Nanobowl-Supported Liposomes Improve Drug Loading and Delivery.

Liposomal drug delivery for cancer therapy can be limited due to drug leakage in circulation. Here, we develop a new method to enhance the stability of actively loaded liposomal doxorubicin (DOX) through embedding a stiff nanobowl in the liposomal water cavity. Nanobowl-supported liposomal DOX (DOX@NbLipo) resists the influence of plasma protein and blood flow shear force to prevent drug leakage. This approach yields improved drug delivery to tumor sites and enhanced antitumor efficacy. Compared to alternative methods of modifying liposome surface and composition for stability, this approach designs a physical support for an all-aqueous nanoliposomal cavity. Nanobowl stabilization of liposomes is a simple and effective method to improve carrier stability and drug delivery.

Mutations in C1orf194, encoding a calcium regulator, cause dominant Charcot-Marie-Tooth disease.

Charcot-Marie-Tooth disease is a hereditary motor and sensory neuropathy exhibiting great clinical and genetic heterogeneity. Here, the identification of two heterozygous missense mutations in the C1orf194 gene at 1p21.2-p13.2 with Charcot-Marie-Tooth disease are reported. Specifically, the p.I122N mutation was the cause of an intermediate form of Charcot-Marie-Tooth disease, and the p.K28I missense mutation predominately led to the demyelinating form. Functional studies demonstrated that the p.K28I variant significantly reduced expression of the protein, but the p.I122N variant increased. In addition, the p.I122N mutant protein exhibited the aggregation in neuroblastoma cell lines and the patient's peroneal nerve. Either gain-of-function or partial loss-of-function mutations to C1ORF194 can specify different causal mechanisms responsible for Charcot-Marie-Tooth disease with a wide range of clinical severity. Moreover, a knock-in mouse model confirmed that the C1orf194 missense mutation p.I121N led to impairments in motor and neuromuscular functions, and aberrant myelination and axonal phenotypes. The loss of normal C1ORF194 protein altered intracellular Ca2+ homeostasis and upregulated Ca2+ handling regulatory proteins. These findings describe a novel protein with vital functions in peripheral nervous systems and broaden the causes of Charcot-Marie-Tooth disease, which open new avenues for the diagnosis and treatment of related neuropathies.

Molecular systematics of Goodyerinae (Cranichideae, Orchidoideae, Orchidaceae) based on multiple nuclear and plastid regions.

Goodyerinae are one of the most species-rich and widespread subtribes of Orchidaceae but notorious for their taxonomic difficulty. Here, a comprehensive molecular phylogenetic study of the subtribe is presented based on two nuclear (ITS, Xdh) and five plastid (matK, psaB, rbcL, trnL, trnL-F) regions. A total of 119 species were included representing all clades recovered by previous phylogenetic analyses as well as seven outgroups. Maximum parsimony, maximum likelihood and Bayesian inference methods were used to infer the phylogenetic relationships. The results show that the Goodyerinae subdivided into three major subdivisions and six groupings: Pachyplectron, Goodyera clade (including Goodyera procera, Microchilus subclade and Goodyera subclade) and Cheirostylis clade (including Gonatostylis, Cheirostylis subclade and Ludisia subclade). Four genera, Erythrodes, Goodyera, Myrmechis and Odontochilus, are not monophyletic. The results support Odontochilus s. l. to include Myrmechis and Kuhlhasseltia. The systematic positions of Goodyera procera and two isolated genera, Herpysma and Orchipedum, are difficult to determine.

Soybean Lecithin-Mediated Nanoporous PLGA Microspheres with Highly Entrapped and Controlled Released BMP-2 as a Stem Cell Platform.

