Pioneering Astaxanthin-Tumor Cell Membrane Nanoparticles for Innovative Targeted Drug Delivery on Melanoma.

Background: Recently, the use of the tumor or its secretions as drug carriers has gradually become popular, with the advantages of high biocompatibility and enhanced drug delivery to specific cells. Melanoma is the most malignant tumor of all skin cancers; it is the most metastatic and, therefore, the most difficult to treat. The main purpose of this study is to develop nanovesicles with tumor cell membrane secretion properties to encapsulate target substances to enhance the therapeutic effect of cancer. Methods: Astaxanthin was selected as an anticancer drug due to our previous research finding that astaxanthin has extremely high antioxidant, anti-ultraviolet damage, and anti-tumor properties. The manufacturing method of the astaxanthin nanovesicle carrier is to mix melanoma cells and astaxanthin in an appropriate ratio and then remove the genetic material and inflammatory factors of cancer cells by extrusion. Results: In terms of results, after the co-culture of astaxanthin nanovesicles and melanoma cancer cells, it was confirmed that the ability of astaxanthin nanovesicles to inhibit the growth and metastasis of melanoma cancer cells was significantly better than the same amount of astaxanthin alone, and it had no effect on normal Human cells are also effective. There was no apparent harm on normal cells, indicating the ability of the vesicles to be selectively transported. Conclusion: Our findings illustrated the potential of astaxanthin nanovesicles as an anticancer drug.

Development of Water-Soluble Electrospun Fibers for the Oral Delivery of Cannabinoids.

Cannabidiol (CBD) and cannabigerol (CBG) are two active pharmaceutical ingredients, derived from cannabis plant. In the present study, CBD and CBG were formulated with polyvinyl(pyrrolidone) (PVP) and Eudragit L-100, using electrohydrodynamic atomization (electrospinning). The produced fibers were smooth and uniform in shape, with average fiber diameters in the range of 700-900 nm for PVP fibers and 1-5 µm for Eudragit L-100 fibers. The encapsulation efficiency for both CB and CBG was high (over 90%) for all formulations tested. Both in vitro release and disintegration tests of the formulations in simulated gastric fluids (SGF) and simulated intestinal fluids (SIF) indicated the rapid disintegration and dissolution of the fibers and the subsequent rapid release of the drugs. The study concluded that the electrospinning process is a fast and efficient method to produce drug-loaded fibers suitable for the per os administration of cannabinoids.

Electrospinning/electrospraying coatings for metal microneedles: A design of experiments (DOE) and quality by design (QbD) approach.

The research presented here shows QbD implementation for the optimisation of the key process parameters in electrohydrodynamic atomisation (EHDA). Here, the electrosprayed nanoparticles and electrospun fibers consisting of a polymeric matrix and dye. Eight formulations were assessed consisting of 5% w/v of polycaprolactone (PCL) in dichloromethane (DCM) and 5% w/v polyvinylpyrrolidone (PVP) in ethanol. A full factorial DOE was used to assess the various parameters (applied voltage, deposition distance, flow rate). Further particle and fiber analysis using Scanning Electron Microscopy (SEM), Differential Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA), Fourier Transform Infrared Spectroscopy (FTIR), particle/fiber size distribution. In addition to this in vitro release studied were carried out using fluorescein and Rhodamine B as model dyes and in vitro permeation studies were applied. The results show a significant difference in the morphology of resultant structures as well as a more rapid release profile for the PVP particles and fibers in comparison to the sustained release profiles found with PCL. In vitro drug release studies showed 100% drug release after 7 days for PCL particles and showed 100% drug release within 120 min for PVP particles. The release kinetics and the permeation study showed that the MN successfully pierced the membrane and the electrospun MN coating released a large amount of the loaded drug within 6 h. This study has demonstrated the capability of these robust MNs to encapsulate a diverse range drugs within a polymeric matrix giving rise to the potential of developed personalised medical devices.

Application of mesoporous silica nanoparticles as drug delivery carriers for chemotherapeutic agents.

Recently, remarkable efforts have focused on research towards enhancing and delivering efficacious and advanced therapeutic agents. Even though this involves significant challenges, innovative techniques and materials have been explored to overcome these. The advantageous properties of mesoporous silica nanoparticles (MSNs), such as unique morphologies and geometries, makes then favorable for use for various drug delivery targeting purposes, particularly in cancer therapy. As we discuss here, MSNs have been utilized over the past few decades to improve the efficiency of anticancer drugs by enhancing their solubility to render them suitable for application, reducing adverse effects, and improving their anticancer cytotoxic efficiency.

