Rotary Jet-Spun Polycaprolactone/Hydroxyapatite and Carbon Nanotube Scaffolds Seeded with Bone Marrow Mesenchymal Stem Cells Increase Bone Neoformation.

Clinically, bone tissue replacements and/or bone repair are challenging. Strategies based on well-defined combinations of osteoconductive materials and osteogenic cells are promising to improve bone regeneration but still require improvement. Herein, we combined polycaprolactone (PCL) fibers, carbon nanotubes (CNT), and hydroxyapatite (nHap) nanoparticles to develop the next generation of bone regeneration material. Fibers formed by rotary jet spinning (RJS) instead of traditional electrospinning (ES) with embedded bone marrow mesenchymal stem cells (BMMSCs) showed the best outcomes to repair rat calvarial defects after 6 weeks. To understand this, it was observed that different morphologies were formed depending on the manufacturing method used. RJS fibers presented a particular topography with rough fibers, which allowed for better cellular growth and cell spreading in vitro around and into a three-dimensional (3D) mesh, while fibers made by ES were more smooth and cellular growth was only measured on the 3D mesh surface. The fibers with incorporated nHap/CNT nanoparticles enhanced in vitro cell performance as indicated by more cellular proliferation, alkaline phosphatase activity, proliferation, and deposition of calcium. Greater bone neoformation occurred by combining three characteristics: the presence of nHap and CNT nanoparticles, the topography of the RJS fibers, and the addition of BMMSCs. RJS fibers with nanoparticles and seeded with BMMSCs showed 10 136 mm3 of bone neoformation, meaning a 10-fold increase compared to using RJS only and BMMSCs (0.853 mm3) and a 5-fold increase from using ES only (2054 mm3) after 6 weeks of implantation. Conversely, none of these approaches used individually showed any significant difference for in vivo bone neoformation, suggesting that their combination is essential for optimizing bone formation. In summary, our work generated a potential material composed of well-defined combinations of suitable scaffolds seeded with BMMSCs for enhancing numerous orthopedic tissue engineering applications.

Fumaryl diketopiperazine based effervescent microparticles to escape macrophage phagocytosis for enhanced treatment of pneumonia via pulmonary delivery.

The treatment of pulmonary infections with antibiotics administered via pulmonary delivery provides for higher local therapeutic efficacy rather than through systemic administration. Pneumonia is globally considered a major cause of death due to a lack of proper medication. The treatment of pneumonia with inhalable antibiotics (such as azithromycin (AZM)) can provide a maximum pulmonary therapeutic effect without significant systemic side effects. Compared to non-effervescent microparticles, effervescent microparticles can provide an active driving force to release loaded antibiotics for subsequent distribution deep into the lung by virtue of its smaller size. In this study, N-fumaroylated diketopiperazine (FDKP) was used as a carrier to prepare effervescent inhalable microparticles loaded with AZM (AZM@FDKP-E-MPs). This effervescent dry powder was characterized for both in vitro and in vivo deposition in the lung and the results obtained showed significant improvement in lung deposition and anti-bacterial efficiency, suggesting a strong potential application for pneumonia treatment.

Co-delivery of Poria cocos extract and doxorubicin as an 'all-in-one' nanocarrier to combat breast cancer multidrug resistance during chemotherapy.

Recent studies have indicated that multidrug resistance (MDR) can significantly limit the effects of conventional chemotherapy. In this study, PT (Pachymic acid and dehydrotumulosic acid) are the two major triterpenoid components purified and identified in P. cocos. A liposomal co-delivery system encapsulating doxorubicin (DOX) and PT was prepared. Notably, the mechanism of PT reversed P-glycoprotein (P-gp) mediated MDR mainly relied on the inhibition of the P-gp function, which further decreased the levels of P-gp and caveolin-1 proteins. In drug-resistant MCF cells, co-administration with 5 µg/ml PT significantly enhanced sensitivity of DOX. Finally, liposome-mediated co-delivery with PT significantly improved the anti-tumor effect of DOX in tumor-bearing mice when compared to other single therapy groups. In conclusion, this study showed for the first time that DOX and PT act synergistically as an "all-in-one" treatment to reverse MDR during tumor treatment and, thus, should be studied further for a wide range of anti-cancer applications.

