Multifunctional nanocarriers and intracellular drug delivery

Multifunctional nanocarriers for the delivery and targeting of therapeutic and diagnostic agents in cancer therapy have received significantly increased interest in recent years.Several multifunctional nanocarriers engineered from a wide range of materials with consolidation of various functionalities for long circulation, targetability, stimuli-sensitivity, intracellular delivery for therapy and imaging have been shown to be capable of killing the desired target diseased cells with minimal side effects to provide enhanced contrast during imaging for disease location and monitor both the fate of the nanocarrier and treatment in real time. This review highlights recent advances in the design and engineering of multifunctional nanocarriers, along with the importance of intracellular delivery.     

Polyethyleneimine modified biocompatible poly(N-isopropylacrylamide)-based nanogels for drug delivery

A series of biocompatible and stimuli-sensitive poly(N-isopropylacrylamide-co-propyl acrylic acid) (P(NIPAAm-co-PAAc)) nanogels were synthesized by emulsion polymerization. In addition, polyethyleneimine (PEI) was further grafted to modify the PNIPAAm-based nanogels. The P(NIPAAm-co-PAAc)-g-PEI nanogels exhibited good thermosensitivity as well as pH sensitivity. Transmission electron microscopy (TEM) showed that the P(NIPAAm-co-PAAc)-g-PEI and P(NIPAAm-co-PAAc) nanogels displayed well dispersed spherical morphology. The mean sizes of the nanogels measured by dynamic light scattering (DLS) were from 100 nm to 500 nm at different temperatures. The cytotoxicity study indicated P(NIPAAm-co-PAAc) nanogels exhibited a better biocompatibility than both PNIPAAm nanogel and P(NIPAAm-co-PAAc)-g-PEI nanogel although all the three kinds of nanogels did not exhibit apparent cytotoxicity. The drug-loaded nanogels, especially the PEI-grafted nanogels, showed temperature-trigged controlled release behaviors, indicating the potential applications as an intelligent drug delivery system.     

Multifunctional polymeric micelles as cancer-targeted, MRI-ultrasensitive drug delivery systems

We describe the development of multifunctional polymeric micelles with cancer-targeting capability via alpha(v)beta(3) integrins, controlled drug delivery, and efficient magnetic resonance imaging (MRI) contrast characteristics. Doxorubicin and a cluster of superparamagnetic iron oxide (SPIO) nanoparticles were loaded successfully inside the micelle core. The presence of cRGD on the micelle surface resulted in the cancer-targeted delivery to alpha(v)beta(3)-expressing tumor cells. In vitro MRI and cytotoxicity studies demonstrated the ultrasensitive MRI imaging and alpha(v)beta(3)-specific cytotoxic response of these multifunctional polymeric micelles.     

Recent advances in emerging 2D nanomaterials for biosensing and bioimaging applications

The great success of graphene throws new light on discovering more two-dimensional (2D) layered nanomaterials that stem from atomically thin 2D sheets. Compared with a single element of graphene, emerging graphene-like 2D materials composed of multiple elements that possess more versatility, greater flexibility and better functionality with a wide range of potential applications. In this review, we provide insights into the rapidly emerging 2D materials and their biosensing and bioimaging applications in recent three years, including 2D transition metal nanomaterials, graphitic nitride materials, black phosphorus, and emerging 2D organic polymers. We first briefly highlight their unique 2D morphology and physicochemical properties and then focus on their recent applications in electrochemical biosensing, optical biosensing and bioimaging. The challenges and some thoughts on future perspectives in this field are also addressed.     

Biomimetic self-assembly of calcium phosphate templated by PNIPAAm nanogels for sustained smart drug delivery

Poly(N-isopropylacrylamide) (PNIPAAm)/calcium phosphate (CaP) hybrid nanocomposites with dual-responsive controlled drug delivery property have been prepared by in-situ biomineralization process. Poly(acrylic acid) (PAA) is used as a crystal growth additive to control the morphology of the hybrid nanocomposites. The interaction between PAA and Ca²⁺ contributes to the formation of homogeneous and robust nanocomposites. Vitamin B₂ release behavior is found to be pH- and thermal-responsive. Additionally, the release profiles are sustained with the introduction of CaP, indicating that CaP nanocrystallines could decrease the permeation of the encapsulated drug effectively. The results suggest that the prepared hybrid nanocomposites can be used as “smart” nanoscale materials for sustained dual-responsive drug delivery.     