Extraction of triterpenoid compounds from Ganoderma Lucidum spore powder through a dual-mode sonication process.

Development of drug products from natural sources enable advantageous treatment and therapy options. Bioactive compounds in Ganoderma lucidum spore powder (GLSP) are known for vast antibacterial, antioxidant and anti-cancer properties. Herein, we studied the use of dual-probe ultrasound to extract triterpenoids from GLSP and further investigated the bioactivity of resulting products. FTIR results confirm the presence of key peaks although dual-probe ultrasound varied extraction efficacy. Response surface methodology (RSM) was used to optimize extraction conditions (55:28 for solvent to solid ratio, 10.38 s of ultrasound time and 94% v/v of ethanol concentration). HPLC-Q-TOF-MS confirmed the presence of nine different compounds and in vitro tests confirm good biocompatibility. Extracts are shown to inhibit DPPH radicals, reaching a maximum (61.09 ± 1.38%) at triterpenoid concentrations of 600 µg/mL. Dual-mode assisted extraction provides an enhanced approach for active embedded fiber production on a scale favorable to industry when using optimized process parameters. Furthermore, triterpenoid extracts show antibacterial properties on Staphylococcus aureus and Escherichia coli with potential in antibacterial and anticancer applications.

Production of triterpenoid compounds from Ganoderma lucidum spore powder using ultrasound-assisted extraction.

When ingested as a dietary supplement, Ganoderma lucidum spore powders (GLSP) provide various health benefits such as enhanced immunity, liver protection and anti-cancer effects. In this study, triterpenoid extraction from GLSP was achieved using an ultrasound-assisted process which was optimized using response surface methodology (RSM). Ultrasound-assisted extraction (UAE) was also compared to the most conventional chemical extraction method. For UAE, optimum extraction conditions were found to be ethanol concentration = 95% v/v; solvent to solid ratio = 50:1 mL/g; ultrasound time = 5.4 min; ultrasound power = 564.7 w, and ultrasound probe distance = 8.2 cm. At optimal UAE conditions, no significant differences were found between experimental (0.97 ± 0.04 %) and predicted values (99%); which indicates appreciable correlation at the 97% confidence interval. The findings show the application of Box-Behnken design (BBD) to predict and optimize triterpenoid yield for UAE of triterpenoid from GLSP. Furthermore, glucose consumption was 2.68 times that of control samples when tested with insulin-resistant HepG2 cell, showing potential use in type 2 diabetes. In addition, triterpenoid extracts show good biocompatibility and inhibition of antioxidant activity.

Precision Printing of Customized Cylindrical Capsules with Multifunctional Layers for Oral Drug Delivery.

Advances in personalized medicine will require custom drug formulations and delivery mechanisms. Herein, we demonstrate a new type of personalized capsule comprising of printed concentric cylindrical layers with each layer having a distinctive functional drug component. Poly ε-caprolactone (PCL) with paracetamol (APAP) and chlorpheniramine maleate (CM), synergistic drugs commonly used to alleviate influenza symptoms, are printed as an inner layer and outer layer, respectively, via microscaled electrohydrodynamic (EHD) printing. Polyvinylpyrrolidone (PVP) nanofibers are embedded as interlayers between the two printed PCL-drug layers using electrospinning (ES) techniques. The complete concentric cylindrical capsule with a 6 mm inner diameter and 15 mm length can be swallowed for oral drug delivery. After dissolution of the PVP interlayer, the capsule separates in two, with inner and outer capsules for continuous drug dosing and targeting. Imaging was achieved using a 3T MRI system which allowed temporal observations of the targeted release through the incorporation of nanoparticles (Fe3O4). The morphology and structure, chemical composition, mechanical properties, and biocompatibility of the capsules were studied in vitro. In summary, this new type of custom printed and electrospun capsule that enabled component separation, targeted drug release may advance personalized medicine via multidrug oral delivery.

A novel core-shell nanofiber drug delivery system intended for the synergistic treatment of melanoma.

Here, we introduce core-shell nanofibers based on chitosan (CS)-loaded poly (ε-caprolactone) (PCL) shell and 5-fluorouracil (5-FU)-loaded Poly(N-vinyl-2-pyrrolidone) (PVP) core for synergistic therapy of melanoma skin cancer. The yielded nanofibers exhibited an average diameter of 503 nm with high drug-encapsulating efficiency and good mechanical properties. Moreover, the burst release of 5-FU significantly inhibited melanoma skin cancer cells (B16F10 cells), and the sustained release of CS exhibited "remedying effects" on normal skin cells (L929 cells) after suffering adverse effects from 5-FU treatment. For the B16F10 cells, the early apoptosis cells increased from 0.8% to 62.2% after being treated with blended films loaded with 5-FU (2 wt%) for 24 h; for the L929 cells, the vital cells increased from 68.9% to 77.0%, and the early apoptosis of stage cells decreased from 12.3% to 10.9% after being treated with blended films with CS (8 wt%) for 24 h. In conclusion, the results introduced in this work can be a promising strategy for cancer treatment and possesses synergism potential to broaden an avenue for chemotherapeutic therapy with minimum adverse effects on normal cells.

