Precise Diabetic Wound Therapy: PLS Nanospheres Eliminate Senescent Cells via DPP4 Targeting and PARP1 Activation.

Diabetic ulcers, a difficult problem faced by clinicians, are strongly associated with an increase in cellular senescence. Few empirical studies have focused on exploring a targeted strategy to cure diabetic wounds by eliminating senescent fibroblasts (SFs) and reducing side effects. In this study, poly-l-lysine/sodium alginate (PLS) is modified with talabostat (PT100) and encapsulates a PARP1 plasmid (PARP1@PLS-PT100) for delivery to target the dipeptidyl peptidase 4 (DPP4) receptor and eliminate SFs. PARP1@PLS-PT100 releases encapsulated plasmids, displaying high selectivity for SFs over normal fibroblasts by targeting the DPP4 receptor, decreasing senescence-associated secretory phenotypes (SASPs), and stimulating the secretion of anti-inflammatory factors. Furthermore, the increased apoptosis of SFs and the disappearance of cellular senescence alleviates SASPs, accelerates re-epithelialization and collagen deposition, and significantly induces macrophage M2 polarization, which mediates tissue repair and the inflammatory response. This innovative strategy has revealed the previously undefined role of PARP1@PLS-PT100 in promoting diabetic wound healing, suggesting its therapeutic potential in refractory wound repair.

A facile strategy for synthesis of porous Cu2O nanospheres and application as nanozymes in colorimetric biosensing.

Due to the unique advantages, developing a rapid, simple and economical synthetic strategy for porous nanomaterials is of great interest. In this work, for the first time, using sodium hypochlorite as a green oxidant, urea was oxidized to CO2 as a carbon source to prepare the fine-particle crosslinked Cu-precursors, which could be further reduced by sodium ascorbate into pure Cu2O nanospheres (NPs) with a porous morphology at room temperature. Interestingly, our study reveals that introduction of an appropriate amount of MgCl2 into the raw materials can tune the pore sizes and surface area, but has no influence on the phase purity of the resulting Cu2O NPs. Significantly, all the synthesized Cu2O NPs exhibited intrinsic peroxidase-like activity with higher affinity towards both 3,3,5,5-tetramethylbenzidine (TMB) and H2O2 than horseradish peroxidase (HRP) due to the highly porous morphology and the electrostatic attraction towards TMB. The colorimetric detection of glucose based on the resulting porous Cu2O NPs presented a limit of detection (LOD) of 2.19 µM with a broad linear range from 1-1000 µM, much better than many recently reported composite-based nanozymes. Meanwhile, this nanozyme system was utilized to detect l-cysteine, exhibiting a LOD value as low as 0.81 µM within a linear range from 0 to 10 µM. More interesting, this sensing system shows high sensitivity and excellent selectivity in determining glucose and l-cysteine, which is suitable for detecting serum samples with reliable results. Therefore, the present study not only develops a simple strategy to prepare Cu2O NPs with controllable porous structure, but also indicates its promising applications in bioscience and disease diagnosis.

Nanocarriers for treatment of dermatological diseases: Principle, perspective and practices.

Skin diseases are the fourth leading non-fatal skin conditions that act as a burden and affect the world economy globally. This condition affects the quality of a patient's life and has a pronounced impact on both their physical and mental state. Treatment of these skin conditions with conventional approaches shows a lack of efficacy, long treatment duration, recurrence of conditions, systemic side effects, etc., due to improper drug delivery. However, these pitfalls can be overcome with the applications of nanomedicine-based approaches that provide efficient site-specific drug delivery at the target site. These nanomedicine-based strategies are evolved as potential treatment opportunities in the form of nanocarriers such as polymeric and lipidic nanocarriers, nanoemulsions along with emerging others viz. carbon nanotubes for dermatological treatment. The current review focuses on challenges faced by the existing conventional treatments along with the topical therapeutic perspective of nanocarriers in treating various skin diseases. A total of 213 articles have been reviewed and the application of different nanocarriers in treating various skin diseases has been explained in detail through case studies of previously published research works. The toxicity related aspects of nanocarriers are also discussed.

Magnetic Nanostructure-Loaded Bicontinuous Nanospheres Support Multicargo Intracellular Delivery and Oxidation-Responsive Morphological Transitions.