Glioma-targeted dual functionalized thermosensitive Ferri-liposomes for drug delivery through an in vitro blood-brain barrier.

To date, the delivery of therapeutic agents for malignant brain tumors (such as glioblastoma multiforme (GBM)) remains a significant obstacle due to the existence of the blood-brain barrier (BBB). A multitude of delivery systems (hydrogels, micelles, polymeric nanoparticles, etc.) have been proposed, yet many of them exhibit limited tumor-specific inhibition effects. Herein, a drug-encapsulated dual-functionalized thermosensitive liposomal system (DOX@P1NS/TNC-FeLP) was developed for targeted delivery across the BBB. Specifically, a GBM-specific cell-penetrating peptide (P1NS) and an anti-GBM antibody (TN-C) were conjugated onto the liposome surface for targeted delivery. In addition, superparamagnetic iron oxide nanoparticles (SPIONs) and doxorubicin (DOX) were co-loaded inside the liposomes to achieve thermo-triggered drug release when applying an alternating magnetic field (AMF). Results demonstrated that P1NS/TNC-FeLPs readily transported across an in vitro BBB model and displayed a thermo-responsive and GBM-specific cellular uptake as well as drug release profile. Additionally, results from immunofluorescent (IF) staining and RT-qPCR further demonstrated that DOX@P1NS/TNC-FeLPs specifically entered U-87 human GBM cells and suppressed tumor cell proliferation without causing any significant impact on healthy brain cell function. As such, the novel DOX@P1NS/TNC-FeLPs presented potent and precise anti-GBM capability and, therefore, are suggested here for the first time as a promising DDS to deliver therapeutic agents across the BBB for GBM treatment.

In vitro and ex vivo systems at the forefront of infection modeling and drug discovery.

Bacterial infections and antibiotic resistant bacteria have become a growing problem over the past decade. As a result, the Centers for Disease Control predict more deaths resulting from microorganisms than all cancers combined by 2050. Currently, many traditional models used to study bacterial infections fail to precisely replicate the in vivo bacterial environment. These models often fail to incorporate fluid flow, bio-mechanical cues, intercellular interactions, host-bacteria interactions, and even the simple inclusion of relevant physiological proteins in culture media. As a result of these inadequate models, there is often a poor correlation between in vitro and in vivo assays, limiting therapeutic potential. Thus, the urgency to establish in vitro and ex vivo systems to investigate the mechanisms underlying bacterial infections and to discover new-age therapeutics against bacterial infections is dire. In this review, we present an update of current in vitro and ex vivo models that are comprehensively changing the landscape of traditional microbiology assays. Further, we provide a comparative analysis of previous research on various established organ-disease models. Lastly, we provide insight on future techniques that may more accurately test new formulations to meet the growing demand of antibiotic resistant bacterial infections.

A Status Report on FDA Approval of Medical Devices Containing Nanostructured Materials.

Commercialization has been slow since the FDA approved a medical device containing nanomaterials in 1980. In 2017, the FDA released draft guidance to accelerate approval. We highlight here that geographical and structural separation of researchers, manufacturers, and clinical servicers may slow commercialization more than FDA approval.

Galactosylated chitosan triptolide nanoparticles for overcoming hepatocellular carcinoma: Enhanced therapeutic efficacy, low toxicity, and validated network regulatory mechanisms.

