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The development of nanoparticles (NPs) for use in all facets of oncological disease detection and therapy has shown great progress over the past two decades. NPs have been tailored for use as contrast enhancement agents for imaging, drug delivery vehicles, and most recently as a therapeutic component in initiating tumor cell death in magnetic and photonic ablation therapies. Of the many possible core constituents of NPs, such as gold, silver, carbon nanotubes, fullerenes, manganese oxide, lipids, micelles, etc., iron oxide (or magnetite) based NPs have been extensively investigated due to their excellent superparamagnetic, biocompatible, and biodegradable properties. This review addresses recent applications of magnetite NPs in diagnosis, treatment, and treatment monitoring of cancer. Finally, some views will be discussed concerning the toxicity and clinical translation of iron oxide NPs and the future outlook of NP development to facilitate multiple therapies in a single formulation for cancer theranostics. [ABSTRACT FROM AUTHOR]

Advances in disease treatment and tissue regeneration are buoyed by new, multifaceted materials that emulate and coercively interact with the local microenvironment. Polyblend nanofibers represent an emerging class of biomimetic nanostructures that can act as proxies of the native tissue, while providing topographical and biochemical cues that promote healing. These fibers are prepared with mixtures of synthetically and naturally derived polymers that can behave cooperatively to demonstrate unique combinations of mechanical, biochemical and structural properties. This flexibility has led to the application of polyblend nanofibers in a wide assortment of tissue engineering and drug delivery systems. In this review, we will examine design criteria and properties of polymer-blend nanofibers and their use in tissue engineering and local therapeutic delivery applications. [Copyright &y& Elsevier]

A new technique of combining the gel-casting and polymer sponge methods is introduced in this study to prepare macroporous hydroxyapatite scaffolds, which provides a better control over the microstructures of scaffolds and enhances their mechanical properties. With this technique, we were able to produce scaffolds with mechanical and structural properties that cannot be attained by either the polymer sponge or gel-casting method. The scaffolds prepared have an open, uniform and interconnected porous structure with a pore size of 200–400 μm. A compressive yield strength of ∼5 MPa equivalent to that of cancellous bone and a compressive modulus of ∼8 GPa similar to that of cortical bone were achieved. The pore morphology, size, and distribution of the scaffolds were characterized using a scanning electron microscope. X-ray diffraction and Fourier transform infrared spectroscopy were used to determine the crystal structure and chemical composition of scaffolds, respectively. Scaffolds with desired porosity, pore size, and geometry can be prepared by using polymer sponges of appropriate structures. [Copyright &y& Elsevier]

Cancer is a genetic disease originating from the accumulation of gene mutations in a cellular subpopulation. Although many therapeutic approaches have been developed to treat cancer, recent studies have revealed an irrefutable challenge that tumors evolve defenses against some therapies. Gene therapy may prove to be the ultimate panacea for cancer by correcting the fundamental genetic errors in tumors. The engineering of nanoscale inorganic carriers of cancer therapeutics has shown promising results in the efficacious and safe delivery of nucleic acids to treat oncological diseases in small‐animal models. When these nanocarriers are used for co‐delivery of gene therapeutics along with auxiliary treatments, the synergistic combination of therapies often leads to an amplified health benefit. In this review, an overview of the inorganic nanomaterials developed for combinatorial therapies of gene and other treatment modalities is presented. First, the main principles of using nucleic acids as therapeutics, inorganic nanocarriers for medical applications and delivery of gene/drug payloads are introduced. Next, the utility of recently developed inorganic nanomaterials in different combinations of gene therapy with each of chemo, immune, hyperthermal, and radio therapy is examined. Finally, current challenges in the clinical translation of inorganic nanomaterial‐mediated therapies are presented and outlooks for the field are provided. [ABSTRACT FROM AUTHOR]

Human neural stem cells (hNSCs) possess remarkable potential for regenerative medicine in the treatment of presently incurable diseases. However, a key challenge lies in producing sufficient quantities of hNSCs, which is necessary for effective treatment. Dynamic culture systems are recognized as a powerful approach to producing large quantities of hNSCs required, where microcarriers play a critical role in supporting cell expansion. Nevertheless, the currently available microcarriers have limitations, including a lack of appropriate surface chemistry to promote cell adhesion, inadequate mechanical properties to protect cells from dynamic forces, and poor suitability for mass production. Here, we present the development of three-dimensional (3D) chitosan scaffolds as microcarriers for hNSC expansion under defined conditions in bioreactors. We demonstrate that chitosan scaffolds with a concentration of 4 wt% (4CS scaffolds) exhibit desirable microstructural characteristics and mechanical properties suited for hNSC expansion. Furthermore, they could also withstand degradation in dynamic conditions. The 4CS scaffold condition yields optimal metabolic activity, cell adhesion, and protein expression, enabling sustained hNSC expansion for up to three weeks in a dynamic culture. Our study introduces an effective microcarrier approach for prolonged expansion of hNSCs, which has the potential for mass production in a three-dimensional setting. [ABSTRACT FROM AUTHOR]

mRNA-based therapeutics have emerged as a promising strategy for cancer treatment. However, the effective delivery of mRNA into hard-to-transfect cancer cells remains a significant challenge. This study introduces a novel approach that utilizes iron oxide nanoparticles (NPs) synthesized through a layer-by-layer (LbL) method for safe and efficient mRNA delivery. The developed NPs consist of an iron oxide core modified with a thin charge-bearing layer, an mRNA middle layer, and an outer layer composed of perfluorinated polyethyleneimine with heparin (PPH), which facilitates efficient mRNA delivery. Through a comparative analysis of four nanoparticle delivery formulations, we investigated the effects of the iron oxide core's surface chemistry and surface charge on mRNA complexation, cellular uptake, and mRNA release. We identified an optimal and effective mRNA delivery platform, namely, (IOCCP)-mRNA-PPH, capable of transporting mRNA into various hard-to-transfect cancer cell lines in vitro. The (IOCCP)-mRNA-PPH formulation demonstrated significant enhancements in cellular internalization of mRNA, facilitated endosomal escape, enabled easy mRNA release, and exhibited minimal cytotoxicity. These findings suggest that (IOCCP)-mRNA-PPH holds great promise as a solution for mRNA therapy against hard-to-transfect cancers. [ABSTRACT FROM AUTHOR]

Natural polymer-based porous scaffolds have been investigated to serve as three-dimensional (3D) tumor models for drug screening owing to their structural properties with better resemblance to human tumor microenvironments than two-dimensional (2D) cell cultures. In this study, a 3D chitosan–hyaluronic acid (CHA) composite porous scaffold with tunable pore size (60, 120 and 180 µm) was produced by freeze-drying and fabricated into a 96-array platform for high-throughput screening (HTS) of cancer therapeutics. We adopted a self-designed rapid dispensing system to handle the highly viscous CHA polymer mixture and achieved a fast and cost-effective large-batch production of the 3D HTS platform. In addition, the adjustable pore size of the scaffold can accommodate cancer cells from different sources to better mimic the in vivo malignancy. Three human glioblastoma multiforme (GBM) cell lines were tested on the scaffolds to reveal the influence of pore size on cell growth kinetics, tumor spheroid morphology, gene expression and dose-dependent drug response. Our results showed that the three GBM cell lines showed different trends of drug resistance on CHA scaffolds of varying pore size, which reflects the intertumoral heterogeneity across patients in clinical practice. Our results also demonstrated the necessity to have a tunable 3D porous scaffold for adapting the heterogeneous tumor to generate the optimal HTS outcomes. It was also found that CHA scaffolds can produce a uniform cellular response (CV < 0.15) and a wide drug screening window (Z′ > 0.5) on par with commercialized tissue culture plates, and therefore, can serve as a qualified HTS platform. This CHA scaffold-based HTS platform may provide an improved alternative to traditional 2D-cell-based HTS for future cancer study and novel drug discovery. [ABSTRACT FROM AUTHOR]

