Xueyan Cao

PEGylated dendrimer-entrapped gold nanoparticles for in vivo blood pool and tumor imaging by computed tomography.

We report the synthesis and characterization of dendrimer-entrapped gold nanoparticles (Au DENPs) modified by polyethylene glycol (PEG) with enhanced biocompatibility for computed tomography (CT) imaging applications. In this study, amine-terminated poly(amidoamine) dendrimers of generation 5 (G5.NH(2)) modified by PEG monomethyl ether (G5.NH(2)-mPEG(20)) were used as templates to synthesize Au DENPs, followed by acetylation of the remaining dendrimer terminal amines to generate PEGylated Au DENPs. The partial PEGylation modification of dendrimer terminal amines allows high loading of Au within the dendrimer interior, and consequently by simply varying the Au salt/dendrimer molar ratio, the size of the PEGylated Au DENPs can be controlled at a range of 2-4 nm with a narrow size distribution. The formed PEGylated Au DENPs are water-dispersible, stable in a pH range of 5-8 and a temperature range of 0-50 °C, and non-cytotoxic at a concentration as high as 100 μm. X-ray absorption coefficient measurements show that the attenuation intensity of the PEGylated Au DENPs is much higher than that of Omnipaque with iodine concentration similar to Au. With the sufficiently long half-decay time demonstrated by pharmacokinetics studies, the PEGylated Au DENPs enabled not only X-ray CT blood pool imaging of mice and rats after intravenous injection of the particles, but also effective CT imaging of a xenograft tumor model in nude mice. These findings suggest that the designed PEGylated Au DENPs can be used as a promising contrast agent with enhanced biocompatibility for CT imaging of various biological systems, especially in cancer diagnosis.

Encapsulation of 2-methoxyestradiol within multifunctional poly(amidoamine) dendrimers for targeted cancer therapy

We report here a general approach to using multifunctional poly(amidoamine) (PAMAM) dendrimer-based platform to encapsulate a potential anticancer drug for targeted cancer therapy. In this approach, amine-terminated generation 5 (G5) PAMAM dendrimers were sequentially modified with fluorescein isothiocyanate (FI) and folic acid (FA) via covalent conjugation, followed by an acetylation reaction to neutralize the remaining amines of the dendrimer surfaces. The synthesized multifunctional dendrimers (G5.NHAc-FI-FA) were then used to complex a potential anticancer drug, 2-methoxyestradiol (2-ME) for targeted delivery of the drugs to cancer cells overexpressing high-affinity folic acid receptors (FAR). We show that the formed G5.NHAc-FI-FA/2-ME complexes with each dendrimer encapsulating approximately 3.7 2-ME molecules are water soluble and stable. In vitro release studies show that 2-ME complexed with the multifunctional dendrimers can be released in a sustained manner. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in conjunction with cell morphology observation demonstrates that the G5.NHAc-FI-FA/2-ME complexes can specifically target and display specific therapeutic efficacy to cancer cells overexpressing high-affinity FAR. Findings from this study suggest that multifunctional dendrimers may be used as a general drug carrier to encapsulate various cancer drugs for targeted therapy of different types of cancer.

Electrospun poly(lactic-co-glycolic acid)/halloysite nanotube composite nanofibers for drug encapsulation and sustained release

We report a novel electrospun composite nanofiber-based drug delivery system. In this study, halloysite nanotubes (HNTs) were first used to encapsulate a model drug, tetracycline hydrochloride. Then, the drug-loaded HNTs with an optimized encapsulation efficiency were mixed with poly(lactic-co-glycolic acid) (PLGA) polymer for subsequent electrospinning to form drug-loaded composite nanofibrous mats. The structure, morphology, and mechanical properties of the formed electrospun composite nanofibrous mats were characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, and tensile testing. In vitro drug release behavior was examined using UV-vis spectroscopy. The biocompatibility of HNT-containing PLGA fibers was evaluated through cell culture and MTT assay. We show that the incorporation of HNTs within the nanofibrous mats does not significantly change the morphology of the mats. In addition, our results indicate that this double-container drug delivery system (both PLGA polymer and HNTs are drug carriers) is beneficial to reduce the burst release of the drug and the introduction of HNTs can significantly improve the tensile strength of the polymer nanofibrous mats. Given the proved biocompatibility of the HNT-containing PLGA nanofibers via MTT assay of cell viability and SEM observation of cell morphology, the drug loaded electrospun composite nanofibrous mats developed in this study may find various applications in tissue engineering and pharmaceutical sciences.