Magnetic nanostructures (MNS) have a wide range of biological applications due to their biocompatibility, superparamagnetic properties, and customizable composition that includes iron oxide (Fe3O4), Zn2+, and Mn2+. However, several challenges to the biomedical usage of MNS must still be addressed, such as formulation stability, inability to encapsulate therapeutic payloads, and variable clearance rates in vivo. Here, we enhance the utility of MNS during controlled delivery applications via encapsulation within polymeric bicontinuous nanospheres (BCNs) composed of poly(ethylene glycol)-block-poly(propylene sulfide) (PEG-b-PPS) copolymers. PEG-b-PPS BCNs have demonstrated versatile encapsulation and delivery capabilities for both hydrophilic and hydrophobic payloads due to their unique and highly organized cubic phase nanoarchitecture. MNS-embedded BCNs (MBCNs) were thus coloaded with physicochemically diverse molecular payloads using the technique of flash nanoprecipitation and characterized in terms of their structure and in vivo biodistribution following intravenous administration. Retention of the internal aqueous channels and cubic architecture of MBCNs were verified using cryogenic transmission electron microscopy and small-angle X-ray scattering, respectively. MBCNs demonstrated improvement in magnetic resonance imaging (MRI) contrast enhancement (r2 relaxivity) as compared to free MNS, which in combination with scanning transmission electron microscopy and energy-dispersive X-ray spectroscopy evidenced the clustering and continued access to water of MNS following encapsulation. Furthermore, MBCNs were found to be noncytotoxic and able to deliver their hydrophilic and hydrophobic small-molecule payloads both in vitro and in vivo. Finally, the oxidation sensitivity of the hydrophobic PPS block allowed MBCNs to undergo a unique, triggerable transition in morphology into MNS-bearing micellar nanocarriers. In summary, MBCNs are an attractive platform for the delivery of molecular and nanoscale payloads for diverse on-demand and sustained drug delivery applications.

Metal-DNA coordination based bioinspired hybrid nanospheres for in situ amplification and sensing of microRNA.

Sufficient delivery of biomolecules into cells with high loading efficiency and easy cleavability would be significant for the visualization of biomolecules in living cells. Herein, a facile approach based on nano-wire balls (NWs) for efficient loading, intracellular delivery of nucleic acids and in situ targeted miRNA bioimaging is proposed, by feeding of Zn ions for generating DNA-inorganic hybrid structures with large surface areas and good stability. Given that the versatile and robust hybridization chain reaction (HCR) amplification strategy combines DNA assembly with intracellular assay, the resulting NWs without any complicated modification are capable of enhanced signals for the targeted imaging of cancer cells. This method realized a linear detection range of 100 fM to 10 nM, with a low detection limit of 83.6 fM in vitro, and could be used to effectively differentiate the expression levels of miRNA-21 in living cells. Due to its high loading efficiency, excellent biocompatibility and low toxicity, this system can be used to construct a coordination-based delivery nanoplatform for in situ enzyme-free amplified imaging of miRNAs, expanding the application of DNA-based nanomaterials for cellular delivery and intracellular molecule analysis.

Self-Assembled mRNA-Responsive DNA Nanosphere for Bioimaging and Cancer Therapy in Drug-Resistant Cells.

DNA assembly has provided new opportunities for the development of a novel drug delivery system (DDS) for real-time monitoring and precision treatment of cancer lesions. Herein, we propose mRNA-responsive DNA nanospheres (DNA-NS), whose self-assembly can be triggered by products of rolling circle amplification and functional hairpins and deliver anticancer drug doxorubicin (DOX) for bioimaging and cancer therapy. It has been demonstrated that DNA-NS exhibited good stability in biological environments. Hence, DNA-NS can serve as a universal platform of detections of mRNA related to various tumor cells. DNA-NS can also be applied in the mRNA-dependent DDS. For drug-resistant cells, which are widely present in actual cancer models, DNA-NS can effectively overcome the efflux action of drug-resistant cells to improve the therapeutic efficacy of DOX. In summary, this study provides a potential strategy for constructing the endogenous mRNA-responsive DDS for cancer diagnosis and chemotherapy in vivo.