Compared to conventional treatments, gene therapy offers a variety of advantages for cancer treatment including high potency and specificity, low off-target toxicity, and delivery of multiple genes that concurrently target cancer tumorigenesis, recurrence, and drug resistance. In the past decades, gene therapy has undergone remarkable progress, and is now poised to become a first line therapy for cancer. Among various gene delivery systems, nanoparticles have attracted much attention because of their desirable characteristics including low toxicity profiles, well-controlled and high gene delivery efficiency, and multi-functionalities. This review provides an overview on gene therapeutics and gene delivery technologies, and highlight recent advances, challenges and insights into the design and the utility of nanoparticles in gene therapy for cancer treatment. [ABSTRACT FROM AUTHOR]

[Display omitted] • Different MNZs were developed to study the influence of geometry on the therapeutic effect. • MNZs with dual enzymatic functions to improve the production of •OH. • Self-cascading MNZs exhibit a shape-dependent CDT efficacy at both cellular and tissue levels. Magnetite nanoparticles have demonstrated great potential as nanozymes in chemodynamic therapy (CDT) for cancer. However, their in vivo efficiency remains limited. Here we developed magnetite nanozymes (MNZs) with dual enzyme activities and demonstrated their shape-dependent behavior in eradicating tumors. Three versions of MNZs were constructed of the same materials but largely different geometric forms: spherical (SMNZ), elliptic (EMNZ) and flaky (FMNZ); they were evaluated against each other in treating an aggressive mouse tumor model of triple-negative breast cancer. We showed that the shape influence is substantial, and with the optimal geometric form, FMNZ dramatically outperform its two counterparts and demonstrates phenomenal ability to shrink tumors via systemic injection. We further illustrated that the vastly increased treatment efficiency can be attributed to the large surface area of FMNZ that resulted in increased cellular uptake, greater iron ions release and increased production of highly toxic hydroxyl radicals (•OH), which leads to enhanced self-cascade CDT. [ABSTRACT FROM AUTHOR]

[Display omitted] Radiotherapy is a mainstay adjunctive therapy for glioblastoma (GBM). Despite the outcome improvement achieved with radiation, GBM prognosis remains dismal. Here we introduce a tumor-targeted iron oxide nanoparticle (NP) that intensifies the energy transfer of conventional photon radiotherapy on a selective cellular basis. Several NPs were formulated with systematic architectural variation to study and optimize reactive oxygen species (ROS) production. Selected from this screening stage, a biocompatible tumor-targeted NP was tested in vitro using two models of GBM, and then in vivo , using an orthotopic human primary GBM xenograft mouse model. Animals that received intravenous NP before irradiation demonstrated a 3-fold reduction in tumor growth and a 2-fold increase in survival. Cellular damage was investigated using in vivo magnetic resonance spectroscopy, which demonstrated increased therapeutic cytotoxicity specific to the tumor mass. Our work presents a viable therapeutic strategy to improve radiation therapy for GBM. [ABSTRACT FROM AUTHOR]

Breast cancer has attracted tremendous research interest in treatment development as one of the major threats to public health. The use of non-viral carriers for therapeutic DNA delivery has shown promise in treating various cancer types, including breast cancer, due to their high DNA loading capacity, high cell transfection efficiency, and design versatility. However, cytotoxicity and large sizes of non-viral DNA carriers often raise safety concerns and hinder their applications in the clinic. Here we report the development of a novel nanoparticle formulation (termed NP-Chi-xPEI) that can safely and effectively deliver DNA into breast cancer cells for successful transfection. The nanoparticle is composed of an iron oxide core coated with low molecular weight (800 Da) polyethyleneimine crosslinked with chitosan via biodegradable disulfide bonds. The NP-Chi-xPEI can condense DNA into a small nanoparticle with the overall size of less than 100 nm and offer full DNA protection. Its biodegradable coating of small-molecular weight xPEI and mildly positive surface charge confer extra biocompatibility. NP-Chi-xPEI-mediated DNA delivery was shown to achieve high transfection efficiency across multiple breast cancer cell lines with significantly lower cytotoxicity as compared to the commercial transfection agent Lipofectamine 3000. With demonstrated favorable physicochemical properties and functionality, NP-Chi-xPEI may serve as a reliable vehicle to deliver DNA to breast cancer cells. [ABSTRACT FROM AUTHOR]

[Display omitted] Triple negative breast cancer (TNBC) is difficult to treat effectively, due to its aggressiveness, drug resistance, and lack of the receptors required for hormonal therapy, particularly at the metastatic stage. Here, we report the development and evaluation of a multifunctional nanoparticle formulation containing an iron oxide core that can deliver doxorubicin, a cytotoxic agent, and polyinosinic:polycytidylic acid (Poly IC), a TLR3 agonist, in a targeted and simultaneous fashion to both breast cancer and dendritic cells. Endoglin-binding peptide (EBP) is used to target both TNBC cells and vasculature endothelium. The nanoparticle demonstrates favorable physicochemical properties and a tumor-specific targeting profile. The nanoparticle induces tumor apoptosis through multiple mechanisms including direct tumor cell killing, dendritic cell-initiated innate and T cell-mediated adaptive immune responses. The nanoparticle markedly inhibits tumor growth and metastasis and substantially extends survival in an aggressive and drug-resistant metastatic mouse model of TNBC. This study points to a promising platform that may substantially improve the therapeutic efficacy for treating metastatic TNBC. [ABSTRACT FROM AUTHOR]

Nanotechnology has given scientists new tools for the development of advanced materials for the detection and diagnosis of disease. In particular, superparamagnetic iron oxide nanoparticles (SPIONs) have been extensively investigated as novel magnetic resonance imaging (MRI) contrast agents due to a combination of favorable superparamagnetic properties, biodegradability, and surface properties. This review discusses the basics of MR imaging, the origin of SPION''s unique magnetic properties, recent developments in MRI acquisition methods for detection of SPIONs, synthesis and post-synthesis processes that improve SPION''s imaging characteristics, and the outlook of the translational potential of SPIONs. [Copyright &y& Elsevier]

