enhanced therapeutic effects and reduced side effects. Poly (DL-lactide-co-glycolide) (PLGA), as one of the biodegradable polyesters, has been widely used to fabricate particulate systems because of advantages including controlled and sustained release, biodegradability, and biocompatibility. However, PLGA is known for low encapsulation efficiency (%) and insufficient controlled release of water-soluble drugs. It would result in fluctuation in the plasma levels and unexpected side effects of drugs. Therefore, the purpose of this work was to develop microcapsules loaded with alginate-coated chitosan that can increase the encapsulation efficiency of the hydrophilic drug while exhibiting a controlled and sustained release profile with reduced initial burst release. The encapsulation of nanoparticles in PLGA microcapsules was done by the emulsion solvent evaporation method. The encapsulation of nanoparticles in PLGA microcapsules was confirmed by scanning electron microscopy and confocal microscopy. The release profile of hydrophilic drugs can further be altered by the chitosan coating. The chitosan coating onto alginate exhibited a less initial burst release and sustained release of the hydrophilic drug. In addition, the encapsulation of alginate nanoparticles and alginate nanoparticles coated with chitosan in PLGA microcapsules was shown to enhance the encapsulation efficiency of a hydrophilic drug. Based on the results, this delivery system could be a promising platform for the high encapsulation efficiency and sustained release with reduced initial burst release of the hydrophilic drug.     

Microcapsules prepared from a polycondensate‐based cement dispersant via layer‐by‐layer self‐assembly on melamine‐formaldehyde core templates

Novel microcapsules were prepared from colloidal core–shell particles by acid dissolution of the organic core. Weakly crosslinked, monodisperse and spherical melamine‐formaldehyde polycondensate particles (diameter ～ 1 μm) were synthesized as core template and coated with multilayers of an anionic polyelectrolyte via layer‐by‐layer deposition technique. As polyelectrolytes, an anionic naphthalenesulfonate formaldehyde polycondensate that is a common concrete superplasticizer and thus industrially available, and cationic poly(allylamine hydrochloride) were used. Core removal was achieved by soaking the core–shell particles in aqueous hydrochloric acid at pH 1.6, resulting in hollow microcapsules consisting of the polyelectrolytes. Characterization of the template, the core–shell particles, and the microcapsules plus tracking of the layer‐by‐layer polyelectrolyte deposition was performed by means of zeta potential measurement and scanning electron microscopy. The microcapsules might be useful as microcontainers for cement additives. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Chitosan/calcium–alginate beads for oral delivery of insulin

A mild chitosan/calcium alginate microencapsulation process, as applied to encapsulation of biological macromolecules such as albumin and insulin, was investigated. The microcapsules were derived by adding dropwise a protein-containing sodium alginate mixture into a chitosan–CaCl₂ system. The beads containing a high concentration of entrapped bovine serum albumin (BSA) as more than 70% of the initial concentration were achieved via varying chitosan coat. It was observed that approximately 70% of the content is being released into Tris-HCl buffer, pH 7.4 within 24 h and no significant release of BSA was observed during treatment with 0.1M HCl pH 1.2 for 4 h. But the acid-treated beads had released almost all the entrapped protein into Tris-HCl pH 7.4 media within 24 h. Instead of BSA, the insulin preload was found to be very low in the chitosan/calcium alginate system; the release characteristics were similar to that of BSA. From scanning electron microscopic studies, it appears that the chitosan modifies the alginate microspheres and subsequently the protein loading. The results indicate the possibility of modifying the formulation in order to obtain the desired controlled release of bioactive peptides (insulin), for a convenient gastrointestinal tract delivery system. © 1996 John Wiley & Sons, Inc.     

A novel microencapsulation of neem (Azadirachta Indica A. Juss.) seed oil (NSO) in polyelectrolyte complex of κ‐carrageenan and chitosan

Microcapsules containing neem (Azadirachta Indica A. Juss.) seed oil (NSO) were prepared by encapsulation of natural liquid pesticide NSO in a polyelectrolyte complex of κ‐carrageenan and chitosan. The optimum ratio between carrageenan and chitosan to form a stable polyelectrolyte complex was found as 1 : 0.36. The microencapsulation method for NSO loading was also optimized. SEM study demonstrated that the surface of the microcapsules became more irregular as oil loading increased. The release rates of NSO were studied by varying the percentage of oil loading, concentration of cross‐linking agent, and polymer concentration. Fourier transform infrared spectroscopy (FTIR) study confirmed the complex formation between κ‐carrageenan and chitosan. Differential scanning calorimetry (DSC) and FTIR study indicated the absence of any significant interaction between polyelectrolyte complex of κ‐carrageenan ‐chitosan and NSO. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