Injectable polymer microsphere-based stem cell delivery systems have a severe problem that they do not offer a desirable environment for stem cell adhesion, proliferation, and differentiation because it is difficult to entrap a large number of hydrophilic functional protein molecules into the core of hydrophobic polymer microspheres. In this work, soybean lecithin (SL) is applied to entrap hydrophilic bone morphogenic protein-2 (BMP-2) into nanoporous poly(lactide-co-glycolide) (PLGA)-based microspheres by a two-step method: SL/BMP-2 complexes preparation and PLGA/SL/BMP-2 microsphere preparation. The measurements of their physicochemical properties show that PLGA/SL/BMP-2 microspheres had significantly higher BMP-2 entrapment efficiency and controlled triphasic BMP-2 release behavior compared with PLGA/BMP-2 microspheres. Furthermore, the in vitro and in vivo stem cell behaviors on PLGA/SL/BMP-2 microspheres are analyzed. Compared with PLGA/BMP-2 microspheres, PLGA/SL/BMP-2 microspheres have significantly higher in vitro and in vivo stem cell attachment, proliferation, differentiation, and matrix mineralization abilities. Therefore, injectable nanoporous PLGA/SL/BMP-2 microspheres can be potentially used as a stem cell platform for bone tissue regeneration. In addition, SL can be potentially used to prepare hydrophilic protein-loaded hydrophobic polymer microspheres with highly entrapped and controlled release of proteins.

Tumor-penetrating peptide mediation: an effective strategy for improving the transport of liposomes in tumor tissue.

Currently, the inefficient transport of liposomes in tumor tissue hinders their clinical application. Tumor-penetrating peptides (TPP) are a series of targeting peptides with the function of penetrating tumor blood vessels and tumor stroma. This work aimed to improve the penetration of liposomes in tumor tissues by TPP modification, thereby enhancing the antitumor effect. First, RPARPAR, a TPP, was modified to the surface of liposomes loaded with doxorubicin. The RPARPAR-modified liposomes (RPA-LP) and unmodified liposomes (LP) showed spherical morphology with average sizes about 90 nm. RPA-LP exhibited remarkably increased cellular accumulation by PC-3 tumor cells than LP as evidenced by the cellular uptake test. The in vivo imaging study confirmed that RPARPAR modification significantly increased the liposome accumulation in subcutaneous tumor tissues. RPA-LP could penetrate through tumor blood vessels and tumor stroma and into the deep tumor tissues as evidence by the immunofluorescence staining analysis. The cytotoxicity of RPARPAR-modified doxorubicin liposomes (RPA-LP-DXR) is considerably increased compared with that of doxorubicin liposomes (LP-DXR). The RPA-LP-DXR also showed significantly (p < 0.005) stronger growth-inhibiting effect on tumor than LP-DXR, possibly due to the tumor-penetrating ability of RPA-LP and targeted killing of tumor cells. This study proved that TPP mediation may be an effective strategy for improving the transport of liposomes in tumor tissue.

Fish oil-loaded multicore submillimeter-sized capsules prepared with monoaxial electrospraying, chitosan-tripolyphosphate ionotropic gelation, and Tween blending.

In order to load fish oil for potential encapsulation of fat-soluble functional active substances, fish oil-loaded multicore submillimeter-sized capsules were prepared with a combination method of three strategies (monoaxial electrospraying, chitosan-tripolyphosphate ionotropic gelation, and Tween blending). The chitosan-tripolyphosphate/Tween (20, 40, 60, and 80) capsules had smaller and evener fish oil cores than the chitosan-tripolyphosphate capsules, which resulted from that Tween addition induced smaller and evener fish oil droplets in the emulsions. Tween addition decreased the water contents from 56.6 % to 35.0 %-43.4 %, increased the loading capacities from 10.4 % to 12.7 %-17.2 %, and increased encapsulation efficiencies from 97.4 % to 97.8 %-99.1 %. In addition, Tween addition also decreased the highest peroxide values from 417 meq/kg oil to 173-262 meq/kg oil. These properties' changes might result from the structural differences between the chitosan-tripolyphosphate and chitosan-tripolyphosphate/Tween capsules. All the results suggested that the obtained chitosan-tripolyphosphate/Tween capsules are promising carriers for fish oil encapsulation. This work also provided useful knowledge to understand the preparation, structural, and physicochemical properties of the chitosan-tripolyphosphate capsules.

Tumor-penetrating peptide functionalization enhances the anti-glioblastoma effect of doxorubicin liposomes.