Fabrication of patterned three-dimensional micron scaled core-sheath architectures for drug patches.

Recent advances in selective integration of micro and nano-scaled features towards material design have paved way to enhance desirable properties or functions of biomaterials. For drug delivery applications these include improved active component encapsulation, controlled drug release and managed interaction with the intended host environment. Electrohydrodynamic (EHD) direct-printing technique is a one-step on demand fiber deposition method which enables precise micron-scaled topographic and structural enhancement during material fabrication. In this study, core-sheath composite fibers comprising polycaprolactone, polyvinyl pyrrolidone and the drug tetracycline hydrochloride were prepared using the coaxial format of EHD direct-printing. Once positioned and aligned; multi-stacked fibers gave rise to patches. Coaxial fiber (diameter range ~13-25 µm) optimization (deposition and integrity) involved parameter-structure (e.g. collector speed, flow rate, working distance and applied voltage) impact analysis. Water contact angle measurements, tensile testing and Fourier transform infrared spectroscopy were used to analyze core-sheath integrated patches. In-vitro drug release studies clearly elude to the impact of core-shell and patterned architectures; demonstrating their viability and the forming method as emerging tools for advanced drug delivery system design and fabrication.

Pharmacological effects of natural Ganoderma and its extracts on neurological diseases: A comprehensive review.

Ganoderma, has been used for clinical applications for thousands of years as a highly-nutritious and significantly-effective medicinal herb. The active components and efficacy of Ganoderma are constantly being explored and supplemented every year. In recent years, more and more literature has reported the pharmacological effects of Ganoderma on anti-tumor, liver protection and immunity enhancement, especially on neuroprotection. Numerous research works on the neuroprotective effects of Ganoderma have been documented (e.g., modulation of neurogenesis, amelioration of Alzheimer's disease, therapeutic effect on epilepsy, the protective effect on neural cells in stroke injury, etc.) thus it has drawn increasing attention. However, an integrated and comprehensive review of recent research findings has not been detailed in any great depth. Therefore, the purpose of this review is to summarize and elucidate recent progress of neuroprotective effects of natural Ganoderma and its extracts.

Three-Dimensional Electrohydrodynamic Printing and Spinning of Flexible Composite Structures for Oral Multidrug Forms.

A simple method to rapidly customize and to also mass produce oral dosage forms is arguably a current bottleneck in the development of modern personalized medicine. Specifically, delayed-release mechanisms with well-controlled dosage profiles for combinations of traditional Chinese herbal extracts and Western medications are not well established. Herein, we demonstrate a novel multidrug-loaded membrane sandwich with structures infused with ibuprofen (IBU) and Ganoderma lucidum polysaccharide (GLP) using three-dimensional electrohydrodynamic printing and electrospinning techniques. The resulting flexible membrane consists of microscaled, multilayered cellulose acetate (CA) membranes loaded with IBU in the shape of either concentric squares or circles, as the top and bottom layers of a sandwich structure. In between the CA-IBU layers are randomly electrospun polyvinyl pyrrolidone (PVP) layers loaded with GLP. The complete fibrous membrane sandwich can be folded and embedded into a 0-size capsule to achieve oral compliance. Simulated in vitro testing of gastric and intestinal fluids demonstrated a triphasic release profile. There was an immediate release of GLP after gastric juices dissolved the capsule shell and the PVP, followed by the short-term release of 60% of the IBU within an hour afterward, and the remaining IBU was released in a sustained manner following a Fickian diffusion profile. In summary, this multidrug (both hydrophilic and/or hydrophobic) oral system with precision-designed structures should enable personalized therapeutic dosing.

Stable increased formulation atomization using a multi-tip nozzle device.