UV–Vis irradiation fatigue resistance improvement of azo photochromic compound using polyurethane‐chitosan double shell encapsulation

Double shell photochromic microcapsules were prepared by in situ polymerization with polyurethane and chitosan as inner and outer shell respectively. FT‐IR indicated that chitosan‐glutaraldehyde copolymer formed by imine and combined with polyurethane photochromic nanocapsules. The polyurethane‐chitosan microcapsules exhibited a near‐spherical shape, and the average particle size of nanocapsules was around 1.2 μm. The half‐life of azo compound increased from 135 to 340 min after encapsulated in polyurethane‐chitosan microcapsules. The polyurethane‐chitosan shell delayed the coloration process for 14 s compared with azo compound in ethanol, however, the absorbance of azo compound increased by 17.15% in polyurethane‐chitosan microcapsules. It decreased from 0.3486 to 0.1738 in ethanol during 20 s, however, it decreased from 0.4084 to 0.2625 in polyurethane‐chitosan microcapsules in 55 s when it reached steady state during decoloration process. Polyurethane‐chitosan double shell encapsulation is an effective route for improving the fatigue resistance, increasing the absorbance of azo compound. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40895.     

溶剂挥发法制备聚砜包覆桐油自修复微胶囊

以聚砜为壁材,桐油为芯材,采用溶剂挥发法制备了聚砜（PSF）包覆桐油自修复微胶囊。考查了不同种类的分散剂、搅拌速度、芯壁比（芯材与壁材的质量比）等工艺参数对微胶囊性能的影响,通过扫描电子显微镜、光学显微镜和热重分析仪等对微胶囊的表观形貌、粒径、壁厚、包覆率和热稳定性能等进行表征。采用所合成的微胶囊制备了环氧树脂基防腐蚀涂层,并对其防腐蚀性能进行了评价。结果表明,30 ℃时,以明胶/聚乙烯醇复配体系作为分散剂,芯材与壁材质量比为1.3:1,搅拌速度为700 r/min时制备出的微胶囊表面光滑致密,粒径在130 μm左右,热稳定温度为350 ℃;盐雾实验结果表明,所制备的微胶囊自修复涂层具有良好的防腐蚀性能。     

Microencapsulation of Zanthoxylum limonella oil (ZLO) in genipin crosslinked chitosan–gelatin complex for mosquito repellent application

Essential oil containing chitosan gelatin complex microcapsules crosslinked with genipin were prepared by complex coacervation process. The effects of various parameters such as oil loading, ratio of chitosan to gelatin, degree of crosslinking on oil content, encapsulation efficiency, and the release rate of the essential oil were studied. Scanning electron microscopy study indicated that the surface of the microcapsules were more irregular as the amount of oil loading increased. Thermal stability of microcapsules improved with the increase in the amount of chitosan in chitosan–gelatin matrix as revealed by thermogravimetric analysis. FT‐IR spectroscopy and differential scanning calorimetry study indicated that there was no significant interaction between chitosan–gelatin complex and oil. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Preparation and characterization of a novel thermosensitive hydrogel based on chitosan and gelatin blends

In this study, a novel injectable in situ gelling thermosensitive hydrogel system based on chitosan and gelatin blends was designed and investigated. The addition of gelatin provides the correct buffering and other physicochemical conditions including control of hydrophobic interactions and hydrogen bonding, which are necessary to retain chitosan in solution at neutral pH near 4°C and furthermore to allow gel formation upon heating to body temperature. The chitosan/gelatin hydrogels were studied by FTIR, swelling, and rheological analysis. The rheological analysis evidenced the endothermic gelation of chitosan/gelatin solutions, which indicated their gelation temperatures and reflected the effect of gelatin concentration on the thermosensitive properties of gels. The morphology of this system was examined with laser scanning confocal microscopy and scanning electron microscopy. The images indicated that the gels were quite heterogeneous and porous. The investigation of these gels as vehicles for delivering bovine serum albumin as a model drug of protein showed that the system could sustain the release of the protein drug. These results show that chitosan/gelatin solutions can form gels rapidly at body temperature and have promising perspective for their use in local and sustained delivery of protein drug. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Hyaluronan/Diethylaminoethyl Chitosan Polyelectrolyte Complexes as Carriers for Improved Colistin Delivery