Abstract: The ability to produce aligned sub-micron fibers may open new avenues for the development of scaffolds for application in tissue engineering and regenerative medicine. An area of particular interest is functional restoration of damaged or diseased nerves where the aligned fibers serve to support cell adhesion and proliferation, and guide neurite outgrowth in the direction of fiber orientation. In this study, we developed an aligned chitosan–polycaprolactone (chitosan–PCL) fibrous scaffold and investigated how the fiber alignment influenced nerve cell organization and function in comparison with randomly oriented fibrous scaffolds and cast films of the same material. Schwann cells (SCs) were shown to attach and proliferate on all the substrates regardless of their topography, demonstrating the cellular compatibility of the chitosan–PCL material. SCs grown on the aligned chitosan–PCL fibers exhibited a bipolar morphology that oriented along the fiber alignment direction, while those on the films and randomly oriented fibers had a multipolar morphology. Similarly, the chitosan–PCL material supported neuron-like PC-12 cell adhesion, and the aligned fibers regulated the growth of PC-12 cells along the fiber orientation. Additionally, PC-12 cells cultured on the aligned fibers exhibited enhanced unidirectional neurite extension along fiber orientation and significantly higher β-tubulin gene expression than those grown on chitosan–PCL films and randomly oriented fibers. Our investigation suggested that the aligned chitosan–PCL fibers can serve as a suitable scaffold for improved nerve tissue reconstruction. [Copyright &y& Elsevier]

Abstract: Magnetic nanoparticles (MNPs) represent a class of non-invasive imaging agents that have been developed for magnetic resonance (MR) imaging. These MNPs have traditionally been used for disease imaging via passive targeting, but recent advances have opened the door to cellular-specific targeting, drug delivery, and multi-modal imaging by these nanoparticles. As more elaborate MNPs are envisioned, adherence to proper design criteria (e.g. size, coating, molecular functionalization) becomes even more essential. This review summarizes the design parameters that affect MNP performance in vivo, including the physicochemical properties and nanoparticle surface modifications, such as MNP coating and targeting ligand functionalizations that can enhance MNP management of biological barriers. A careful review of the chemistries used to modify the surfaces of MNPs is also given, with attention paid to optimizing the activity of bound ligands while maintaining favorable physicochemical properties. [Copyright &y& Elsevier]

Abstract: Hydrogels are high-water content materials prepared from cross-linked polymers that are able to provide sustained, local delivery of a variety of therapeutic agents. Use of the natural polymer, chitosan, as the scaffold material in hydrogels has been highly pursued thanks to the polymer''s biocompatibility, low toxicity, and biodegradability. The advanced development of chitosan hydrogels has led to new drug delivery systems that release their payloads under varying environmental stimuli. In addition, thermosensitive hydrogel variants have been developed to form a chitosan hydrogel in situ, precluding the need for surgical implantation. The development of these intelligent drug delivery devices requires a foundation in the chemical and physical characteristics of chitosan-based hydrogels, as well as the therapeutics to be delivered. In this review, we investigate the newest developments in chitosan hydrogel preparation and define the design parameters in the development of physically and chemically cross-linked hydrogels. [Copyright &y& Elsevier]

Abstract: Magnetic nanoparticles (MNPs) possess unique magnetic properties and the ability to function at the cellular and molecular level of biological interactions making them an attractive platform as contrast agents for magnetic resonance imaging (MRI) and as carriers for drug delivery. Recent advances in nanotechnology have improved the ability to specifically tailor the features and properties of MNPs for these biomedical applications. To better address specific clinical needs, MNPs with higher magnetic moments, non-fouling surfaces, and increased functionalities are now being developed for applications in the detection, diagnosis, and treatment of malignant tumors, cardiovascular disease, and neurological disease. Through the incorporation of highly specific targeting agents and other functional ligands, such as fluorophores and permeation enhancers, the applicability and efficacy of these MNPs have greatly increased. This review provides a background on applications of MNPs as MR imaging contrast agents and as carriers for drug delivery and an overview of the recent developments in this area of research. [Copyright &y& Elsevier]

Clean silicon and gold-patterned silicon platforms were modified with methoxy-polyethylene glycol (M-PEG silane) via a self-assembly technique, which significantly improved their plasma protein resistance capability and cell patterning selectivity. Fibrinogen and IgG were used as model plasma proteins to study the efficacy of PEG layers in resisting protein adsorption. Selective cell patterning on the gold regions of a gold-patterned silicon substrate and tissue compatibility were studied with macrophage and fibroblast cells. The research also revealed how the presence of gold electrodes on a silicon substrate would influence the cell patterning selectivity. Our experimental results showed that the PEG-modified silicon surfaces had a high resistivity to protein and cell attachment and that the PEG-modified gold-patterned silicon surfaces nearly completely eliminated the protein adsorption and cell attachment on silicon. This study provides a new approach to developing biocompatible surfaces for silicon-based BioMEMS devices, particularly for biosensors where a metal-insulator format must be enforced. [Copyright &y& Elsevier]

Superparamagnetic magnetite nanoparticles were surface-modified with poly (ethylene glycol) (PEG) and folic acid, respectively, to improve their intracellular uptake and ability to target specific cells. PEG and folic acid were successfully immobilized on the surfaces of magnetite nanoparticles and characterized using fourier transform infrared spectra. The nanoparticle internalization into mouse macrophage (RAW 264.7) and human breast cancer (BT20) cells was visualized using both fluorescence and confocal microscopy, and quantified by inductively coupled plasma emission spectroscopy (ICP). After the cells were cultured for 48 h in the medium containing the nanoparticles modified with PEG or folic acid, the results of fluorescence and confocal microscopy showed that the nanoparticles were internalized into the cells. The ICP measurements indicated that the uptake amount of PEG-modified nanoparticles into macrophage cells was much lower than that of unmodified nanoparticles, while folic acid modification did not change the amount of the uptake. However, for breast cancer cells, both PEG and folic acid modification facilitated the nanoparticle internalization into the cells. Therefore, PEG and folic acid modification of magnetite nanoparticles could be used to resist the protein adsorption and thus avoid the particle recognition by macrophage cells, and to facilitate the nanoparticle uptake to specific cancer cells for cancer therapy and diagnosis. [Copyright &y& Elsevier]

Temozolomide (TMZ) is the standard‐of‐care chemotherapy drug for treating glioblastomas (GBMs), the most aggressive cancer that affects people of all ages. However, its therapeutic efficacy is limited by the drug‐resistance mediated by a DNA repair protein, O6‐methylguanine‐DNA methyltransferase (MGMT), which eliminates the TMZ‐induced DNA lesions. Here, the development of an iron oxide nanoparticle (NP) system for targeted delivery of small interfering RNAs to suppress the TMZ‐resistance gene (MGMT) is reported. The NPs are able to overcome biological barriers, bind specifically to tumor cells, and reduce MGMT expression in tumors of mice bearing orthotopic GBM serially passaged patient‐derived xenografts. The treatment with sequential administration of this NP and TMZ result in increased apoptosis of GBM stem‐like cells, reduced tumor growth, and significantly prolonged survival as compared to mice treated with TMZ alone. This study introduces an approach that holds great promise to improve the outcomes of GBM patients. [ABSTRACT FROM AUTHOR]