Enzyme Instructed Self-assembly of Naphthalimide-dipeptide: Spontaneous Transformation from Nanosphere to Nanotubular Structures that Induces Hydrogelation.

Understanding the structure-morphology relationships of self-assembled nanostructures is crucial for developing materials with the desired chemical and biological functions. Here, phosphate-based naphthalimide (NI) derivatives have been developed for the first time to study the enzyme-instructed self-assembly process. Self-assembly of simple amino acid derivative NI-Yp resulted in non-specific amorphous aggregates in the presence of alkaline phosphatase enzyme. On the other hand, NI-FYp dipeptide forms spherical nanoparticles under aqueous conditions which slowly transformed into partially unzipped nanotubular structures during the enzymatic catalytic process through multiple stages which subsequently resulted in hydrogelation. The self-assembly is driven by the formation of ß-sheet type structures stabilized by offset aromatic stacking of NI core and hydrogen bonding interactions which is confirmed with PXRD, Congo-red staining and molecular mechanical calculations. We propose a mechanism for the self-assembly process based on TEM and spectroscopic data. The nanotubular structures of NI-FYp precursor exhibited higher cytotoxicity to human breast cancer cells and human cervical cancer cells when compared to the nanofiber structures of the similar Fmoc-derivative. Overall this study provides a new understanding of the supramolecular self-assembly of small-molecular-weight hydrogelators.

Nanospheres Loaded with Curcumin Improve the Bioactivity of Umbilical Cord Blood-Mesenchymal Stem Cells via c-Src Activation During the Skin Wound Healing Process.

Curcumin, a hydrophobic polyphenol derived from turmeric, has been used a food additive and as a herbal medicine for the treatment of various diseases, but the clinical application of curcumin is restricted by its poor aqueous solubility and its low permeability and bioavailability levels. In the present study, we investigate the functional role of a nanosphere loaded with curcumin (CN) in the promotion of the motility of human mesenchymal stem cells (MSCs) during the skin wound healing process. CN significantly increased the motility of umbilical cord blood (UCB)-MSCs and showed 10000-fold greater migration efficacy than curcumin. CN stimulated the phosphorylation of c-Src and protein kinase C which are responsible for the distinctive activation of the MAPKs. Interestingly, CN significantly induced the expression levels of α-actinin-1, profilin-1 and filamentous-actin, as regulated by the phosphorylation of nuclear factor-kappa B during its promotion of cell migration. In a mouse skin excisional wound model, we found that transplantation of UCB-MSCs pre-treated with CN enhanced wound closure, granulation, and re-epithelialization at mouse skin wound sites. These results indicate that CN is a functional agent that promotes the mobilization of UCB-MSCs for cutaneous wound repair.

First steps in the formulation of praziquantel nanosuspensions for pharmaceutical applications.

Praziquantel (PZQ), a broad spectrum anthelmintic drug, cannot be found in acceptable dosage forms for elderly patients, paediatric patients, and for veterinary use. In fact, very little has been done up to now in the formulation of liquid dosage forms, being they always formulated for parenteral administration. To beat this important challenge, it was accomplished a comprehensive analysis of the influence of two elementary physicochemical aspects, i.e. surface thermodynamic and electrokinetic properties, on the colloidal stability of PZQ nanosuspensions. The hydrophobic character of the drug, intensely determining the flocculation curves, was confirmed by the thermodynamic characterization. The electrophoretic characterization, in combination with the sedimentation and relative absorbance versus time curves, highlighted that the electrical double layer thickness and the surface charge can play an essential role in the stability of the pharmaceutical colloid. Finally, it was demonstrated that controlling the pH values and the incorporation of electrolytes can help in formulating PZQ aqueous nanosuspensions with appropriate stability and redispersibility behaviours for pharmaceutical use.

Selective Entropy Gain-Driven Adsorption of Nanospheres onto Spherical Bacteria Endows Photodynamic Treatment with Narrow-Spectrum Activity.