The targeted therapeutic effect of nano drug delivery system for glioblastoma has been hampered by the weak enhanced permeability and retention (EPR) effect of glioblastoma and the low delivering efficiency of NDDS in glioblastoma tissue. In this study, a tumor-penetrating peptide (RGERPPR), the specific ligand of neuropilin-1 overexpressed on glioblastoma and endothelial cells, was used as a targeting moiety to enhance the anti-glioblastoma effect of doxorubicin liposomes. Firstly, RGERPPR-PEG-DSPE was synthesized and used to prepare the RGERPPR peptide-functionalized liposomes (RGE-LS), which showed vesicle sizes of around 90 nm and narrow size distributions. The cellular uptake and in vivo near-infrared fluorescence imaging test displayed that RGE-LS exhibited increased uptake by glioblastoma cells and intracranial glioblastoma tissues. The cytotoxicity assay and anti-glioblastoma study proved that RGERPPR functionalization significantly enhanced the in vitro inhibitory effect of doxorubicin liposomes on glioblastoma cells and prolonged the median survival time of nude mice bearing intracranial glioblastoma. Finally, the immunofluorescence analysis evidenced that RGE-LS were able to penetrate through tumor vessels and stroma and deep into the whole tumor tissue. The results indicated that tumor-penetrating peptide functionalization is an effective strategy for enhancing the anti-glioblastoma effect of doxorubicin liposomes.

Novel biodegradable electrospun nanofibrous P(DLLA-CL) balloons for the treatment of vertebral compression fractures.

Significant interest has been expressed by the spinal surgeon community for the use of calcium phosphate cement (CPC) in the treatment of vertebral compression fractures (VCFs), but the water-induced collapsibility and poor mechanical properties limit its clinical use. Here we fabricated novel electrospun nanofibrous P(DLLA-CL) balloons (ENPBs) using the nanotechnique of electrospinning. The ENPBs could separate the cements from the surrounding environment, and therefore can prevent the water-induced collapsibility of CPC and eliminate cement leakage. The ENPBs filling with CPC had enough load-bearing ability to restore the height of the fractured vertebral body and had no obvious effects on the initial strength and stiffness of natural bones. Further, the ENPBs had good biodegradability and cell proliferation ability. Calcium can be released from ENPBs filling with CPC. All these results strongly demonstrate ENPBs can be potentially used as CPC filling containers that keep the advantages and eliminate the disadvantages of CPC. FROM THE CLINICAL EDITOR: Calcium phosphate cement (CPC) is a promising modality in vertebral compression fracture treatment, but its water-induced collapsibility limits clinical applications. This team of investigators fabricated novel nanofibrous balloons using electrospinning, which enabled the separation of CPC from its surrounding environment, and therefore prevented water-induced collapsibility of CPC and eliminated cement leakage while maintaining all the advantages of CPC treatment.

Symmetric and asymmetric Au-AgCdSe hybrid nanorods.

This paper describes a facile method for synthesis of Au-AgCdSe hybrid nanorods with controlled morphologies and spatial distributions. The synthesis involved deposition of Ag tips at the ends of Au nanorod seeds, followed by selenization of the Ag tips and overgrowth of CdSe on these sites. By simply manipulating the pH value of the system, the AgCdSe could selectively grow at one end, at both the ends or on the side surface of a Au nanorod, generating a mike-like, dumbbell-like, or toothbrush-like hybrid nanorod, respectively. These three types of Au-AgCdSe hybrid nanorods displayed distinct localized surface plasmon resonance and photoluminescence properties, demonstrating an effective pathway for maneuvering the optical properties of nanocrystals.

Gelatin as a bioactive nanodelivery system for functional food applications.

Gelatin has long been used as an encapsulant agent in the pharmaceutical and biomedical industries because of its low cost, wide availability, biocompatibility, and degradability. However, the exploitation of gelatin for nanodelivery application is not fully achieved in the functional food filed. In this review article, we highlight the latest work being performed for gelatin-based nanocarriers, including polyelectrolyte complexes, nanoemulsions, nanoliposomes, nanogels, and nanofibers. Specifically, we discuss the applications and challenges of these nanocarriers for stabilization and controlled release of bioactive compounds. To achieve better efficacy, gelatin is frequently used in combination with other biomaterials such as polysaccharides. The fabrication and synergistic effects of the newly developed gelatin composite nanocarriers are also present.

Pegylated arginine deiminase downregulates colitis in murine models.