Systemic delivery of hydrophobic anti-cancer drugs with nanocarriers, particularly for drug-resistant and metastatic cancer, remain a challenge because of the difficulty to achieve high drug loading, while maintaining a small hydrodynamic size and colloid stability in blood to ensure delivery of an efficacious amount of drug to tumor cells. Here we introduce a new approach to address this challenge. In this approach, nanofibers of larger size with good drug loading capacity are first constructed by a self-assembly process, and upon intravascular injection and interacting with serum proteins in vivo, these nanofibers break down into ultra-fine nanoparticles of smaller size that inherit the drug loading property from their parent nanofibers. We demonstrate the efficacy of this approach with a clinically available anti-cancer drug: paclitaxel (PTX). In vitro, the PTX-loaded nanoparticles enter cancer cells and induce cellular apoptosis. In vivo, they demonstrate prolonged circulation in blood, induce no systemic toxicity, and show high potency in inhibiting tumor growth and metastasis in both mouse models of aggressive, drug-resistant breast cancer and melanoma. This study points to a new strategy toward improved anti-cancer drug delivery and therapy. [ABSTRACT FROM AUTHOR]

A portable, handheld electrospinning apparatus is designed and constructed using off-the-shelf components and 3D-printed parts. The portable electrospinner is used to generate nanofibers with diameters ranging from 85 to 600 nm; examination of these fibers is achieved with scanning electron microscopy. This portable electrospinner has similar capabilities to standard stationary benchtop electrospinners in terms of the diversity of polymers the device is able to spin into nanofibers and their resulting size and morphology. However, it provides much more ambulatory flexibility, employs current-limiting measures that allow for safer operation and is cost effective. As a demonstration of the device's unique application space afforded by its portability, the device is applied in direct-to-skin electrospinning to improve the aesthetics of simulated hair loss in a mouse model by electrospinning dyed polyacrylonitrile nanofibers that mimic hair. The superficial nanofiber treatment for thinning hair is able to achieve an improvement in appearance similar to that of a commercially available powder product but outperforms the powder in the nanofiber's superior adherence to the affected area. The portable electrospinning apparatus overcomes many limitations of immobile benchtop electrospinners and holds promise for applications in consumer end-use scenarios such as the treatment of alopecia via cosmetic hair thickening. [ABSTRACT FROM AUTHOR]

Abstract Nanostructured materials that have low tissue toxicity, multi-modal imaging capability and high photothermal conversion efficiency have great potential to enable image-guided near infrared (NIR) photothermal therapy (PTT). Here, we report a bifunctional nanoparticle (BFNP, ∼16 nm) comprised of a magnetic Fe 3 O 4 core (∼9.1 nm) covered by a fluorescent carbon shell (∼3.4 nm) and prepared via a one-pot solvothermal synthesis method using ferrocene as the sole source. The BFNP exhibits excitation wavelength-tunable, upconverted and near-infrared (NIR) fluorescence property due to the presence of the carbon shell, and superparamagnetic behavior resulted from the Fe 3 O 4 core. BFNPs demonstrate dual-modal imaging capacity both in vitro and in vivo with fluorescent imaging excited under a varying wavelength from 405 nm to 820 nm and with T 2 -weighted magnetic resonance imaging (r 2 = 264.76 mM−1 s−1). More significantly, BFNPs absorb and convert NIR light to heat enabling photothermal therapy as demonstrated mice bearing C6 glioblastoma. These BFNPs show promise as an advanced nanoplatform to provide imaging guided photothermal therapy. Graphical abstract Unlabelled Image Highlights • A bifunctional theranostic nanoparticle (BFNP) is developed. • BFNP is a contrast agent for both magnetic resonance imaging and fluorescence imaging. • BFNP converts NIR light into heat enabling photothermal therapy. • BFNP is benign to organs and healthy tissue. • Imaging and therapeutic efficacy of BFNP is demonstrated in a mouse model. [ABSTRACT FROM AUTHOR]

A uniaxially-aligned nanofibrous matrix was produced by electrospinning and it demonstrates strong piezoelectric property. The matrix is composed of highly-aligned polyvinylidene fluoride (PVDF) polymer nanofibers embedded with barium titanate (BaTiO 3 ) nanoparticles. Measurements of piezoelectrically generated voltages in response to mechanical deformation of BaTiO 3 –PVDF composite nanofibers with various concentrations of BaTiO 3 nanoparticles showed that the magnitude of the resultant voltage increases as the nanoparticle concentration increases. PVDF nanofibers with 16 wt% BaTiO 3 nanoparticles exhibit piezoelectric output voltages that are 1.7 times greater than PVDF fibers without BaTiO 3 nanoparticles when subjected to the same degree of deformation. Scanning electron microscopy confirmed the uniaxial alignment of PVDF nanofibers with individual fiber diameters of ~200 nm as well as the presence of BaTiO 3 deposits. The increased piezoelectric response of uniaxially-aligned BaTiO 3 –PVDF nanofibers in comparison with randomly oriented PVDF fibers and thin films suggests possible uses for such nanofibers in energy harvesting and as power sources in miniaturized electronic devices like wearable smart textiles and implantable biosensors. [ABSTRACT FROM AUTHOR]

The inability to produce large quantities of nanofibers has been a primary obstacle in advancement and commercialization of electrospinning technologies, especially when aligned nanofibers are desired. Here, we present a high-throughput centrifugal electrospinning (HTP-CES) system capable of producing a large number of highly-aligned nanofiber samples with high-yield and tunable diameters. The versatility of the design was revealed when bead-less nanofibers were produced from copolymer chitosan/polycaprolactone (C-PCL) solutions despite variations in polymer blend composition or spinneret needle gauge. Compared to conventional electrospinning techniques, fibers spun with the HTP-CES not only exhibited superior alignment, but also better diameter uniformity. Nanofiber alignment was quantified using Fast Fourier Transform (FFT) analysis. In addition, a concave correlation between the needle diameter and resultant fiber diameter was identified. This system can be easily scaled up for industrial production of highly-aligned nanofibers with tunable diameters that can potentially meet the requirements for various engineering and biomedical applications. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop chitosan composite scaffolds with high strength and controlled pore structures by homogenously dispersed nano-sized hydroxyapatite (nano-HAp) powders. In the fabrication of composite scaffolds, nano-HAp powders distributed in an alginate (AG) solution with a pH higher than 10 were mixed with a chitosan (CS) solution and then freeze dried. While the HAp content increased up to 70 wt.%, the compressive strength and the elastic modulus of the composite scaffolds significantly increased from 0.27 MPa and 4.42 MPa to 0.68 MPa and 13.35 MPa, respectively. Higher content of the HAp also helped develop more differentiation and mineralization of the MC3T3-E1 cells on the composite scaffolds. The uniform pore structure and the excellent mechanical properties of the HAp/CS composite scaffolds likely resulted from the use of the AG solution at pH 10 as a dispersant for the nano-HAp powders. [ABSTRACT FROM AUTHOR]