Improving the therapeutic characteristics of antibiotics is an effective strategy for controlling the growth of multidrug-resistant Gram-negative microorganisms. The purpose of this study was to develop a colistin (CT) delivery system based on hyaluronic acid (HA) and the water-soluble cationic chitosan derivative, diethylaminoethyl chitosan (DEAECS). The CT delivery system was a polyelectrolyte complex (PEC) obtained by interpolymeric interactions between the HA polyanion and the DEAECS polycation, with simultaneous inclusion of positively charged CT molecules into the resulting complex. The developed PEC had a hydrodynamic diameter of 210–250 nm and a negative surface charge (ζ-potential = −19 mV); the encapsulation and loading efficiencies were 100 and 16.7%, respectively. The developed CT delivery systems were characterized by modified release (30–40% and 85–90% of CT released in 15 and 60 min, respectively) compared to pure CT (100% CT released in 15 min). In vitro experiments showed that the encapsulation of CT in polysaccharide carriers did not reduce its antimicrobial activity, as the minimum inhibitory concentrations against Pseudomonas aeruginosa of both encapsulated CT and pure CT were 1 μg/mL.     

Synthesis and characterization of chitosan/urea‐formaldehyde shell microcapsules containing dicyclopentadiene

Microcapsules containing healing agent have been used to develop the self‐healing composites. These microcapsules must possess special properties during the use of composites such as stability in surrounding, appropriate mechanical strength, and lower permeability. A new series of microcapsules containing dicyclopentadiene with chitosan/urea‐formaldehyde copolymer as shell materials were synthesized by in situ copolymerization technology. The microencapsulating mechanism was discussed and the process was explained. Also, the factors influencing the preparation of microcapsules were analyzed. The morphology and shell wall thickness of microcapsules were observed by using scanning electron microscopy. The size of microcapsules was measured using optical microscope and the size distribution was investigated based on data sets of at least 200 measurements. The chemical structure and thermal properties of microcapsules were characterized by Fourier transform infrared spectroscopy and thermogravimetric analysis, respectively. The storage stability and isothermal aging experiment of microcapsules were also investigated. Results indicted that the chitosan/urea‐formaldehyde microcapsules containing dicyclopentadiene were synthesized successfully; the copolymerization occurred between chitosan and urea‐formaldehyde prepolymer. The microcapsule size is in the range of 10–160 μm with an average of 45 μm. The shell thickness of microcapsules is in the range of 1–7 μm and the core content of microcapsules is 67%. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Application of the biodegradable diblock copolymer poly(L‐lactide)‐block‐poly(L‐cysteine): Drug delivery and protein conjugation

A novel approach to self‐assembled and shell‐crosslinked (SCL) micelles from the diblock copolymer poly(L ‐lactide)‐block‐poly(L ‐cysteine) to be used as drug and protein delivery carriers is described. Rifampicin was used as a model drug. The drug‐loaded SCL micelles were obtained by self‐assembly of the copolymer in the presence of the drug in aqueous media. Their morphology and size were studied with dynamic light scattering and field emission scanning electron microscopy. The rifampicin loading capacity and encapsulation efficiency were studied with ultraviolet–visible spectrophotometry. The drug‐release rate in vitro depended on the oxidizing and reducing environment. Moreover, a straightforward approach to the conjugation of the copolymer with bovine serum albumin (BSA) was developed, and a gel electrophoresis test demonstrated that this conjugated BSA could be reversibly released from the copolymer substrate under reducing conditions. In conclusion, this L ‐cysteine copolymer can be used in drug delivery and in protein fixation and recovery. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Synthesis and characterization of cross-linked poly(acrylic acid)-poly(styrene-alt-maleic anhydride) core-shell microcapsule absorbents for cement mortar

We synthesized core-shell microcapsule absorbents with cPAA (cross-linked poly(acrylic acid)) as the core and PSMA (poly(styrene-alt-maleic anhydride)) as the shell by precipitation polymerization, where the shell served to delay the absorption of excess water in cement mortars. To control shell thickness, the cPAA-PSMA capsules were synthesized with core monomer mass to shell monomer mass ratios of 1/0.5, 1/1, and 1/1.5. We observed the hydrolysis of the PSMA polymer in a cement-saturated aqueous solution by Fourier transform infrared (FT-IR) spectroscopy. Furthermore, core-shell structures were observed for 1/1 (cPAA-PSMA #3) and 1/1.5 (cPAA-PSMA #4) core/shell monomer mass ratios, whereas no core-shell structures were observed for the 1/0.5 (cPAA-PSMA #2) microcapsules by transmission electron microscopy (TEM).     