Electrohydrodynamic atomization (EHDA) is an emerging technique for the production of micron and nano-scaled particles. The process often involves Taylor cone enablement, which results in a fine spray yielding formulated droplets, which then undergo drying during deposition. In this work, novel multi-tip emitter (MTE) devices were designed, engineered and utilized for potential up-scaled EHDA, by comparison with a conventional single-needle system. To demonstrate this, the active ketoprofen (KETO) was formulated using polyvinylpyrrolidone (PVP) polymer as the matrix material. Here, PVP polymer (5% w/v) solution was prepared using ethanol and distilled water (80:20) as the vehicle. KETO was incorporated as 5% w/w of PVP. Physical properties of resulting solutions (viscosity, electrical conductivity, density and surface tension) were obtained. Formulations were electrosprayed through both single and novel MTEs under EHDA conditions at various flow rates (5-300 µl/min) and applied voltages (0-30 kV). The atomization process using MTEs and single nozzle was monitored at using various process parameters via a digital optical camera. Resulting particles were collected 200 mm below processing heads and were analyzed using differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), X-ray diffraction (XRD) and scanning electron microscopy (SEM). Digital recordings confirmed stable MTE jetting at higher flow rates. Electron micrographs confirmed particle size variation arising due to nozzle head design and evidenced stable jetting derived greater near-uniform particles. DSC, XRD and TGA confirm KETO molecules were encapsulated and dispersed into PVP polymer particles. In conclusion, novel MTE devices enabled stable atomization even at higher flow rates when compared to conventional single-needle device. This indicates an exciting approach for scaling up (EHDA) in contrast to current efforts focusing on multiple-nozzle and pore-based processing outlets.

Electrosprayed mesoporous particles for improved aqueous solubility of a poorly water soluble anticancer agent: in vitro and ex vivo evaluation.

Encapsulation of poorly water-soluble drugs into mesoporous materials (e.g. silica) has evolved as a favorable strategy to improve drug solubility and bioavailability. Several techniques (e.g. spray drying, solvent evaporation, microwave irradiation) have been utilized for the encapsulation of active pharmaceutical ingredients (APIs) into inorganic porous matrices. In the present work, a novel chalcone (KAZ3) with anticancer properties was successfully synthesized by Claisen-Schmidt condensation. KAZ3 was loaded into mesoporous (SBA-15 and MCM-41) and non-porous (fumed silica, FS) materials via two techniques; electrohydrodynamic atomization (EHDA) and solvent impregnation. The effect of both loading methods on the physicochemical properties of the particles (e.g. size, charge, entrapment efficiency, crystallinity, dissolution and permeability) was investigated. Results indicated that EHDA technique can load the active in a complete amorphous form within the pores of the silica particles. In contrast, reduced crystallinity (~79%) was obtained for the solvent impregnated formulations. EHDA engineered formulations significantly improved drug dissolution up to 30-fold, compared to the crystalline drug. Ex vivo studies showed EHDA formulations to exhibit higher permeability across rat intestine than their solvent impregnated counterparts. Cytocompatibility studies on Caco-2 cells demonstrated moderate toxicity at high concentrations of the anticancer agent. The findings of the present study clearly show the immense potential of EHDA as a loading technique for mesoporous materials to produce poorly water-soluble API carriers of high payload at ambient conditions. Furthermore, the scale up potential in EHDA technologies indicate a viable route to enhance drug encapsulation and dissolution rate of loaded porous inorganic materials.

Essential Oil Bioactive Fibrous Membranes Prepared via Coaxial Electrospinning.

A novel antimicrobial composite material was prepared by encapsulating orange essential oil (OEO) in zein prolamine (ZP) via the coaxial electrospinning (ES) technique. By manipulating process parameters, the morphological features of ZP/OEO fibers were modulated. Fine fibers with diameters ranging from 0.7 to 2.3 µm were obtained by regulating ZP solution concentration and process parameters during the ES process. Optimal loading capacity (LC) and encapsulation efficiency (EE) of OEO in fibrous ZP mats were determined to be 22.28% and 53.68%, respectively, and were achieved using a 35 w/v% ZP ES solution. The encapsulation of OEO was found to be reliant on ZP solution concentration (the enveloping medium). SEM analysis indicates the surface morphology of ZP/OEO electrospun fibers is dependent on ZP solution loading volume, with lower ZP concentrations yielding defective fibrous structures (for example, beaded and spindled-string like morphologies). Furthermore, this loading volume also influences OEO LC, EE, mat water contact angle and oil retention. CCK-8 assay and cell morphology assessment (HEK293T cells) indicate no significant change with electrospun ZP and ZP/OEO fibrous membranes over an 8 h period. Antimicrobial activity assessment using Escherichia coli, suggests composite nonwovens possess sterilization properties; elucidating potential application in active food packaging, food preservation and therefore sustainability.

Development and characterisation of cellulose based electrospun mats for buccal delivery of non-steroidal anti-inflammatory drug (NSAID).