Narrow-spectrum antimicrobials specifically eradicate the target pathogens but suffer from significantly lagging development. Photodynamic therapy eliminates cells with reactive oxygen species (ROS) generated upon light irradiation but is intrinsically a wide-spectrum modality. We herein converted photodynamic therapy into a narrow-spectrum modality by taking advantage of a previously unnoticed physics recognition pathway. We found that negatively charged nanospheres undergo selective entropy gain-driven adsorption onto spherical bacteria, but not onto rod-like bacteria. This bacterial morphology-targeting selectivity, combined with the extremely limited effective radii of action of ROS, enabled photodynamic nanospheres to kill >99% of inoculated spherical bacteria upon light irradiation and <1% of rod-like bacteria under comparable conditions, indicative of narrow-spectrum activity against spherical bacteria. This work unveils the bacterial morphology selectivity in the adsorption of negatively charged nanospheres and suggests a new approach for treating infections characterized by overthriving spherical bacteria in niches naturally dominated by rod-like bacteria.

Distribution of benzalkonium chloride into the aqueous phases of submicron dispersed systems: emulsions, aqueous lecithin dispersion and nanospheres.

Partitioning of benzalkonium chloride (BAC) into the aqueous phases of submicron dispersed systems such as submicron emulsions, aqueous lecithin dispersion (WLD), and suspension of nanospheres (NLC) was studied. The aqueous phases of the investigated systems were obtained by ultracentrifugation and subsequently were subjected to ultrafiltration, which procedure allowed distinguishing between the fractions of free benzalkonium chloride (w) and those incorporated in the liposomal and micellar region (wlm). The fractions present in the oily phase and in the interphase of submicron emulsions were calculated. Despite the various composition of the investigated formulations and the initial concentration of BAC, w values were very small at 0.2-8.0%. The wlm value in submicron emulsions was increased by increasing the total concentration of preservative from 29.0 to 42.0%. Using polysorbate 80 instead of lecithin resulted in a distribution of BAC to aqueous-liposomal-micellar phase that was twice as high. The very low concentration of antimicrobial active form of benzalkonium chloride was analyzed in the aqueous phase of emulsions stabilized with lecithin as well as in aqueous lecithin dispersion and nanospheres (below 3%). Replacement of lecithin with polysorbate 80 in emulsions with polysorbate significantly increase (up to 8%) the fraction of benzalkonium chloride in the aqueous phase where microbial growth occurs.

Core-shell nanospheres behind the blue eyes of the bay scallop Argopecten irradians.

The bay scallop Argopecten irradians (Mollusca: Bivalvia) has dozens of iridescent blue eyes that focus light using mirror-based optics. Here, we test the hypothesis that these eyes appear blue because of photonic nanostructures that preferentially scatter short-wavelength light. Using transmission electron microscopy, we found that the epithelial cells covering the eyes of A. irradians have three distinct layers: an outer layer of microvilli, a middle layer of random close-packed nanospheres and an inner layer of pigment granules. The nanospheres are approximately 180 nm in diameter and consist of electron-dense cores approximately 140 nm in diameter surrounded by less electron-dense shells 20 nm thick. They are packed at a volume density of approximately 60% and energy-dispersive X-ray spectroscopy indicates that they are not mineralized. Optical modelling revealed that the nanospheres are an ideal size for producing angle-weighted scattering that is bright and blue. A comparative perspective supports our hypothesis: epithelial cells from the black eyes of the sea scallop Placopecten magellanicus have an outer layer of microvilli and an inner layer of pigment granules but lack a layer of nanospheres between them. We speculate that light-scattering nanospheres help to prevent UV wavelengths from damaging the internal structures of the eyes of A. irradians and other blue-eyed scallops.

Structure of Rat Testicles after Intravenous Injection of Nanosized Magnetite Particles.

The structure of the testicles was studied in adult rats in 120 days after a single intravenous injection of chitosan-modified (magnetic nanospheres) and lipid-modified (magnetoliposomes) nanosized magnetite particles. Perls histochemical reaction detected in the testicular interstitial connective tissue the cells which absorbed and accumulated magnetite nanoparticles. The dynamics of spermatogenesis index and the count of Perls+ cells in the rat testicles were traced throughout the experiment. The studied modified nanosized magnetite particles did not penetrate through the blood-testicle barrier in rats.

Role of Endocytosis in Nanoparticle Penetration of 3D Pancreatic Cancer Spheroids.