Fabrication of Core–Shell Novel Polyurea Microcapsules Using Isophorone Diisocyanate (IPDI) Trimer for Release System

Isophorone diisocyanate (IPDI) trimer based novel polyurea core shell structures were developed by interfacial polymerization. Different operating conditions have been used to fabricate shell to encapsulate core. Characterizations of prepared microcapsules were done by Fourier transform infrared spectroscopy, thermogravimetric analysis, and particle size analyzer. The surface morphology of microcapsules was examined by optical microscopy, scanning electron microscopy, and transmission electron microscopy. The release rate of core from microcapsules was estimated by UV and gas chromatography. The results revealed that tailor made release can be adjusted by varying operational protocol for shell and fabricated shell can be extended to other applications such as self-healing coatings and drug delivery.     

Polymer hollow particles: Encapsulation of phosphoric acid partial esters and morphology manipulation

The present work demonstrates the possible use of emulsion polymerization for fabricating structured-polymer particles which can store active materials. The hollow polymer particles were synthesized by multi-stage emulsion polymerization consisting of four main stages, (1) the preparation of alkali-swellable core latexes containing active materials, (2) first core-shell polymerization of a monomer mixture of methyl methacrylate (MMA), butyl acrylate (BA) and methacrylic acid (MAA), (3) second core-shell polymerization of styrene and (4) a neutralization stage. The morphology of synthesized capsules was observed by transmission electron microscopy (TEM). The size of the capsule prepared by standard recipe was around 300 nm and the polydispersity index was 0.024 representing that size distribution was highly monodisperse. The specific target material of encapsulation was the phosphoric acid partial ester. The amount of phosphoric acid partial ester encapsulated was determined by thermogravimetric analysis (TGA). From studies of encapsulation behaviors, it was found that the encapsulation efficiency and location of phosphoric acid partial ester in the interior of the particles were mainly dependant on its partition coefficient. In addition, the morphology of polymer capsule was manipulated by varying process parameters. The morphology changes, such as those of pore size and roughness of polymer shell, were characterized by scanning electron microscopy (SEM) and analysis of nitrogen adsorption and desorption isotherm. When neutralized with N,N′-dimethylethanolamine simultaneously during the styrene polymerization, the surface area of polymer capsule was increased drastically by about 5 times due to the formation of mesopores and the roughening of the surface on the hollow polymer shell.     

Arachidonic acid-encapsulated microcapsules with core-shell structure prepared by coaxial electrospray

Arachidonic acid (AA) is an n-6 polyunsaturated 20-carbon fatty acid taking important immunomodulatory roles in infant growth, brain development, and health. However, one of the major drawbacks of polyunsaturated fatty acids is their high susceptibility to oxidation followed by unpleasant odors. Microencapsulation has been proposed as an excellent strategy to solve these problems. In this work, coaxial electrospray was utilized to fabricate AA/zein microcapsules with core-shell structures which were verified by scanning electron microscopy, transmission electron microscopy, and confocal laser scanning microscopy. Microcapsules morphology was conveniently controlled through electrospray parameters and AA was centralized in the core. The microcapsule diameter ranged from 1 to 7 μm. Results of peroxide values and GC–MS demonstrated that AA in microcapsules possess improved oxidative stability and the unpleasant odors of AA oil was inhibited. The results indicate coaxial electrospray is a useful and convenient tool to fabricate core-shell microcapsules with immiscible materials, which will broaden the potential use of microcapsules as nutrition fortifiers in the food industry.     

Mechanical properties and drug release of microcapsules containing quaternized‐chitosan‐modified reduced graphene oxide in the capsular wall

Reduced graphene oxide (rGO) sheets were first modified with 2‐hydroxypropyltrimethyl ammonium chloride chitosan (HACC), and these modified rGO sheets (named HACC–rGO) were used as reinforcement materials and introduced to the walls of chitosan (CS) microcapsules. All of the monodisperse microcapsules were conveniently generated by a gas–liquid microfluidic technique. The results of scanning electron microscopy, X‐ray diffraction, and thermogravimetric analysis all demonstrate that the HACC–rGO sheets existed and were dispersed in the capsular shell. The HACC–rGO‐reinforced CS microcapsules showed better mechanical strength and better chemical stability with an α‐cyclodextrin solution than the CS microcapsules without HACC–rGO. Importantly, the HACC–rGO‐reinforced CS microcapsules exhibited a slower drug‐release behavior and provide a method for the control of the release rate of drug‐loaded microcapsules. In an in vitro cytotoxicity evaluation by a 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide assay, the Schwann cells still showed good cell viability after they were treated by extracts of the CS/HACC–rGO microcapsules with concentrations ranging from 0.02 to 2000 μg/mL. Therefore, the HACC–rGO‐reinforced CS microcapsules are promising for applications in the fields of drug delivery and controlled release. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44549.