UNLABELLED: Arginine deiminase (ADI), an arginine-metabolizing enzyme involved in cell signaling, is dysregulated in multiple inflammatory diseases and cancers. We hypothesized that pegylated ADI (ADI-PEG) provide protection against colitis. METHODS: Dextran sodium sulfate colitis was induced in IL-10-deficient and BALB/c (WT) mice. ADI-PEG was administered i.p., and inflammatory mediators and pathology were evaluated. RESULTS: Acute colitis in mice was manifested by increases in inflammatory biomarkers, such as serum amyloid A (SAA, P < 0.001), IL-12 p40, and disease index (3-Fold). In contrast, ADI-PEG significantly decreased clinical disease index, SAA levels, and inflammatory cytokines in blood as well as in colonic explants. Animals developed moderate (2.2 ± 0.3 WT) to severe (3.6 ± 0.5 IL-10 deficient) colonic pathology; and ADI-PEG treatment significantly improved the severity of colitis (P < 0.05). Marked infiltration of CD68+ macrophages and iNOS expression were detected in colonic submucosa in colitic animals but not detected in ADI-PEG-treated animals. CONCLUSION: ADI-PEG attenuated inflammatory responses by suppression of macrophage infiltration and iNOS expression in colitic animals. ADI-PEG can serve as a potential therapeutic value in IBD.

Electromagnetically-actuated reciprocating pump for high-flow-rate microfluidic applications.

This study presents an electromagnetically-actuated reciprocating pump for high-flow-rate microfluidic applications. The pump comprises four major components, namely a lower glass plate containing a copper microcoil, a middle PMMA plate incorporating a PDMS diaphragm with a surface-mounted magnet, upper PMMA channel plates, and a ball-type check valve located at the channel inlet. When an AC current is passed through the microcoil, an alternating electromagnetic force is established between the coil and the magnet. The resulting bi-directional deflection of the PDMS diaphragm causes the check-valve to open and close; thereby creating a pumping effect. The experimental results show that a coil input current of 0.4 A generates an electromagnetic force of 47 mN and a diaphragm deflection of 108 µm. Given an actuating voltage of 3 V and a driving frequency of 15 Hz, the flow rate is found to be 13.2 mL/min under zero head pressure conditions.

Highly Efficient Fractionation of Cornstalk into Noncondensed Lignin, Xylose, and Cellulose in Formic Acid.

Traditional pretreatment of lignocellulose is usually conducted under higher acidic and high temperature conditions, which leads to both the degradation of sugar and the condensation of lignin, hindering the subsequent conversion. An effective approach to fractionate lignocellulose into 93.9% of noncondensed lignin, 99.4% of cellulose, 17.8% of xylose, and 66.7% of xylooligosaccharides under mild conditions was developed using the formic acid solution at 80 °C for 100 min. The ß-O-4 bond content of lignin fractionated with formic acid (54.6 per 100 C9 units) was higher than dioxasolv lignin (48.4 per 100 C9 units), indicating that formic acid pretreatment well protected the ether bonds in lignin. Therefore, the hydrogenolysis of fractionated lignin contributed to 28.0% of aromatic monomer yield, which was comparable to dioxasolv lignin. As cellulose possesses a large amount of porosity because lignin was separated from lignocellulose, the hydrolysis of fractionated cellulose by molten salt hydrates gave a 96.4% of glucose yield.

Extraction of Noncondensed Lignin from Poplar Sawdusts with p-Toluenesulfonic Acid and Ethanol.

The traditional pretreatment leads to the recalcitration of C-C bonds during lignin fractionation, thus hindering their depolymerization into aromatic monomers. It is essential to develop an applicable approach to extract noncondensed lignin for its high-value applications. In this work, noncondensed lignins were extracted from poplar sawdust using recyclable p-toluenesulfonic acid for cleaving lignin-carbohydrate complex bonds effectively and ethanol as a stabilization reagent to inhibit lignin condensation. Lignin yield of 83.74% was recovered by 3 mol/L acid in ethanol at 85 °C for 5 h, and carbohydrates were well preserved (retaining 98.97% cellulose and 50.01% hemicelluloses). During lignin fractionation, the acid concentration and extraction time were the major drivers of condensation. Ethanol reacted with lignin at the α-position to prevent the formation of the condensed structure. The extracted lignin depolymerized over the Pd/C catalysts gave a yield of 50.35% of aromatic monomers, suggesting that the novel extraction process provided a promising way for noncondensed lignin production.