Smart hydrogels play an increasingly important role in biomedical applications, since materials that are both biocompatible and multi-stimuli-responsive are highly desirable. A simple, organic solvent-free method is presented to synthesize a biocompatible hydrogel that undergoes a sol-gel transition in response to multiple stimuli. Methoxy-poly(ethylene glycol) (mPEG) is modified into carboxylic-acid-terminated-methoxy-poly(ethylene glycol) (mPEG-acid), which is then grafted onto chitosan via amide linkages yielding mPEG- g-chitosan. Grafting of mPEG onto hydrophobic chitosan imparts hydrophilic properties to the resultant polymer. The mPEG- g-chitosan gel exhibits a controllable multi-stimuli-responsive property. The balance between hydrophilicity and hydrophobicity is believed to confer mPEG-g-chitosan with stimuli-responsive behavior. The effect of salt concentration, solute concentration, temperature, and pH on the sol-gel transition of mPEG- g-chitosan is evaluated and the underlying mechanisms of mPEG- g-chitosan polymer packing and gelation property is discussed. [ABSTRACT FROM AUTHOR]

Thiswork describes the preparation and validation of single-chainsemiconducting polymer dots (sPdots), which weregenerated using a method based on surface immobilization, washing,and cleavage. The sPdots have an ultrasmall sizeof ∼3.0 nm as determined by atomic force microscopy, a sizethat is consistent with the anticipated diameter calculated from themolecular weight of the single-chain semiconducting polymer. sPdots should find use in biology and medicine as a newclass of fluorescent probes. The FRET assay this work presents isa simple and rapid test to ensure methods developed for preparing sPdot indeed produced single-chain Pdots as designed. [ABSTRACT FROM AUTHOR]

Emerging evidence implicates cancer stem cells (CSCs) as primary determinants of the clinical behavior of human cancers, representing an ideal target for next-generation anti-cancer therapies. However CSCs are difficult to propagate in vitro , severely limiting the study of CSC biology and drug development. Here we report that growing cells from glioblastoma (GBM) cell lines on three dimensional (3D) porous chitosan-alginate (CA) scaffolds dramatically promotes the proliferation and enrichment of cells possessing the hallmarks of CSCs. CA scaffold-grown cells were found more tumorigenic in nude mouse xenografts than cells grown from monolayers. Growing in CA scaffolds rapidly promoted expression of genes involved in the epithelial-to-mesenchymal transition that has been implicated in the genesis of CSCs. Our results indicate that CA scaffolds have utility as a simple and inexpensive means to cultivate CSCs in vitro in support of studies to understand CSC biology and develop more effective anti-cancer therapies. [ABSTRACT FROM AUTHOR]

Breast cancer has attracted tremendous research interest in treatment development as one of the major threats to public health. The use of non-viral carriers for therapeutic DNA delivery has shown promise in treating various cancer types, including breast cancer, due to their high DNA loading capacity, high cell transfection efficiency, and design versatility. However, cytotoxicity and large sizes of non-viral DNA carriers often raise safety concerns and hinder their applications in the clinic. Here we report the development of a novel nanoparticle formulation (termed NP-Chi-xPEI) that can safely and effectively deliver DNA into breast cancer cells for successful transfection. The nanoparticle is composed of an iron oxide core coated with low molecular weight (800 Da) polyethyleneimine crosslinked with chitosan via biodegradable disulfide bonds. The NP-Chi-xPEI can condense DNA into a small nanoparticle with the overall size of less than 100 nm and offer full DNA protection. Its biodegradable coating of small-molecular weight xPEI and mildly positive surface charge confer extra biocompatibility. NP-Chi-xPEI-mediated DNA delivery was shown to achieve high transfection efficiency across multiple breast cancer cell lines with significantly lower cytotoxicity as compared to the commercial transfection agent Lipofectamine 3000. With demonstrated favorable physicochemical properties and functionality, NP-Chi-xPEI may serve as a reliable vehicle to deliver DNA to breast cancer cells. [ABSTRACT FROM AUTHOR]

The outcome for glioblastoma patientsremains dismal for its invariablyrecrudesces within 2 cm of the resection cavity. Local immunotherapyhas the potential to eradicate the residual infiltrative componentof these tumors. Here, we report the development of a biodegradablehydrogel containing therapeutic T lymphocytes for localized deliveryto glioblastoma cells for brain tumor immunotherapy. Thermoreversiblepoly(ethylene glycol)-g-chitosan hydrogels (PCgels)were optimized for steady T lymphocyte release. Nuclear magnetic resonancespectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-geltransition of the PCgel occurred around ≥32 °C. T lymphocyteinvasion through the PCgel and subsequent cytotoxicity to glioblastomawere assessed in vitro. The PCgel was shown to becellular compatible with T lymphocytes, and the T lymphocytes retaintheir anti-glioblastoma activity after being encapsulated in the PCgel.T lymphocytes in the PCgel were shown to be more effective in killingglioblastoma than those in the Matrigel control. This may be attributedto the optimal pore size of the PCgel allowing better invasion ofT lymphocytes. Our study suggests that this unique PCgel depot mayoffer a viable approach for localized immunotherapy for glioblastoma. [ABSTRACT FROM AUTHOR]

Abstract: Cancer therapeutics are developed through extensive screening; however, many therapeutics evaluated with 2D in vitro cultures during pre-clinical trials suffer from lower efficacy in patients. Replicating the in vivo tumor microenvironment in vitro with three-dimensional (3D) porous scaffolds offers the possibility of generating more predictive pre-clinical models to enhance cancer treatment efficacy. We developed a chitosan and hyaluronic acid (HA) polyelectrolyte complex 3D porous scaffold and evaluated its physical properties. Chitosan-HA (C-HA) scaffolds had a highly porous network. C-HA scaffolds were compared to 2D surfaces for in vitro culture of U-118 MG human glioblastoma (GBM) cells. C-HA scaffold cultures promoted tumor spheroid formation and increased stem-like properties of GBM cells as evidenced by the upregulation of CD44, Nestin, Musashi-1, GFAP, and HIF-1α as compared with 2D cultures. Additionally, the invasiveness of GBM cells cultured in C-HA scaffolds was significantly enhanced compared to those grown in 2D cultures. C-HA scaffold cultures were also more resistant to chemotherapy drugs, which corresponded to the increased expression of ABCG2 drug efflux transporter. These findings suggest that C-HA scaffolds offer promise as an in vitro GBM platform for study and screening of novel cancer therapeutics. [Copyright &y& Elsevier]

Myogenicprogenitor cells derived from human embryonic stem cells(hESCs) can provide unlimited sources of cells in muscle regenerationbut their clinical uses are largely hindered by the lack of efficientmethods to induce differentiation of stem cells into myogenic cells.We present a novel approach to effectively enhance myogenic differentiationof human embryonic stem cells using aligned chitosan-polycaprolactone(C-PCL) nanofibers constructed to resemble the microenvironment ofthe native muscle extracellular matrix (ECM) in concert with Wnt3aprotein. The myogenic differentiation was assessed by cell morphology,gene activities, and protein expression. hESCs grown on C-PCL uniaxiallyaligned nanofibers in media containing Wnt3a displayed an elongatedmorphology uniformly aligned in the direction of fiber orientation,with increased expressions of marker genes and proteins associatedwith myogenic differentiation as compared to control substrates. Thecombination of Wnt3a signaling and aligned C-PCL nanofibers resultedin high percentages of myogenic-protein expressing cells over totaltreated hESCs (83% My5, 91% Myf6, 83% myogenin, and 63% MHC) after2 days of cell culture. Significantly, this unprecedented high-leveland fast myogenic differentiation of hESC was demonstrated in a culturemedium containing no feeder cells. This study suggests that chitosan-basedaligned nanofibers combined with Wnt3a can potentially act as a modelsystem for embryonic myogenesis and muscle regeneration. [ABSTRACT FROM AUTHOR]