Multifunctional FeCo-graphitic carbon nanocrystals for combined imaging, drug delivery and tumor-specific photothermal therapy in mice

Ultrasmall FeCo-graphitic carbon shell nanocrystals (FeCo/GC) are promising multifunctional materials capable of highly efficient drug delivery in vitro and magnetic resonance imaging in vivo. In this work, we demonstrate the use of FeCo/GC for highly effective cancer therapy through combined drug delivery, tumor-selective near-infrared photothermal therapy, and cancer imaging of a 4T1 syngeneic breast cancer model. The graphitic carbon shell of the ∼4 nm FeCo/GC readily loads doxorubicin (DOX) via π-π stacking and absorbs near-infrared light giving photothermal heating. When used for cancer treatment, intravenously administrated FeCo/GC-DOX led to complete tumor regression in 45% of mice when combined with 20 min of near-infrared laser irradiation selectively heating the tumor to 43–45 °C. In addition, the use of FeCo/GC-DOX results in reduced systemic toxicity compared with free DOX and appears to be safe in mice monitored for over 1 yr. FeCo/GC-DOX is shown to be a highly integrated nanoparticle system for synergistic cancer therapy leading to tumor regression of a highly aggressive tumor model.      

Recent progress in engineering near-infrared persistent luminescence nanoprobes for time-resolved biosensing/bioimaging

Persistent luminescence nanoprobes (PLNPs) can remain luminescent after ceasing excitation. Due to the ultra-long decay time of persistent luminescence (PersL), autofluorescence interference can be efficiently eliminated by collecting PersL signal after autofluorescence decays completely, thus the imaging contrast and sensing sensitivity can be significantly improved. Since near-infrared (NIR) light shows reduced scattering and absorption coefficient in penetrating biological organs or tissues, near-infrared persistent luminescence nanoprobes (NIR PLNPs) possess deep tissue penetration and offer a bright prospect in the areas of in vivo biosensing/bioimaging. In this review, we firstly summarize the design of different types of NIR PLNPs for biosensing/bioimaging, such as transition metal ions-doped NIR PLNPs, lanthanide ions-doped NIR PLNPs, organic molecules-based NIR PLNPs, and semiconducting polymer self-assembled NIR PLNPs. Notably, organic molecules-based NIR PLNPs and semiconductor self-assembled NIR PLNPs, for the first time, were introduced to the review of PLNPs. Secondly, the effects of different types of charge carriers on NIR PersL and luminescence decay of NIR PLNPs are significantly emphasized so as to build up an in-depth understanding of their luminescence mechanism. It includes the regulation of valence band and conduction band of different host materials, alteration of defect types, depth and concentration changes caused by ion doping, effective radiation transitions and energy transfer generated by different luminescence centers. Given the design and potential of NIR PLNPs as long-lived luminescent materials, the current challenges and future perspective in this rapidly growing field are also discussed.

     

Multifunctional nanorods for gene delivery

The goal of gene therapy is to introduce foreign genes into somatic cells to supplement defective genes or provide additional biological functions, and can be achieved using either viral or synthetic non-viral delivery systems. Compared with viral vectors, synthetic gene-delivery systems, such as liposomes and polymers, offer several advantages including ease of production and reduced risk of cytotoxicity and immunogenicity, but their use has been limited by the relatively low transfection efficiency. This problem mainly stems from the difficulty in controlling their properties at the nanoscale. Synthetic inorganic gene carriers have received limited attention in the gene-therapy community, the only notable example being gold nanoparticles with surface-immobilized DNA applied to intradermal genetic immunization by particle bombardment. Here we present a non-viral gene-delivery system based on multisegment bimetallic nanorods that can simultaneously bind compacted DNA plasmids and targeting ligands in a spatially defined manner. This approach allows precise control of composition, size and multifunctionality of the gene-delivery system. Transfection experiments performed in vitro and in vivo provide promising results that suggest potential in genetic vaccination applications.     

BioMEMS for drug delivery

Recent research into methods of using microelectromechanical systems (MEMS) technology for medical and biological applications has developed several interesting devices. This paper reviews various approaches to the use of MEMS for drug therapy, including devices based on microporous silicon, microneedles, micropumps, and microreservoirs. Microdevices can improve drug therapy because they allow precise and complex dosing, induce less pain, or increase compliance. Microneedles have been tested on humans, and the other drug delivery MEMS have shown promise in vitro and in vivo. Investigations into the use of microelectromechanical systems (MEMS) technology to produce microdevices for drug delivery have expanded recently. We present several different approaches to the use of microdevices for drug therapy and the current state of the field.     