Robust deposition of extracellular matrix is a significant barrier for delivery of nanotherapeutics and small-molecule anticancer drugs to different tumors including pancreatic ductal adenocarcinoma. Here, we investigated permeation and total uptake of polystyrene nanoparticles of different diameters in 3D multicellular spheroid models of pancreatic tumors. Special attention was given to analysis of the impact of endocytic processes on nanoparticle accumulation and distribution in spheroids. We generated spheroids of BxPC3 or PANC-1 cells that were able to internalize 20, 100, and 500 nm fluorescent polystyrene beads with different efficacies, resulting in 20 ≫100 > 500 nm and 100 > 500 > 20 nm trends, respectively. It was found that endocytosis and transcytosis increased overall nanoparticle uptake and facilitated permeation of 20 nm beads in BxPC3 spheroids, whereas 100 and 500 nm particles did not penetrate. In PANC-1 spheroids, penetration of nanoparticles also decreased with the increase of size but was not significantly affected by endocytic processes. Thus, our study showed that passive diffusion and endocytic processes may have a different contribution to nanoparticle accumulation and distribution in spheroid models of pancreatic cancer.

Cellular uptake, intracellular distribution and degradation of Her2-targeting silk nanospheres.

BACKGROUND: The development of nanocarrier technology has attracted great interest in the last decade. Biodegradable spheres made of functionalized silk have considerable potential to be used as drug delivery systems for cancer treatment. A targeting ligand displayed at the surface of a carrier, with a specific affinity towards a particular receptor, can further enhance the accumulation and uptake of nanoparticles at the site of a tumor. MATERIALS AND METHODS: The hybrid constructs were obtained by adding a Her2-binding peptide (H2.1) to MS1 and MS2 bioengineered silks based on the MaSp1 and MaSp2 proteins from N. clavipes, respectively. The H2.1MS1 and H2.1MS2 proteins were blended at a weight ratio of 8:2. Stable silk particles were formed by mixing a soluble protein with potassium phosphate using a micromixing technique. We used specific inhibitors of endocytosis to determine the cellular uptake pathway of the silk nanoparticles in human Her2-positive breast cancer cells. The subcellular distribution of silk particles was investigated by evaluating the signal colocalization with organelle-specific tracker. Moreover, lysosomal and exosomal inhibitors were implemented to evaluate their impact on the silk spheres behavior and degradation. RESULTS: The functionalized spheres were specifically taken up by Her2-positive cancer cells. Silk particles facilitated the entry into cells through both the clathrin- and caveola-dependent pathways of endocytosis. Upon entering the cells, the particles accumulated in the lysosomes, where intracellular degradation occurred. CONCLUSIONS: The present study demonstrated directly that the lysosomal function was essential for silk-based carrier elimination. The degradation of the carrier is of great importance to develop an optimal drug delivery system.

On the advancement of polymeric bicontinuous nanospheres toward biomedical applications.

Self-assembled soft nanocarriers that are capable of simultaneous encapsulation of both lipophilic and water soluble payloads have significantly enhanced controlled delivery applications in biomedicine. These nanoarchitectures, such as liposomes, polymersomes and cubosomes, are primarily composed of either amphiphilic polymers or lipids, with the polymeric variants generally possessing greater stability and control over biodistribution and bioresponsive release. Polymersomes have long demonstrated such advantages over their lipid analogs, liposomes, but only recently have bicontinuous nanospheres emerged as a polymeric cubic phase alternative to lipid cubosomes. In this review, we summarize the current state of the field for bicontinuous nanosphere formulation and characterization and suggest future directions for this nascent delivery platform as it is adopted for biomedical applications.

Systematic in vivo study of NiO nanowires and nanospheres: biodegradation, uptake and biological impacts.