Biotelemetry has become an important part of medical research for patient care by remotely monitoring continuing biological processes and physiological functions. However, current biotelemetry systems are complex requiring multiple electronic components to function: a battery, a sensor, and a transmitter, and a receiver. Another paramount concern of biotelemetry is the coupling of its in vivo portion to external supporting equipment. Here we report a novel biotelemetry device made primarily of a coiled bundle of uniaxially-aligned biocompatible polyvinylidene fluoride (PVDF) nanofibers of ∼200 nm in diameter and with piezoelectric properties that can serve concurrently as a power source, sensor, and transmitter. We tested this device on a cantilever beam that was periodically deflected at its free end. Without a power supply the coil of a nanofiber bundle is shown to generate and transmit an electrical signal wirelessly in response to the beam deflection which was received by an external receiver. The coil of a nanofiber bundle was encapsulated in a thin biocompatible polymer shell for device integrity and moisture isolation. Our results suggest that the device can potentially serve as a mechanical sensor and biotelemeter for various in vitro and in vivo biomedical applications. [ABSTRACT FROM AUTHOR]

Background: At present there is no standard of care for patients with primary central nervous system lymphoma (PCNSL) because of the difficulty in delivering therapeutically effective doses of drugs to the intracellular site of the target PCNSL. Here we report the use of an iron oxide nanoparticle to promote the internalization of a PCNSL targeting antibody by target cells. Methods: Iron oxide nanoparticles coated with a copolymer of chitosan-grafted polyethylene glycol (NPs) were conjugated with an anti-CD20 single-chain variable fragment-streptavidin fusion protein (FP), and optically activated with Oregon Green 488. The ability of NP-FP to target PCNSL cells was assessed using flow cytometry and the ferrozine assay. Cell internalization of NP-FP was examined by confocal fluorescence microscopy. Results: The antibody-conjugated NPs had a near-neutral zeta potential and remained stable in biological media for more than 1 week, which may minimizes nonspecific cell uptake. The diameter of the NPs was about 70 nm, which is in an optimal range for maximizing cell uptake. The selective binding of these NPs was demonstrated with binding to PCNSL cells 3- to 4-fold higher than binding to control cells. Z-stack imaging by confocal microscopy revealed the NPs were internalized by PCNSL cells. Conclusions: The high-degree specific binding and cell uptake of NP-FP in PCNSL suggests this NP formulation can be further developed to improve therapy of PCNSL. [ABSTRACT FROM AUTHOR]

In the tumor microenvironment, thesignals from tumor-associatedfibroblasts (TAF) that suppress antitumor immunity remain unclear.Here, we develop and investigate an in vitro three-dimensional (3D)scaffold model for the novel evaluation of TAF interaction with breasttumor cells and breast specific, neu antigen (p98) reactive T cells.Breast cancer cells seeded on 3D chitosan-alginate (CA) scaffoldsshowed productive growth and formed distinct tumor spheroids. Antigenspecific p98 T cells, but not naïve T cells, bound significantlybetter to tumor cells on scaffolds. The p98 T cells induced potenttumor cell killing but T helper cell cytokine function was impairedin the presence of TAF coseeding on scaffolds. We found that the immunosuppressionwas mediated, in part, by transforming growth factor beta (TGF-b)and interleukin-10 (IL-10). Therefore, TAF appear capable of inducingpotent T cell suppression. CA scaffolds can provide clinically relevantfindings prior to preclinical testing of novel immunotherapies. [ABSTRACT FROM AUTHOR]

Abstract: Nanofibrous membranes have drawn considerable interest for filtration applications due to their ability to withstand high fluid flux while removing micro- and nano-sized particulates from solution. The desire to introduce an antibacterial function into water filter applications presents a challenge to widespread application of fibrous membranes because the addition of chemicals or biocides may produce harmful byproducts downstream. Here, we report the development of chitosan–polycaprolactone (PCL) nanofibrous membranes to utilize the natural antibacterial property of chitosan for antibacterial water filtration. Chitosan–PCL fibers with diameters of 200–400nm and chitosan contents of 25, 50 and 75wt% were prepared by electrospinning. In a series of bacterial challenge tests, chitosan–PCL fibrous membranes significantly reduced Staphylococcus aureus adhesion compared to PCL fibrous membranes. In water permeability and particulate size removal tests, fibrous membranes with 25% chitosan supported the greatest water flux (∼7000L/h/m2) with 100% removal of 300-nm particulates, while maintaining the membrane integrity. This study demonstrates the potential of chitosan–PCL nanofibrous membranes as pre-filters for water filtration systems that demonstrate combinatorial filtration and intrinsic antibacterial advantages. [Copyright &y& Elsevier]

Abstract: Nanoparticle-based cancer therapeutics promises to improve drug delivery safety and efficacy. However, fabrication of consistent theranostic nanoparticles with high and controllable drug loading remains a challenge, primarily due to the cumbersome, multi-step synthesis processes conventionally applied. Here, we present a simple and highly controllable method for assembly of theranostic nanoparticles, which may greatly reduce batch-to-batch variation. The major components of this nanoparticle system include a superparamagnetic iron oxide nanoparticle (SPION), a biodegradable and pH-sensitive poly (beta-amino ester) (PBAE) copolymer, a chemotherapeutic agent doxorubicin (DOX). Here the polymer pre-loaded with drug is directly assembled to the surface of SPIONs forming a drug loaded nanoparticle (NP-DOX). NP-DOX demonstrated a high drug loading efficiency of 679μg DOX per mg iron, sustained stability in cell culture media up to 7days, and a strong r2 relaxivity of 146mM−1•s−1 for magnetic resonance imaging (MRI). The drug release analysis of NP-DOX showed fast DOX release at pH 5.5 and 6.4 (as in endosomal environment) and slow release at pH 7.4 (physiological condition), demonstrating pH-sensitive drug release kinetics. In vitro evaluation of NP-DOX efficacy using drug-resistant C6 glioma cells showed a 300% increase in cellular internalization at 24h post-treatment and 65% reduction of IC50 at 72h post-treatment when compared to free DOX. These nanoparticles could serve as a foundation for building smart theranostic formulations for sensitive detection through MRI and effective treatment of cancer by controlled drug release. [Copyright &y& Elsevier]