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths worldwide. Current therapies present significant limitations. Triptolide (TP) is highly effective against multiple cancers including HCC. However, high toxicity, low water solubility, and unknown therapeutic targets limit its clinical application. Herein, we designed galactosylated-chitosan-TP-nanoparticles (GC-TP-NPs) with high drug loading capacities for targeted delivery to HCC. In addition to a sustained release pattern, an efficient asialoglycoprotein receptor mediated cellular uptake in vitro, and high liver tumor accumulation in vivo, GC-TP-NPs showed lower systemic and male reproductive toxicities than free TP. Importantly, GC-TP-NPs retained the anti-cancer activities of the free TP, exerting the same pro-apoptotic and anti-proliferative effects on HCC cells in vitro, and displayed higher efficacies in reducing tumor sizes in vivo. Further investigation revealed that GC-TP-NPs induced cancer cell apoptosis via blocking TNF/NF-κB/BCL2 signaling. Collectively, GC-TP-NP represents a promising candidate in halting liver cancer progression while minimizing systemic toxicity.

Acid-Induced Activated Cell-Penetrating Peptide-Modified Cholesterol-Conjugated Polyoxyethylene Sorbitol Oleate Mixed Micelles for pH-Triggered Drug Release and Efficient Brain Tumor Targeting Based on a Charge Reversal Mechanism.

Glioblastoma multiforme is the most devastating malignant brain tumor in adults. Even with the standard care of therapy, the prognosis remains dismal due to tumor heterogeneity, tumor infiltration, and, more importantly, the restrictive nature of the blood-brain barrier (BBB). To overcome the challenge of effectively delivering therapeutic cargo into the brain, herein a "smart", multifunctional polymeric micelle was developed using a cholesterol-conjugated polyoxyethylene sorbitol oleate. A cell-penetrating peptide, arginine-glycine repeats (RG)5, was incorporated into the micelles to improve cellular uptake, while a pH-sensitive masking sequence, histidine-glutamic acid repeats (HE)5, was introduced for charge shielding to minimize nonspecific binding and uptake at physiological pH. Results demonstrated that (RG)5- and (HE)5-modified mixed micelles were optimized using this strategy to effectively mask the cationic charges of the activated cell-penetrating peptide (RG)5 at physiological pH, i.e., limiting internalization, and were selectively triggered in response to a mildly acidic microenvironment in vitro based on a charge reversal mechanism. In vivo results further confirmed that such micelles preferentially accumulated in both brain and tumor tissues in both xenograft and orthotropic glioma mouse models. Furthermore, micelles significantly inhibited tumor growth with limited toxicity to peripheral tissues. The combination of BBB penetration, tumor targeting, potent efficacy, and high tolerance of these micelles strongly suggests that they could be a promising candidate for safe and effective drug delivery to the brain.

Reducing Bacterial Infections and Biofilm Formation Using Nanoparticles and Nanostructured Antibacterial Surfaces.

With the rapid spreading of resistance among common bacterial pathogens, bacterial infections, especially antibiotic-resistant bacterial infections, have drawn much attention worldwide. In light of this, nanoparticles, including metal and metal oxide nanoparticles, liposomes, polymersomes, and solid lipid nanoparticles, have been increasingly exploited as both efficient antimicrobials themselves or as delivery platforms to enhance the effectiveness of existing antibiotics. In addition to the emergence of widespread antibiotic resistance, of equal concern are implantable device-associated infections, which result from bacterial adhesion and subsequent biofilm formation at the site of implantation. The ineffectiveness of conventional antibiotics against these biofilms often leads to revision surgery, which is both debilitating to the patient and expensive. Toward this end, micro- and nanotopographies, especially those that resemble natural surfaces, and nonfouling chemistries represent a promising combination for long-term antibacterial activity. Collectively, the use of nanoparticles and nanostructured surfaces to combat bacterial growth and infections is a promising solution to the growing problem of antibiotic resistance and biofilm-related device infections.

Azithromycin-loaded respirable microparticles for targeted pulmonary delivery for the treatment of pneumonia.