High aspect ratio nanomaterials (NM) have a promising future in medicine and industry as a unique category of NM. Consequently, it is important to evaluate their potential biological side-effects before crediting their use. To understand the mechanisms of degradation, internalisation, and interaction with different biological targets, we used the in vivo model Drosophila melanogaster to obtain a systematic and complete study on high aspect ratio Ni nanowires (NiNW), compared with low aspect ratio Ni nanospheres (NiNS), and NiSO4 as a model of agent releasing nickel ions. The distinguished shape of nanowires showed changes in their characteristics after oral administration until they reached the intestinal lumen, where their diameter decreased significantly. For the first time, we confirmed the internalization of needle-shaped materials via perforation of the intestinal barrier. Moreover, the results showed that D. melanogaster is a valid and effective tool in studies related to magnetic resonance imaging (MRI). Additionally, NiNM induced DNA damage and molecular changes at the gene expression level, in association with increase in oxidative stress levels. Notably, the observed negative effects were related to nickel as a metal rather than to its shape, since the effects induced by the three Ni forms were notably similar. In addition, independent of their form, Ni compounds did not induce toxic or mutagenic impacts. Our Drosophila model can be used to understand different phenomena related to high aspect ratio NM exposure, such as degradation, internalization and interaction with different targets.

Modifying Self-Assembled Peptide Cages To Control Internalization into Mammalian Cells.

Nanoparticles can be used to transport a variety of biological cargoes into eukaryotic cells. Polypeptides provide a versatile material for constructing such systems. Previously, we have assembled nanoscale peptide cages (SAGEs) from de novo designed coiled-coil modules. Here, we show that the modules can be extended with short charged peptides to alter endocytosis of the assembled SAGE particles by cultured human cells in a tunable fashion. First, we find that the peptide extensions affect coiled-coil stability predictably: N-terminal polylysine and C-terminal polyglutamate tags are destabilizing; whereas, the reversed arrangements have little impact. Second, the cationic assembled particles are internalized faster and to greater extents by cells than the parent SAGEs. By contrast, anionic decorations markedly inhibit both aspects of uptake. These studies highlight how the modular SAGE system facilitates rational peptide design to fine-tune the bioactivity of nanoparticles, which should allow engineering of tailored cell-delivery vehicles.

Improvement in dissolution rate and photodynamic efficacy of chlorin e6 by sucrose esters as drug carrier in nanosuspension formulation: optimisation and in vitro characterisation.

OBJECTIVES: Chlorin e6 is a poorly water-soluble photoactive drug. Its monomers form aggregates at the tumour physiological pH, which drastically reduces its photodynamic efficacy. This study aimed to improve the dissolution rate and photodynamic efficacy of chlorin e6 by nanosuspension formulation using biodegradable sucrose esters as drug carrier. METHODS: A modified emulsion-solvent diffusion method was used to prepare the nanosuspension, where amount of Ce6, ratio of sucrose monopalmitate to sucrose monolaurate as carrier and ratio of dichloromethane to acetone as solvent, were varied using central composite design. Particle size, zeta potential, encapsulation efficiency and in vitro drug release characteristics of the nanosuspensions were evaluated. The formulation was optimised by response surface methodology and its photodynamic efficacy evaluated. KEY FINDINGS: The optimised nanosuspension had mean particle size of ~200 nm, 88% drug encapsulation efficiency and faster drug release compared to pure Ce6. Spectroscopic studies showed that Ce6 exists in monomeric form in the carrier, which facilitated a remarkable increase in cellular uptake, in vitro singlet oxygen generation and cytotoxicity to oral squamous carcinoma cells. CONCLUSIONS: The dissolution rate and photodynamic efficacy of Ce6 were markedly improved by formulating the drug as a nanosuspension with sucrose esters as drug carrier.

In vivo guiding nitrogen-doped carbon nanozyme for tumor catalytic therapy.

Nanomaterials with intrinsic enzyme-like activities (nanozymes), have been widely used as artificial enzymes in biomedicine. However, how to control their in vivo performance in a target cell is still challenging. Here we report a strategy to coordinate nanozymes to target tumor cells and selectively perform their activity to destruct tumors. We develop a nanozyme using nitrogen-doped porous carbon nanospheres which possess four enzyme-like activities (oxidase, peroxidase, catalase and superoxide dismutase) responsible for reactive oxygen species regulation. We then introduce ferritin to guide nitrogen-doped porous carbon nanospheres into lysosomes and boost reactive oxygen species generation in a tumor-specific manner, resulting in significant tumor regression in human tumor xenograft mice models. Together, our study provides evidence that nitrogen-doped porous carbon nanospheres are powerful nanozymes capable of regulating intracellular reactive oxygen species, and ferritinylation is a promising strategy to render nanozymes to target tumor cells for in vivo tumor catalytic therapy.