Recombinant human insulin-like growth factor-1 promotes osteoclast formation and accelerates orthodontic tooth movement in rats.

BACKGROUND: IGF-1 may be an important factor in bone remodeling, but its mechanism of action on osteoclasts during orthodontic tooth movement is complex and unclear. METHODOLOGY: The closed-coil spring was placed between the left maxillary first molar and upper incisors with a force of 50 g to establish an orthodontic movement model. Eighty SD rats were randomized to receive phosphate buffer saline or 400 ng rhIGF-1 in the lateral buccal mucosa of the left maxillary first molar every two days. Tissue sections were stained for tartrate-resistant acidic phosphatase (TRAP), the number of TRAP-positive cells was estimated and tooth movement measured. RESULTS: The rhIGF-1 group exhibited evidential bone resorption and lacuna appeared on the alveolar bone compared to the control group. Moreover, the number of osteoclasts in compression side of the periodontal ligament in the rhIGF-1 group peaked at day 4 (11.37±0.95 compared to 5.28±0.47 in the control group) after the orthodontic force was applied and was significantly higher than that of the control group (p<0.01). Furthermore, the distance of tooth movement in the rhIGF-1 group was significantly larger than that of the control group from day 4 to day 14 (p<0.01), suggesting that rhIGF-1 accelerated orthodontic tooth movement. CONCLUSION: Our study has showed that rhIGF-1 could stimulate the formation of osteoclasts in the periodontal ligament, and accelerate bone remodeling and orthodontic tooth movement.

A20-OVA Nanoparticles Inhibit Allergic Asthma in a Murine Model.

The skewed T helper (Th) 2 response plays a critical role in the pathogenesis of allergic asthma. Regulatory T (Treg) cells and the regulatory cytokines are required in maintaining the homeostasis in the body. This study aims to determine the effects of a poly(lactic-co-glycolic) acid (PLGA)-ovalbumin (OVA)+A20 (a ubiquitin E3 ligase) nanovaccine on inhibiting allergic asthma in a murine model. In this study, A20 and OVA (a model antigen) were encapsulated into PLGA to be a nanovaccine (PLGA-OVA+A20). An allergic asthma murine model was developed with OVA as the specific antigen to test the role of PLGA-OVA+A20 nanovaccine in maintaining the immune homeostasis in the airway tissues. The results showed that PLGA-OVA+A20 nanovaccine inhibited the asthma responses in mice by suppressing Th2 inflammatory responses, promoting the generation of Treg cells in the airway tissues. We conclude that the PLGA-OVA+A20 nanovaccine has a marked inhibitory effect on the airway allergic response in sensitized mice by significantly promoting the generation of Treg cell and IL-10. The data suggest that PLGA-OVA+A20 has translational potential in the treatment of allergic asthma.

Effects of lipid composition and phase on the membrane interaction of the prion peptide 106-126 amide.

Lipid rafts are specialized liquid-ordered (L(o)) phases of the cell membrane that are enriched in sphingolipids and cholesterol (Chl), and surrounded by a liquid-disordered (L(d)) phase enriched in glycerophospholipids. Lipid rafts are involved in the generation of pathological forms of proteins that are associated with neurodegenerative diseases. To investigate the effects of lipid composition and phase on the generation of pathological forms of proteins, we constructed an L(d)-gel phase-separated 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/sphingomyelin (from bovine brain (BSM))-supported lipid bilayer (SLB) and an L(d)-L(o) phase-separated POPC/BSM/Chl SLB. We used in situ time-lapse atomic force microscopy to study the interactions between these SLBs and the prion peptide K(106)TNMKHMAGAAAAGAVVGGLG(126) (PrP106-126) amide, numbered according to the human prion-peptide sequence. Our results show that: 1), with the presence of BSM in the L(d) phase, the PrP106-126 amide induces fully penetrated porations in the L(d) phase of POPC/BSM SLB and POPC/BSM/Chl SLB; 2), with the presence of both BSM and Chl in the L(d) phase, the PrP106-126 amide induces the disintegration of the L(d) phase of POPC/BSM/Chl SLB; and 3), with the presence of both BSM and Chl in the L(o) phase, PrP106-126 amide induces membrane thinning in the L(o) phase of POPC/BSM/Chl SLB. These results provide comprehensive insight into the process by which the PrP106-126 amide interacts with lipid membranes.