Pneumonia is a major contributor to infection-based hospitalizations and deaths in the United States. Antibiotics such as azithromycin (AZM), although effective at managing pneumonia, often suffer from off-target diffusion and poor bioavailability when administered orally or via intravenous injection. The formation of biofilms at the disease sites makes the treatment more complicated by protecting bacteria from antimicrobial agents and thus necessitating a much higher dosage of antibiotics to eradicate the biofilms. As such, targeted pulmonary delivery of antibiotics has emerged as a promising alternative by providing direct access to the lung while also allowing higher local therapeutic concentrations but minimal systemic exposure. In this study, AZM was encapsulated in N-fumaroylated diketopiperazine (FDKP) microparticles for efficient pulmonary delivery. Both in vitro and in vivo results demonstrated that AZM@FDKP-MPs administered via intratracheal insufflation achieved at least a 3.4 times higher local concentration and prolonged retention times compared to intravenous injection and oral administration, suggesting their potential to better manage bacterial pneumonia.

Synthesis and characterization of biogenic selenium nanoparticles with antimicrobial properties made by Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, and Pseudomonas aeruginosa.

Antimicrobial resistance is a global concern that affects more than two million people each year. Therefore, new approaches to kill bacteria are needed. One of the most promising methodologies may come from metallic nanoparticles, since bacteria may not develop a resistance to these nanostructures as they do for antibiotics. While metallic nanoparticle synthesis methods have been well studied, they are often accompanied by significant drawbacks such as cost, extreme processing conditions, and toxic waste production since they use harsh chemicals such as corrosive agents (hydrazine) or strong acids (hydrochloride acid). In this work, we explored the environmentally safe synthesis of selenium nanoparticles, which have shown promise in killing bacteria. Using Escherichia coli, Pseudomonas aeruginosa, Methicillin-resistance Staphylococcus aureus, and S. aureus, 90-150 nm average diameter selenium nanoparticles were synthesized using an environmentally safe approach. Nanoparticles were characterized using transmission electron microscopy, energy dispersive X-ray spectroscopy to determine the chemical composition, and inductively coupled plasma mass spectrometry to validate chemistry. Nanoparticles were also characterized and tested for their ability to inhibit bacterial growth. A decay in bacterial growth after 24 h was achieved against both S. aureus and E. coli at biogenic selenium nanoparticle concentrations from 25 to 250 µg/mL and showed no significant cytotoxicity effect against human dermal fibroblasts for 24 h. Bacteria were able to synthesize selenium nanoparticles through the use of different functional structures within the organisms, mainly enzymes such as selenite reductases. Therefore, biogenic selenium nanoparticles made by bacteria represent a viable approach to reduce bacteria growth without antibiotics overcoming the drawbacks of synthetic methods that employ toxic chemicals. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1400-1412, 2018.

Ion-paired pirenzepine-loaded micelles as an ophthalmic delivery system for the treatment of myopia.

Myopia is one of the most common ocular disorders for which standard treatments, such as refractive surgery, often involve invasive procedures. Pirenzepine (PRZ), a muscarinic receptor antagonist, has been recognized as a promising candidate for the treatment of myopia, but possesses poor ocular bioavailability. The overall objective of this study was to prepare PRZ-sorbic acid complexes suitable to be encapsulated into micelles with high efficiency for optimal ophthalmic delivery. The results demonstrated that sorbic acid, used as the counter ion, had the most significant effects in increasing the octanol-water distribution coefficient of PRZ as well as improving its corneal permeability in vitro among various counter ions tested. In vivo absorption results showed that a 1.5 times higher bioavailability was achieved by the addition of sorbic acid at a 1:1 ratio. Cytotoxicity studies in vitro and biocompatibility studies in vivo indicated that the micelles did not cause significant toxicities to the eyes.

Self-assembled arginine-rich peptides as effective antimicrobial agents.