Stealth dendrimers for drug delivery: correlation between PEGylation,cytocompatibility, and drug payload

It is advantageous to utilize low generation polyamidoamine (PAMAM) dendrimers for drug delivery because low generations (generation 4.0 or below) have more biologically favorable properties as compared to high generations. Nevertheless, modification of low generation dendrimers with PEG to create stealth dendrimers is still necessary to avoid potential side effects by long term accumulation. However, low generation dendrimers have much fewer surface sites for drug loading as compared to higher generations. To efficiently utilize low generation dendrimer-based stealth dendrimers for drug loading, PEGylation needs to be optimized. In this study, we synthesized a series of stealth dendrimers based on low generation Starburst™ PAMAM dendrimers (i.e., G2.5, G3.0, G3.5, and G4.0) and quantitatively assessed PEGylation efficacy in modulating cytocompatibility of low generation PAMAM dendrimers. Cytocompatibility of stealth dendrimers was examined using endothelial cells. The results showed that PEGylation degree on low generation dendrimers could be dramatically reduced to leave as many unoccupied surface groups as possible for drug loading, while maintaining the drug carrier cytocompatibility. 3PEGs-G3.0 and 10PEGs-G4.0 were considered initially optimized stealth dendrimers that would be further modified to deliver drugs of interest. Correlation of PEGylation, cytocompatibility, and drug payload allowed us to optimize low generation dendrimer-based stealth dendrimers for drug delivery and advance the understanding of structure-property relationship of stealth dendrimers.     

Thermosensitive P(NIPAAm-co-PAAc-co-HEMA) nanogels conjugated with transferrin for tumor cell targeting delivery

Multifunctional and thermosensitive poly(N-isopropylacrylamide-co-propyl acrylic acid-co-hydroxyethyl methacrylate) (P(NIPAAm-co-PAAc-co-HEMA)) nanogels were prepared by miniemulsion polymerization. The mean sizes of the nanogels measured by dynamic light scattering (DLS) varied from 120 to 400?nm with an increase in temperature. Transmission electron microscopy (TEM) showed that the nanogels displayed well-dispersed spherical morphology. The nanogels were conjugated by human transferrin (Tf) and the coupling of transferrin molecules with nanogels was verified by UV-vis spectroscopy. The?cytotoxicity study indicated that the nanogels did not exhibit apparent cytotoxicity. Fluorescence spectroscopy analysis as well as confocal laser scanning microscopy (CLSM) was used to confirm that the Tf-conjugated nanogels could specifically bind to A549 tumor cells. In addition, the Tf-conjugated nanogels loaded with Doxorubicin (Dox) could efficiently release the drug inside the cell, suggesting that the Tf-conjugated nanogels are useful drug carriers for tumor cell targeting.     

Electrospun fibers for tissue engineering, drug delivery, and wound dressing

Electrospinning, a technique well known for fabricating nanoscale fibers, has recently been studied extensively due to its various advantages such as high surface-to-volume ratio, tunable porosity, and ease of surface functionalization. The resulting fibers are extremely useful for applications in the fields of tissue engineering, drug delivery, and wound dressing. Since electrospun fiber mimic extracellular matrix of tissue in terms of scale and morphology, its potential to be used as scaffold is continuously explored by researchers, especially in the field of vascular, nerve, bone, and tendon/ligament tissue engineering. Besides morphology, physical, and chemical properties, electrospun scaffolds are often evaluated through various cell studies. Researchers have adopted approaches such as surface modification and drug loading to enhance the property and function of scaffold. This review gives an overview of some current aspects of various applications of electrospun fibers, particularly in biomedical fields, how researchers have enhanced electrospun fibers with different methods and attempted to overcome the inherent limitation of electrospinning by using novel techniques.     

Bioeliminable nanohydrogels for drug delivery

One of the most significant obstacles for systematic delivery of nanopayloads is the foreign particle clearance by the mononuclear phagocyte system (MPS). The majority of biocompatible nanopayloads with charged groups on their surface cannot fully evade the clearance by MPS during systemic circulation. For safe and effective targeted nanodrug delivery in vivo, we describe a novel approach for evading the macrophage clearance. We demonstrate that neutral and hydrophilic materials can effectively evade the macrophage uptake and also quickly degrade into bioeliminable fragments. We show that there is no opsonization effect and no toxic effect on living cells. In addition, the payloads are stable in an aqueous environment, and they can release drugs in a cellular environment. These results suggest that the unique properties of this kind of payloads may make them useful for designing new drug delivery systems.     

Hydroxypatite-polysaccharide granules for drug delivery

The formation of hydroxyapatite by co-precipitation from sodium alginate and sodium carboxymethylcellulose aqueous solutions with the use of dibasic calcium phosphate dihydrate and calcium hydroxide as starting reagents is studied. A technique to prepare the hydroxyapatite/polysaccharide (micro)granules is developed. An introduction of an antimicrobial Biocide 1 agent in proper amount into the granules is provided, and the behavior of the granules is evaluated.     