In this study conventional electrospinning (ESp) was used to prepare a series of buccal films containing indomethacin (INDO, a nonsteroidal anti-inflammatory drug), Ethocel (10), hydroxypropylmethylcellulose (HPMC) and Tween® 80 at various concentrations. The films were characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM), fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, differential scanning calorimetry (DSC) and X-ray diffraction (XRD). Drug release behaviour was assessed in vitro (buffer pH6.8). SEM revealed film morphology and mean fibre diameter was dependent on the process formulation. INDO was encapsulated in the amorphous state once electrospun as evidenced from DSC and XRD studies. The presence of other excipients within fibrous matrices was confirmed using FTIR and Raman spectroscopy. Loading and release of INDO from filamentous structures was influenced by formulation composition; indicating potential to 'fine-tune' dosage forms. Given that ESp is a one-step preparation method and operational at ambient conditions; an attractive route for engineering tailored film type dosage forms is presented. This is a valuable approach for optimizing dosage forms as needed in a single step for various age groups.

Hollow polycaprolactone composite fibers for controlled magnetic responsive antifungal drug release.

Hollow magnetic fibers for trigger based drug release were synthesized using one-step co-axial electrospinning (COX-ES). This was achieved by encapsulating the antifungal active 'ketoconazole' (KCZ) and iron oxide (Fe3O4) nanoparticles (NPs) in composite form within the core shell polymeric matrix material (polycaprolactone, PCL) during the COX-ES process. Dimethyl silicone oil was used as the inner core (liquid) of co-flowing solutions, which subsequently perfused out of the two-phase electrospun microstructures to form hollow fibers. Resulting drug-loaded magnetic hollow fibers were characterized using optical microscopy, scanning electron microscopy and Fourier Transform Infra-Red. The tensile strength and magnetization properties of composite fibers were also assessed. KCZ drug concentration in electrospinning solutions strongly influenced resulting fiber morphology, drug loading efficiency and release. Expedited drug release during a slow-sustained phase was demonstrated through the application of an auxiliary magnetic field. Variations in tensile strength (∼1.3-6.3MPa) were due to composite fiber components compromising polymer chain integrity. In-vitro cell studies (using human cervical carcinoma cell lines) demonstrated fiber biocompatibility. The present study demonstrates the potential application of magnetic hollow fibers for controlled treatment of fungal infections and antimicrobial indications.

Combination of RGD compound and low-dose paclitaxel induces apoptosis in human glioblastoma cells.

BACKGROUND: Integrins are a family of transmembrane adhesion proteins that mediate cell adhesion and intracellular signaling. Integrin-αvß3 is expressed on the surface of human glioblastoma cells, and can be further induced by chemical stress. The Arg-Gly-Asp (RGD) motif-containing peptides are specifically bound to integrin-αvß3, and to inhibit neovasculature underlying competition to normal extracellular matrix proteins. This study employed two types of RGD peptides, cyclic RGD (c(RGDyK)) and bi-cyclic RGD (E[c(RGDyK)](2)) peptide, to human glioblastoma U87MG cells with combination of low dose Paclitaxel (PTX) pre-treatment to augment therapeutic activity for RGD peptide-induced apoptosis. PRINCIPAL FINDINGS: Human glioblastoma U87MG cells were treated with RGD peptides in the absence or presence of initial exposure to low-dose 10 nM PTX. Results showed that integrin-αvß3 expressing on the surface of U87MG cells was induced by 10 nM PTX pre-treatment for 12 hrs. Additionally, the U87MG cells pre-treated with PTX and followed by RGD peptides exhibited greater expression of caspases-3, -8 and -9 than those merely treated with single agent of PTX or RGD peptide. Furthermore, the caspase-3, -8 and -9 inhibitor presented significant protection against E[c(RGDyK)](2) peptide induced U87MG programmed cell death. The increased expression of PTX-induced integrin-αvß3 was correlated with the enhanced apoptosis in U87MG cells. CONCLUSIONS: This study provides a novel concept of targeting integrin-αvß3 with RGD peptides in combination with low-dose PTX pre-treatment to improve efficiency in human glioblastoma treatment.

Spontaneous rupture with pseudoaneurysm formation in a nodular goiter presenting as a large neck mass.

Acute airway obstruction caused by goiter hemorrhage is a rare but potentially life-threatening condition. We report a case of nodular goiter in which spontaneous rupture with active bleeding developed suddenly, presenting as a large neck mass with dyspnea. Sonographic examination revealed a large inhomogeneous soft tissue mass in the left thyroid lobe containing some hypoechoic areas. Color Doppler and spectral Doppler analysis were consistent with a large hematoma and pseudoaneurysm, which was subsequently confirmed at surgery.