Gene delivery using dendrimer-entrapped gold nanoparticles as nonviral vectors

Development of highly efficient nonviral gene delivery vectors still remains a great challenge. In this study, we report a new gene delivery vector based on dendrimer-entrapped gold nanoparticles (Au DENPs) with significantly higher gene transfection efficiency than that of dendrimers without AuNPs entrapped. Amine-terminated generation 5 poly(amidoamine) (PAMAM) dendrimers (G5.NH(2)) were utilized as templates to synthesize AuNPs with different Au atom/dendrimer molar ratios (25:1, 50:1, 75:1, and 100:1, respectively). The formed Au DENPs were used to complex two different pDNAs encoding luciferase (Luc) and enhanced green fluorescent protein (EGFP), respectively for gene transfection studies. The Au DENPs/pDNA polyplexes with different N/P ratios and compositions of Au DENPs were characterized by gel retardation assay, light scattering, zeta potential measurements, and atomic force microscopic imaging. We show that the Au DENPs can effectively compact the pDNA, allowing for highly efficient gene transfection into the selected cell lines as demonstrated by both Luc assay and fluorescence microscopic imaging of the EGFP expression. The transfection efficiency of Au DENPs with Au atom/dendrimer molar ratio of 25:1 was at least 100 times higher than that of G5.NH(2) dendrimers without AuNPs entrapped at the N/P ratio of 2.5:1. The higher gene transfection efficiency of Au DENPs is primarily due to the fact that the entrapment of AuNPs helps preserve the 3-dimensional spherical morphology of dendrimers, allowing for more efficient interaction between dendrimers and DNA. With the less cytotoxicity than that of G5.NH(2) dendrimers demonstrated by thiazoyl blue tetrazolium bromide assay and higher gene transfection efficiency, it is expected that Au DENPs may be used as a new gene delivery vector for highly efficient transfection of different genes for various biomedical applications.

Facile immobilization of gold nanoparticles into electrospun polyethyleneimine/polyvinyl alcohol nanofibers for catalytic applications

We report a facile approach to immobilizing gold nanoparticles (AuNPs) into electrospun polyethyleneimine (PEI)/polyvinyl alcohol (PVA) nanofibers for catalytic applications. In this study, electrospun PEI/PVA nanofibers with a mean diameter of 490 nm were first crosslinked with glutaraldehyde vapor to render them water stable. Then, the water-insoluble nanofibrous mats were used as nanoreactors to complex AuCl4− anions via binding with the free amine groups of PEI for subsequent formation and immobilization of AuNPs. The formed AuNPs with a diameter of 11.8 nm within the nanofibers do not significantly change the morphology of the nanofibers; while importantly the mechanical property of the fibers was greatly improved compared to the crosslinked fibers without AuNPs. Scanning electron microscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, energy dispersive spectroscopy, and thermogravimetric analysis were used to characterize these hybrid nanofibers. Furthermore, we show that the AuNP-containing nanofibers display an excellent catalytic activity and reusability for the transformation of 4-nitrophenol to 4-aminophenol. The present approach to fabricating AuNP-containing nanofibers may be extended for producing other nanoparticle-containing composite nanofibrous materials for various applications in catalysis, sensing, and biomedical sciences.

Efficient Catalytic Reduction of Hexavalent Chromium Using Palladium Nanoparticle-Immobilized Electrospun Polymer Nanofibers

We report a facile and economic approach to fabricating catalytic active palladium (Pd) nanoparticle (NP)-immobilized electrospun polyethyleneimine (PEI)/polyvinyl alcohol (PVA) nanofibers for catalytic reduction of hexavalent chromium (Cr(VI)) to trivalent chromium (Cr(III)). In this study, PEI/PVA nanofibrous mats were first electrospun from homogeneous mixture solution of PEI and PVA, followed by cross-linking with glutaraldehyde vapor to render the fibers with good water stability. The nanofibrous mats were then alternatively soaked in potassium tetrachloropallidate (K2PdCl4) and sodium borohydride solution, and the PdCl4(2-) anions complexed with the free amine groups of PEI were able to be reduced to form zero-valent Pd NPs. The formed Pd NP-containing PEI/PVA nanofibers were characterized by different techniques. We show that the immobilization of Pd NPs does not significantly change the morphology of the PEI/PVA nanofibers; instead the mechanical durability of the fibers is significantly improved. The formed Pd NPs with a mean diameter of 2.6 nm are quite uniformly distributed within the fibers with a small portion of particles having a denser distribution at the outer surface of the fibers. The catalytic activity and reusability of the fabricated Pd NP-containing fibrous mats were evaluated by transformation of Cr(VI) to Cr(III) in aqueous solution in the presence of a reducing agent. Our results reveal that the Pd NP-containing nanofibrous mats display an excellent catalytic activity and reusability for the reduction of Cr(VI) to Cr(III). The facile approach to fabricating metal NP-immobilized polymer nanofibers with a high surface area to volume ratio, enhanced mechanical durability, and uniform NP distribution may be extended to prepare different NP-immobilized fibrous systems for various applications in catalysis, sensing, environmental sciences, and biomedicine.