Bacteria can adapt to their ever-changing environment to develop a resistance to commonly used antibiotics. This escalating evolution of bacteria coupled with a diminished number of effective antibiotics has caused a global healthcare crisis. New antimicrobials and novel approaches to tackle this problem are urgently needed. Antimicrobial peptides are of particular interest in this endeavor due to their broad spectrum antimicrobial properties as well as ability to combat multi-drug resistant bacteria. Most peptides have both hydrophobic and hydrophilic regions that enable them to be soluble in an aqueous solution, yet can insert into and subsequently disintegrate lipid rich membranes through diverse mechanisms. In this study, a novel class of cationic nanoparticles (formed by the self-assembly of an amphiphilic peptide) were shown to have strong antimicrobial properties against gram-positive bacteria, specifically Staphylococcus aureus, Staphylococcus epidermidis, and methicillin-resistant Staphylococcus aureus (MRSA) with minimal toxicity to human dermal fibroblasts. The particular self-assembled structure tested here included an arginine rich nanoparticle (C17 H35 GR7RGDS or amphiphilic peptide nanoparticles, APNPs) which incorporated seven arginine residues (imparting a positive charge to improve membrane interactions), a hydrophobic block which drove the self-assembly process, and the presence of an amino acid quadruplet arginine-glycine-aspartic acid-serine (RGDS) which may render these nanoparticles capable of attracting healthy cells while competing bacterial adherence to fibronectin, an adhesive protein found on cell surfaces. As such, this in vitro study demonstrated that the presently formulated APNPs should be further studied for a wide range of antibacterial applications where antibiotics are no longer useful. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1046-1054, 2017.

Optimizing superparamagnetic iron oxide nanoparticles as drug carriers using an in vitro blood-brain barrier model.

In the current study, an optimized in vitro blood-brain barrier (BBB) model was established using mouse brain endothelial cells (b.End3) and astrocytes (C8-D1A). Before measuring the permeability of superparamagnetic iron oxide nanoparticle (SPION) samples, the BBB was first examined and confirmed by an immunofluorescent stain and evaluating the transendothelial electrical resistance. After such confirmation, the permeability of the following five previously synthesized SPIONs was determined using this optimized BBB model: 1) GGB (synthesized using glycine, glutamic acid, and bovine serum albumin [BSA]), 2) GGC (glycine, glutamic acid, and collagen), 3) GGP (glycine, glutamic acid, and polyvinyl alcohol), 4) BPC (BSA, polyethylene glycol, and collagen), and 5) CPB (collagen, polyvinyl alcohol, and BSA). More importantly, after the permeability test, transmission electron microscopy thin section technology was used to investigate the mechanism behind this process. Transmission electron microscopy thin section images supported the hypothesis that collagen-coated CPB SPIONs displayed better cellular uptake than glycine and glutamine acid-coated GGB SPIONs. Such experimental data demonstrated how one can modify SPIONs to better deliver drugs to the brain to treat a wide range of neurological disorders.

Controlling ferrofluid permeability across the blood­brain barrier model.

In the present study, an in vitro blood­brain barrier model was developed using murine brain endothelioma cells (b.End3 cells). Confirmation of the blood­brain barrier model was completed by examining the permeability of FITCDextran at increasing exposure times up to 96 h in serum-free medium and comparing such values with values from the literature. After such confirmation, the permeability of five novel ferrofluid (FF) nanoparticle samples, GGB (ferrofluids synthesized using glycine, glutamic acid and BSA), GGC (glycine, glutamic acid and collagen), GGP (glycine, glutamic acid and PVA), BPC (BSA, PEG and collagen) and CPB (collagen, PVA and BSA), was determined using this blood­brain barrier model. All of the five FF samples were characterized by zeta potential to determine their charge as well as TEM and dynamic light scattering for determining their hydrodynamic diameter. Results showed that FF coated with collagen passed more easily through the blood­brain barrier than FF coated with glycine and glutamic acid based on an increase of 4.5% in permeability. Through such experiments, diverse magnetic nanomaterials (such as FF) were identified for: (1) MRI use since they were less permeable to penetrate the blood­brain barrier to avoid neural tissue toxicity (e.g. GGB) or (2) brain drug delivery since they were more permeable to the blood­brain barrier (e.g. CPB).