Nanoparticles for intracellular-targeted drug delivery

Nanoparticles (NPs) are very promising for the intracellular delivery of anticancer and immunomodulatory drugs, stem cell differentiation biomolecules and cell activity modulators. Although initial studies in the area of intracellular drug delivery have been performed in the delivery of DNA, there is an increasing interest in the use of other molecules to modulate cell activity. Herein, we review the latest advances in the intracellular-targeted delivery of short interference RNA, proteins and small molecules using NPs. In most cases, the drugs act at different cellular organelles and therefore the drug-containing NPs should be directed to precise locations within the cell. This will lead to the desired magnitude and duration of the drug effects. The spatial control in the intracellular delivery might open new avenues to modulate cell activity while avoiding side-effects.     

Nanocarrier-based topical drug delivery for an antifungal drug

Objective: The conventional liposomal amphotericin B causes many unwanted side effects like blood disorder, nephrotoxicity, dose-dependent side effects, highly variable oral absorption and formulation-related instability. The objective of the present investigation was to develop cost-effective nanoemulsion as nanocarreir for enhanced and sustained delivery of amphotericin B into the skin.

Methods and characterizations: Different oil-in-water nanoemulsions were developed by varying the composition of hydrophilic (Tween® 80) surfactants and co-surfactant by the spontaneous titration method. The developed formulation were characterized, optimized, evaluated and compared for the skin permeation with commercial formulation (fungisome 0.01% w/w). Optimized formulations loaded with amphotericin B were screened using varied concentrations of surfactants and co-surfactants as decided by the ternary phase diagram.

Results and discussion: The maximum % transmittance obtained were 96.9?±?1.0%, 95.9?±?3.0% and 93.7?±?1.2% for the optimized formulations F-I, F-III and F-VI, respectively. These optimized nanoemulsions were subjected to thermodynamic stability study to get the most stable nanoemulsions (F-I). The results of the particle size and zeta potential value were found to be 67.32?±?0.8 nm and –3.7?±?1.2?mV for the final optimized nanoemulsion F-I supporting transparency and stable nanoemulsion for better skin permeation. The steady state transdermal flux for the formulations was observed between 5.89?±?2.06 and 18.02?±?4.3?µg/cm²/h whereas the maximum enhancement ratio were found 1.85- and 3.0-fold higher than fungisome and drug solution, respectively, for F-I. The results of the skin deposition study suggests that 231.37?±?3.6?µg/cm² drug deposited from optimized nanoemulsion F-I and 2.11-fold higher enhancement ratio as compared to fungisome. Optimized surfactants and co-surfactant combination-mediated transport of the drug through the skin was also tried and the results were shown to have facilitated drug permeation and skin perturbation (SEM).

Conclusion: The combined results suggested that amphotericin B nanoemulsion could be a better option for localized topical drug delivery and have greater potential as an effective, efficient and safe approach.     

Multifunctional hydroxyapatite and poly(d,l-lactide-co-glycolide) nanoparticles for the local delivery of cholecalciferol

Cholecalciferol, vitamin D₃, plays an important role in bone metabolism by regulating extracellular levels of calcium. Presented here is a study on the effects of the local delivery of cholecalciferol (D₃) using nanoparticulate carriers composed of hydroxyapatite (HAp) and poly(d,l-lactide-co-glycolide) (PLGA). Multifunctional nanoparticulate HAp-based powders were prepared for the purpose of: (a) either fast or sustained, local delivery of cholecalciferol, and (b) the secondary, osteoconductive and defect-filling effect of the carrier itself. Two types of HAp-based powders with particles of narrowly dispersed sizes in the nano range were prepared and tested in this study: HAp nanoparticles as direct cholecalciferol delivery agents and HAp nanoparticles coated with cholecalciferol-loaded poly(d,l)-lactide-co-glycolide (HAp/D₃/PLGA).Satisfying biocompatibility of particulate systems, when incubated in contact with MC3T3-E1 osteoblastic cells in vitro, was observed for HAp/D₃/PLGA and pure HAp. In contrast, an extensively fast release of cholecalciferol from the system comprising HAp nanoparticles coated with cholecalciferol (HAp/D3) triggered necrosis of the osteoblastic cells in vitro. Artificial defects induced in the osteoporotic bone of the rat mandible were successfully reconstructed following implantation of cholecalciferol-coated HAp nanoparticles as well as those comprising HAp nanoparticles coated with cholecalciferol-loaded PLGA (HAp/D₃/PLGA). The greatest levels of enhanced angiogenesis, vascularization, osteogenesis and bone structure differentiation were achieved upon the implementation of HAp/D₃/PLGA systems.