Targeted delivery of doxorubicin into cancer cells using a folic acid–dendrimer conjugate

We report here a general approach to using multifunctional poly(amidoamine) (PAMAM) dendrimer-based platform to encapsulate an anticancer drug doxorubicin (DOX) for targeted cancer therapy. In this approach, generation 5 (G5) PAMAM dendrimers modified with fluorescein isothiocyanate (FI) and folic acid (FA) via covalent conjugation, and with remaining terminal amines being acetylated (G5.NHAc-FI-FA) were used to complex DOX for targeted delivery of the drug to cancer cells overexpressing high-affinity folic acid receptors (FAR). We show that the formed G5.NHAc-FI-FA/DOX complexes with each dendrimer encapsulating approximately one DOX molecule are water soluble and stable. In vitro release studies show that DOX complexed with the multifunctional dendrimers can be released in a sustained manner. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability assay in conjunction with cell morphology observation demonstrates that the G5.NHAc-FI-FA/DOX complexes can specifically target and display specific therapeutic efficacy to cancer cells overexpressing high-affinity FAR. Findings from this study suggest that multifunctional dendrimers may be used as a general drug carrier to encapsulate various cancer drugs for targeting therapy of different types of cancer.

Improved cellular response on multiwalled carbon nanotube-incorporated electrospun polyvinyl alcohol/chitosan nanofibrous scaffolds

We report the fabrication of multiwalled carbon nanotube (MWCNT)-incorporated electrospun polyvinyl alcohol (PVA)/chitosan (CS) nanofibers with improved cellular response for potential tissue engineering applications. In this study, smooth and uniform PVA/CS and PVA/CS/MWCNTs nanofibers with water stability were formed by electrospinning, followed by crosslinking with glutaraldehyde vapor. The morphology, structure, and mechanical properties of the formed electrospun fibrous mats were characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, and mechanical testing, respectively. We showed that the incorporation of MWCNTs did not appreciably affect the morphology of the PVA/CS nanofibers; importantly the protein adsorption ability of the nanofibers was significantly improved. In vitro cell culture of mouse fibroblasts (L929) seeded onto the electrospun scaffolds showed that the incorporation of MWCNTs into the PVA/CS nanofibers significantly promoted cell proliferation. Results from this study hence suggest that MWCNT-incorporated PVA/CS nanofibrous scaffolds with small diameters (around 160 nm) and high porosity can mimic the natural extracellular matrix well, and potentially provide many possibilities for applications in the fields of tissue engineering and regenerative medicine.

Biocompatibility of Electrospun Halloysite Nanotube-Doped Poly(Lactic-co-Glycolic Acid) Composite Nanofibers

Organic/inorganic hybrid nanofiber systems have generated great interest in the area of tissue engineering and drug delivery. In this study, halloysite nanotube (HNT)-doped poly(lactic-co-glycolic acid) (PLGA) composite nanofibers were fabricated via electrospinning and the influence of the incorporation of HNTs within PLGA nanofibers on their in vitro biocompatibility was investigated. The morphology, mechanical and thermal properties of the composite nanofibers were characterized by scanning electron microscopy (SEM), tensile test, differential scanning calorimetry and thermogravimetric analysis. The adhesion and proliferation of mouse fibroblast cells cultured on both PLGA and HNT-doped PLGA fibrous scaffolds were compared through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay of cell viability and SEM observation of cell morphology. We show that the morphology of the PLGA nanofibers does not appreciably change with the incorporation of HNTs, except that the mean diameter of the fibers increased with the increase of HNT incorporation in the composite. More importantly, the mechanical properties of the nanofibers were greatly improved. Similar to electrospun PLGA nanofibers, HNT-doped PLGA nanofibers were able to promote cell attachment and proliferation, suggesting that the incorporation of HNTs within PLGA nanofibers does not compromise the biocompatibility of the PLGA nanofibers. In addition, we show that HNT-doped PLGA scaffolds allow more protein adsorption than those without HNTs, which may provide sufficient nutrition for cell growth and proliferation. The developed electrospun HNT-doped composite fibrous scaffold may find applications in tissue engineering and pharmaceutical sciences.

Carbon nanotube-incorporated multilayered cellulose acetate nanofibers for tissue engineering applications.

We report the fabrication of a novel carbon nanotube-containing nanofibrous polysaccharide scaffolding material via the combination of electrospinning and layer-by-layer (LbL) self-assembly techniques for tissue engineering applications. In this approach, electrospun cellulose acetate (CA) nanofibers were assembled with positively charged chitosan (CS) and negatively charged multiwalled carbon nanotubes (MWCNTs) or sodium alginate (ALG) via a LbL technique. We show that the 3-dimensional fibrous structures of the CA nanofibers do not appreciably change after the multilayered assembly process except that the surface of the fibers became much rougher than that before assembly. The incorporation of MWCNTs in the multilayered CA fibrous scaffolds tends to endow the fibers with improved mechanical property and promote fibroblast attachment, spreading, and proliferation when compared with CS/ALG multilayer-assembled fibrous scaffolds. The approach to engineering the nanofiber surfaces via LbL assembly likely provides many opportunities for new scaffolding materials design in various tissue engineering applications.