Abstract: We developed a new instrumental method by which human melanoma cells (LU1205) are sonoporated via radiation pressures exerted by highly-confined ultrasonic waves produced by high lateral-resolution ultrasonic micro-transducer arrays (UMTAs). The method enables cellular-level site-specific sonoporation within the cell monolayer due to UMTAs and can be applicable in the delivery of drugs and gene products in cellular assays. In this method, cells are seeded on the biochip that employs UMTAs for high spatial resolution and specificity. UMTAs are driven by 30-MHz sinusoidal signals and the resulting radiation pressures induce sonoporation in the targeted cells. The sonoporation degree and the effective lateral resolution of UMTAs are determined by performing fluorescent microscopy and analysis of carboxylic-acid-derivatized CdSe/ZnS quantum dots passively transported into the cells. Models representing the transducer-generated ultrasound radiation pressure, the ultrasound-inflicted cell membrane wound, and the transmembrane transport through the wound are developed to determine the ultrasound-pressure-dependent wound size and enhanced cellular uptake of nanoparticles. Model-based calculations show that the effective wound size and cellular uptake of nanoparticles increase linearly with increasing ultrasound pressure (i.e., at applied radiation pressures of 0.21, 0.29, and 0.40MPa, the ultrasound-induced initial effective wound radii are 150, 460, and 650nm, respectively, and the post-sonoporation intracellular quantum-dot concentrations are 7.8, 22.8, and 29.9nM, respectively) and the threshold pressure required to induce sonoporation in LU1205 cells is ∼0.12MPa. [Copyright &y& Elsevier]

Abstract: Lack of safe and effective carriers for delivery of RNA therapeutics remains a barrier to its broad clinical application. We report the development of a cell tanscytosing magnetic nanovector engineered as an siRNA carrier. Iron oxide nanoparticles were modified with poly(ethylene glycol) (PEG), small interfering RNA (siRNA), and a cationic polymer layer. Three nanovector formulations with cationic polymer coatings of poly-arginine (pArg), polylysine (pLys), and polyethylenimine (PEI), respectively, were prepared. The three nanovector formulations where evaluated for safety and ability to promote gene silencing in three types of cancer cells C6/GFP+, MCF7/GFP+, and TC2/GFP+, mimicking human cancers of the brain, breast, and prostate, respectively. Cell viability and fluorescence quantification assays revealed that pArg-coated nanovectors were most effective in promoting gene knockdown and least toxic of the three nanovector formulations tested. Transmission electron microscopy (TEM) imaging of nanovector treated cells further demonstrated that pArg-coated nanovectors enter cells through cell transcytosis, while pLys and PEI coated nanovectors enter cells endocytosis. Our findings suggest that NPs engineered to exploit the cell transcytosis intracellular trafficking pathway may offer a more safe and efficient route for siRNA delivery. [Copyright &y& Elsevier]

Abstract: Cell invasion is an intrinsic cellular pathway whereby cells respond to extracellular stimuli to migrate through and modulate the structure of their extracellular matrix (ECM) in order to develop, repair, and protect the body''s tissues. In cancer cells this process can become aberrantly regulated and lead to cancer metastasis. This cellular pathway contributes to the vast majority of cancer related fatalities, and therefore has been identified as a critical therapeutic target. Researchers have identified numerous potential molecular therapeutic targets of cancer cell invasion, yet delivery of therapies remains a major hurdle. Nanomedicine is a rapidly emerging technology which may offer a potential solution for tackling cancer metastasis by improving the specificity and potency of therapeutics delivered to invasive cancer cells. In this review we examine the biology of cancer cell invasion, its role in cancer progression and metastasis, molecular targets of cell invasion, and therapeutic inhibitors of cell invasion. We then discuss how the field of nanomedicine can be applied to monitor and treat cancer cell invasion. We aim to provide a perspective on how the advances in cancer biology and the field of nanomedicine can be combined to offer new solutions for treating cancer metastasis. [Copyright &y& Elsevier]

Abstract: Multidrug resistance (MDR) is characterized by the overexpression of ATP-binding cassette (ABC) transporters that actively pump a broad class of hydrophobic chemotherapeutic drugs out of cancer cells. MDR is a major mechanism of treatment resistance in a variety of human tumors, and clinically applicable strategies to circumvent MDR remain to be characterized. Here we describe the fabrication and characterization of a drug-loaded iron oxide nanoparticle designed to circumvent MDR. Doxorubicin (DOX), an anthracycline antibiotic commonly used in cancer chemotherapy and substrate for ABC-mediated drug efflux, was covalently bound to polyethylenimine via a pH sensitive hydrazone linkage and conjugated to an iron oxide nanoparticle coated with amine terminated polyethylene glycol. Drug loading, physiochemical properties and pH lability of the DOX-hydrazone linkage were evaluated in vitro. Nanoparticle uptake, retention, and dose-dependent effects on viability were compared in wild-type and DOX-resistant ABC transporter over-expressing rat glioma C6 cells. We found that DOX release from nanoparticles was greatest at acidic pH, indicative of cleavage of the hydrazone linkage. DOX-conjugated nanoparticles were readily taken up by wild-type and drug-resistant cells. In contrast to free drug, DOX-conjugated nanoparticles persisted in drug-resistant cells, indicating that they were not subject to drug efflux. Greater retention of DOX-conjugated nanoparticles was accompanied by reduction of viability relative to cells treated with free drug. Our results suggest that DOX-conjugated nanoparticles could improve the efficacy of chemotherapy by circumventing MDR. [Copyright &y& Elsevier]

Abstract: Ribonucleic acid interference (RNAi) is a powerful molecular tool that has potential to revolutionize the treatment of cancer. One major challenge of applying this technology for clinical application is the lack of site-specific carriers that can effectively deliver short interfering RNA (siRNA) to cancer cells. Here we report the development and assessment of a cancer-cell specific magnetic nanovector construct for efficient siRNA delivery and non-invasive monitoring through magnetic resonance imaging (MRI). The base of the nanovector construct is comprised of a superparamagnetic iron oxide nanoparticle core coated with polyethylene glycol (PEG)-grafted chitosan, and polyethylenimine (PEI). The construct was then further functionalized with siRNA and a tumor-targeting peptide, chlorotoxin (CTX), to improve tumor specificity and potency. Flow cytometry, quantitative RT-PCR, and fluorescence microscopy analyses confirmed receptor-mediated cellular internalization of nanovectors and enhanced gene knockdown through targeted siRNA delivery. The ability of this nanovector construct to generate specific contrast enhancement of glioblastoma cells was demonstrated through MR imaging. These findings suggest that this CTX enabled nanoparticle carrier may be well suited for delivery of RNAi therapeutics to brain cancer cells. [ABSTRACT FROM AUTHOR]

Abstract: Despite recent advances in the understanding of its cell biology, glioma remains highly lethal. Development of effective therapies requires a cost-effective in vitro tumor model that more accurately resembles the in vivo tumor microenvironment as standard two-dimensional (2D) tissue culture conditions do so poorly. Here we report on the use of a three-dimensional (3D) chitosan–alginate (CA) scaffold to serve as an extracellular matrix that promotes the conversion of cultured cancer cells to a more malignant in vivo-like phenotype. Human U-87 MG and U-118 MG glioma cells and rat C6 glioma cells were chosen for the study. In vitro tumor cell proliferation and secretion of factors that promote tumor malignancy, including VEGF, MMP-2, fibronectin, and laminin, were assessed. The scaffolds pre-cultured with U-87 MG and C6 cells were then implanted into nude mice to evaluate tumor growth and blood vessel recruitment compared to the standard 2D cell culture and 3D Matrigel matrix xenograft controls. Our results indicate that while the behavior of C6 cells showed minimal differences due to their highly malignant and invasive nature, U-87 MG and U-118 MG cells exhibited notably higher malignancy when cultured in CA scaffolds. CA scaffolds provide a 3D microenvironment for glioma cells that is more representative of the in vivo tumor, thus can serve as a more effective platform for development and study of anticancer therapeutics. This unique CA scaffold platform may offer a valuable alternative strategy to the time-consuming and costly animal studies for a wide variety of experimental designs. [Copyright &y& Elsevier]