Allogenic chondrocyte/osteoblast-loaded ß-tricalcium phosphate bioceramic scaffolds for articular cartilage defect treatment.

The medical community has expressed significant interest in the treatment of cartilage defect. Successful repair of articular cartilage defects remains a challenge in clinics. Due to the huge advantages of 3D micro/nanomaterials, 3D artificial micro/nano scaffolds have been widely developed and explored in the tissue repair of articular joints. In this study, chondrocyte/osteoblast-loaded ß-tricalcium phosphate (ß-TCP) bioceramic scaffold and chondrocyte-loaded ß-TCP bioceramic scaffold were prepared by micromass stem cell culture and bioreactor-based cells-loaded scaffold culture for articular cartilage defect treatment. The results demonstrate chondrocyte and osteoblast can be successfully induced from allogeneic bone marrow stromal cells using micromass stem cell culture. Further, chondrocyte-loaded ß-TCP scaffold and osteoblast-loaded ß-TCP scaffold can be successfully prepared by bioreactor-based cells-loaded scaffold culture. Finally, the scaffolds are applied for Beagle articular cartilage defect treatment. The relative cartilage regeneration abilities on Beagle femoral trochleae were as follows: Chondrocyte/osteoblast-loaded ß-TCP bioceramic scaffold group > chondrocyte-loaded ß-TCP bioceramic scaffold group > ß-TCP bioceramic scaffold. Therefore, micromass stem cell culture and bioreactor-based cells-loaded scaffold culture can be applied to prepare chondrocyte/osteoblast-loaded ß-TCP bioceramic scaffold for articular cartilage defect treatment. It suggests allogenic chondrocyte/osteoblast-loaded ß-TCP bioceramic scaffold could be potentially used in the treatment of patients with cartilage defects.

PrP106-126 amide causes the semi-penetrated poration in the supported lipid bilayers.

A major hallmark of prion diseases is the cerebral amyloid accumulation of the pathogenic PrP(Sc), an abnormally misfolded, protease-resistant, and beta-sheet rich protein. PrP106-126 is the key domain responsible for the conformational conversion and aggregation of PrP. It shares important physicochemical characteristics with PrP(Sc) and presents similar neurotoxicity as PrP(Sc). By combination of fluorescence polarization, dye release assay and in situ time-lapse atomic force microscopy (AFM), we investigated the PrP106-126 amide interacting with the large unilamellar vesicles (LUVs) and the supported lipid bilayers (SLBs). The results suggest that the interactions involve a poration-mediated process: firstly, the peptide binding results in the formation of pores in the membranes, which penetrate only half of the membranes; subsequently, PrP106-126 amide undergoes the poration-mediated diffusion in the SLBs, represented by the formation and expansion of the flat high-rise domains (FHDs). The possible mechanisms of the interactions between PrP106-126 amide and lipid membranes are proposed based on our observations.

Gambogic Acid-Loaded Polymeric Micelles for Improved Therapeutic Effect in Breast Cancer.

Gambogic acid (GA) possesses good anti-tumor efficacy in preclinical studies, however, its poor hydrophilicity, short blood circulation time and side effect limited its clinical application. In this work, monomethyl poly(ethylene glycol)-poly(ε-caprolactone) (MPEG-PCL) copolymer was synthesized and used to encapsulate GA by a facile one-step solid dispersion and form nano-sized micelles (GA micelles). The GA micelles exhibited small average particle size (29±2 nm), high encapsulation efficiency (92.1±0.3%), and long drug release time-in vitro. Compared to free GA, GA micelles showed superior aqueous dispersity, better tumor cellular uptake, enhanced cytotoxicity and apoptosis induction effect against MCF-7 cells. Furthermore, in vivo studies demonstrated that GA micelles have better antitumor effect in the MCF-7 subcutaneous xenograft tumor model. Histopathological analysis of Ki-67 and TUNEL staining further proved that GA micelles could significantly suppress proliferation as well as increase the apoptosis of tumor cells. These results suggested that GA micelles could potentially improve therapeutic outcomes for breast cancer therapy.