Abstract: The underlying sensing mechanism of single-cell-based integrated microelectrode array (IMA) biosensors was investigated via experimental and modeling studies. IMA chips were microfabricated and single-cell-level manipulation was achieved through surface chemistry modification of IMA chips. Individual fibroblast cells (NIH3T3) were immobilized on either lysine–arginine–glycine–aspartic acid (KRGD) short peptide-modified or fibronectin extracellular-cell-adhesion-molecule-modified microelectrodes to record the impedance variations of cell–electrode heterostructure over a frequency range of 1–10kHz. By fitting experimental data to an application-specific single-cell-level equivalent circuit model, important sensing parameters, including specific cell membrane capacity, cell membrane resistivity, and averaged cell-to-substrate separation, were determined. It was demonstrated that biofunctionalization of planar microelectrode surface by covalently linking short peptides or fibronectin molecules could achieve strong or tight cell adhesion (with an estimated averaged cell-to-substrate separation distance of 11–16nm), which, in turn, improves the transduced electrical signal from IMA chips. Analyses on frequency-dependent characteristics of single-cell-covered microelectrode impedance and of IMA sensor circuitry response have revealed an addressable frequency band wherein electrical properties of single cells can be distinctively determined and monitored for cellular biosensing applications. The presented work addresses some major limitations in single-cell-based biosensing schemes, i.e., the manipulation of a single cell, the transduction of weak biological signals, and the implementation of a proper model for data analysis, and demonstrates the potential of IMA devices as single-cell biosensors. [Copyright &y& Elsevier]

Abstract: Human embryonic stem cells (hESCs) are routinely cultured on fibroblast feeder layers or in fibroblast-conditioned medium, which requires continued supply of feeder cells and poses the risks of xenogenic contamination and other complications such as feeder-dependent outcome. Here, we demonstrate a strategy that supports sustained self-renewal of hESCs in a three-dimensional porous natural polymer scaffold, comprised of chitosan and alginate, without the support of feeder cells or conditioned medium. We assessed the pluripotency of the renewed hESCs both in vitro by evaluation of cellular proliferation, functionality, and gene activities for 21 days, and in vivo by implantation of the stem cell populated scaffolds in an immunodeficient mouse model to induce teratoma formation. The self-renewed stem cells can be easily recovered for subculture by decomposing the scaffold under a mild condition. We further subcultured recovered hESCs for 14 days and verified their pluripotency. In addition to providing a clean environment for stem cell renewal, this strategy, with the demonstrated biocompatibility and biodegradability of chitosan and alginate, may potentially allow for the direct implantation of stem cell populated scaffolds for a broad spectrum of applications in tissue engineering and regenerative medicine. [Copyright &y& Elsevier]

Abstract: As conventional cancer therapies struggle with toxicity issues and irregular remedial efficacy, the preparation of novel gene therapy vectors could offer clinicians the tools for addressing the genetic errors of diseased tissue. The transfer of gene therapy to the clinic has proven difficult due to safety, target specificity, and transfection efficiency concerns. Polyethylenimine (PEI) nanoparticles have been identified as promising gene carriers that induce gene transfection with high efficiency. However, the inherent toxicity of the material and non-selective delivery are the major concerns in applying these particles clinically. Here, a non-viral nanovector has been developed by PEGylation of DNA-complexing PEI in nanoparticles functionalized with an Alexa Fluor 647 near infrared fluorophore, and the chlorotoxin (CTX) peptide which binds specifically to many forms of cancer. With this nanovector, the potential toxicity to healthy cells is minimized by both the reduction of the toxicity of PEI with the biocompatible copolymer and the targeted delivery of the nanovector to cancer cells, as evaluated by viability studies. The nanovector demonstrated high levels of targeting specificity and gene transfection efficiency with both C6 glioma and DAOY medulloblastoma tumor cells. Significantly, with the CTX as the targeting ligand, the nanovector may serve as a widely applicable gene delivery system for a broad array of cancer types. [Copyright &y& Elsevier]

Abstract: Impedance measurements of cell-based sensors are a primary characterization route for detection and analysis of cellular responses to chemical and biological agents in real time. The detection sensitivity and limitation depend on sensor impedance characteristics and thus on cell patterning techniques. This study introduces a cell patterning approach to bind cells on microarrays of gold electrodes and demonstrates that single-cell patterning can substantially improve impedance characteristics of cell-based sensors. Mouse fibroblast cells (NIH3T3) are immobilized on electrodes through a lysine–arginine–glycine–aspartic acid (KRGD) peptide-mediated natural cell adhesion process. Electrodes are made of three sizes and immobilized with either covalently bound or physically adsorbed KRGD (c-electrodes or p-electrodes). Cells attached to c-electrodes increase the measurable electrical signal strength by 48.4%, 24.2%, and 19.0% for three electrode sizes, respectively, as compared to cells attached to p-electrodes, demonstrating that both the electrode size and surface chemistry play a key role in cell adhesion and spreading and thus the impedance characteristics of cell-based sensors. Single cells patterned on c-electrodes with dimensions comparable to cell size exhibit well-spread cell morphology and substantially outperform cells patterned on electrodes of other configurations. [Copyright &y& Elsevier]

Abstract: Microarrays of single macrophage cell-based sensors were developed and demonstrated for potential real-time bacterium detection by synchrotron FTIR microscopy. The cells were patterned on gold electrodes of silicon oxide substrates by a surface engineering technique, in which the gold electrodes were immobilized with fibronectin to mediate cell adhesion and the silicon oxide background was passivated with polyethylene glycol (PEG) to resist protein adsorption and cell adhesion. Cell morphology and IR spectra of single, double, and triple cells on gold electrodes exposed to lipopolysaccharide (LPS) of different concentrations were compared to reveal the detection capability of this cell-based sensing platform. The single-cell-based system was found to generate the most significant and consistent IR spectrum shifts upon exposure to LPS, thus providing the highest detection sensitivity. Changes in cell morphology and IR shifts upon cell exposure to LPS were found to be dependent on the LPS concentration and exposure time, which established a method for the identification of LPS concentration and infected cell population. Possibility of using this single-cell system with conventional IR spectroscopy as well as its limitation was investigated by comparing IR spectra of single-cell arrays with gold electrode surface areas of 25, 100, and 400μm2 using both synchrotron and conventional FTIR spectromicroscopes. This cell-based platform may potentially provide real-time, label-free, and rapid bacterial detection, and allow for high-throughput statistical analyses, and portability. [Copyright &y& Elsevier]